Methods and compositions for re-dosing AAV using Anti-CD40 antagonistic antibody to suppress host Anti-AAV antibody response

Abstract

Provided herein are methods of inserting a nucleic acid encoding a polypeptide of interest into a target genomic locus in a cell or a population of cells in a subject, methods of expressing a polypeptide of interest from a target genomic locus in a cell or a population of cells in a subject, methods of treating an enzyme deficiency in a subject in need thereof, and methods of preventing or reducing the onset of a sign or symptom of an enzyme deficiency in a subject in need thereof. The methods use CD40 inhibitors (e.g., CD40 antigen-binding molecules) to mitigate immune response and facilitate redosing of nucleic acid constructs encoding a polypeptide of interest and nuclease agents targeting a target genomic locus to achieve, for example, a step-wise increase in expression of a polypeptide of interest in a subject following insertion of the nucleic acid construct without overshooting.

Description

Attorney Docket No.057766 / 625782 METHODS AND COMPOSITIONS FOR RE-DOSING AAV USING ANTI-CD40 ANTAGONISTIC ANTIBODY TO SUPPRESS HOST ANTI-AAV ANTIBODY RESPONSE CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of US Application No.63 / 560,295, filed March 1, 2024, US Application No.63 / 639,227, filed April 26, 2024, US Application No.63 / 660,014, filed June 14, 2024, and US Application No.63 / 733,542, filed December 13, 2024, each of which is herein incorporated by reference in its entirety for all purposes. REFERENCE TO A SEQUENCE LISTING SUBMITTED AS AN XML FILE

[0002] The Sequence Listing written in file 625782SEQLIST.xml is 477,278 bytes, was created on February 28, 2025, and is hereby incorporated by reference in its entirety. BACKGROUND

[0003] CD40 is a cell surface receptor that is part of the tumor necrosis factor (TNF) receptor superfamily. CD40 is expressed on antigen-presenting cells such as B cells, macrophages, and dendritic cells, as well as some non-immune cells and tumors. The interaction of CD40 with its ligand CD40L provides a co-stimulatory signal that is essential to the survival of many cell types and is required for functions of immune response such as B cell activation (particularly in response to T-dependent antigens), germinal center formation and selection, development of long-lived plasma cells and memory B cells responses, IgG class switching, and "licensing" of dendritic cells to mature and become more potent inducers of T-cell immunity.

[0004] In AAV gene therapies, seronegative / naive patients are dosed with AAV and develop antibody responses to the AAV capsid antigen. This antibody response prevents future re-dosing of AAV because the antibodies are neutralizing, and the antibody response is sustained for 10+ years. SUMMARY

[0005] Provided herein are methods of inserting a nucleic acid encoding a polypeptide of interest into a target genomic locus in a cell or a population of cells in a subject, methods of expressing a polypeptide of interest from a target genomic locus in a cell or a population of cellsAttorney Docket No.057766 / 625782 in a subject, methods of treating an enzyme deficiency in a subject in need thereof, and methods of preventing or reducing the onset of a sign or symptom of an enzyme deficiency in a subject in need thereof. Also provided are compositions, combinations, or kits, e.g., for use in such methods.

[0006] In one aspect, provided are methods of inserting a nucleic acid encoding a polypeptide of interest into a target genomic locus in a cell or a population of cells in a subject. Such methods can comprise administering to the subject: (a) a nucleic acid construct comprising a coding sequence for the polypeptide of interest; (b) a nuclease agent or one or more nucleic acids encoding the nuclease agent, wherein the nuclease agent targets a nuclease target site in the target genomic locus; and (c) a CD40 inhibitor (e.g., CD40 antigen-binding molecule), wherein the nuclease agent cleaves the nuclease target site, and the nucleic acid construct is inserted into the target genomic locus. In another aspect, provided are methods of expressing a polypeptide of interest from a target genomic locus in a cell or a population of cells in a subject. Such methods can comprise administering to the subject: (a) a nucleic acid construct comprising a coding sequence for the polypeptide of interest; and (b) a nuclease agent or one or more nucleic acids encoding the nuclease agent, wherein the nuclease agent targets a nuclease target site in the target genomic locus, (c) a CD40 inhibitor (e.g., CD40 antigen-binding molecule), wherein the nuclease agent cleaves the nuclease target site, the nucleic acid construct is inserted into the target genomic locus to create a modified target genomic locus, and the polypeptide of interest is expressed from the modified target genomic locus. In another aspect, provided are methods of treating an enzyme deficiency in a subject in need thereof. Such methods can comprise administering to the subject: (a) a nucleic acid construct comprising a coding sequence for a polypeptide of interest, wherein the polypeptide of interest comprises an enzyme to treat the enzyme deficiency; (b) a nuclease agent or one or more nucleic acids encoding the nuclease agent, wherein the nuclease agent targets a nuclease target site in a target genomic locus; and (c) a CD40 inhibitor (e.g., CD40 antigen-binding molecule), wherein the nuclease agent cleaves the nuclease target site, the nucleic acid construct is inserted into the target genomic locus to create a modified target genomic locus, and the polypeptide of interest is expressed from the modified target genomic locus, thereby treating the enzyme deficiency. In another aspect, provided are methods of preventing or reducing the onset of a sign or symptom of an enzyme deficiency in a subject in need thereof. Such methods can comprise administering to the subject: (a) a nucleicAttorney Docket No.057766 / 625782 acid construct comprising a coding sequence for a polypeptide of interest, wherein the enzyme deficiency is characterized by a loss-of-function of the polypeptide of interest; (b) a nuclease agent or one or more nucleic acids encoding the nuclease agent, wherein the nuclease agent targets a nuclease target site in a target genomic locus; and (c) a CD40 inhibitor (e.g., CD40 antigen-binding molecule), wherein the nuclease agent cleaves the nuclease target site, the nucleic acid construct is inserted into the target genomic locus to create a modified target genomic locus, and the polypeptide of interest is expressed from the modified target genomic locus, thereby preventing or reducing the onset of the sign or symptom of the enzyme deficiency. In some such methods, the subject has a disease of a bleeding disorder characterized by the enzyme deficiency, a disease of an inborn error of metabolism characterized by the enzyme deficiency, or a lysosomal storage disease characterized by the enzyme deficiency. In some such methods, wherein the disease is hemophilia B and the polypeptide of interest is a factor IX protein, the disease is hemophilia A and the polypeptide of interest is a factor VIII protein, or the disease is Pompe disease and the polypeptide of interest is a multidomain therapeutic protein comprising a delivery domain fused to a lysosomal alpha-glucosidase. In some such methods, the subject does not have preexisting immunity against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or a delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent (e.g., at the time the CD40 inhibitor is administered).

[0007] In some such methods, the method further comprises a subsequent administration step. In some such methods, the further administration step comprises administering to the subject at one or more subsequent times: (a) the nucleic acid construct; (b) the nuclease agent or the one or more nucleic acids encoding the nuclease agent; and optionally (c) the CD40 inhibitor (e.g., CD40 antigen-binding molecule), until a desired level of expression and / or activity of the polypeptide of interest is achieved in the subject. In some such methods, the further administration step comprises administering to the subject at one or more subsequent times: (a) the nucleic acid construct; (b) a second nuclease agent or one or more nucleic acids encoding the second nuclease agent, wherein the second nuclease agent targets a second nuclease target site in the target genomic locus, wherein the second nuclease target site is different from the first nuclease target site; and optionally (c) the CD40 inhibitor (e.g., CD40 antigen-binding molecule), until a desired level of expression and / or activity of the polypeptide of interest isAttorney Docket No.057766 / 625782 achieved in the subject. In one example, the first nuclease target site can be a first location in ALB, and the second nuclease target site can be a second location in ALB. For example, the first nuclease target site can be a first location in intron 1 of ALB, and the second nuclease target site can be a second location in intron 1 of ALB. In a specific example, the first nuclease agent targets SEQ ID NO: 108 (e.g., G009860). For example, the first nuclease agent can target SEQ ID NO: 108 (e.g., G009860), and the second nuclease agent can target SEQ ID NO: 106 (e.g., G009844) (or vice versa). For example, the first nuclease agent can target SEQ ID NO: 108 (e.g., G009860), and the second nuclease agent can target SEQ ID NO: 107 (e.g., G009857) (or vice versa). For example, the first nuclease agent can target SEQ ID NO: 108 (e.g., G009860), and the second nuclease agent can target SEQ ID NO: 109 (e.g., G009874) (or vice versa). In some such methods, the further administration step comprises administering to the subject at one or more subsequent times: (a) the nucleic acid construct; (b) a second nuclease agent or one or more nucleic acids encoding the second nuclease agent, wherein the second nuclease agent targets a second nuclease target site in a second target genomic locus that is different from the first target genomic locus (e.g., ALB is the first target genomic locus, and the second target genomic locus is different (e.g., TTR)); and optionally (c) the CD40 inhibitor (e.g., CD40 antigen-binding molecule), until a desired level of expression and / or activity of the polypeptide of interest is achieved in the subject. In one example, the first target genomic locus can be ALB (e.g., intron 1 of ALB). For example, the first target genomic locus can be ALB (e.g., intron 1 of ALB), and the second target genomic locus can be TTR (e.g., intron 1 of TTR). In some such methods, the further administration step comprises administering to the subject at one or more subsequent times: (a) a second nucleic acid construct comprising a second coding sequence for the polypeptide of interest, wherein the second coding sequence is different from the first coding sequence; (b) (i) the first nuclease agent or the one or more nucleic acids encoding the first nuclease agent; (ii) a second nuclease agent or one or more nucleic acids encoding the second nuclease agent, wherein the second nuclease agent targets a second nuclease target site in the target genomic locus, wherein the second nuclease target site is different from the first nuclease target site; or (iii) a second nuclease agent or one or more nucleic acids encoding the second nuclease agent, wherein the second nuclease agent targets a second nuclease target site in a second target genomic locus that is different from the first target genomic locus (e.g., ALB is the first target genomic locus, and the second target genomic locus is different (e.g., TTR)); andAttorney Docket No.057766 / 625782 optionally (c) the CD40 inhibitor (e.g., CD40 antigen-binding molecule), until a desired level of expression and / or activity of the polypeptide of interest is achieved in the subject. In one example, the first nuclease target site can be a first location in ALB, and the second nuclease target site can be a second location in ALB. For example, the first nuclease target site can be a first location in intron 1 of ALB, and the second nuclease target site can be a second location in intron 1 of ALB. In a specific example, the first nuclease agent targets SEQ ID NO: 108 (e.g., G009860). For example, the first nuclease agent can target SEQ ID NO: 108 (e.g., G009860), and the second nuclease agent can target SEQ ID NO: 106 (e.g., G009844) (or vice versa). For example, the first nuclease agent can target SEQ ID NO: 108 (e.g., G009860), and the second nuclease agent can target SEQ ID NO: 107 (e.g., G009857) (or vice versa). For example, the first nuclease agent can target SEQ ID NO: 108 (e.g., G009860), and the second nuclease agent can target SEQ ID NO: 109 (e.g., G009874) (or vice versa). In one example, the first target genomic locus can be ALB (e.g., intron 1 of ALB). For example, the first target genomic locus can be ALB (e.g., intron 1 of ALB), and the second target genomic locus can be TTR (e.g., intron 1 of TTR). In some such methods, the method comprises the following steps prior to the subsequent administration step: (i) measuring expression and / or activity of the polypeptide of interest in the subject; and (ii) determining the dose of the nucleic acid construct and the nuclease agent or the one or more nucleic acids encoding the nuclease agent for the subsequent administration step in order to achieve the desired level of expression and / or activity of the polypeptide of interest is achieved in the subject. In some such methods, the dose of the nucleic acid construct for the subsequent administration step is higher than the dose of the nucleic acid construct in the first administration step and / or the dose of the nuclease agent or one or more nucleic acids encoding the nuclease agent for the subsequent step is higher than the dose of the nuclease agent or one or more nucleic acids encoding the nuclease agent in the first administration step. In some such methods, the dose of the nucleic acid construct for the subsequent administration step is higher than the dose of the nucleic acid construct in the first administration step. In some such methods, the dose of the nucleic acid construct for the subsequent administration step is lower than the dose of the nucleic acid construct in the first administration step and / or the dose of the nuclease agent or one or more nucleic acids encoding the nuclease agent for the subsequent step is lower than the dose of the nuclease agent or one or more nucleic acids encoding the nuclease agent in the first administration step. In some such methods, the dose of the nucleic acid construct for theAttorney Docket No.057766 / 625782 subsequent administration step is lower than the dose of the nucleic acid construct in the first administration step. In some such methods, the dose of the nucleic acid construct for the subsequent administration step is the same as the dose of the nucleic acid construct in the first administration step and / or the dose of the nuclease agent or one or more nucleic acids encoding the nuclease agent for the subsequent step is the same as the dose of the nuclease agent or one or more nucleic acids encoding the nuclease agent in the first administration step. In some such methods, the dose of the nucleic acid construct for the subsequent administration step is the same as the dose of the nucleic acid construct in the first administration step. In some such methods, the polypeptide of interest is a factor IX protein, and the desired expression level of the factor IX protein in the subject is a serum level of at least about 3 ^g / mL or about 3-5 ^g / mL. In some such methods, the polypeptide of interest is a multidomain therapeutic protein comprising a delivery domain fused to a lysosomal alpha-glucosidase, and the desired expression level of the multidomain therapeutic protein in the subject is a serum level of at least about 2 ^g / mL or at least about 5 ^g / mL. In some such methods, the further administration step comprises administering to the subject at one or more subsequent times: (a) a second nucleic acid construct comprising a coding sequence for a second polypeptide of interest that is different from the first polypeptide of interest; (b) (i) the first nuclease agent or the one or more nucleic acids encoding the first nuclease agent; (ii) a second nuclease agent or one or more nucleic acids encoding the second nuclease agent, wherein the second nuclease agent targets a second nuclease target site in the target genomic locus, wherein the second nuclease target site is different from the first nuclease target site; or (iii) a second nuclease agent or one or more nucleic acids encoding the second nuclease agent, wherein the second nuclease agent targets a second nuclease target site in a second target genomic locus that is different from the first target genomic locus (e.g., ALB is the first target genomic locus, and the second target genomic locus is different (e.g., TTR)); and optionally (c) the CD40 inhibitor (e.g., CD40 antigen-binding molecule), wherein the second nuclease agent cleaves the second nuclease target site, and the second nucleic acid construct is inserted into the second target genomic locus. In one example, the first nuclease target site can be a first location in ALB, and the second nuclease target site can be a second location in ALB. For example, the first nuclease target site can be a first location in intron 1 of ALB, and the second nuclease target site can be a second location in intron 1 of ALB. In a specific example, the first nuclease agent targets SEQ ID NO: 108 (e.g., G009860). For example, the first nuclease agentAttorney Docket No.057766 / 625782 can target SEQ ID NO: 108 (e.g., G009860), and the second nuclease agent can target SEQ ID NO: 106 (e.g., G009844) (or vice versa). For example, the first nuclease agent can target SEQ ID NO: 108 (e.g., G009860), and the second nuclease agent can target SEQ ID NO: 107 (e.g., G009857) (or vice versa). For example, the first nuclease agent can target SEQ ID NO: 108 (e.g., G009860), and the second nuclease agent can target SEQ ID NO: 109 (e.g., G009874) (or vice versa). In one example, the first target genomic locus can be ALB (e.g., intron 1 of ALB). For example, the first target genomic locus can be ALB (e.g., intron 1 of ALB), and the second target genomic locus can be TTR (e.g., intron 1 of TTR). In some such methods, the one or more subsequent administration steps is one subsequent administration step. In some such methods, the one or more subsequent administration steps is two subsequent administration steps or comprises at least two subsequent administration steps. In some such methods, the CD40 inhibitor (e.g., CD40 antigen-binding molecule) is administered in the one or more subsequent administration steps if there is no preexisting CD40 inhibitor (e.g., CD40 antigen-binding molecule) in the subject or if preexisting CD40 inhibitor (e.g., CD40 antigen-binding molecule) expression and / or activity levels are below a desired threshold level, In some such methods, the method comprises measuring CD40 inhibitor (e.g., CD40 antigen-binding molecule) expression and / or activity levels prior to the one or more subsequent administration steps.

[0008] In some such methods, the CD40 inhibitor is a CD40 antigen-binding molecule. In some such methods, the CD40 antigen-binding molecule is a monospecific CD40 antigen- binding molecule. In some such methods, the CD40 antigen-binding molecule is a bispecific antigen-binding molecule comprising: (a) a first antigen-binding domain (D1) that binds a first epitope of human CD40; and (b) a second antigen-binding domain (D2) that binds a second epitope of human CD40. In some such methods, the bispecific antigen-binding molecule: (i) binds human CD40 with a KD of less than 25 nM as measured by surface plasmon resonance at 25°C; (ii) binds human CD40 with a KD of less than 70 nM as measured by surface plasmon resonance at 37°C; (iii) binds human CD40 with a dissociative half-life (t1 / 2) of greater than 75 minutes as measured by surface plasmon resonance at 25°C; (iv) binds a human CD40- expressing cell with an EC50 value of about 10 nM or less; (v) inhibits binding of human CD40 dimer to CD40L; (vi) inhibits CD40 ligand (CD40L)-induced activation; and / or (vii) does not significantly agonize CD40 in the absence of CD40L. In some such methods, the bispecific antigen-binding molecule: (i) binds human CD40 with a KD of less than 25 nM as measured byAttorney Docket No.057766 / 625782 surface plasmon resonance at 25°C; (ii) binds human CD40 with a KD of less than 70 nM as measured by surface plasmon resonance at 37°C; (iii) binds human CD40 with a dissociative half-life (t1 / 2) of greater than 75 minutes as measured by surface plasmon resonance at 25°C; (iv) binds a human CD40-expressing cell with an EC50 value of about 10 nM or less; (v) inhibits binding of human CD40 monomer to CD40L; (vi) inhibits CD40 ligand (CD40L)-induced activation; and / or (vii) does not significantly agonize CD40 in the absence of CD40L. In some such methods, the bispecific antigen-binding molecule inhibits CD40L-induced activation. In some such methods, the bispecific antigen-binding molecule inhibits CD40L-induced activation and does not significantly agonize CD40 in the absence of CD40L. In some such methods, the D1 domain and the D2 domain each comprise a heavy chain immunoglobulin variable region comprising a set of three heavy chain complementarity determining region sequences HCDR1, HCDR2, and HCDR3 independently selected from the group consisting of: (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8; (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28; and (c) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38. In some such methods, the D1 domain and the D2 domain each comprise a light chain immunoglobulin variable region comprising a set of three light chain complementarity determining region sequences LCDR1, LCDR2, and LCDR3, wherein the LCDR1 comprises the amino acid sequence of SEQ ID NO: 12, the LCDR2 comprises the amino acid sequence AAS (SEQ ID NO: 14), and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 16.

[0009] In some such methods, the D1 domain comprises: (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising theAttorney Docket No.057766 / 625782 amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some such methods, the D1 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some such methods, the D1 domain comprises a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 2. In some such methods, the D1 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some such methods, the D1 domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 22.

[0010] In some such methods, the D1 domain comprises a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 10. In some such methods, the D2 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some such methods, the D2 domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 32. In some such methods, the D2 domain comprises an LCVR comprising the amino acid sequence of SEQ ID NO: 10.

[0011] In some such methods, the bispecific antigen-binding molecule comprises: a D1 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3Attorney Docket No.057766 / 625782 comprising the amino acid sequence of SEQ ID NO: 16; and a D2 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some such methods, the D1 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and the D2 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 32 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10.

[0012] In some such methods, the bispecific antigen-binding molecule comprises: a D1 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; and a D2 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some such methods, the D1 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 22 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and the D2 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 32 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10.

[0013] In some such methods, the D1 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some such methods, the D1 domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 32. In some such methods, the D1 domain comprises an LCVR comprising the amino acid sequence of SEQ ID NO: 10.Attorney Docket No.057766 / 625782

[0014] In some such methods, the D2 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some such methods, the D2 domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2. In some such methods, the D2 domain comprises an LCVR comprising the amino acid sequence of SEQ ID NO: 10.

[0015] In some such methods, the bispecific antigen-binding molecule comprises: a D1 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; and a D2 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16.

[0016] In some such methods, the D1 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 32 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and the D2 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10.

[0017] In some such methods, the bispecific antigen-binding molecule is a bispecific antibody. In some such methods, the bispecific antibody comprises a human IgG heavy chain constant region. In some such methods, the human IgG heavy chain constant region comprises one or more modifications that increase binding to a neonatal Fc receptor (FcRn) and / or the human IgG heavy chain constant region comprises one or more modifications that decrease binding to an Fc-gamma receptor (FcγR). In some such methods, the human IgG heavy chain constant region is isotype IgG4 or IgG1. In some such methods, the human IgG heavy chain constant region comprises one or more modifications in a hinge region. In some such methods,Attorney Docket No.057766 / 625782 the human IgG heavy chain constant region comprises one or more modifications that reduce binding to an Fc receptor (e.g., one or more modifications in a hinge region and / or CH region).

[0018] In some such methods, the D1 comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 42 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; or a heavy chain comprising the amino acid sequence of SEQ ID NO: 46 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; or a heavy chain comprising the amino acid sequence of SEQ ID NO: 48 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; or a heavy chain comprising the amino acid sequence of SEQ ID NO: 52 and a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some such methods, the D1 comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 283 and a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some such methods, the D2 comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 44 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; or a heavy chain comprising the amino acid sequence of SEQ ID NO: 50 and a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some such methods, the D2 comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 285 and a light chain comprising the amino acid sequence of SEQ ID NO: 20.

[0019] In some such methods, the bispecific antigen-binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 42 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 44 and a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some such methods, the bispecific antigen-binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 46 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 44 and a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some such methods, the bispecific antigen-binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 48 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 50 and a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some suchAttorney Docket No.057766 / 625782 methods, the bispecific antigen-binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 52 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 50 and a light chain comprising the amino acid sequence of SEQ ID NO: 20.

[0020] In some such methods, the D1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 283 and a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some such methods, the D2 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 285 and a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some such methods, the bispecific antigen-binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 283 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 285 and a light chain comprising the amino acid sequence of SEQ ID NO: 20.

[0021] In some such methods, the bispecific antigen-binding molecule is a bispecific antibody having the amino acid sequences of REGN16334. In some such methods, the bispecific antigen-binding molecule is a bispecific antibody having the amino acid sequences of REGN16335. In some such methods, the bispecific antigen-binding molecule is a bispecific antibody having the amino acid sequences of REGN16431. In some such methods, the bispecific antigen-binding molecule is a bispecific antibody having the amino acid sequences of REGN16432. In some such methods, the bispecific antigen-binding molecule is a bispecific antibody having the amino acid sequences of REGN20484.

[0022] In some such methods, the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1- LCDR2-LCDR3 amino acid sequences set contained within an HCVR / LCVR amino acid sequence pair selected from the group consisting of SEQ ID NOS: 2 / 10, 22 / 10, 32 / 10, 183 / 184, 141 / 142, 151 / 268, 231 / 232, 160 / 161, 170 / 171, 173 / 174, 188 / 189, 199 / 200, 183 / 212, 183 / 215, 183 / 217, 211 / 184, 211 / 212, 211 / 215, 211 / 217, 213 / 184, 213 / 212, 213 / 215, 213 / 217, 214 / 184, 214 / 212, 214 / 215, 214 / 217, 216 / 184, 216 / 212, 216 / 215, 216 / 217, 224 / 225, 244 / 245, 251 / 252, 259 / 260, 264 / 265, and 259 / 265; (II) the HCVR and LCVR amino acid sequences contained within an HCVR / LCVR amino acid sequence pair selected from the group consisting of SEQ IDAttorney Docket No.057766 / 625782 NOS: 2 / 10, 22 / 10, 32 / 10, 183 / 184, 141 / 142, 151 / 268, 231 / 232, 160 / 161, 170 / 171, 173 / 174, 188 / 189, 199 / 200, 183 / 212, 183 / 215, 183 / 217, 211 / 184, 211 / 212, 211 / 215, 211 / 217, 213 / 184, 213 / 212, 213 / 215, 213 / 217, 214 / 184, 214 / 212, 214 / 215, 214 / 217, 216 / 184, 216 / 212, 216 / 215, 216 / 217, 224 / 225, 244 / 245, 251 / 252, 259 / 260, 264 / 265, and 259 / 265; or (III) the heavy chain and light chain amino acid sequences contained within a heavy chain / light chain amino acid sequence pair selected from the group consisting of SEQ ID NOS: 18 / 20, 30 / 20, 40 / 20, 185 / 186, 143 / 144, 152 / 153, 236 / 237, 162 / 163, 172 / 144, 175 / 176, 190 / 192, 191 / 192, 201 / 202, 218 / 153, 229 / 230, 233 / 237, 234 / 237, 235 / 237, 246 / 250, 247 / 250, 248 / 250, 249 / 250, 261 / 262, 266 / 267, and 261 / 267.

[0023] In some such methods, the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1- LCDR2-LCDR3 amino acid sequences set contained within an HCVR / LCVR amino acid sequence pair selected from the group consisting of SEQ ID NOS: 2 / 10, 22 / 10, and 32 / 10; (II) (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or (c) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; (III) (a) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 10; (b) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 22 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 10; (c) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 32 and an LCVRAttorney Docket No.057766 / 625782 comprising or consisting of the amino acid sequence of SEQ ID NO: 10; or (IV) (a) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 18 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 20; (b) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 30 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 20; or (c) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 40 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 20.

[0024] In some such methods, the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1- LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 183 / 184; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 177, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 178, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 179, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 180, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 181, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 182; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 183 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 184; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 185 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 186.

[0025] In some such methods, the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1- LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 141 / 142; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 135, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 136, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 137, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 138, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 139, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 140; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 141 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 142; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 143 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 144.Attorney Docket No.057766 / 625782

[0026] In some such methods, the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1- LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 151 / 268; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 145, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 146, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 147, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 148, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 149, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 150; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 151 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 268; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 152 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 153.

[0027] In some such methods, the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1- LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 231 / 232; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 269, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 220, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 221, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 222, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 223, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 150; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 231 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 232; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 236 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 237.

[0028] In some such methods, the CD40 inhibitor is a CD40L binding protein. In some such methods, (I) the CD40L binding protein comprises two Tn3 proteins fused to human serum albumin; or (II) the CD40L binding protein comprises the sequence set forth in SEQ ID NO: 280. In some such methods, the CD40L binding protein is an anti-CD40L antibody or antigen- binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 276 / 277; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO:Attorney Docket No.057766 / 625782 270, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 271, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 272, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 273, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 274, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 275; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 276 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 277; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 278 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 279.

[0029] In some such methods, the CD40 inhibitor (e.g., CD40 antigen-binding molecule) is administered simultaneously with the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent. In some such methods, the CD40 inhibitor (e.g., CD40 antigen-binding molecule) is administered prior to the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent. In some such methods, the CD40 inhibitor (e.g., CD40 antigen-binding molecule) is administered prior to and after the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent. In some such methods, the CD40 inhibitor (e.g., CD40 antigen- binding molecule) is administered within about 6 months after the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent. In some such methods, the CD40 inhibitor (e.g., CD40 antigen-binding molecule) is administered about 1 week prior to or within about 1 week prior to the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent.

[0030] In some such methods, the CD40 inhibitor (e.g., CD40 antigen-binding molecule) is administered simultaneously with the nucleic acid construct. In some such methods, the CD40 inhibitor (e.g., CD40 antigen-binding molecule) is administered prior to the nucleic acid construct. In some such methods, the CD40 inhibitor (e.g., CD40 antigen-binding molecule) is administered prior to and after the nucleic acid construct. In some such methods, the CD40 inhibitor (e.g., CD40 antigen-binding molecule) is administered within about 6 months after the nucleic acid construct, optionally wherein the nucleic acid construct is in a viral vector, and the CD40 inhibitor (e.g., CD40 antigen-binding molecule) is administered if the viral vector is still present in the subject. In some such methods, the CD40 inhibitor (e.g., CD40 antigen-binding molecule) is administered about 1 week prior to or within about 1 week prior to the nucleic acidAttorney Docket No.057766 / 625782 construct.

[0031] In some such methods, the nucleic acid construct is administered simultaneously with the nuclease agent or the one or more nucleic acids encoding the nuclease agent. In some such methods, the nucleic acid construct is administered prior to or after the nuclease agent or the one or more nucleic acids encoding the nuclease agent.

[0032] In some such methods, the nucleic acid construct is in the nucleic acid vector, optionally wherein the nucleic acid vector is a viral vector. In some such methods, the viral vector is administered at a dose of about 3E11 vg / kg to about 5E13 vg / kg. In some such methods, the nucleic acid vector is an adeno-associated viral (AAV) vector. In some such methods, the nucleic acid construct is flanked by inverted terminal repeats (ITRs) on each end, optionally wherein the ITR on at least one end comprises, consists essentially of, or consists of SEQ ID NO: 112, and optionally wherein the ITR on each end comprises, consists essentially of, or consists of SEQ ID NO: 112, or optionally wherein the ITR on at least one end comprises, consists essentially of, or consists of SEQ ID NO: 110, and optionally wherein the ITR on each end comprises, consists essentially of, or consists of SEQ ID NO: 110. In some such methods, the AAV vector is a single-stranded AAV (ssAAV) vector. In some such methods, the AAV vector is a recombinant AAV8 (rAAV8) vector.

[0033] In some such methods, the polypeptide of interest is a factor IX protein. In some such methods, the factor IX protein coding sequence encodes a factor IX protein comprising SEQ ID NO: 63. In some such methods, the factor IX protein coding sequence comprises or consists of SEQ ID NO: 61, or wherein the factor IX protein coding sequence comprises or consists of SEQ ID NO: 60. In some such methods, the nucleic acid construct is a bidirectional construct, wherein the factor IX protein coding sequence is a first factor IX protein coding sequence, and the bidirectional construct further comprises a reverse complement of a second factor IX protein coding sequence, wherein the first factor IX protein coding sequence and the second factor IX protein coding sequence are different but encode the same factor IX protein sequence. In some such methods, the nucleic acid construct comprises from 5’ to 3’: a first splice acceptor, the first factor IX protein coding sequence, a first polyadenylation signal, a reverse complement of a second polyadenylation signal, the reverse complement of the second factor IX protein coding sequence, and a reverse complement of a second splice acceptor, wherein: (i) the first factor IX protein coding sequence comprises SEQ ID NO: 60 and the second factor IX protein codingAttorney Docket No.057766 / 625782 sequence comprises SEQ ID NO: 61; or (ii) the first factor IX protein coding sequence comprises SEQ ID NO: 61 and the second factor IX protein coding sequence comprises SEQ ID NO: 60, wherein the nucleic acid construct does not comprise a promoter that drives the expression of the factor IX protein, and wherein the nucleic acid construct does not comprise homology arms. In some such methods, the nucleic acid construct comprises SEQ ID NO: 64 or 62 or the reverse complement thereof.

[0034] In some such methods, the polypeptide of interest is a multidomain therapeutic protein comprising a delivery domain fused to a lysosomal alpha-glucosidase. In some such methods, the lysosomal alpha-glucosidase comprises or consists of the sequence set forth in SEQ ID NO: 117. In some such methods, the lysosomal alpha-glucosidase coding sequence comprises or consist of the sequence set forth in SEQ ID NO: 127.

[0035] In some such methods, the delivery domain is a CD63-binding delivery domain. In some such methods, the CD63-binding delivery domain comprises an anti-CD63 antigen-binding protein. In some such methods, the CD63-binding delivery domain is a single-chain variable fragment (scFv). In some such methods, the scFv comprises or consists of the sequence set forth in SEQ ID NO: 119. In some such methods, the scFv coding sequence comprises or consists of the sequence set forth in SEQ ID NO: 129. In some such methods, the multidomain therapeutic protein comprises or consists of the sequence set forth in SEQ ID NO: 120. In some such methods, the coding sequence for the multidomain therapeutic protein comprises or consists of the sequence set forth in SEQ ID NO: 128, optionally wherein the nucleic acid construct comprises the sequence set forth in SEQ ID NO: 134 or 132. In some such methods, the nucleic acid construct comprises from 5’ to 3’: a splice acceptor, the coding sequence for the multidomain therapeutic protein, and a polyadenylation signal or sequence, wherein the coding sequence for the multidomain therapeutic protein comprises the sequence set forth in SEQ ID NO: 128, optionally wherein the nucleic acid construct comprises the sequence set forth in SEQ ID NO: 134 or 132, wherein the polyadenylation signal comprises a BGH polyadenylation signal and a unidirectional SV40 late polyadenylation signal, wherein the nucleic acid construct does not comprise a promoter that drives the expression of the multidomain therapeutic protein, and wherein the nucleic acid construct does not comprise a homology arm.

[0036] In some such methods, the delivery domain is a TfR-binding delivery domain. In some such methods, the TfR-binding delivery domain comprises an anti-TfR antigen-bindingAttorney Docket No.057766 / 625782 protein. In some such methods, the TfR-binding delivery domain comprises a single-chain variable fragment (scFv). In some such methods, the scFv comprises or consists of the sequence set forth in SEQ ID NO: 121. In some such methods, the scFv coding sequence comprises or consists of the sequence set forth in SEQ ID NO: 122. In some such methods, the multidomain therapeutic protein comprises or consists of the sequence set forth in SEQ ID NO: 126. In some such methods, the coding sequence for the multidomain therapeutic protein comprises or consists of the sequence set forth in SEQ ID NO: 125, optionally wherein the nucleic acid construct comprises the sequence set forth in SEQ ID NO: 133 or 130. In some such methods, the nucleic acid construct comprises from 5’ to 3’: a splice acceptor, the coding sequence for the multidomain therapeutic protein, and a polyadenylation signal or sequence, wherein the coding sequence for the multidomain therapeutic protein comprises the sequence set forth in SEQ ID NO: 125, optionally wherein the nucleic acid construct comprises the sequence set forth in SEQ ID NO: 133 or 130, wherein the polyadenylation signal comprises a BGH polyadenylation signal and a unidirectional SV40 late polyadenylation signal, wherein the nucleic acid construct does not comprise a promoter that drives the expression of the multidomain therapeutic protein, and wherein the nucleic acid construct does not comprise a homology arm.

[0037] In some such methods, the polypeptide of interest is a factor VIII protein. In some such methods, the polypeptide of interest is an antigen-binding protein, optionally wherein the antigen-binding protein is an antibody.

[0038] In some such methods, the target genomic locus is an albumin gene, optionally wherein the albumin gene is a human albumin gene. In some such methods, the nuclease target site is in intron 1 of the albumin gene.

[0039] In some such methods, the nuclease agent comprises: (a) a zinc finger nuclease (ZFN); (b) a transcription activator-like effector nuclease (TALEN); or (c) (i) a Cas protein or a nucleic acid encoding the Cas protein; and (ii) a guide RNA or one or more DNAs encoding the guide RNA, wherein the guide RNA comprises a DNA-targeting segment that targets a guide RNA target sequence, and wherein the guide RNA binds to the Cas protein and targets the Cas protein to the guide RNA target sequence. In some such methods, the nuclease agent comprises: (a) a Cas protein or a nucleic acid encoding the Cas protein; and (b) a guide RNA or one or more DNAs encoding the guide RNA, wherein the guide RNA comprises a DNA-targeting segment that targets a guide RNA target sequence, and wherein the guide RNA binds to the Cas proteinAttorney Docket No.057766 / 625782 and targets the Cas protein to the guide RNA target sequence.

[0040] In some such methods, the DNA-targeting segment comprises or consists of SEQ ID NO: 96. In some such methods, the guide RNA comprises SEQ ID NO: 100 or 104. In some such methods, the method comprises administering the guide RNA in the form of RNA. In some such methods, the guide RNA comprises at least one modification. In some such methods, the at least one modification comprises: (i) phosphorothioate bonds between the first four nucleotides at the 5’ end of the guide RNA; (ii) phosphorothioate bonds between the last four nucleotides at the 3’ end of the guide RNA; (iii) 2’-O-methyl-modified nucleotides at the first three nucleotides at the 5’ end of the guide RNA; and (iv) 2’-O-methyl-modified nucleotides at the last three nucleotides at the 3’ end of the guide RNA. In some such methods, the method comprises administering the guide RNA in the form of RNA, the guide RNA comprises SEQ ID NO: 104, and the guide RNA comprises: (i) phosphorothioate bonds between the first four nucleotides at the 5’ end of the guide RNA; (ii) phosphorothioate bonds between the last four nucleotides at the 3’ end of the guide RNA; (iii) 2’-O-methyl-modified nucleotides at the first three nucleotides at the 5’ end of the guide RNA; and (iv) 2’-O-methyl-modified nucleotides at the last three nucleotides at the 3’ end of the guide RNA.

[0041] In some such methods, the Cas protein is a Cas9 protein, optionally wherein the Cas protein is derived from a Streptococcus pyogenes Cas9 protein. In some such methods, the Cas protein comprises the sequence set forth in SEQ ID NO: 75. In some such methods, the method comprises administering the nucleic acid encoding the Cas protein, wherein the nucleic acid comprises an mRNA encoding the Cas protein. In some such methods, the mRNA encoding the Cas protein comprises at least one modification. In some such methods, the mRNA encoding the Cas protein is fully substituted with N1-methyl-pseudouridine. In some such methods, the mRNA encoding the Cas protein comprises the sequence set forth in SEQ ID NO: 65 or 66. In some such methods, the method comprises administering the nucleic acid encoding the Cas protein, wherein the nucleic acid comprises an mRNA encoding the Cas protein, the mRNA encoding the Cas protein comprises the sequence set forth in SEQ ID NO: 65 or 66, and the mRNA encoding the Cas protein is fully substituted with N1-methyl-pseudouridine, comprises a 5’ cap, and comprises a poly(A) tail. In some such methods, the method comprises administering the guide RNA in the form of RNA, and the guide RNA comprises SEQ ID NO: 100 or 104, and wherein the method comprises administering the nucleic acid encoding the Cas protein, whereinAttorney Docket No.057766 / 625782 the nucleic acid comprises an mRNA encoding the Cas protein, and the mRNA encoding the Cas protein comprises the sequence set forth in SEQ ID NO: 65 or 66. In some such methods, the method comprises administering the guide RNA in the form of RNA, the guide RNA comprises SEQ ID NO: 104, and the guide RNA comprises: (i) phosphorothioate bonds between the first four nucleotides at the 5’ end of the guide RNA; (ii) phosphorothioate bonds between the last four nucleotides at the 3’ end of the guide RNA; (iii) 2’-O-methyl-modified nucleotides at the first three nucleotides at the 5’ end of the guide RNA; and (iv) 2’-O-methyl-modified nucleotides at the last three nucleotides at the 3’ end of the guide RNA, and wherein the method comprises administering the nucleic acid encoding the Cas protein, wherein the nucleic acid comprises an mRNA encoding the Cas protein, the mRNA encoding the Cas protein comprises the sequence set forth in SEQ ID NO: 65 or 66, and the mRNA encoding the Cas protein is fully substituted with N1-methyl-pseudouridine, comprises a 5’ cap, and comprises a poly(A) tail.

[0042] In some such methods, the Cas protein or the nucleic acid encoding the Cas protein and the guide RNA or the one or more DNAs encoding the guide RNA are associated with a lipid nanoparticle. In some such methods, the lipid nanoparticle comprises a cationic lipid, a neutral lipid, a helper lipid, and a stealth lipid. In some such methods, the cationic lipid is Lipid A ((9Z,12Z)-3-((4,4-bis(octyloxy)butanoyl)oxy)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propyl octadeca-9,12-dienoate), and / or the neutral lipid is distearoylphosphatidylcholine or 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), and / or wherein the helper lipid is cholesterol, and / or wherein the stealth lipid is 1,2-dimyristoyl- rac-glycero-3-methoxypolyethylene glycol-2000. In some such methods, the cationic lipid is Lipid A, the neutral lipid is DSPC, the helper lipid is cholesterol, and the stealth lipid is PEG2k- DMG. In some such methods, the lipid nanoparticle comprises four lipids at the following molar ratios: about 50 mol% Lipid A, about 9 mol% DSPC, about 38 mol% cholesterol, and about 3 mol% PEG2k-DMG.

[0043] In some such methods, the cell is a liver cell or a hepatocyte, or the population of cells is a population of liver cells or hepatocytes. In some such methods, the subject is a human subject. In some such methods, the subject is a neonatal subject. In some such methods, the method further comprises determining whether the subject has immunity against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or the delivery vehicle for the nucleic acid construct, the nuclease agent, orAttorney Docket No.057766 / 625782 the one or more nucleic acids encoding the nuclease agent prior to the administering, optionally wherein the determining comprises determining the presence of neutralizing antibodies against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or the delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent. In some such methods, the nucleic acid construct is in an adeno-associated viral (AAV) vector, and wherein the subject does not have preexisting AAV immunity (e.g., at the time the CD40 inhibitor is administered). In some such methods, the subject does not have preexisting AAV immunity (i.e., naïve or seronegative). In some such methods, the subject has preexisting AAV immunity.

[0044] In some such methods, the method further comprises administering to the subject a plasma cell depleting agent at a time when the subject has preexisting immunity against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or a delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent when the plasma cell depleting agent is administered, optionally wherein the subject has preexisting AAV immunity when the plasma cell depleting agent is administered. In some such methods, the plasma cell depleting agent is capable of depleting long-lived plasma cells (LLPC). In some such methods, the plasma cell depleting agent is a B cell maturation antigen (BCMA) targeting agent. In some such methods, the BCMA targeting agent is a chimeric antigen receptor against BCMA or an anti-BCMA antibody or a functional fragment thereof. In some such methods, the anti- BCMA antibody or functional fragment thereof is conjugated to a cytotoxic agent. In some such methods, the anti-BCMA antibody is a multispecific antibody or a functional fragment thereof.

[0045] In some such methods, the multispecific anti-BCMA antibody or functional fragment thereof targets BCMA and CD3. In some such methods, the multispecific anti-BCMA antibody or functional fragment thereof is anti-BCMAxCD3 bispecific antibody or functional fragment thereof. In some such methods, the anti-BCMAxCD3 bispecific antibody is selected from linvoseltamab (REGN5458), REGN5459, pacanalotamab (AMG420), teclistamab (JNJ- 64007957), AMG701, alnuctamab (CC-93269), EM801, EM901, elranatamab (PF-06863135), TNB383B (ABBV-383), and TNB384B. In some such methods, the anti-BCMAxCD3 bispecific antibody or functional fragment thereof comprises a first antigen-binding domain that specifically binds to BCMA comprising three heavy chain CDRs (HCDR1, HCDR2 andAttorney Docket No.057766 / 625782 HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 287, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 303. In some such methods, the first antigen-binding domain that specifically binds to BCMA comprises a HCDR1 comprising the amino acid sequence of SEQ ID NO: 289, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 291, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 293, a LCDR1 comprising the amino acid sequence of SEQ ID NO: 305, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 307, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 309. In some such methods, the anti- BCMAxCD3 bispecific antibody or functional fragment thereof comprises a second antigen- binding domain that specifically binds to CD3 comprising three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence selected from the group consisting of SEQ ID NOS: 311 and 319, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 303. In some such methods, the second antigen-binding domain that specifically binds to CD3 comprises a HCDR1 comprising the amino acid sequence of SEQ ID NO: 313 or 321, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 315 or 323, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 317 or 325, a LCDR1 comprising the amino acid sequence of SEQ ID NO: 305, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 307, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 309. In some such methods, the anti-BCMAxCD3 bispecific antibody or functional fragment thereof comprises: (a) a first antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 289, 291, and 293, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 305, 307, and 309, respectively; and (b) a second antigen- binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 313, 315, and 317, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 305, 307, and 319, respectively. In some such methods, the anti-BCMAxCD3 bispecific antibody or functional fragment thereof comprises: (a) a first antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 289, 291, and 293, respectively, andAttorney Docket No.057766 / 625782 LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 305, 307, and 309, respectively; and (b) a second antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 321, 323, and 325, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 305, 307, and 319, respectively. In some such methods, the anti-BCMAxCD3 bispecific antibody or functional fragment thereof comprises a human IgG heavy chain constant region. In some such methods, the human IgG heavy chain constant region is isotype IgG4 or IgG1. In some such methods, the human IgG heavy chain constant region comprises one or more modifications that increase binding to a neonatal Fc receptor (FcRn) and / or the human IgG heavy chain constant region comprises one or more modifications that decrease binding to an Fc- gamma receptor (FcγR).

[0046] In some such methods, the method further comprises administering to the subject an effective amount of a B cell depleting agent (i.e., in combination with the plasma cell depleting agent) and / or an immunoglobulin depleting agent (i.e., in combination with the plasma cell depleting agent) at a time when the subject has preexisting immunity against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or a delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent when the B cell depleting agent and / or the immunoglobulin depleting agent is administered, optionally wherein the subject has preexisting AAV immunity when the B cell depleting agent and / or the immunoglobulin depleting agent is administered. In some such methods, the method further comprises administering to the subject an effective amount of a B cell depleting agent (i.e., in combination with the plasma cell depleting agent) and an immunoglobulin depleting agent (i.e., in combination with the plasma cell depleting agent) at a time when the subject has preexisting immunity against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or a delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent when the B cell depleting agent and the immunoglobulin depleting agent are administered, optionally wherein the subject has preexisting AAV immunity when the B cell depleting agent and the immunoglobulin depleting agent are administered. In some such methods, the B cell depleting agent is administered before, at the same time as, or after the plasma cell depleting agent. In some such methods, theAttorney Docket No.057766 / 625782 immunoglobulin depleting agent is administered after the plasma cell depleting agent. In some such methods, the B cell depleting agent is administered prior to and after the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent. In some such methods, the immunoglobulin depleting agent is administered prior to and after the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent. In some such methods, the B cell depleting agent is administered prior to and after the nucleic acid construct. In some such methods, the immunoglobulin depleting agent is administered prior to and after the nucleic acid construct. In some such methods, the immunoglobulin depleting agent is administered after an initial dose of the plasma cell depleting agent, or wherein the immunoglobulin depleting agent is administered after an initial dose of the plasma cell depleting agent and after an initial dose of the B cell depleting agent. In some such methods, the B cell depleting agent is capable of depleting B cells and plasma cells that express low levels of BCMA. In some such methods, the B cell depleting agent is an agent that binds to a B cell surface molecule. In some such methods, the B cell depleting agent is selected from an anti-CD19 antibody, an anti-CD20 antibody, an anti-CD19 antibody and an anti-CD20 antibody, an anti-CD22 antibody, an anti-CD79 antibody, an anti-CD20xCD3 bispecific antibody, an anti- CD19xCD3 bispecific antibody, an anti-CD22xCD3 bispecific antibody, an anti-CD79xCD3 bispecific antibody, functional fragments of any of said antibodies, and any combinations thereof. In some such methods, the B cell depleting agent is an anti-CD20 antibody or a functional fragment thereof, wherein the anti-CD20 antibody is a multispecific antibody or a functional fragment thereof. In some such methods, the multispecific anti-CD20 antibody or functional fragment thereof targets CD20 and CD3. In some such methods, the multispecific anti-CD20 antibody or functional fragment thereof is anti-CD20xCD3 bispecific antibody or functional fragment thereof. In some such methods, the anti-CD20xCD3 bispecific antibody or functional fragment thereof comprises a first antigen-binding domain that specifically binds to CD20 comprising three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 329, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 330. In some such methods, the first antigen-binding domain that specifically binds to CD20 comprises a HCDR1 comprising the amino acid sequence of SEQ ID NO: 332, a HCDR2 comprising the amino acidAttorney Docket No.057766 / 625782 sequence of SEQ ID NO: 333, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 334, a LCDR1 comprising the amino acid sequence of SEQ ID NO: 335, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 336, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 337. In some such methods, the anti-CD20xCD3 bispecific antibody or functional fragment thereof comprises a second antigen-binding domain that specifically binds to CD3 comprising three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 331, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 330. In some such methods, the second antigen-binding domain that specifically binds to CD3 comprises a HCDR1 comprising the amino acid sequence of SEQ ID NO: 338, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 339, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 340, a LCDR1 comprising the amino acid sequence of SEQ ID NO: 335, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 336, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 337. In some such methods, the anti-CD20xCD3 bispecific antibody or functional fragment thereof comprises: (a) a first antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 332, 333, and 334, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 335, 336, and 337, respectively; and (b) a second antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 338, 339, and 340, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 335, 336, and 337, respectively. In some such methods, the anti-CD20xCD3 bispecific antibody or functional fragment thereof comprises a human IgG heavy chain constant region. In some such methods, the human IgG heavy chain constant region is isotype IgG4 or IgG1. In some such methods, the human IgG heavy chain constant region comprises one or more modifications that increase binding to a neonatal Fc receptor (FcRn) and / or the human IgG heavy chain constant region comprises one or more modifications that decrease binding to an Fc-gamma receptor (FcγR). In some such methods, the B cell depleting agent is an agent targeting a B cell survival factor. In some such methods, the B cell depleting agent is a BLyS / BAFF inhibitor, an APRIL inhibitor, a BLyS receptor 3 / BAFF receptor inhibitor, or any combination thereof.Attorney Docket No.057766 / 625782

[0047] In some such methods, the immunoglobulin depleting agent is capable of accelerating IgG clearance. In some such methods, the immunoglobulin depleting agent is a neonatal Fc receptor (FcRn) blocker. In some such methods, the FcRn blocker is selected from Efgartigimod (ARGX-113), Rozanolixizumab (UCB7665), Batoclimab (RVT-1401), IMVT-1402, Nipocalimab (M281), Orilanolimab (SYNT001), and any combinations thereof.

[0048] In some such methods, the immunoglobulin depleting agent is an immunoglobulin degrading enzyme. In some such methods, the CD40 inhibitor is resistant to the immunoglobulin degrading enzyme. In some such methods, the immunoglobulin degrading enzyme is selected from Imlifidase / IdeS / Fabricator, IdeZ, IdeXork, IceMG, CYR-212, CYR-241, S-1117, HNSA- 5487, and IdeE / KJ103. In some such methods, the immunoglobulin degrading enzyme is Imlifidase / IdeS / Fabricator. In some such methods, the CD40 inhibitor is administered before administration of the immunoglobulin depleting agent to the subject, and the immunoglobulin depleting agent is administered before the administration of the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent to the subject. In some such methods, the subject does not have preexisting immunity against the immunoglobulin degrading enzyme. In some such methods, the subject has preexisting immunity against the immunoglobulin degrading enzyme.

[0049] In some such methods, the method further comprises plasmapheresis, therapeutic plasma exchange, or immunoadsorption.

[0050] In some such methods, the plasma cell depleting agent is administered simultaneously with the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent. In some such methods, the plasma cell depleting agent is administered prior to the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent. In some such methods, the plasma cell depleting agent is administered prior to and after the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent. In some such methods, the plasma cell depleting agent is administered within about 6 months after the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent. In some such methods, the plasma cell depleting agent is administered about 1 week prior to or within about 1 week prior to the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent.Attorney Docket No.057766 / 625782

[0051] In some such methods, the plasma cell depleting agent is administered simultaneously with the nucleic acid construct. In some such methods, the plasma cell depleting agent is administered prior to the nucleic acid construct. In some such methods, the plasma cell depleting agent is administered prior to and after the nucleic acid construct. In some such methods, the plasma cell depleting agent is administered within about 6 months after the nucleic acid construct, optionally wherein the nucleic acid construct is in a viral vector, and the plasma cell depleting agent is administered if the viral vector is still present in the subject. In some such methods, the plasma cell depleting agent is administered about 1 week prior to or within about 1 week prior to the nucleic acid construct.

[0052] In some such methods, the nucleic acid construct is administered simultaneously with the nuclease agent or the one or more nucleic acids encoding the nuclease agent. In some such methods, the nucleic acid construct is administered prior to or after the nuclease agent or the one or more nucleic acids encoding the nuclease agent.

[0053] In some such methods, the subject has preexisting immunity against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or a delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent, wherein the plasma cell depleting agent and the CD40 inhibitor are both administered prior to the nucleic acid construct and the nuclease agent or the one or more nucleic acids encoding the nuclease agent, and the plasma cell depleting agent is administered prior to the CD40 inhibitor. In some such methods, the subject has preexisting immunity against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or a delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent, wherein the plasma cell depleting agent and the CD40 inhibitor are both administered prior to the nucleic acid construct and the nuclease agent or the one or more nucleic acids encoding the nuclease agent, and the plasma cell depleting agent and the CD40 inhibitor are administered simultaneously, optionally wherein the subject has ongoing B cell responses to the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or the delivery vehicle when the plasma cell depleting agent and the CD40 inhibitor are administered. In some such methods, the subject does not have preexisting immunity against the nucleic acid construct, the polypeptide of interest, the nucleaseAttorney Docket No.057766 / 625782 agent, the one or more nucleic acids encoding the nuclease agent, or a delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent before administration of the nucleic acid construct and the nuclease agent or the one or more nucleic acids encoding the nuclease agent, the CD40 inhibitor is administered prior to the nucleic acid construct and the nuclease agent or the one or more nucleic acids encoding the nuclease agent, the subject develops immunity against the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or the delivery vehicle from the administration of the nucleic acid construct and the nuclease agent or the one or more nucleic acids encoding the nuclease agent, and the plasma cell depleting agent is administered after the administration of the nucleic acid construct and the nuclease agent or the one or more nucleic acids encoding the nuclease agent but prior to a second dose of the nucleic acid construct, a second dose of the nuclease agent or the one or more nucleic acids encoding the nuclease agent, a second nucleic acid construct, and / or a second nuclease agent or one or more nucleic acids encoding the second nuclease agent. Optionally, the plasma cell depleting agent is administered in combination with a B cell depleting agent and / or an immunoglobulin depleting agent (i.e., at a time when the subject has preexisting immunity against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or the delivery vehicle).

[0054] In some such methods, the nucleic acid construct is in an AAV vector, the subject has preexisting immunity against the AAV vector, and the plasma cell depleting agent and the CD40 inhibitor are both administered prior to the nucleic acid construct, wherein the plasma cell depleting agent is administered prior to the CD40 inhibitor. Optionally, the plasma cell depleting agent is administered in combination with a B cell depleting agent and / or an immunoglobulin depleting agent (i.e., when the subject has preexisting immunity against the AAV vector). In some such methods, the nucleic acid construct is in an AAV vector, the subject has preexisting immunity against the AAV vector, and the plasma cell depleting agent and the CD40 inhibitor are both administered prior to the nucleic acid construct, wherein the plasma cell depleting agent and the CD40 inhibitor are administered simultaneously, optionally wherein the subject has ongoing B cell responses to the AAV vector when the plasma cell depleting agent and the CD40 inhibitor are administered. Optionally, the plasma cell depleting agent is administered in combination with a B cell depleting agent and / or an immunoglobulin depleting agent (i.e., whenAttorney Docket No.057766 / 625782 the subject has preexisting immunity against the AAV vector). In some such methods, the nucleic acid construct is in an AAV vector, the subject does not have preexisting immunity against the AAV vector before administration of a first dose of the nucleic acid construct, the CD40 inhibitor is administered prior to the first dose of the nucleic acid construct, the subject develops immunity against the AAV vector from the first dose of the nucleic acid construct, and the plasma cell depleting agent is administered after the first dose of the nucleic acid construct but prior to a second dose of the nucleic acid construct. Optionally, the plasma cell depleting agent is administered in combination with a B cell depleting agent and / or an immunoglobulin depleting agent (i.e., when the subject has preexisting immunity against the AAV vector). In some such methods, the nucleic acid construct is in an AAV vector, the subject does not have preexisting immunity against the AAV vector before administration of the nucleic acid construct, the CD40 inhibitor is administered prior to the nucleic acid construct, the subject develops immunity against the AAV vector from administration of the nucleic acid construct, and the plasma cell depleting agent is administered after the nucleic acid construct but prior to administration of a second nucleic acid construct (e.g., in an AAV vector, e.g., an AAV vector of the same serotype as the first nucleic acid construct). Optionally, the plasma cell depleting agent is administered in combination with a B cell depleting agent and / or an immunoglobulin depleting agent (i.e., when the subject has preexisting immunity against the AAV vector).

[0055] In another aspect, provided are compositions or combinations comprising a CD40 inhibitor (e.g., CD40 antigen-binding molecule) in combination with: (a) a nucleic acid construct comprising a coding sequence for the polypeptide of interest; and (b) a nuclease agent or one or more nucleic acids encoding the nuclease agent, wherein the nuclease agent targets a nuclease target site in a target genomic locus.

[0056] In some such compositions or combinations, the CD40 inhibitor is a CD40 antigen- binding molecule. In some such compositions or combinations, the CD40 antigen-binding molecule is a monospecific antigen-binding molecule. In some such compositions or combinations, the CD40 antigen-binding molecule is a bispecific antigen-binding molecule comprising: (a) a first antigen-binding domain (D1) that binds a first epitope of human CD40; and (b) a second antigen-binding domain (D2) that binds a second epitope of human CD40. In some such compositions or combinations, the bispecific antigen-binding molecule: (i) binds human CD40 with a KD of less than 25 nM as measured by surface plasmon resonance at 25°C;Attorney Docket No.057766 / 625782 (ii) binds human CD40 with a KD of less than 70 nM as measured by surface plasmon resonance at 37°C; (iii) binds human CD40 with a dissociative half-life (t1 / 2) of greater than 75 minutes as measured by surface plasmon resonance at 25°C; (iv) binds a human CD40-expressing cell with an EC50 value of about 10 nM or less; (v) inhibits binding of human CD40 dimer to CD40L; (vi) inhibits CD40 ligand (CD40L)-induced activation; and / or (vii) does not significantly agonize CD40 in the absence of CD40L. In some such compositions or combinations, the bispecific antigen-binding molecule: (i) binds human CD40 with a KDof less than 25 nM as measured by surface plasmon resonance at 25°C; (ii) binds human CD40 with a KD of less than 70 nM as measured by surface plasmon resonance at 37°C; (iii) binds human CD40 with a dissociative half-life (t1 / 2) of greater than 75 minutes as measured by surface plasmon resonance at 25°C; (iv) binds a human CD40-expressing cell with an EC50 value of about 10 nM or less; (v) inhibits binding of human CD40 monomer to CD40L; (vi) inhibits CD40 ligand (CD40L)-induced activation; and / or (vii) does not significantly agonize CD40 in the absence of CD40L. In some such compositions or combinations, the bispecific antigen-binding molecule inhibits CD40L- induced activation. In some such compositions or combinations, the bispecific antigen-binding molecule inhibits CD40L-induced activation and does not significantly agonize CD40 in the absence of CD40L. In some such compositions or combinations, the D1 domain and the D2 domain each comprise a heavy chain immunoglobulin variable region comprising a set of three heavy chain complementarity determining region sequences HCDR1, HCDR2, and HCDR3 independently selected from the group consisting of: (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8; (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28; and (c) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38. In some such compositions or combinations, the D1 domain and the D2 domain each comprise a light chain immunoglobulin variable region comprising a set of three light chain complementarity determining region sequences LCDR1, LCDR2, and LCDR3, wherein the LCDR1 comprises the amino acid sequence of SEQ ID NO: 12, the LCDR2 comprises the amino acid sequence AAS (SEQ ID NO: 14), and the LCDR3 comprisesAttorney Docket No.057766 / 625782 the amino acid sequence of SEQ ID NO: 16.

[0057] In some such compositions or combinations, the D1 domain comprises: (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some such compositions or combinations, the D1 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some such compositions or combinations, the D1 domain comprises a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 2. In some such compositions or combinations, the D1 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some such compositions or combinations, the D1 domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 22.

[0058] In some such compositions or combinations, the D1 domain comprises a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 10. In some such compositions or combinations, the D2 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequenceAttorney Docket No.057766 / 625782 AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some such compositions or combinations, the D2 domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 32. In some such compositions or combinations, the D2 domain comprises an LCVR comprising the amino acid sequence of SEQ ID NO: 10.

[0059] In some such compositions or combinations, the bispecific antigen-binding molecule comprises: a D1 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; and a D2 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some such compositions or combinations, the D1 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and the D2 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 32 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10. In some such compositions or combinations, the bispecific antigen- binding molecule comprises: a D1 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; and a D2 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some such compositions or combinations, the D1 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 22 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and the D2 comprisesAttorney Docket No.057766 / 625782 an HCVR comprising the amino acid sequence of SEQ ID NO: 32 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10.

[0060] In some such compositions or combinations, the D1 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some such compositions or combinations, the D1 domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 32. In some such compositions or combinations, the D1 domain comprises an LCVR comprising the amino acid sequence of SEQ ID NO: 10.

[0061] In some such compositions or combinations, the D2 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some such compositions or combinations, the D2 domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2. In some such compositions or combinations, the D2 domain comprises an LCVR comprising the amino acid sequence of SEQ ID NO: 10.

[0062] In some such compositions or combinations, the bispecific antigen-binding molecule comprises: a D1 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; and a D2 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16.Attorney Docket No.057766 / 625782

[0063] In some such compositions or combinations, the D1 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 32 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and the D2 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10.

[0064] In some such compositions or combinations, the bispecific antigen-binding molecule is a bispecific antibody. In some such compositions or combinations, the bispecific antibody comprises a human IgG heavy chain constant region. In some such compositions or combinations, the human IgG heavy chain constant region comprises one or more modifications that increase binding to a neonatal Fc receptor (FcRn) and / or the human IgG heavy chain constant region comprises one or more modifications that decrease binding to an Fc-gamma receptor (FcγR). In some such compositions or combinations, the human IgG heavy chain constant region is isotype IgG4 or IgG1. In some such compositions or combinations, the human IgG heavy chain constant region comprises one or more modifications in a hinge region. In some such compositions or combinations, the human IgG heavy chain constant region comprises one or more modifications that reduce binding to an Fc receptor (e.g., one or more modifications in a hinge region and / or CH region). In some such compositions or combinations, the D1 comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 42 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; or a heavy chain comprising the amino acid sequence of SEQ ID NO: 46 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; or a heavy chain comprising the amino acid sequence of SEQ ID NO: 48 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; or a heavy chain comprising the amino acid sequence of SEQ ID NO: 52 and a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some such compositions or combinations, the D1 comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 283 and a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some such compositions or combinations, the D2 comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 44 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; or a heavy chain comprising the amino acid sequence of SEQ ID NO: 50 and a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some such compositions or combinations, the D2 comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 285 and a light chain comprising the amino acid sequence of SEQ ID NO: 20.Attorney Docket No.057766 / 625782

[0065] In some such compositions or combinations, the bispecific antigen-binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 42 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 44 and a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some such compositions or combinations, the bispecific antigen-binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 46 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 44 and a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some such compositions or combinations, the bispecific antigen-binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 48 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 50 and a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some such compositions or combinations, the bispecific antigen-binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 52 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 50 and a light chain comprising the amino acid sequence of SEQ ID NO: 20.

[0066] In some such compositions or combinations, the D1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 283 and a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some such compositions or combinations, the D2 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 285 and a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some such compositions or combinations, the bispecific antigen-binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 283 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 285 and a light chain comprising the amino acid sequence of SEQ ID NO: 20.

[0067] In some such compositions or combinations, the bispecific antigen-binding moleculeAttorney Docket No.057766 / 625782 is a bispecific antibody having the amino acid sequences of REGN16334. In some such compositions or combinations, the bispecific antigen-binding molecule is a bispecific antibody having the amino acid sequences of REGN16335. In some such compositions or combinations, the bispecific antigen-binding molecule is a bispecific antibody having the amino acid sequences of REGN16431. In some such compositions or combinations, the bispecific antigen-binding molecule is a bispecific antibody having the amino acid sequences of REGN16432. In some such compositions or combinations, the bispecific antigen-binding molecule is a bispecific antibody having the amino acid sequences of REGN20484.

[0068] In some such compositions or combinations, the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2- HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within an HCVR / LCVR amino acid sequence pair selected from the group consisting of SEQ ID NOS: 2 / 10, 22 / 10, 32 / 10, 183 / 184, 141 / 142, 151 / 268, 231 / 232, 160 / 161, 170 / 171, 173 / 174, 188 / 189, 199 / 200, 183 / 212, 183 / 215, 183 / 217, 211 / 184, 211 / 212, 211 / 215, 211 / 217, 213 / 184, 213 / 212, 213 / 215, 213 / 217, 214 / 184, 214 / 212, 214 / 215, 214 / 217, 216 / 184, 216 / 212, 216 / 215, 216 / 217, 224 / 225, 244 / 245, 251 / 252, 259 / 260, 264 / 265, and 259 / 265; (II) the HCVR and LCVR amino acid sequences contained within an HCVR / LCVR amino acid sequence pair selected from the group consisting of SEQ ID NOS: 2 / 10, 22 / 10, 32 / 10, 183 / 184, 141 / 142, 151 / 268, 231 / 232, 160 / 161, 170 / 171, 173 / 174, 188 / 189, 199 / 200, 183 / 212, 183 / 215, 183 / 217, 211 / 184, 211 / 212, 211 / 215, 211 / 217, 213 / 184, 213 / 212, 213 / 215, 213 / 217, 214 / 184, 214 / 212, 214 / 215, 214 / 217, 216 / 184, 216 / 212, 216 / 215, 216 / 217, 224 / 225, 244 / 245, 251 / 252, 259 / 260, 264 / 265, and 259 / 265; or (III) the heavy chain and light chain amino acid sequences contained within a heavy chain / light chain amino acid sequence pair selected from the group consisting of SEQ ID NOS: 18 / 20, 30 / 20, 40 / 20, 185 / 186, 143 / 144, 152 / 153, 236 / 237, 162 / 163, 172 / 144, 175 / 176, 190 / 192, 191 / 192, 201 / 202, 218 / 153, 229 / 230, 233 / 237, 234 / 237, 235 / 237, 246 / 250, 247 / 250, 248 / 250, 249 / 250, 261 / 262, 266 / 267, and 261 / 267.

[0069] In some such compositions or combinations, the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2- HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within an HCVR / LCVR amino acid sequence pair selected from the group consisting of SEQ ID NOS: 2 / 10, 22 / 10, and 32 / 10; (II) (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2Attorney Docket No.057766 / 625782 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or (c) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; (III) (a) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 10; (b) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 22 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 10; (c) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 32 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 10; or (IV) (a) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 18 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 20; (b) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 30 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 20; or (c) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 40 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 20.

[0070] In some such compositions or combinations, the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2- HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 183 / 184; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 177, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 178, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 179, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 180, an LCDR2 comprising theAttorney Docket No.057766 / 625782 amino acid sequence of SEQ ID NO: 181, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 182; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 183 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 184; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 185 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 186.

[0071] In some such compositions or combinations, the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2- HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 141 / 142; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 135, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 136, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 137, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 138, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 139, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 140; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 141 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 142; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 143 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 144.

[0072] In some such compositions or combinations, the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2- HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 151 / 268; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 145, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 146, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 147, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 148, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 149, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 150; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 151 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 268; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 152 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 153.

[0073] In some such compositions or combinations, the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-Attorney Docket No.057766 / 625782 HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 231 / 232; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 269, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 220, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 221, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 222, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 223, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 150; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 231 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 232; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 236 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 237.

[0074] In some such compositions or combinations, the CD40 inhibitor is a CD40L binding protein. In some such compositions or combinations, (I) the CD40L binding protein comprises two Tn3 proteins fused to human serum albumin; or (II) the CD40L binding protein comprises the sequence set forth in SEQ ID NO: 280. In some such compositions or combinations, the CD40L binding protein is an anti-CD40L antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 276 / 277; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 270, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 271, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 272, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 273, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 274, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 275; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 276 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 277; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 278 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 279.

[0075] In some such compositions or combinations, the nucleic acid construct is in the nucleic acid vector. In some such compositions or combinations, the nucleic acid vector is a viral vector. In some such compositions or combinations, the nucleic acid vector is an adeno- associated viral (AAV) vector. In some such compositions or combinations, the nucleic acid construct is flanked by inverted terminal repeats (ITRs) on each end, optionally wherein the ITRAttorney Docket No.057766 / 625782 on at least one end comprises, consists essentially of, or consists of SEQ ID NO: 112, and optionally wherein the ITR on each end comprises, consists essentially of, or consists of SEQ ID NO: 112, or optionally wherein the ITR on at least one end comprises, consists essentially of, or consists of SEQ ID NO: 110, and optionally wherein the ITR on each end comprises, consists essentially of, or consists of SEQ ID NO: 110. In some such compositions or combinations, the AAV vector is a single-stranded AAV (ssAAV) vector. In some such compositions or combinations, the AAV vector is a recombinant AAV8 (rAAV8) vector.

[0076] In some such compositions or combinations, the polypeptide of interest is a factor IX protein. In some such compositions or combinations, the factor IX protein coding sequence encodes a factor IX protein comprising SEQ ID NO: 63. In some such compositions or combinations, the factor IX protein coding sequence comprises or consists of SEQ ID NO: 61, or wherein the factor IX protein coding sequence comprises or consists of SEQ ID NO: 60. In some such compositions or combinations, the nucleic acid construct is a bidirectional construct, wherein the factor IX protein coding sequence is a first factor IX protein coding sequence, and the bidirectional construct further comprises a reverse complement of a second factor IX protein coding sequence, wherein the first factor IX protein coding sequence and the second factor IX protein coding sequence are different but encode the same factor IX protein sequence. In some such compositions or combinations, the nucleic acid construct comprises from 5’ to 3’: a first splice acceptor, the first factor IX protein coding sequence, a first polyadenylation signal, a reverse complement of a second polyadenylation signal, the reverse complement of the second factor IX protein coding sequence, and a reverse complement of a second splice acceptor, wherein: (i) the first factor IX protein coding sequence comprises SEQ ID NO: 60 and the second factor IX protein coding sequence comprises SEQ ID NO: 61; or (ii) the first factor IX protein coding sequence comprises SEQ ID NO: 61 and the second factor IX protein coding sequence comprises SEQ ID NO: 60, wherein the nucleic acid construct does not comprise a promoter that drives the expression of the factor IX protein, and wherein the nucleic acid construct does not comprise homology arms. In some such compositions or combinations, the nucleic acid construct comprises SEQ ID NO: 64 or 62 or the reverse complement thereof.

[0077] In some such compositions or combinations, the polypeptide of interest is a multidomain therapeutic protein comprising a delivery domain fused to a lysosomal alpha- glucosidase. In some such compositions or combinations, the lysosomal alpha-glucosidaseAttorney Docket No.057766 / 625782 comprises or consists of the sequence set forth in SEQ ID NO: 117. In some such compositions or combinations, the lysosomal alpha-glucosidase coding sequence comprises or consist of the sequence set forth in SEQ ID NO: 127.

[0078] In some such compositions or combinations, the delivery domain is a CD63-binding delivery domain. In some such compositions or combinations, the CD63-binding delivery domain comprises an anti-CD63 antigen-binding protein. In some such compositions or combinations, the CD63-binding delivery domain is a single-chain variable fragment (scFv). In some such compositions or combinations, the scFv comprises or consists of the sequence set forth in SEQ ID NO: 119. In some such compositions or combinations, the scFv coding sequence comprises or consists of the sequence set forth in SEQ ID NO: 129. In some such compositions or combinations, the multidomain therapeutic protein comprises or consists of the sequence set forth in SEQ ID NO: 120. In some such compositions or combinations, the coding sequence for the multidomain therapeutic protein comprises or consists of the sequence set forth in SEQ ID NO: 128, optionally wherein the nucleic acid construct comprises the sequence set forth in SEQ ID NO: 134 or 132. In some such compositions or combinations, the nucleic acid construct comprises from 5’ to 3’: a splice acceptor, the coding sequence for the multidomain therapeutic protein, and a polyadenylation signal or sequence, wherein the coding sequence for the multidomain therapeutic protein comprises the sequence set forth in SEQ ID NO: 128, optionally wherein the nucleic acid construct comprises the sequence set forth in SEQ ID NO: 134 or 132, wherein the polyadenylation signal comprises a BGH polyadenylation signal and a unidirectional SV40 late polyadenylation signal, wherein the nucleic acid construct does not comprise a promoter that drives the expression of the multidomain therapeutic protein, and wherein the nucleic acid construct does not comprise a homology arm.

[0079] In some such compositions or combinations, the delivery domain is a TfR-binding delivery domain. In some such compositions or combinations, the TfR-binding delivery domain comprises an anti-TfR antigen-binding protein. In some such compositions or combinations, the TfR-binding delivery domain comprises a single-chain variable fragment (scFv). In some such compositions or combinations, the scFv comprises or consists of the sequence set forth in SEQ ID NO: 121. In some such compositions or combinations, the scFv coding sequence comprises or consists of the sequence set forth in SEQ ID NO: 122. In some such compositions or combinations, the multidomain therapeutic protein comprises or consists of the sequence setAttorney Docket No.057766 / 625782 forth in SEQ ID NO: 126. In some such compositions or combinations, the coding sequence for the multidomain therapeutic protein comprises or consists of the sequence set forth in SEQ ID NO: 125, optionally wherein the nucleic acid construct comprises the sequence set forth in SEQ ID NO: 133 or 130. In some such compositions or combinations, the nucleic acid construct comprises from 5’ to 3’: a splice acceptor, the coding sequence for the multidomain therapeutic protein, and a polyadenylation signal or sequence, wherein the coding sequence for the multidomain therapeutic protein comprises the sequence set forth in SEQ ID NO: 125, optionally wherein the nucleic acid construct comprises the sequence set forth in SEQ ID NO: 133 or 130, wherein the polyadenylation signal comprises a BGH polyadenylation signal and a unidirectional SV40 late polyadenylation signal, wherein the nucleic acid construct does not comprise a promoter that drives the expression of the multidomain therapeutic protein, and wherein the nucleic acid construct does not comprise a homology arm.

[0080] In some such compositions or combinations, the polypeptide of interest is a factor VIII protein. In some such compositions or combinations, the polypeptide of interest is an antigen-binding protein, optionally wherein the antigen-binding protein is an antibody.

[0081] In some such compositions or combinations, the target genomic locus is an albumin gene, optionally wherein the albumin gene is a human albumin gene. In some such compositions or combinations, the nuclease target site is in intron 1 of the albumin gene.

[0082] In some such compositions or combinations, the nuclease agent comprises: (a) a zinc finger nuclease (ZFN); (b) a transcription activator-like effector nuclease (TALEN); or (c) (i) a Cas protein or a nucleic acid encoding the Cas protein; and (ii) a guide RNA or one or more DNAs encoding the guide RNA, wherein the guide RNA comprises a DNA-targeting segment that targets a guide RNA target sequence, and wherein the guide RNA binds to the Cas protein and targets the Cas protein to the guide RNA target sequence. In some such compositions or combinations, the nuclease agent comprises: (a) a Cas protein or a nucleic acid encoding the Cas protein; and (b) a guide RNA or one or more DNAs encoding the guide RNA, wherein the guide RNA comprises a DNA-targeting segment that targets a guide RNA target sequence, and wherein the guide RNA binds to the Cas protein and targets the Cas protein to the guide RNA target sequence.

[0083] In some such compositions or combinations, the DNA-targeting segment comprises or consists of SEQ ID NO: 96. In some such compositions or combinations, the guide RNAAttorney Docket No.057766 / 625782 comprises SEQ ID NO: 100 or 104. In some such compositions or combinations, the composition or combination comprises the guide RNA in the form of RNA. In some such compositions or combinations, the guide RNA comprises at least one modification. In some such compositions or combinations, the at least one modification comprises: (i) phosphorothioate bonds between the first four nucleotides at the 5’ end of the guide RNA; (ii) phosphorothioate bonds between the last four nucleotides at the 3’ end of the guide RNA; (iii) 2’-O-methyl- modified nucleotides at the first three nucleotides at the 5’ end of the guide RNA; and (iv) 2’-O- methyl-modified nucleotides at the last three nucleotides at the 3’ end of the guide RNA. In some such compositions or combinations, the composition or combination comprises the guide RNA in the form of RNA, the guide RNA comprises SEQ ID NO: 104, and the guide RNA comprises: (i) phosphorothioate bonds between the first four nucleotides at the 5’ end of the guide RNA; (ii) phosphorothioate bonds between the last four nucleotides at the 3’ end of the guide RNA; (iii) 2’-O-methyl-modified nucleotides at the first three nucleotides at the 5’ end of the guide RNA; and (iv) 2’-O-methyl-modified nucleotides at the last three nucleotides at the 3’ end of the guide RNA.

[0084] In some such compositions or combinations, the Cas protein is a Cas9 protein, optionally wherein the Cas protein is derived from a Streptococcus pyogenes Cas9 protein. In some such compositions or combinations, the Cas protein comprises the sequence set forth in SEQ ID NO: 75. In some such compositions or combinations, the composition or combination comprises the nucleic acid encoding the Cas protein, wherein the nucleic acid comprises an mRNA encoding the Cas protein. In some such compositions or combinations, the mRNA encoding the Cas protein comprises at least one modification. In some such compositions or combinations, the mRNA encoding the Cas protein is fully substituted with N1-methyl- pseudouridine. In some such compositions or combinations, the mRNA encoding the Cas protein comprises the sequence set forth in SEQ ID NO: 65 or 66. In some such compositions or combinations, the composition or combination comprises the nucleic acid encoding the Cas protein, wherein the nucleic acid comprises an mRNA encoding the Cas protein, the mRNA encoding the Cas protein comprises the sequence set forth in SEQ ID NO: 65 or 66, and the mRNA encoding the Cas protein is fully substituted with N1-methyl-pseudouridine, comprises a 5’ cap, and comprises a poly(A) tail. In some such compositions or combinations, the composition or combination comprises the guide RNA in the form of RNA, and the guide RNAAttorney Docket No.057766 / 625782 comprises SEQ ID NO: 100 or 104, and wherein the composition or combination comprises administering the nucleic acid encoding the Cas protein, wherein the nucleic acid comprises an mRNA encoding the Cas protein, and the mRNA encoding the Cas protein comprises the sequence set forth in SEQ ID NO: 65 or 66. In some such compositions or combinations, the composition or combination comprises the guide RNA in the form of RNA, the guide RNA comprises SEQ ID NO: 104, and the guide RNA comprises: (i) phosphorothioate bonds between the first four nucleotides at the 5’ end of the guide RNA; (ii) phosphorothioate bonds between the last four nucleotides at the 3’ end of the guide RNA; (iii) 2’-O-methyl-modified nucleotides at the first three nucleotides at the 5’ end of the guide RNA; and (iv) 2’-O-methyl-modified nucleotides at the last three nucleotides at the 3’ end of the guide RNA, and wherein the composition or combination comprises the nucleic acid encoding the Cas protein, wherein the nucleic acid comprises an mRNA encoding the Cas protein, the mRNA encoding the Cas protein comprises the sequence set forth in SEQ ID NO: 65 or 66, and the mRNA encoding the Cas protein is fully substituted with N1-methyl-pseudouridine, comprises a 5’ cap, and comprises a poly(A) tail.

[0085] In some such compositions or combinations, the Cas protein or the nucleic acid encoding the Cas protein and the guide RNA or the one or more DNAs encoding the guide RNA are associated with a lipid nanoparticle. In some such compositions or combinations, the lipid nanoparticle comprises a cationic lipid, a neutral lipid, a helper lipid, and a stealth lipid. In some such compositions or combinations, the cationic lipid is Lipid A ((9Z,12Z)-3-((4,4- bis(octyloxy)butanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)propyl octadeca-9,12-dienoate), and / or wherein the neutral lipid is distearoylphosphatidylcholine or 1,2- distearoyl-sn-glycero-3-phosphocholine (DSPC), and / or wherein the helper lipid is cholesterol, and / or wherein the stealth lipid is 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol- 2000. In some such compositions or combinations, the cationic lipid is Lipid A, the neutral lipid is DSPC, the helper lipid is cholesterol, and the stealth lipid is PEG2k-DMG. In some such compositions or combinations, the lipid nanoparticle comprises four lipids at the following molar ratios: about 50 mol% Lipid A, about 9 mol% DSPC, about 38 mol% cholesterol, and about 3 mol% PEG2k-DMG.

[0086] In some such compositions or combinations, the CD40 inhibitor is further in combination with a plasma cell depleting agent. In some such compositions or combinations, theAttorney Docket No.057766 / 625782 plasma cell depleting agent is capable of depleting long-lived plasma cells (LLPC). In some such compositions or combinations, the plasma cell depleting agent is a B cell maturation antigen (BCMA) targeting agent. In some such compositions or combinations, the BCMA targeting agent is a chimeric antigen receptor against BCMA or an anti-BCMA antibody or a functional fragment thereof. In some such compositions or combinations, the anti-BCMA antibody or functional fragment thereof is conjugated to a cytotoxic agent. In some such compositions or combinations, the anti-BCMA antibody is a multispecific antibody or a functional fragment thereof.

[0087] In some such compositions or combinations, the multispecific anti-BCMA antibody or functional fragment thereof targets BCMA and CD3. In some such compositions or combinations, the multispecific anti-BCMA antibody or functional fragment thereof is anti- BCMAxCD3 bispecific antibody or functional fragment thereof. In some such compositions or combinations, the anti-BCMAxCD3 bispecific antibody is selected from linvoseltamab (REGN5458), REGN5459, pacanalotamab (AMG420), teclistamab (JNJ-64007957), AMG701, alnuctamab (CC-93269), EM801, EM901, elranatamab (PF-06863135), TNB383B (ABBV-383), and TNB384B. In some such compositions or combinations, the anti-BCMAxCD3 bispecific antibody or functional fragment thereof comprises a first antigen-binding domain that specifically binds to BCMA comprising three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 287, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 303. In some such compositions or combinations, the first antigen-binding domain that specifically binds to BCMA comprises a HCDR1 comprising the amino acid sequence of SEQ ID NO: 289, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 291, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 293, a LCDR1 comprising the amino acid sequence of SEQ ID NO: 305, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 307, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 309. In some such compositions or combinations, the anti-BCMAxCD3 bispecific antibody or functional fragment thereof comprises a second antigen-binding domain that specifically binds to CD3 comprising three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence selected from the group consistingAttorney Docket No.057766 / 625782 of SEQ ID NOS: 311 and 319, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 303. In some such compositions or combinations, the second antigen-binding domain that specifically binds to CD3 comprises a HCDR1 comprising the amino acid sequence of SEQ ID NO: 313 or 321, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 315 or 323, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 317 or 325, a LCDR1 comprising the amino acid sequence of SEQ ID NO: 305, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 307, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 309. In some such compositions or combinations, the anti-BCMAxCD3 bispecific antibody or functional fragment thereof comprises: (a) a first antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 289, 291, and 293, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 305, 307, and 309, respectively; and (b) a second antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 313, 315, and 317, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 305, 307, and 309, respectively. In some such compositions or combinations, the anti-BCMAxCD3 bispecific antibody or functional fragment thereof comprises: (a) a first antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 289, 291, and 293, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 305, 307, and 309, respectively; and (b) a second antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 321, 323, and 325, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 305, 307, and 309, respectively. In some such compositions or combinations, the anti- BCMAxCD3 bispecific antibody or functional fragment thereof comprises a human IgG heavy chain constant region. In some such compositions or combinations, the human IgG heavy chain constant region is isotype IgG4 or IgG1. In some such compositions or combinations, the human IgG heavy chain constant region comprises one or more modifications that increase binding to a neonatal Fc receptor (FcRn) and / or the human IgG heavy chain constant region comprises one or more modifications that decrease binding to an Fc-gamma receptor (FcγR).

[0088] In some such compositions or combinations, the plasma cell depleting agent is furtherAttorney Docket No.057766 / 625782 in combination with an effective amount of a B cell depleting agent and / or an immunoglobulin depleting agent. In some such compositions or combinations, the plasma cell depleting agent is further in combination with an effective amount of a B cell depleting agent and an immunoglobulin depleting agent. In some such compositions or combinations, the B cell depleting agent is capable of depleting B cells and plasma cells that express low levels of BCMA. In some such compositions or combinations, the B cell depleting agent is an agent that binds to a B cell surface molecule. In some such compositions or combinations, the B cell depleting agent is selected from an anti-CD19 antibody, an anti-CD20 antibody, an anti-CD19 antibody and an anti-CD20 antibody, an anti-CD22 antibody, an anti-CD79 antibody, an anti- CD20xCD3 bispecific antibody, an anti-CD19xCD3 bispecific antibody, an anti-CD22xCD3 bispecific antibody, an anti-CD79xCD3 bispecific antibody, functional fragments of any of said antibodies, and any combinations thereof. In some such compositions or combinations, the B cell depleting agent is an anti-CD20 antibody or a functional fragment thereof, wherein the anti- CD20 antibody is a multispecific antibody or a functional fragment thereof. In some such compositions or combinations, the multispecific anti-CD20 antibody or functional fragment thereof targets CD20 and CD3. In some such compositions or combinations, the multispecific anti-CD20 antibody or functional fragment thereof is anti-CD20xCD3 bispecific antibody or functional fragment thereof. In some such compositions or combinations, the anti-CD20xCD3 bispecific antibody or functional fragment thereof comprises a first antigen-binding domain that specifically binds to CD20 comprising three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 329, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 330. In some such compositions or combinations, the first antigen-binding domain that specifically binds to CD20 comprises a HCDR1 comprising the amino acid sequence of SEQ ID NO: 332, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 333, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 334, a LCDR1 comprising the amino acid sequence of SEQ ID NO: 335, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 336, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 337. In some such compositions or combinations, the anti-CD20xCD3 bispecific antibody or functional fragment thereof comprises a second antigen-binding domain that specifically binds to CD3 comprising three heavy chainAttorney Docket No.057766 / 625782 CDRs (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 331, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 330. In some such compositions or combinations, the second antigen-binding domain that specifically binds to CD3 comprises a HCDR1 comprising the amino acid sequence of SEQ ID NO: 338, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 339, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 340, a LCDR1 comprising the amino acid sequence of SEQ ID NO: 335, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 336, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 337. In some such compositions or combinations, the anti-CD20xCD3 bispecific antibody or functional fragment thereof comprises: (a) a first antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 332, 333, and 334, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 335, 336, and 337, respectively; and (b) a second antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 338, 339, and 340, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 335, 336, and 337, respectively. In some such compositions or combinations, the anti-CD20xCD3 bispecific antibody or functional fragment thereof comprises a human IgG heavy chain constant region. In some such compositions or combinations, the human IgG heavy chain constant region is isotype IgG4 or IgG1. In some such compositions or combinations, the human IgG heavy chain constant region comprises one or more modifications that increase binding to a neonatal Fc receptor (FcRn) and / or the human IgG heavy chain constant region comprises one or more modifications that decrease binding to an Fc-gamma receptor (FcγR). In some such compositions or combinations, the B cell depleting agent is an agent targeting a B cell survival factor. In some such compositions or combinations, the B cell depleting agent is a BLyS / BAFF inhibitor, an APRIL inhibitor, a BLyS receptor 3 / BAFF receptor inhibitor, or any combination thereof.

[0089] In some such compositions or combinations, the immunoglobulin depleting agent is capable of accelerating IgG clearance. In some such compositions or combinations, the immunoglobulin depleting agent is a neonatal Fc receptor (FcRn) blocker. In some such compositions or combinations, the FcRn blocker is selected from Efgartigimod (ARGX-113),Attorney Docket No.057766 / 625782 Rozanolixizumab (UCB7665), Batoclimab (RVT-1401), IMVT-1402, Nipocalimab (M281), Orilanolimab (SYNT001), and any combinations thereof.

[0090] In some such compositions or combinations, the composition or combination is for use in a method of inserting a nucleic acid encoding a polypeptide of interest into a target genomic locus in a cell or a population of cells in a subject. In some such compositions or combinations, the composition or combination is for use in a method of expressing a polypeptide of interest from a target genomic locus in a cell or a population of cells in a subject. In some such compositions or combinations, the composition or combination is for use in a method of treating an enzyme deficiency in a subject in need thereof. In some such compositions or combinations, the composition or combination is for use in a method of preventing or reducing the onset of a sign or symptom of an enzyme deficiency in a subject in need thereof.

[0091] In another aspect, provided are kits comprising any of the above compositions or combinations. In another aspect, provided are any of the above CD40 inhibitors (e.g., CD40 antigen-binding molecules) for use in any of the above methods. BRIEF DESCRIPTION OF THE FIGURES

[0092] Figures 1A-1C show the effect of anti-CD40xCD40 bispecific antibodies on IL6 production in the presence of constant CD40L in human B cells from three different donors.

[0093] Figures 1D-1E show the effect of anti-CD40xCD40 bispecific antibodies on IL6 production in the presence of constant CD40L in human B cells from two different donors.

[0094] Figures 2A-2C show the effect of anti-CD40xCD40 bispecific antibodies on IL10 production in the presence of constant CD40L in human B cells from three different donors.

[0095] Figures 2D-2E show the effect of anti-CD40xCD40 bispecific antibodies on IL10 production in the presence of constant CD40L in human B cells from two different donors.

[0096] Figures 3A-3C show the effect of anti-CD40xCD40 bispecific antibodies on TNFα production in the presence of constant CD40L in human B cells from three different donors.

[0097] Figures 3D-3E show the effect of anti-CD40xCD40 bispecific antibodies on TNFα production in the presence of constant CD40L in human B cells from two different donors.

[0098] Figures 3F-3G show analysis of agonist activity of anti-CD40xCD40 bispecific antibodies as measured by IL6 production in human B cells from two different donors.Attorney Docket No.057766 / 625782

[0099] Figures 4A-4B show the effect of anti-CD40xCD40 bispecific antibodies on IL- 12 / IL-23p40 production in the presence of constant CD40L from human monocyte derived dendritic cells from two different donors.

[0100] Figures 5A-5B show analysis of agonist activity of anti-CD40xCD40 bispecific antibodies as measured by IL6 production in human B cells from two different donors.

[0101] Figures 6A-6B show analysis of agonist activity of anti-CD40xCD40 bispecific antibodies as measured by IL10 production in human B cells from two different donors.

[0102] Figure 7 shows an experiment timeline using a NP-KLH immunization model as disclosed in Example 9.

[0103] Figure 8A shows the effect of anti-CD40xCD40 bispecific antibodies on the frequency of NP positive germinal center B cells. * p<0.05.

[0104] Figure 8B shows the effect of anti-CD40xCD40 bispecific antibodies on the frequency of NP IgG1 titers in mouse serum. * p<0.05; ** p<0.005.

[0105] Figure 9A shows the effect of anti-CD40xCD40 bispecific antibodies on the frequency of NP positive germinal center B cells.

[0106] Figure 9B shows the effect of anti-CD40xCD40 bispecific antibodies on the frequency of NP IgG1 titers in mouse serum.

[0107] Figure 10 shows an experiment timeline using a mouse EAE model as disclosed in Example 10.

[0108] Figure 11A shows mean EAE symptom scores for REGN16431- or REGN16432- treated mice.

[0109] Figure 11B shows mean EAE symptom scores for REGN16334- or REGN16335- treated mice.

[0110] Figure 11C shows percent of initial body weight in REGN16431- or REGN16432- treated mice.

[0111] Figure 11D shows percent of initial body weight in REGN16334- or REGN16335- treated mice.

[0112] Figure 12A shows mean EAE symptom scores for anti-CD40xCD40 bispecific antibody-treated mice.

[0113] Figure 12B shows percent of initial body weight in anti-CD40xCD40 bispecific antibody-treated mice.Attorney Docket No.057766 / 625782

[0114] Figure 13 shows an experiment timeline for the study described in Example 11.

[0115] Figure 14 shows the effect of antibody-mediated CD40L blockade on the development of anti-AAV IgG titers following treatment with an AAV8 vector.

[0116] Figures 15A-15C show the effect of antibody-mediated CD40L blockade on transduction by a second AAV8 vector.

[0117] Figure 16 shows an experiment timeline for the study described in Examples 12, 13, and 16.

[0118] Figures 17A-17B show the effect of antagonistic anti-CD40 antibodies on the development of anti-AAV IgG and IgM titers following treatment with an AAV8 vector.

[0119] Figures 18A-18C show the effect of antagonistic anti-CD40 antibodies on transduction by a second AAV8 vector.

[0120] Figure 19 shows an experiment timeline for the study described in Examples 14 and 15.

[0121] Figures 20A-20D show the effect of antagonistic anti-CD40 antibodies on steady- state polyclonal and AAV-specific germinal center B cell responses.

[0122] Figures 21A-21E show the effect of antagonistic anti-CD40 antibodies on follicular T helper cell (TFH) responses and AAV-specific T cell responses.

[0123] Figure 22 shows the effect of antagonistic anti-CD40 antibodies on the development of anti-transgene IgG following treatment with an AAV8 vector.

[0124] Figure 23 shows a cryoEM reconstruction of CD40 in complex with the Fab arms 30027P2, 21519P2, and 21520P2.

[0125] Figure 24 shows an experimental timeline for the study described in Examples 28, 29, 30, and 31.

[0126] Figure 25 shows the effect of plasma cell depletion with anti-BCMAxCD3 bispecific antibody, FcRn blockade via efgartigimod alfa, B cell depletion with anti-CD19 and anti-CD20 antibodies (anti-CD19 / CD20 antibodies), or combination thereof, on anti-AAV8 capsid IgG titers over time in mice previously treated with recombinant AAV8 vector.

[0127] Figure 26 shows the effect of plasma cell depletion with anti-BCMAxCD3 bispecific antibody, FcRn blockade via efgartigimod alfa, B cell depletion with anti-CD19 / CD20 antibodies, or combination thereof, on liver transduction 10 days following administration of a second AAV8 vector in mice previously treated with recombinant AAV8 vector, as measured byAttorney Docket No.057766 / 625782 Taqman quantitative real-time polymerase chain reaction (PCR) of green fluorescent protein (GFP) transgene DNA.

[0128] Figure 27 shows the effect of plasma cell depletion with anti-BCMAxCD3 bispecific antibody, FcRn blockade via efgartigimod alfa, B cell depletion with anti-CD19 / CD20 antibodies, or combination thereof, on liver transduction 10 days following administration of a second recombinant AAV8 vector in mice previously treated with a first recombinant AAV8 vector, as measured by Taqman quantitative real-time reverse-transcription PCR of GFP transgene RNA.

[0129] Figures 28A-28B show the effect of plasma cell depletion with anti-BCMAxCD3 bispecific antibody, FcRn blockade via efgartigimod alfa, B cell depletion with anti-CD19 / CD20 antibodies, or combination thereof, on liver transduction 10 days following administration of a second recombinant AAV8 vector in mice previously treated with a first recombinant AAV8 vector, as measured by GFP immunohistochemical (IHC) staining of formalin-fixed paraffin embedded liver sections. Figure 28A shows GFP-positive area quantified using HALO software (Indica labs). Figure 28B shows representative images.

[0130] Figures 29A-29J show flow cytometry analysis of B cell and plasma cell frequencies and counts in bone marrow and spleen following treatment with anti-BCMAxCD3 bispecific antibody, FcRn blockade, anti-CD19 / CD20 antibodies, or combinations thereof. Figure 29A shows bone marrow plasma cell frequencies, Figure 29B shows spleen plasma cell frequencies, Figure 29C shows spleen naïve B cell frequencies, Figure 29D shows spleen total memory B cell frequencies, Figure 29E shows spleen AAV-specific memory B cell frequencies, Figure 29F shows bone marrow plasma cell counts, Figure 29G shows spleen plasma cell counts, Figure 29H shows spleen naïve B cell counts, Figure 29I shows spleen total memory B cell counts, and Figure 29J shows spleen AAV-specific memory B cell counts.

[0131] Figure 30 shows the effect of efgartigimod on serum drug concentration of REGN5458 (BCMAxCD3).

[0132] Figure 31 shows an experimental timeline for the study described in Example 32.

[0133] Figures 32A-32B show the effect of plasma cell depletion, B cell depletion, neonatal Fc receptor blockade, and combinations thereof, on naturally-occurring anti-AAV antibody titers in cynomolgus macaques. AAV8 neutralizing antibody (NAb) titer levels are presented for eachAttorney Docket No.057766 / 625782 treatment group over the duration of the study (Figure 32A) and specifically at Study Day 29 (Figure 32B).

[0134] Figure 33 shows an experimental diagram for the non-human primate studies described in Examples 36 and 37.

[0135] Figures 34A-34F show the effect of prophylactic CD40 blockade on serum anti- AAV8 IgM, IgG, and neutralizing antibody titers in cynomolgus macaques.

[0136] Figures 35A-35C show the effect of prophylactic CD40 blockade on ability to systemically re-administer a second AAV8 vector in cynomolgus macaques.

[0137] Figures 36A-36C shows the effect of prophylactic CD40 blockade on T cell-induced liver injury and expression of an immunogenic GFP transgene in liver. DEFINITIONS

[0138] The terms “protein,” “polypeptide,” and “peptide,” used interchangeably herein, include polymeric forms of amino acids of any length, including coded and non-coded amino acids and chemically or biochemically modified or derivatized amino acids. The terms also include polymers that have been modified, such as polypeptides having modified peptide backbones. The term “domain” refers to any part of a protein or polypeptide having a particular function or structure.

[0139] The terms “nucleic acid” and “polynucleotide,” used interchangeably herein, include polymeric forms of nucleotides of any length, including ribonucleotides, deoxyribonucleotides, or analogs or modified versions thereof. They include single-, double-, and multi-stranded DNA or RNA, genomic DNA, cDNA, DNA-RNA hybrids, and polymers comprising purine bases, pyrimidine bases, or other natural, chemically modified, biochemically modified, non-natural, or derivatized nucleotide bases.

[0140] The term “expression vector” or “expression construct” or “expression cassette” refers to a recombinant nucleic acid containing a desired coding sequence operably linked to appropriate nucleic acid sequences necessary for the expression of the operably linked coding sequence in a particular host cell or organism. Nucleic acid sequences necessary for expression in prokaryotes usually include a promoter, an operator (optional), and a ribosome binding site, as well as other sequences. Eukaryotic cells are generally known to utilize promoters, enhancers,Attorney Docket No.057766 / 625782 and termination and polyadenylation signals, although some elements may be deleted and other elements added without sacrificing the necessary expression.

[0141] The term “viral vector” refers to a recombinant nucleic acid that includes at least one element of viral origin and includes elements sufficient for or permissive of packaging into a viral vector particle. The vector and / or particle can be utilized for the purpose of transferring DNA, RNA, or other nucleic acids into cells either ex vivo or in vivo. Numerous forms of viral vectors are known.

[0142] The term “isolated” with respect to proteins, nucleic acids, and cells includes proteins, nucleic acids, and cells that are relatively purified with respect to other cellular or organism components that may normally be present in situ, up to and including a substantially pure preparation of the protein, nucleic acid, or cell. The term “isolated” may include proteins and nucleic acids that have no naturally occurring counterpart or proteins or nucleic acids that have been chemically synthesized and are thus substantially uncontaminated by other proteins or nucleic acids. The term “isolated” may include proteins, nucleic acids, or cells that have been separated or purified from most other cellular components or organism components with which they are naturally accompanied (e.g., but not limited to, other cellular proteins, nucleic acids, or cellular or extracellular components).

[0143] The term “wild type” or “wild-type” includes entities having a structure and / or activity as found in a normal (as contrasted with mutant, diseased, altered, or so forth) state or context. Wild type genes and polypeptides often exist in multiple different forms (e.g., alleles).

[0144] The term “endogenous sequence” refers to a nucleic acid sequence that occurs naturally within a cell or animal. For example, an endogenous APP sequence of an animal refers to a native APP sequence that naturally occurs at the APP locus in the animal.

[0145] “Exogenous” molecules or sequences include molecules or sequences that are not normally present in a cell in that form or that are introduced into a cell from an outside source. Normal presence includes presence with respect to the particular developmental stage and environmental conditions of the cell. An exogenous molecule or sequence, for example, can include a mutated version of a corresponding endogenous sequence within the cell, such as a humanized version of the endogenous sequence, or can include a sequence corresponding to an endogenous sequence within the cell but in a different form (i.e., not within a chromosome). In contrast, endogenous molecules or sequences include molecules or sequences that are normallyAttorney Docket No.057766 / 625782 present in that form in a particular cell at a particular developmental stage under particular environmental conditions.

[0146] The term “heterologous” when used in the context of a nucleic acid or a protein indicates that the nucleic acid or protein comprises at least two segments that do not naturally occur together in the same molecule. For example, the term “heterologous,” when used with reference to segments of a nucleic acid or segments of a protein, indicates that the nucleic acid or protein comprises two or more sub-sequences that are not found in the same relationship to each other (e.g., joined together) in nature. As one example, a “heterologous” region of a nucleic acid vector is a segment of nucleic acid within or attached to another nucleic acid molecule that is not found in association with the other molecule in nature. For example, a heterologous region of a nucleic acid vector could include a coding sequence flanked by a heterologous promoter not found in association with the coding sequence in nature. Likewise, a “heterologous” region of a protein is a segment of amino acids within or attached to another peptide molecule that is not found in association with the other peptide molecule in nature (e.g., a fusion protein, or a protein with a tag). Similarly, a nucleic acid or protein can comprise a heterologous label or a heterologous secretion or localization sequence.

[0147] “Codon optimization” takes advantage of the degeneracy of codons, as exhibited by the multiplicity of three-base pair codon combinations that specify an amino acid, and generally includes a process of modifying a nucleic acid sequence for enhanced expression in particular host cells by replacing at least one codon of the native sequence with a codon that is more frequently or most frequently used in the genes of the host cell while maintaining the native amino acid sequence. For example, a nucleic acid encoding a protein can be modified to substitute codons having a higher frequency of usage in a given prokaryotic or eukaryotic cell, including a bacterial cell, a yeast cell, a human cell, a non-human cell, a mammalian cell, a rodent cell, a mouse cell, a rat cell, a hamster cell, or any other host cell, as compared to the naturally occurring nucleic acid sequence. Codon usage tables are readily available, for example, at the “Codon Usage Database.” These tables can be adapted in a number of ways. See Nakamura et al. (2000) Nucleic Acids Research 28:292, herein incorporated by reference in its entirety for all purposes. Computer algorithms for codon optimization of a particular sequence for expression in a particular host are also available (see, e.g., Gene Forge).Attorney Docket No.057766 / 625782

[0148] A “promoter” is a regulatory region of DNA usually comprising a TATA box capable of directing RNA polymerase II to initiate RNA synthesis at the appropriate transcription initiation site for a particular polynucleotide sequence. A promoter may additionally comprise other regions which influence the transcription initiation rate. The promoter sequences disclosed herein modulate transcription of an operably linked polynucleotide. A promoter can be active in one or more of the cell types disclosed herein (e.g., a eukaryotic cell, a non-human mammalian cell, a human cell, a rodent cell, a pluripotent cell, a one-cell stage embryo, a differentiated cell, or a combination thereof). A promoter can be, for example, a constitutively active promoter, a conditional promoter, an inducible promoter, a temporally restricted promoter (e.g., a developmentally regulated promoter), or a spatially restricted promoter (e.g., a cell-specific or tissue-specific promoter). Examples of promoters can be found, for example, in WO 2013 / 176772, herein incorporated by reference in its entirety for all purposes.

[0149] A constitutive promoter is one that is active in all tissues or particular tissues at all developing stages. Examples of constitutive promoters include the human cytomegalovirus immediate early (hCMV), mouse cytomegalovirus immediate early (mCMV), human elongation factor 1 alpha (hEF1α), mouse elongation factor 1 alpha (mEF1α), mouse phosphoglycerate kinase (PGK), chicken beta actin hybrid (CAG or CBh), SV40 early, and beta 2 tubulin promoters.

[0150] Examples of inducible promoters include, for example, chemically regulated promoters and physically-regulated promoters. Chemically regulated promoters include, for example, alcohol-regulated promoters (e.g., an alcohol dehydrogenase (alcA) gene promoter), tetracycline (tet)-regulated promoters (e.g., a tetracycline-responsive promoter, a tetracycline operator sequence (tetO), a tet-On promoter, or a tet-Off promoter), steroid-regulated promoters (e.g., a rat glucocorticoid receptor, a promoter of an estrogen receptor, or a promoter of an ecdysone receptor), or metal-regulated promoters (e.g., a metalloprotein promoter). Physically regulated promoters include, for example, temperature-regulated promoters (e.g., a heat shock promoter) and light-regulated promoters (e.g., a light-inducible promoter or a light-repressible promoter).

[0151] Tissue-specific promoters can be, for example, neuron-specific promoters or glial- specific promoters or muscle-specific promoters.Attorney Docket No.057766 / 625782

[0152] Developmentally regulated promoters include, for example, promoters active only during an embryonic stage of development, or only in an adult cell.

[0153] “Operable linkage” or being “operably linked” includes juxtaposition of two or more components (e.g., a promoter and another sequence element) such that both components function normally and allow the possibility that at least one of the components can mediate a function that is exerted upon at least one of the other components. For example, a promoter can be operably linked to a coding sequence if the promoter controls the level of transcription of the coding sequence in response to the presence or absence of one or more transcriptional regulatory factors. Operable linkage can include such sequences being contiguous with each other or acting in trans (e.g., a regulatory sequence can act at a distance to control transcription of the coding sequence).

[0154] The term “in vitro” includes artificial environments and to processes or reactions that occur within an artificial environment (e.g., a test tube or an isolated cell or cell line). The term “in vivo” includes natural environments (e.g., a cell, organism, or body) and to processes or reactions that occur within a natural environment. The term “ex vivo” includes cells that have been removed from the body of an individual and processes or reactions that occur within such cells.

[0155] The term “CD40,” as used herein, refers to cluster of differentiation 40 (CD40 / TNFRSF5), a co-stimulatory cell surface receptor that is part of the tumor necrosis factor (TNF) receptor superfamily. In some embodiments, the CD40 is a human CD40. In some embodiments, the CD40 protein comprises the amino acid sequence of human CD40 set forth in UniProt Accession No. Q09LL4.

[0156] The term “antigen-binding molecule” includes antibodies and antigen-binding fragments of antibodies, including multispecific antibodies, e.g., bispecific antibodies.

[0157] The term “antibody,” as used herein, refers to an antigen-binding molecule or molecular complex comprising a set of complementarity determining regions (CDRs) that specifically bind to or interact with a particular antigen (e.g., CD40, BCMA, CD20, CD3). The term “antibody,” as used herein, includes immunoglobulin molecules comprising four polypeptide chains, two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds, as well as multimers thereof (e.g., IgM). In a typical antibody, each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region comprises three domains, CH1, CH2 and CH3.Attorney Docket No.057766 / 625782 Each light chain comprises a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region comprises one domain (CL1). The VHand VLregions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FR). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In some embodiments, the FRs of the antibody (or antigen-binding portion thereof) may be identical to the human germline sequences, or may be naturally or artificially modified. An amino acid consensus sequence may be defined based on a side-by-side analysis of two or more CDRs.

[0158] Methods and techniques for identifying CDRs within HCVR and LCVR amino acid sequences are well known in the art and can be used to identify CDRs within the specified HCVR and / or LCVR amino acid sequences disclosed herein. Exemplary conventions that can be used to identify the boundaries of CDRs include, but are not limited to, the Kabat definition, the Chothia definition, the AbM definition (enhanced Chothia or Martin), the IMGT definition, and the Honneger definition (AHo). In general terms, the Kabat definition is based on sequence variability, the Chothia definition is based on the location of the structural loop regions, and the AbM definition is a compromise between the Kabat and Chothia approaches. See, e.g., Kabat et al., “Sequences of Proteins of Immunological Interest,” National Institutes of Health, Bethesda, Md. (1991); Chothia et al., J Mol Biol (1987), 4:901–17; Al-Lazikani et al., J. Mol. Biol. 273:927-948 (1997); and Martin et al., Proc. Natl. Acad. Sci. USA 86:9268-9272 (1989); see also Dondelinger et al., Front. Immunol. (2018), 9:2278, doi:10.3389 / fimmu.2018.02278. Public databases are also available for identifying CDR sequences within an antibody.

[0159] The term “antibody,” as used herein, also includes antigen-binding fragments of full antibody molecules. The terms “antigen-binding portion” of an antibody, “antigen-binding fragment” of an antibody, “antigen-binding domain,” and the like, as used herein, include any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds an antigen to form a complex. Antigen-binding fragments of an antibody may be derived, e.g., from full antibody molecules using any suitable standard techniques such as proteolytic digestion or recombinant genetic engineering techniques involving the manipulation and expression of DNA encoding antibody variable and optionally constantAttorney Docket No.057766 / 625782 domains. Such DNA is known and / or is readily available from, e.g., commercial sources, DNA libraries (including, e.g., phage-antibody libraries), or can be synthesized. The DNA may be sequenced and manipulated chemically or by using molecular biology techniques, for example, to arrange one or more variable and / or constant domains into a suitable configuration, or to introduce codons, create cysteine residues, modify, add, or delete amino acids, etc.

[0160] Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single-chain Fv (scFv) molecules; (vi) dAb fragments; and (vii) minimal recognition units consisting of the amino acid residues that mimic the hypervariable region of an antibody (e.g., an isolated complementarity determining region (CDR) such as a CDR3 peptide), or a constrained FR3- CDR3-FR4 peptide. Other engineered molecules, such as domain-specific antibodies, single domain antibodies, domain-deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains, are also encompassed within the expression “antigen-binding fragment,” as used herein.

[0161] An antigen-binding fragment of an antibody will typically comprise at least one variable domain. The variable domain may be of any size or amino acid composition and will generally comprise at least one CDR which is adjacent to or in frame with one or more framework sequences. In antigen-binding fragments having a VHdomain associated with a VLdomain, the VH and VL domains may be situated relative to one another in any suitable arrangement. For example, the variable region may be dimeric and contain VH-VH, VH-VLor VL- VLdimers. Alternatively, the antigen-binding fragment of an antibody may contain a monomeric VH or VL domain.

[0162] In certain embodiments, an antigen-binding fragment of an antibody may contain at least one variable domain covalently linked to at least one constant domain. Non-limiting, exemplary configurations of variable and constant domains that may be found within an antigen- binding fragment of an antibody include: (i) VH-CH1; (ii) VH-CH2; (iii) VH-CH3; (iv) VH-CH1- CH2; (v) VH-CH1-CH2-CH3; (vi) VH-CH2-CH3; (vii) VH-CL; (viii) VL-CH1; (ix) VL-CH2; (x) VL- CH3; (xi) VL-CH1-CH2; (xii) VL-CH1-CH2-CH3; (xiii) VL-CH2-CH3; and (xiv) VL-CL. In any configuration of variable and constant domains, including any of the exemplary configurationsAttorney Docket No.057766 / 625782 listed above, the variable and constant domains may be either directly linked to one another or may be linked by a full or partial hinge or linker region. A hinge region may consist of at least 2 (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids which result in a flexible or semi-flexible linkage between adjacent variable and / or constant domains in a single polypeptide molecule. Moreover, an antigen-binding fragment of an antibody may comprise a homo-dimer or hetero- dimer (or other multimer) of any of the variable and constant domain configurations listed above in non-covalent association with one another and / or with one or more monomeric VHor VLdomain (e.g., by disulfide bond(s)).

[0163] The term “antibody,” as used herein, also includes multispecific (e.g., bispecific) antibodies. A multispecific antibody or antigen-binding fragment of an antibody will typically comprise at least two different variable domains, wherein each variable domain is capable of specifically binding to a separate antigen or to a different epitope on the same antigen. In some embodiments, a multispecific antibody (e.g., bispecific antibody) has an arm that binds to a first epitope of an antigen and an arm that binds to a second epitope of the same antigen.

[0164] Any multispecific antibody format may be adapted for use in the context of an antibody or antigen-binding fragment of an antibody of the present disclosure using routine techniques available in the art. For example, the present disclosure includes bispecific antibodies wherein one arm of an immunoglobulin is specific for a first epitope of CD40, and the other arm of the immunoglobulin is specific for a second epitope of CD40. As another example, the present disclosure includes bispecific antibodies wherein one arm of an immunoglobulin is specific for a first epitope of BCMA or CD20, and the other arm of the immunoglobulin is specific for a second epitope of CD3. Exemplary bispecific formats that can be used in the context of the present disclosure include, without limitation, e.g., scFv-based or diabody bispecific formats, IgG-scFv fusions, dual variable domain (DVD)-Ig, Quadroma, knobs-into-holes, common light chain (e.g., common light chain with knobs-into-holes, etc.), CrossMab, CrossFab, (SEED) body, leucine zipper, Duobody, IgG1 / IgG2, dual acting Fab (DAF)-IgG, and Mab2bispecific formats (see, e.g., Klein et al.2012, mAbs 4:6, 1-11, and references cited therein, for a review of the foregoing formats). Bispecific antibodies can also be constructed using peptide / nucleic acid conjugation, e.g., wherein unnatural amino acids with orthogonal chemical reactivity are used to generate site-specific antibody-oligonucleotide conjugates which then self-assemble into multimeric complexes with defined composition, valency, and geometry. See, e.g., Kazane et al.,Attorney Docket No.057766 / 625782 J. Am. Chem. Soc. (Epub: Dec.4, 2012).

[0165] The term “human antibody,” as used herein, is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. The human antibodies of the disclosure may nonetheless include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site- specific mutagenesis in vitro or by somatic mutation in vivo), for example, in the CDRs and in particular CDR3. However, the term “human antibody,” as used herein, is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.

[0166] The term “recombinant antibody,” as used herein, is intended to include all antibodies that are prepared, expressed, created, or isolated by recombinant means. The term includes, but is not limited to, antibodies expressed using a recombinant expression vector transfected into a host cell (e.g., Chinese hamster ovary (CHO) cell) or cellular expression system, antibodies isolated from a recombinant, combinatorial human antibody library, and antibodies isolated from a non- human animal (e.g., a mouse, such as a mouse that is transgenic for human immunoglobulin genes (see e.g., Taylor et al. (1992) Nucl. Acids Res.20:6287-6295). In some embodiments, the recombinant antibody is a recombinant human antibody. In some embodiments, recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. In certain embodiments, however, such recombinant human antibodies are subjected to in vitro mutagenesis (or, when an animal transgenic for human Ig sequences is used, in vivo somatic mutagenesis) and thus the amino acid sequences of the VH and VLregions of the recombinant antibodies are sequences that, while derived from and related to human germline VHand VLsequences, may not naturally exist within the human antibody germline repertoire in vivo.

[0167] An “isolated antibody” refers to an antibody that has been identified and separated and / or recovered from at least one component of its natural environment. For example, an antibody that has been separated or removed from at least one component of an organism, or from a tissue or cell in which the antibody naturally exists or is naturally produced, is an “isolated antibody.” An isolated antibody also includes an antibody in situ within a recombinant cell. Isolated antibodies are antibodies that have been subjected to at least one purification or isolation step. According to certain embodiments, an isolated antibody may be substantially freeAttorney Docket No.057766 / 625782 of other cellular material and / or chemicals.

[0168] The term “specifically binds,” or the like, means that an antibody or antigen-binding fragment thereof forms a complex with an antigen that is relatively stable under physiologic conditions. Specific binding can be characterized by an equilibrium dissociation constant of at least about 1x10-6M or less, e.g., 10-7M, 10-8M, 10-9M, 10-10M, 10-11M, or 10-12M (a smaller KDdenotes a tighter binding). Methods for determining whether an antibody specifically binds to an antigen are known in the art and include, for example, equilibrium dialysis, surface plasmon resonance (e.g., BIACORE™), bio-layer interferometry assay (e.g., Octet® HTX biosensor), solution-affinity ELISA, and the like. In some embodiments, specific binding is measured in a surface plasmon resonance assay, e.g., at 25°C or 37°C. An antibody or antigen-binding fragment that specifically binds an antigen from one species may or may not have cross- reactivity to other antigens, such as an orthologous antigen from another species.

[0169] The term “KD,” as used herein, refers to the equilibrium dissociation constant of a particular antibody-antigen interaction.

[0170] The term “surface plasmon resonance,” as used herein, refers to an optical phenomenon that allows for the analysis of real-time biomolecular interactions by detection of alterations in protein concentrations within a biosensor matrix, for example, using the BIACORE™ system (Cytiva, Marlborough, MA).

[0171] The term “epitope,” as used herein, refers to an antigenic determinant that interacts with a specific antigen binding site in the variable region of an antibody molecule known as a paratope. A single antigen may have more than one epitope. Thus, different antibodies may bind to different areas on an antigen and may have different biological effects. The term “epitope” also refers to a site on an antigen to which B and / or T cells respond. It also refers to a region of an antigen that is bound by an antibody. Epitopes may be either linear or discontinuous (e.g., conformational). A linear epitope is one produced by adjacent amino acid residues in a polypeptide chain. A conformational epitope is produced by spatially juxtaposed amino acids from different segments of the linear polypeptide chain. In certain embodiments, epitopes may include determinants that are chemically active surface groupings of molecules such as amino acids, sugar side chains, phosphoryl groups, or sulfonyl groups, and, in certain embodiments, may have specific three-dimensional structural characteristics, and / or specific charge characteristics. Epitopes may also be defined as structural or functional. Functional epitopes areAttorney Docket No.057766 / 625782 generally a subset of the structural epitopes and have those residues that directly contribute to the affinity of the interaction. An epitope typically includes at least 3, and more usually, e.g., at least 5 or at least 8-10 amino acids in a unique spatial conformation.

[0172] Methods for determining the epitope of an antigen-binding protein, e.g., an antibody or antigen-binding fragment, include alanine scanning mutational analysis, peptide blot analysis (Reineke, Methods Mol Biol 2004, 248:443-463), peptide cleavage analysis, crystallographic studies, and nuclear magnetic resonance (NMR) analysis. In addition, methods such as epitope exclusion, epitope extraction, and chemical modification of antigens can be employed (Tomer, Prot Sci 2000, 9:487-496). Another method that can be used to identify the amino acids within a polypeptide with which an antigen-binding protein (e.g., an antibody or antigen-binding fragment) interacts is hydrogen / deuterium exchange detected by mass spectrometry (HDX). See, e.g., Ehring, Analytical Biochemistry 1999, 267:252-259; Engen and Smith, Anal Chem 2001, 73:256A-265A.

[0173] The term “competes,” as used in reference to competing for binding, refers to an antigen-binding protein (e.g., antibody or antigen-binding fragment) that binds to an antigen and inhibits or blocks the binding of another antigen-binding protein (e.g., antibody or antigen- binding fragment) to the antigen. Unless otherwise stated, the term also includes competition between two antigen-binding proteins (e.g., antibodies) in both orientations, i.e., a first antigen that binds an antigen and blocks binding of the antigen by a second antibody and vice versa. Thus, in some embodiments, competition occurs in one such orientation. In some embodiments, the first antigen-binding protein (e.g., antibody) and second antigen-binding protein (e.g., antibody) may bind to the same epitope. Alternatively, the first and second antigen-binding proteins (e.g., antibodies) may bind to different epitopes, which may be overlapping or non- overlapping, wherein binding of one antigen-binding protein inhibits or blocks the binding of the second antigen-binding protein, e.g., via steric hindrance. Competition between antigen-binding proteins may be measured by methods known in the art, e.g., by a real-time, label-free bio-layer interferometry assay.

[0174] In the context of viruses in the present disclosure (e.g., AAV), the term “neutralizing antibody” or “nAb” refers to an antibody that binds to a virus and interferes with its ability to infect a cell. Non-limiting examples of neutralizing antibodies include antibodies that bind to a viral particle and inhibit successful transduction, e.g., one or more steps selected from binding,Attorney Docket No.057766 / 625782 entry, trafficking to the nucleus, and transcription of the viral genome. Some neutralizing antibodies may block a virus at the post-entry step.

[0175] The terms “substantial identity” and “substantially identical,” as used with reference to a nucleic acid or fragment thereof, indicates that, when optimally aligned with appropriate nucleotide insertions or deletions with another nucleic acid (or its complementary strand), there is nucleotide sequence identity in at least about 90%, e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, of the nucleotide bases, as measured by any well-known algorithm of sequence identity, such as FASTA, BLAST or GAP, as discussed below. A nucleic acid molecule having substantial identity to a reference nucleic acid molecule may, in certain instances, encode a polypeptide having the same or substantially similar amino acid sequence as the polypeptide encoded by the reference nucleic acid molecule.

[0176] As applied to polypeptides, the terms “substantial identity” and “substantially identical” mean that two peptide sequences, when optimally aligned, share at least about 90% sequence identity, e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity. In some embodiments, residue positions that are not identical differ by conservative amino acid substitutions. A “conservative amino acid substitution” is one in which an amino acid residue is substituted by another amino acid residue having a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity). In general, a conservative amino acid substitution will not substantially change the functional properties of a protein.

[0177] Sequence similarity for polypeptides is typically measured using sequence analysis software. Protein analysis software matches similar sequences using measures of similarity assigned to various substitutions, deletions, and other modifications, including conservative amino acid substitutions. For instance, GCG software contains programs such as GAP and BESTFIT which can be used with default parameters to determine sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different species of organisms or between a wild-type protein and a mutein thereof. See, e.g., GCG Version 6.1. Polypeptide sequences also can be compared using FASTA with default or recommended parameters; a program in GCG Version 6.1. FASTA (e.g., FASTA2 and FASTA3) provides alignments and percent sequence identity of the regions of the best overlap between the query and search sequences (Pearson, 2000 supra). Another preferred algorithm when comparing a sequence of the disclosure to a database containing a large number ofAttorney Docket No.057766 / 625782 sequences from different organisms is the computer program BLAST, especially BLASTP or TBLASTN, using default parameters. See, e.g., Altschul et al., 1990, J. Mol. Biol.215: 403-410 and 1997 Nucleic Acids Res.25:3389-3402.

[0178] A “variant” of a polypeptide, such an immunoglobulin, VH, VL, heavy chain, light chain, or CDR comprising an amino acid sequence specifically set forth herein, refers to a polypeptide comprising an amino acid sequence that is at least about 70%-99.9% (e.g., at least 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.5, or 99.9%) identical to the reference polypeptide sequence (e.g., as set forth in the sequence listing below), when the comparison is performed by a BLAST algorithm wherein the parameters of the algorithm are selected to give the largest match between the respective sequences over the entire length of the respective reference sequences. In some embodiments, a variant of a polypeptide includes a polypeptide having the amino acid sequence of a reference polypeptide sequence (e.g., as set forth in the sequence listing below) but for one or more (e.g., 1 to 10, or less than 20, or less than 10) missense mutations (e.g., conservative substitutions), nonsense mutations, deletions, or insertions.

[0179] Compositions or methods “comprising” or “including” one or more recited elements may include other elements not specifically recited. For example, a composition that “comprises” or “includes” a protein may contain the protein alone or in combination with other ingredients. The transitional phrase “consisting essentially of” means that the scope of a claim is to be interpreted to encompass the specified elements recited in the claim and those that do not materially affect the basic and novel characteristic(s) of the claimed invention. Thus, the term “consisting essentially of” when used in a claim of this invention is not intended to be interpreted to be equivalent to “comprising.”

[0180] “Optional” or “optionally” means that the subsequently described event or circumstance may or may not occur and that the description includes instances in which the event or circumstance occurs and instances in which the event or circumstance does not.

[0181] Designation of a range of values includes all integers within or defining the range, and all subranges defined by integers within the range. For example, 5-10 nucleotides is understood as 5, 6, 7, 8, 9, or 10 nucleotides, whereas 5-10% is understood to contain 5% and all possible values through 10%.Attorney Docket No.057766 / 625782

[0182] At least 17 nucleotides of a 20 nucleotide sequence is understood to include 17, 18, 19, or 20 nucleotides of the sequence provided, thereby providing an upper limit even if one is not specifically provided as it would be clearly understood. Similarly, up to 3 nucleotides would be understood to encompass 0, 1, 2, or 3 nucleotides, providing a lower limit even if one is not specifically provided. When “at least,” “up to,” or other similar language modifies a number, it can be understood to modify each number in the series.

[0183] As used herein, “no more than” or “less than” is understood as the value adjacent to the phrase and logical lower values or integers, as logical from context, to zero. For example, a duplex region of “no more than 2 nucleotide base pairs” has a 2, 1, or 0 nucleotide base pairs. When “no more than” or “less than” is present before a series of numbers or a range, it is understood that each of the numbers in the series or range is modified.

[0184] As used herein, it is understood that when the maximum amount of a value is represented by 100% (e.g., 100% inhibition) that the value is limited by the method of detection. For example, 100% inhibition is understood as inhibition to a level below the level of detection of the assay.

[0185] Unless otherwise apparent from the context, the term “about” encompasses values ± 5% of a stated value. In certain embodiments, the term “about” is understood to encompass tolerated variation or error within the art, e.g., 2 standard deviations from the mean, or the sensitivity of the method used to take a measurement, or a percent of a value as tolerated in the art, e.g., with age. When “about” is present before the first value of a series, it can be understood to modify each value in the series.

[0186] The term “and / or” refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative (“or”).

[0187] The term “or” refers to any one member of a particular list and also includes any combination of members of that list.

[0188] The singular forms of the articles “a,” “an,” and “the” include plural references unless the context clearly dictates otherwise. For example, the term “a protein” or “at least one protein” can include a plurality of proteins, including mixtures thereof.

[0189] Statistically significant means p ≤0.05.Attorney Docket No.057766 / 625782

[0190] In the event of a conflict between a sequence in the application and an indicated accession number or position in an accession number, the sequence in the application predominates. DETAILED DESCRIPTION I. Overview

[0191] Provided herein are methods of inserting a nucleic acid encoding a polypeptide of interest into a target genomic locus in a cell or a population of cells in a subject, methods of expressing a polypeptide of interest from a target genomic locus in a cell or a population of cells in a subject, methods of treating an enzyme deficiency (e.g., FIX deficiency or GAA deficiency) in a subject in need thereof, and methods of preventing or reducing the onset of a sign or symptom of an enzyme deficiency in a subject in need thereof.

[0192] The methods use CD40 inhibitors (e.g., CD40 antigen-binding molecules) (e.g., CD40xCD40 bispecific antigen-binding molecules disclosed herein) to mitigate immune response and facilitate redosing of nucleic acid constructs encoding a polypeptide of interest and nuclease agents targeting a target genomic locus (e.g., in a subject without preexisting immunity to the nucleic acid construct, the polypeptide of interest encoded by the nucleic acid construct, the nuclease agent or one or more nucleic acids encoding the nuclease agent, or a delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent, such as, e.g., AAV). The CD40 inhibitors (e.g., CD40 antigen- binding molecules) can be used for inhibiting CD40L-induced activation of CD40, e.g., in or on a cell that expresses CD40 (e.g., a B cell, dendritic cell, monocyte, platelet, or macrophage). The CD40 inhibitors (e.g., CD40 antigen-binding molecules) are able to suppress host B cell responses to new antigens. In AAV gene therapies, seronegative / naive patients are dosed with AAV and develop antibody responses to the AAV capsid antigen. This antibody response prevents future re-dosing of AAV because the antibodies are neutralizing, and the antibody response is sustained for 10+ years. When AAV is co-administered with CD40 inhibitors (e.g., CD40 antigen-binding molecules), the B cell response is suppressed and anti-AAV IgG responses are significantly suppressed. When CD40 inhibitor (e.g., CD40 antigen-binding molecule) is given during the period of AAV antigen exposure, the anti-AAV antibody response can be suppressed in animals. This allows re-dosing of any AAV gene therapy product. ForAttorney Docket No.057766 / 625782 example, for CRISPR-mediated gene insertion platforms consisting of an AAV and LNP, the AAV and / or LNP can be re-dosed multiple times when CD40 inhibitor (e.g., CD40 antigen- binding molecule) is co-administered. The CD40 inhibitor (e.g., CD40 antigen-binding molecule) prevents antibody formation against the AAV. The CD40 inhibitor (e.g., CD40 antigen-binding molecule) can also prevent antibody formation against certain LNP components (e.g., anti-PEG IgG), which can improve efficacy of LNP redosing.

[0193] Using CD40 inhibitors (e.g., CD40 antigen-binding molecules) to mitigate an anti- AAV antibody response can allow for repeated dosing of an identical gene insertion therapeutic cargo. This allows targeted cells in a subject to produce a polypeptide of interest in a step-wise, increasing fashion due to increased gene insertion in additional targeted cells following repeated dosing until a desired level of expression and / or activity of the polypeptide of interest is achieved in a subject without overshooting. This can be particularly advantageous in situations in which overshooting (i.e., achieving higher than desired levels of expression and / or activity of the polypeptide of interest) would result in undesired side effects (e.g., toxicity). Likewise, using CD40 inhibitors (e.g., CD40 antigen-binding molecules) to mitigate an anti-AAV antibody response can allow for gene insertion of an AAV template into two separate genomic locations from two discrete dosings (e.g., two discrete dosings of AAV and LNP). Similarly, using CD40 inhibitors (e.g., CD40 antigen-binding molecules) to mitigate an anti-AAV antibody response can allow for gene insertion of two different AAV templates (encoding different polypeptides of interest or the same polypeptide of interest) from two discrete dosings (e.g., two discrete dosings of AAV and LNP).

[0194] Other broad-spectrum immunosuppression methodologies have not been shown to be effective at enabling AAV vector re-administration at levels equivalent to naïve individuals. Similarly, CD40L blockade has also been shown to enable only partial AAV re-transduction. The CD40 antigen-binding molecules disclosed herein, which show superior binding characteristics and performance with low agonism as compared to other CD40 antigen-binding molecules, can achieve levels of re-transduction similar to naïve animals. In addition, single agent CD40 blockade is more targeted than other immunosuppression methods because it partially preserves de novo T-cell-independent antibody responses, which may provide patients full or partial protection from opportunistic infections in the absence of a de novo T-cell- dependent antibody response.Attorney Docket No.057766 / 625782

[0195] The methods using CD40 inhibitors can also use plasma cell depleting agents or combinations comprising plasma cell depleting agents when a subject has preexisting immunity against an immunogen to be administered to the subject (e.g., an immunogenic delivery vehicle such as, e.g., AAV). For example, the immunogen can be a nucleic acid construct, a polypeptide of interest, a nuclease agent, one or more nucleic acids encoding the nuclease agent, or a delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent. Examples of such methods include the following: (1) a plasma cell depleting agent or a combination comprising a plasma cell depleting agent as disclosed herein [e.g., in combination with a B cell depleting agent (e.g., a CD20xCD3 antigen-binding molecule) and / or an immunoglobulin depleting agent and / or plasmapheresis, therapeutic plasma exchange, or immunoadsorption] is used to eliminate preexisting immunity to an immunogen (e.g., AAV), while a CD40 inhibitor is used to prevent any new antibody response to the immunogen on subsequent immunogen exposure; (2) a plasma cell depleting agent or a combination comprising a plasma cell depleting agent as disclosed herein is used to eliminate potential residual antibody responses generated following immunogen (e.g., AAV) exposure in the presence of CD40 blockade (e.g., if CD40 blockade is not completely effective); or (3) CD40 blockade is used concurrently with a plasma cell depleting agent or a combination comprising a plasma cell depleting agent as disclosed herein to block ongoing antibody responses to an immunogen (e.g., AAV) from recent exposure. The present disclosure provides, for example, a distinct B cell immunosuppression approach that enables AAV vector re-transduction in subjects with preexisting AAV immunity at levels equal to seronegative subjects, by depleting pre-existing nAbs (e.g., via combined plasma cell and immunoglobulin depletion). Long-lived plasma cells (LLPC) mediate constitutive antibody production to most antigens and are the likely reservoir of persistent anti-AAV antibody immunity. It was discovered that pre-existing AAV nAbs in subjects with preexisting AAV immunity could be directly eliminated in vivo by LLPC depletion with linvoseltamab, a fully-human T cell-bridging bispecific antibody targeting B cell maturation antigen and CD3 (anti-BCMAxCD3 bispecific antibody), either alone or in combination with B cell depletion (to eliminate non-LLPC sources of anti-AAV nAbs) and / or FcRn blockade (to accelerate serum IgG clearance). Such methods can use plasma cell depleting agents or combinations comprising plasma cell depleting agents to mitigate immune response in subjects with preexisting immunity and facilitate redosing of nucleic acid constructs encoding aAttorney Docket No.057766 / 625782 polypeptide of interest and nuclease agents targeting a target genomic locus. Optionally, the plasma cell depleting agents (e.g., BCMAxCD3 antigen-binding molecules) are used in combination with other immunosuppression methodologies, such as immunoglobulin depleting agents (e.g., FcRn blockers or IgG degrading enzymes), B cell depleting agents, plasmapheresis, therapeutic plasma exchange, immunoadsorption, broad spectrum immunosuppression, or combinations thereof. In one example, plasma cell depleting agents (e.g., BCMAxCD3 bispecific antigen-binding molecules) are used in combination with immunoglobulin depleting agents (e.g., IgG half-life reducers, such as FcRn blockers). In another example, plasma cell depleting agents (e.g., BCMAxCD3 bispecific antigen-binding molecules) are used in combination with B cell depleting agents (e.g., CD20xCD3 antigen-binding molecules). In another example, BCMAxCD3 bispecific antigen-binding molecules are used in combination with immunoglobulin depleting agents (e.g., FcRn blockers) and B cell depleting agents (e.g., CD20xCD3 antigen-binding molecules). This allows re-dosing of any AAV gene therapy product in subjects with preexisting AAV immunity. For example, for CRISPR-mediated gene insertion platforms consisting of an AAV and LNP, the AAV and / or LNP can be re-dosed multiple times in subjects with preexisting AAV immunity when plasma cell depleting agents or combinations comprising plasma cell depleting agents are co-administered.

[0196] Using plasma cell depleting agents or combinations comprising plasma cell depleting agents to mitigate an anti-AAV antibody response in subjects with preexisting AAV immunity can allow for repeated dosing of an identical gene insertion therapeutic cargo. This allows targeted cells in a subject to produce a polypeptide of interest in a step-wise, increasing fashion due to increased gene insertion in additional targeted cells following repeated dosing until a desired level of expression and / or activity of the polypeptide of interest is achieved in a subject without overshooting. This can be particularly advantageous in situations in which overshooting (i.e., achieving higher than desired levels of expression and / or activity of the polypeptide of interest) would result in undesired side effects (e.g., toxicity). Likewise, using plasma cell depleting agents or combinations comprising plasma cell depleting agents to mitigate an anti- AAV antibody response in subjects with preexisting AAV immunity can allow for gene insertion of an AAV template into two separate genomic locations from two discrete dosings (e.g., two discrete dosings of AAV and LNP). Similarly, using plasma cell depleting agents or combinations comprising plasma cell depleting agents to mitigate an anti-AAV antibodyAttorney Docket No.057766 / 625782 response in subjects with preexisting AAV immunity can allow for gene insertion of two different AAV templates (encoding different polypeptides of interest or the same polypeptide of interest) from two discrete dosings (e.g., two discrete dosings of AAV and LNP).

[0197] Also provided are compositions, combinations, or kits comprising a CD40 inhibitor (e.g., CD40 antigen-binding molecule) in combination with: (a) a nucleic acid construct comprising a coding sequence for the polypeptide of interest; and (b) a nuclease agent or one or more nucleic acids encoding the nuclease agent, wherein the nuclease agent targets a nuclease target site in a target genomic locus. As used herein, the term “in combination with” means that additional component(s) may be administered prior to, concurrent with, or after the administration of the CD40 inhibitor (e.g., CD40 antigen-binding molecule). The different components of the combination can be formulated into a single composition, e.g., for simultaneous delivery, or formulated separately into two or more compositions (e.g., a kit including each component, for example, wherein the further agent is in a separate formulation).

[0198] Also provided are compositions or combinations or kits comprising a plasma cell depleting agent or combination comprising a plasma cell depleting agent in combination with: (a) a nucleic acid construct comprising a coding sequence for the polypeptide of interest; and (b) a nuclease agent or one or more nucleic acids encoding the nuclease agent, wherein the nuclease agent targets a nuclease target site in a target genomic locus. Such compositions, combinations, or kits comprise the plasma cell depleting agent in combination with a CD40 inhibitor. As used herein, the term “in combination with,” for example, a plasma cell depleting agent means that additional component(s) may be administered prior to, concurrent with, or after the administration of the plasma cell depleting agent. The different components of the combination can be formulated into a single composition, e.g., for simultaneous delivery, or formulated separately into two or more compositions (e.g., a kit including each component, for example, wherein the further agent is in a separate formulation). II. CD40 Inhibitors and CD40 Antigen-Binding Molecules

[0199] In one aspect, the present disclosure relates to CD40 inhibitors. CD40 inhibitors can block or suppress CD40 and / or CD40-mediated cellular signaling pathways and / or mechanisms, e.g., cellular signaling pathways and / or mechanisms associated with an immune response. CD40 inhibitors can include antigen-binding molecules such as monospecific, bispecific, andAttorney Docket No.057766 / 625782 multispecific antibodies that bind to CD40 (e.g., antagonist antigen-binding molecules, including monospecific, bispecific, and multispecific antibodies, that bind to CD40) or other CD40-CD40L inhibitors such as CD40L antigen-binding molecules (e.g., antibodies), peptide inhibitors, or small molecule inhibitors. Examples of such inhibitors are disclosed elsewhere herein. In some embodiments, the antigen-binding molecule is a monospecific anti-CD40 antibody. In some embodiments, the antigen-binding molecule is a multispecific (e.g., bispecific) antibody. Multispecific antibodies may be specific for different epitopes of one target polypeptide or may contain antigen-binding domains specific for more than one target polypeptide. See, e.g., Tutt et al., 1991, J. Immunol.147:60-69; Kufer et al., 2004, Trends Biotechnol.22:238-244. In some embodiments, the CD40 antibodies of the present disclosure can be linked to or co-expressed with another functional molecule, e.g., another peptide or protein. For example, an antibody or fragment thereof can be functionally linked (e.g., by chemical coupling, genetic fusion, noncovalent association or otherwise) to one or more other molecular entities, such as another antibody or antibody fragment to produce a bispecific or a multispecific antibody with a second binding specificity. In some embodiments, the multispecific antibody contains an antigen- binding domain that is specific for CD40 and an antigen-binding domain that is specific for another antigen (i.e., not CD40). In some embodiments, the multispecific antibody contains an antigen-binding domain that is specific for a first epitope of CD40 and an antigen-binding domain that is specific for a second epitope of CD40.

[0200] A CD40 inhibitor can be used for inhibiting CD40L-induced activation of CD40, e.g., in or on a cell that expresses CD40 (e.g., a B cell, dendritic cell, monocyte, platelet, or macrophage). The CD40 inhibitors are able to suppress host B cell responses to new antigens. In AAV gene therapies, seronegative / naive subjects are dosed with AAV and develop antibody responses to the AAV capsid antigen. This antibody response prevents future re-dosing of AAV because the antibodies are neutralizing, and the antibody response is sustained for 10+ years. When AAV is co-administered with a CD40 inhibitor, the B cell response is suppressed and anti- AAV IgG responses are significantly suppressed. When a CD40 inhibitor is given during the period of AAV antigen exposure, the anti-AAV antibody response can be suppressed in animals. This allows re-dosing of any AAV gene therapy product.

[0201] In some embodiments, a CD40 inhibitor is administered to a subject not having a pre- existing immunity against an immunogen (e.g., an immunogenic delivery vehicle such as, e.g.,Attorney Docket No.057766 / 625782 AAV (e.g., AAV comprising a nucleic acid construct described herein)). In some embodiments, a CD40 inhibitor is administered to a subject not having a pre-existing immunity against a nucleic acid construct described herein, a polypeptide of interest encoded by a nucleic acid construct described herein, a nuclease agent or one or more nucleic acids encoding the nuclease agent as described herein, or a delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent as described herein. In some embodiments, a CD40 inhibitor is administered to a subject not having a pre-existing immunity against an AAV vector comprising a nucleic acid construct described herein.

[0202] In some embodiments, CD40 blockade using a CD40 inhibitor can prevent antibody and T cell responses to Cas proteins (e.g., Cas9 proteins) that might lead to transduced cell clearance and / or inflammation associated with a T cell response. A. Monospecific Anti-CD40 Antibodies

[0203] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within an HCVR / LCVR amino acid sequence pair selected from the group consisting of SEQ ID NOS: 2 / 10, 22 / 10, and 32 / 10. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.

[0204] In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises: (a) an HCDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 4, 24, and 34; (b) an HCDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 6, 26, and 36; (c) an HCDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 8, 28, and 38; (d) an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12; (e) an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14); and (f) an LCDR3 comprising the amino acid sequence ofAttorney Docket No.057766 / 625782 SEQ ID NO: 16.

[0205] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises: (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or (c) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16.

[0206] In some embodiments, the anti-CD40 antibody comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some embodiments, the anti-CD40 antibody comprises an HCDR1 consisting of the amino acid sequence of SEQ ID NO: 4, an HCDR2 consisting of the amino acid sequence of SEQ ID NO: 6, an HCDR3 consisting of the amino acid sequence of SEQ ID NO: 8, an LCDR1 consisting of the amino acid sequence of SEQ ID NO: 12, an LCDR2 consisting of the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 consisting of the amino acid sequence of SEQ ID NO: 16.

[0207] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 2; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, atAttorney Docket No.057766 / 625782 least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 10. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 2. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 10.

[0208] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 18; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 20. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 18. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 20.

[0209] In some embodiments, the anti-CD40 antibody comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some embodiments, the anti-CD40 antibody comprises an HCDR1 consisting of the amino acid sequence of SEQ ID NO: 24, an HCDR2 consisting of the amino acid sequence of SEQ ID NO: 26, an HCDR3 consisting of the amino acid sequence of SEQ ID NO: 28, an LCDR1 consisting of the amino acid sequence of SEQ ID NO: 12, an LCDR2 consisting of the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 consisting of the amino acid sequence of SEQ ID NO: 16.

[0210] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 22; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ IDAttorney Docket No.057766 / 625782 NO: 10. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 22. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 10.

[0211] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 30; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 20. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 30. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 20.

[0212] In some embodiments, the anti-CD40 antibody comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16. In some embodiments, the anti-CD40 antibody comprises an HCDR1 consisting of the amino acid sequence of SEQ ID NO: 34, an HCDR2 consisting of the amino acid sequence of SEQ ID NO: 36, an HCDR3 consisting of the amino acid sequence of SEQ ID NO: 38, an LCDR1 consisting of the amino acid sequence of SEQ ID NO: 12, an LCDR2 consisting of the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 consisting of the amino acid sequence of SEQ ID NO: 16.

[0213] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 32; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 10. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising orAttorney Docket No.057766 / 625782 consisting of the amino acid sequence of SEQ ID NO: 32. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 10.

[0214] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 40; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 20. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 40. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 20.

[0215] In some embodiments, the anti-CD40 antibody has at least 85% sequence identity, e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity, to 30027P2. In some embodiments, the anti-CD40 antibody has the amino acid sequence of 30027P2.

[0216] In some embodiments, the anti-CD40 antibody has at least 85% sequence identity, e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity, to 21519P2. In some embodiments, the anti-CD40 antibody has the amino acid sequence of 21519P2.

[0217] In some embodiments, the anti-CD40 antibody has at least 85% sequence identity, e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity, to 21520P2. In some embodiments, the anti-CD40 antibody comprises or consists of the amino acid sequence of 21520P2.

[0218] Other CD40 antigen-binding molecules that can be used in the compositions, combinations, kits, or methods disclosed herein include other known anti-CD40 antibodies. Non- limiting examples of additional anti-CD40 antibodies include CD40 Monoclonal Antibody (1C10) (eBioscience™), CD40 Monoclonal Antibody (HM40-3) (eBioscience™), CD40 Monoclonal Antibody (5C3) (eBioscience™), CD40 Monoclonal Antibody (9G10) (Invitrogen),Attorney Docket No.057766 / 625782 CD40 Monoclonal Antibody (3 / 23) (Invitrogen), CD40 Monoclonal Antibody (HB14) (Invitrogen), CD40 Polyclonal Antibody (Invitrogen), CD40 Monoclonal Antibody (OTI1F12), TrueMAB™ (OriGene), CD40 Recombinant Rabbit Monoclonal Antibody (5V8X7) (Invitrogen), CD40 Monoclonal Antibody (LOB7 / 6) (Invitrogen), CD40 Monoclonal Antibody (5C3) (Invitrogen), CD40 Monoclonal Antibody (HI40a) (Invitrogen), CD40 Monoclonal Antibody (IL-A156) (Invitrogen), CD40 Monoclonal Antibody (C7) (Invitrogen), CD40 Polyclonal Antibody (Invitrogen, e.g., Cat # PA5-111025, Cat #PA5-111024, Cat #PA5-109301, Cat #PA5-117850, Cat #PA5-27419, Cat #PA5-78980, Cat #PA1-31075), CD40 Monoclonal Antibody (3 / 23) (Invitrogen), CD40 Monoclonal Antibody (HB14) (Invitrogen), CD40 Recombinant Rabbit Monoclonal Antibody (Bethyl Laboratories), CD40 Monoclonal Antibody (OTI8B8), TrueMAB™ (OriGene), CD40 Monoclonal Antibody (OTI1F12), TrueMAB™ (OriGene), CD40 Monoclonal Antibody (OTI5C9), TrueMAB™ (OriGene), CD40 Monoclonal Antibody (UMAB255), UltraMAB™ (OriGene), CD40 Monoclonal Antibody (2A8G5) (Proteintech), CD40 Monoclonal Antibody (G28.5) (Proteintech), CD40 Monoclonal Antibody (1C10) (Proteintech), CD40 Monoclonal Antibody (G28.5) (Proteintech), CD40 Monoclonal Antibody (2A8G5) (Proteintech), CD40 Monoclonal Antibody (UMAB183), UltraMAB™ (OriGene), CD40 Monoclonal Antibody (UMAB183), UltraMAB™ (OriGene), CD40 Monoclonal Antibody (HB14) (AbboMax), CD40 Monoclonal Antibody (1G1) (Abnova), CD40 Polyclonal Antibody (AbboMax, Cat #500-3704), CD40 Monoclonal Antibody (FGK45) (Leinco Technologies), CD40 Monoclonal Antibody (3D9) (Abnova), and CD40 Monoclonal Antibody (2H8) (Abnova).

[0219] In some embodiments, the anti-CD40 antibody is an antagonistic anti-CD40 antibody or a functional fragment thereof. The antagonistic anti-CD40 antibody may comprise any antagonistic anti-CD40 antibody described above or known in the art. Non-limiting examples of additional antagonistic anti-CD40 antibodies include iscalimab (also known as CFZ533) disclosed in Kahaly et al. (2019) J. Endocr. Sco.3:doi.org / 10.1210 / js.2019-0R19-6, Fisher et al. (2017) Arthritis Rheumatol.69:1784, Farkash et al. (2019) Am. J. Transplant.19:632, U.S. Pat. No. US 8,828,396, International Patent Application Publication No. WO 2012 / 075111, and clinical trial NCT02291029 sponsored by Novartis Pharmaceuticals; ravagalimab (also known as ABBV-323 and Ab102) disclosed in International Patent Application Publication No. WO 2016 / 196314 and U.S. Patent Application Publication No. US 2022 / 0289858; BI-655064Attorney Docket No.057766 / 625782 disclosed in Visvannathan et al. (2016) Arthritis Rheumatol.68:1588, U.S. Pat. No.8,591,900, and clinical trial NCT03385564 sponsored by Boehringer Ingelheim; bleselumab (also known as ASKP1240 or 341G2) disclosed in Anil et al. (2018) Biopharm. Drug Dispos.39:245-255, Harland et al. (2017) Am. J. Transplant.17:159-171, U.S. Pat. Nos.8,716,451 and 8,568,725, and clinical trials NCT01585233 and NCT02921789 sponsored by Astellas Pharma; ch5D12 disclosed in Kasran et al. (2005) Aliment. Pharmacol. Ther.22:111-122 and U.S. Patent Application Publication No. US 2008 / 0085531; lucatumumab (also known as HCD122 or CHIR- 12.12) disclosed in Bensinger et al. (2012) British J. Haematology 159:58-66, Byrd et al. (2012) Leuk. Lymphoma 53:10.3109 / 10428194.2012.681655, International Patent Application Publication No. WO 2005 / 044854, U.S. Patent Application Publication No. US 2007 / 0110754 and U.S. Pat. No. US 8,828,396; CHIR-5.9 disclosed in International Patent Application Publication No. WO 2005 / 044854 and U.S. Pat. No.8,637,032; abiprubart (KPL-404) disclosed in clinical trial NCT04497662 sponsored by Kiniksa Pharmaceuticals, Ltd. as well as in U.S. Patent Application Publication Nos. US 2023 / 0287132, US 2023 / 0203179, US 2023 / 0183367, and US 2023 / 0279135; BIIB063 disclosed in Musselli et al. (2017) 2017 ACR / ARHP Annual Meeting Abstract and International Patent Application Publication No. WO 2016028810; V19 and V15 disclosed in U.S. Patent Application Publication No. US 2022 / 0135694; h2C10 and variants thereof disclosed in U.S. Pat. No.11,439,706; FFP104 (also known as PG102) disclosed in U.S. Pat. Nos.8,669,352 and 11,396,552, International Patent Application Publication No. WO 2001 / 024823, U.S. Patent Application Publication No. US 2008 / 0085531, Bankert et al. (2015) J. Immunol.194:4319-4327, and clinical trials NCT02193360 and NCT02465944 sponsored by Fast Forward Pharmaceuticals; Ab101 disclosed in U.S. Patent Application Publication No. US 2022 / 0289858; Antibody A, antibody B, Antibody C, disclosed in U.S. Pat. No.11,242,394; G28.5 disclosed in International Patent Application Publication No. WO 2016028810; BMS3h-37, BMS3h-38, BMS3h-56, and BMS3h-198 disclosed in International Patent Application Publication No. WO2012145673A1,and Y12XX-hz28 (Vh-hzl4;Vk-hz2), Y12XX-hz40 (Vh-hzl2;Vk-hz3), and Y12XX-hz42 (Vh-hzl4;Vk-hz3) disclosed in International Patent Application Publication No. WO2020 / 106620A1, the contents of each of which are herein incorporated by reference in their entirety. An additional CD40 antibody useful in certain embodiments of the methods and compositions provided herein is teneliximab. Additional CD40 antagonist antibodies useful in certain embodiments of the methods and compositions providedAttorney Docket No.057766 / 625782 herein are disclosed in, for example, International Patent Application Publication Nos. WO 02 / 11763, WO 02 / 28481, WO 03 / 045978, WO 03 / 029296, WO 03 / 028809, WO 2005 / 044854, WO 2006 / 073443, WO 2007 / 124299, WO 2011 / 123489, WO 2016 / 196,314, WO 2017 / 040566, WO 2017 / 060242, WO 2018 / 217976, WO2019 / 156565, WO 2020 / 144605, WO 2020 / 106620, WO 2020 / 006347, U.S. Patent Application Publication Nos. US 2020 / 0291123, US 2017 / 0158771, US 2008 / 0057070, and U.S. Pat. Nos.5,874,082, 7,063,845, 9,125,893, 8,669,352, 9,598,494, 11,254,750, 11,780,927, 11,220,550, 11,202,827, 10,111,958, 11,242,397, 8,591,900, 9,475,879, 10,174,121, the contents of each of which are herein incorporated by reference in their entirety.

[0220] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 183 / 184. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.

[0221] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 177, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 178, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 179, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 180, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 181, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 182.

[0222] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 183; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 184. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising orAttorney Docket No.057766 / 625782 consisting of the amino acid sequence of SEQ ID NO: 183. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 184.

[0223] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 185; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 186. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 185. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 186. In some embodiments, the anti-CD40 antibody is abiprubart (also known as KPL-404). Abiprubart (KPL-404) HCDR1 = SEQ ID NO: 177 Abiprubart (KPL-404) HCDR2 = SEQ ID NO: 178 Abiprubart (KPL-404) HCDR3 = SEQ ID NO: 179 Abiprubart (KPL-404) LCDR1 = SEQ ID NO: 180 Abiprubart (KPL-404) LCDR2 = SEQ ID NO: 181 Abiprubart (KPL-404) LCDR3 = SEQ ID NO: 182 Abiprubart (KPL-404) HCVR = SEQ ID NO: 183 Abiprubart (KPL-404) LCVR = SEQ ID NO: 184 Abiprubart (KPL-404) HC = SEQ ID NO: 185 Abiprubart (KPL-404) LC = SEQ ID NO: 186

[0224] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 141 / 142. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.Attorney Docket No.057766 / 625782

[0225] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 135, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 136, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 137, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 138, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 139, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 140.

[0226] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 141; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 142. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 141. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 142.

[0227] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 143; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 144. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 143. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 144. In some embodiments, the anti-CD40 antibody is iscalimab. Iscalimab (CFZ533) HCDR1 = SEQ ID NO: 135 Iscalimab (CFZ533) HCDR2 = SEQ ID NO: 136 Iscalimab (CFZ533) HCDR3 = SEQ ID NO: 137 Iscalimab (CFZ533) LCDR1 = SEQ ID NO: 138 Iscalimab (CFZ533) LCDR2 = SEQ ID NO: 139 Iscalimab (CFZ533) LCDR3 = SEQ ID NO: 140 Iscalimab (CFZ533) HCVR = SEQ ID NO: 141Attorney Docket No.057766 / 625782 Iscalimab (CFZ533) LCVR = SEQ ID NO: 142 Iscalimab (CFZ533) HC = SEQ ID NO: 143 Iscalimab (CFZ533) LC = SEQ ID NO: 144

[0228] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 151 / 268. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.

[0229] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 145, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 146, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 147, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 148, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 149, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 150.

[0230] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 151; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 268. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 151. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 268.

[0231] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%Attorney Docket No.057766 / 625782 sequence identity to SEQ ID NO: 152; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 153. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 152. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 153. In some embodiments, the anti-CD40 antibody is ravagalimab. Ravagalimab (ABBV-323; Ab102) HCDR1 = SEQ ID NO: 145 Ravagalimab (ABBV-323; Ab102) HCDR2 = SEQ ID NO: 146 Ravagalimab (ABBV-323; Ab102) HCDR3 = SEQ ID NO: 147 Ravagalimab (ABBV-323; Ab102) LCDR1 = SEQ ID NO: 148 Ravagalimab (ABBV-323; Ab102) LCDR2 = SEQ ID NO: 149 Ravagalimab (ABBV-323; Ab102) LCDR3 = SEQ ID NO: 150 Ravagalimab (ABBV-323; Ab102) HCVR = SEQ ID NO: 151 Ravagalimab (ABBV-323; Ab102) LCVR = SEQ ID NO: 268 Ravagalimab (ABBV-323; Ab102) HC = SEQ ID NO: 152 Ravagalimab (ABBV-323; Ab102) LC = SEQ ID NO: 153

[0232] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 231 / 232. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.

[0233] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 269, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 220, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 221, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 222, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 223, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 150.Attorney Docket No.057766 / 625782

[0234] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 231; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 232. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 231. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 232.

[0235] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 236; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 237. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 236. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 237. In some embodiments, the anti-CD40 antibody is BI-655064. BI-655064 HCDR1 = SEQ ID NO: 269 BI-655064 HCDR2 = SEQ ID NO: 220 BI-655064 HCDR3 = SEQ ID NO: 221 BI-655064 LCDR1 = SEQ ID NO: 222 BI-655064 LCDR2 = SEQ ID NO: 223 BI-655064 LCDR3 = SEQ ID NO: 150 BI-655064 HCVR = SEQ ID NO: 231 BI-655064 LCVR = SEQ ID NO: 232 BI-655064 HC = SEQ ID NO: 236 BI-655064 LC = SEQ ID NO: 237

[0236] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set containedAttorney Docket No.057766 / 625782 within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 160 / 161. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.

[0237] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 154, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 155, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 156, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 157, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 158, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 159.

[0238] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 160; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 161. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 160. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 161.

[0239] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 162; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 163. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 162. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 163. In some embodiments, the anti-CD40 antibody is bleselumab.Attorney Docket No.057766 / 625782 Bleselumab HCDR1 = SEQ ID NO: 154 Bleselumab HCDR2 = SEQ ID NO: 155 Bleselumab HCDR3 = SEQ ID NO: 156 Bleselumab LCDR1 = SEQ ID NO: 157 Bleselumab LCDR2 = DAS (SEQ ID NO: 158) Bleselumab LCDR3 = SEQ ID NO: 159 Bleselumab HCVR = SEQ ID NO: 160 Bleselumab LCVR = SEQ ID NO: 161 Bleselumab HC = SEQ ID NO: 162 Bleselumab LC = SEQ ID NO: 163

[0240] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 170 / 171. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.

[0241] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 164, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 165, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 166, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 167, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 168, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 169.

[0242] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 170; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 171. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 170. In some embodiments, the anti-CD40Attorney Docket No.057766 / 625782 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 171. In some embodiments, the anti-CD40 antibody is ch5D12. ch5D12 HCDR1 = SEQ ID NO: 164 ch5D12 HCDR2 = SEQ ID NO: 165 ch5D12 HCDR3 = SEQ ID NO: 166 ch5D12 LCDR1 = SEQ ID NO: 167 ch5D12 LCDR2 = SEQ ID NO: 168 ch5D12 LCDR3 = SEQ ID NO: 169 ch5D12 HCVR = SEQ ID NO: 170 ch5D12 LCVR = SEQ ID NO: 171

[0243] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 141 / 142. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.

[0244] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 135, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 136, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 137, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 138, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 139, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 140.

[0245] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 141; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 142. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising orAttorney Docket No.057766 / 625782 consisting of the amino acid sequence of SEQ ID NO: 141. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 142.

[0246] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 172; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 144. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 172. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 144. In some embodiments, the anti-CD40 antibody is lucatumumab. Lucatumumab (HCD122 or CHIR-12.12) HCDR1 = SEQ ID NO: 135 Lucatumumab (HCD122 or CHIR-12.12) HCDR2 = SEQ ID NO: 136 Lucatumumab (HCD122 or CHIR-12.12) HCDR3 = SEQ ID NO: 137 Lucatumumab (HCD122 or CHIR-12.12) LCDR1 = SEQ ID NO: 138 Lucatumumab (HCD122 or CHIR-12.12) LCDR2 = SEQ ID NO: 139 Lucatumumab (HCD122 or CHIR-12.12) LCDR3 = SEQ ID NO: 140 Lucatumumab (HCD122 or CHIR-12.12) HCVR = SEQ ID NO: 141 Lucatumumab (HCD122 or CHIR-12.12) LCVR = SEQ ID NO: 142 Lucatumumab (HCD122 or CHIR-12.12) HC = SEQ ID NO: 172 Lucatumumab (HCD122 or CHIR-12.12) LC = SEQ ID NO: 144

[0247] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 173 / 174. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.Attorney Docket No.057766 / 625782

[0248] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 173; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 174. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 173. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 174.

[0249] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 175; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 176. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 175. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 176. In some embodiments, the anti-CD40 antibody is CHIR-5.9. CHIR-5.9 HCVR = SEQ ID NO: 173 CHIR-5.9 LCVR = SEQ ID NO: 174 CHIR-5.9 HC = SEQ ID NO: 175 CHIR-5.9 LC = SEQ ID NO: 176

[0250] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 188 / 189. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / orAttorney Docket No.057766 / 625782 LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.

[0251] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 164, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 281, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 166, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 187, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 168, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 169.

[0252] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 188; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 189. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 188. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 189.

[0253] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 190 or 191; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 192. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 190 or 191. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 192. In some embodiments, the anti-CD40 antibody is FFP104 (also known as PG102). PG102 / / FFP104 HCDR1 = SEQ ID NO: 164 PG102 / / FFP104 HCDR2 = SEQ ID NO: 281 PG102 / / FFP104 HCDR3 = SEQ ID NO: 166Attorney Docket No.057766 / 625782 PG102 / / FFP104 LCDR1 = SEQ ID NO: 187 PG102 / / FFP104 LCDR2 = SEQ ID NO: 168 PG102 / / FFP104 LCDR3 = SEQ ID NO: 169 PG102 / / FFP104 HCVR = SEQ ID NO: 188 PG102 / / FFP104 LCVR = SEQ ID NO: 189 PG102 / / FFP104 HC = SEQ ID NO: 190 or 191 PG102 / / FFP104 LC = SEQ ID NO: 192

[0254] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 199 / 200. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.

[0255] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 193, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 194, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 195, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 196, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 197, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 198.

[0256] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 199; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 200. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 199. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 200.

[0257] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprisingAttorney Docket No.057766 / 625782 an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 201; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 202. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 201. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 202. In some embodiments, the anti-CD40 antibody is BIIB063. BIIB063 HCDR1 = SEQ ID NO: 193 BIIB063 HCDR2 = SEQ ID NO: 194 BIIB063 HCDR3 = SEQ ID NO: 195 BIIB063 LCDR1 = SEQ ID NO: 196 BIIB063 LCDR2 = SEQ ID NO: 197 BIIB063 LCDR3 = SEQ ID NO: 198 BIIB063 HCVR = SEQ ID NO: 199 BIIB063 LCVR = SEQ ID NO: 200 BIIB063 HC = SEQ ID NO: 201 BIIB063 LC = SEQ ID NO: 202

[0258] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3 amino acid sequences set contained within the HCVR amino acid sequence set forth in SEQ ID NO: 206. In some embodiments, the CDRs within the HCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR are identified according to the IMGT definition.

[0259] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 203, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 204, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 205.

[0260] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at leastAttorney Docket No.057766 / 625782 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 206. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 206. In some embodiments, the anti-CD40 antibody is V19. V19 HCDR1 = SEQ ID NO: 203 V19 HCDR2 = SEQ ID NO: 204 V19 HCDR3 = SEQ ID NO: 205 V19 HCVR = SEQ ID NO: 206

[0261] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3 amino acid sequences set contained within the HCVR amino acid sequence set forth in SEQ ID NO: 210. In some embodiments, the CDRs within the HCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR are identified according to the IMGT definition.

[0262] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 207, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 208, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 209.

[0263] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 210. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 210. In some embodiments, the anti-CD40 antibody is V15. V15 HCDR1 = SEQ ID NO: 207 V15 HCDR2 = SEQ ID NO: 208 V15 HCDR3 = SEQ ID NO: 209 V15 HCVR = SEQ ID NO: 210Attorney Docket No.057766 / 625782

[0264] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 183 / 184, 183 / 212, 183 / 215, 183 / 217, 211 / 184, 211 / 212, 211 / 215, 211 / 217, 213 / 184, 213 / 212, 213 / 215, 213 / 217, 214 / 184, 214 / 212, 214 / 215, 214 / 217, 216 / 184, 216 / 212, 216 / 215, or 216 / 217. In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 183 / 184, 211 / 212, 213 / 212, 214 / 215, or 216 / 217. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.

[0265] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 177, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 178, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 179, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 180, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 181, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 182.

[0266] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 183, 211, 213, 214, or 216; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 184, 212, 215, or 217. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 183, 211, 213, 214, or 216. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 184, 212, 215, orAttorney Docket No.057766 / 625782 217. In some embodiments, the anti-CD40 antibody is h2C10 or a variant thereof. 2C10 HCDR1 = SEQ ID NO: 177 2C10 HCDR2 = SEQ ID NO: 178 2C10 HCDR3 = SEQ ID NO: 179 2C10 LCDR1 = SEQ ID NO: 180 2C10 LCDR2 = SEQ ID NO: 181 2C10 LCDR3 = SEQ ID NO: 182 2C10_h1 HCVR = SEQ ID NO: 211 2C10_h2 HCVR = SEQ ID NO: 213 2C10_h3 HCVR = SEQ ID NO: 183 2C10_l1 LCVR = SEQ ID NO: 212 2C10_l2 LCVR = SEQ ID NO: 184 2C10HP HCVR = SEQ ID NO: 183 2C10HB1 HCVR = SEQ ID NO: 214 2C10HB2 HCVR = SEQ ID NO: 216 2C10KP LCVR = SEQ ID NO: 184 2C10KB1 LCVR = SEQ ID NO: 215 2C10KB2 LCVR = SEQ ID NO: 217

[0267] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 151 / 268. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.

[0268] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 145, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 146, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 147, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 148, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 149, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 150.

[0269] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at leastAttorney Docket No.057766 / 625782 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 151; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 268. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 151. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 268.

[0270] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 218; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 153. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 218. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 153. In some embodiments, the anti-CD40 antibody is Ab101. Ab101 HCDR1 = SEQ ID NO: 145 Ab101 HCDR2 = SEQ ID NO: 146 Ab101 HCDR3 = SEQ ID NO: 147 Ab101 LCDR1 = SEQ ID NO: 148 Ab101 LCDR2 = SEQ ID NO: 149 Ab101 LCDR3 = SEQ ID NO: 150 Ab101 HCVR = SEQ ID NO: 151 Ab101 LCVR = SEQ ID NO: 268 Ab101 HC = SEQ ID NO: 218 Ab101 LC = SEQ ID NO: 153

[0271] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 224 / 225. In someAttorney Docket No.057766 / 625782 embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.

[0272] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 219, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 220, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 221, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 222, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 223, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 150.

[0273] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 224; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 225. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 224. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 225.

[0274] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 226, 227, 228, or 229; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 230. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 226, 227, 228, or 229. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 230. In some embodiments, the anti-CD40 antibody is Antibody A.Attorney Docket No.057766 / 625782 Antibody A HCDR1 = SEQ ID NO: 219 Antibody A HCDR2 = SEQ ID NO: 220 Antibody A HCDR3 = SEQ ID NO: 221 Antibody A LCDR1 = SEQ ID NO: 222 Antibody A LCDR2 = SEQ ID NO: 223 Antibody A LCDR3 = SEQ ID NO: 150 Antibody A HCVR = SEQ ID NO: 224 Antibody A LCVR = SEQ ID NO: 225 Antibody A HC = SEQ ID NO: 226 or 227 or 228 or 229 Antibody A LC = SEQ ID NO: 230

[0275] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 231 / 232. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.

[0276] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 219, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 220, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 221, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 222, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 223, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 150.

[0277] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 231; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 232. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 231. In some embodiments, the anti-CD40Attorney Docket No.057766 / 625782 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 232.

[0278] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 233, 234, 235, or 236; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 237. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 233, 234, 235, or 236. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 237. In some embodiments, the anti-CD40 antibody is Antibody B. Antibody B HCDR1 = SEQ ID NO: 219 Antibody B HCDR2 = SEQ ID NO: 220 Antibody B HCDR3 = SEQ ID NO: 221 Antibody B LCDR1 = SEQ ID NO: 222 Antibody B LCDR2 = SEQ ID NO: 223 Antibody B LCDR3 = SEQ ID NO: 150 Antibody B HCVR = SEQ ID NO: 231 Antibody B LCVR = SEQ ID NO: 232 Antibody B HC = SEQ ID NO: 233 or 234 or 235 or 236 Antibody B LC = SEQ ID NO: 237

[0279] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 244 / 245. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.Attorney Docket No.057766 / 625782

[0280] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 238, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 239, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 240, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 241, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 242, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 243.

[0281] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 244; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 245. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 244. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 245.

[0282] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 246, 247, 248, or 249; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 250. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 246, 247, 248, or 249. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 250. In some embodiments, the anti-CD40 antibody is Antibody C. Antibody C HCDR1 = SEQ ID NO: 238 Antibody C HCDR2 = SEQ ID NO: 239 Antibody C HCDR3 = SEQ ID NO: 240 Antibody C LCDR1 = SEQ ID NO: 241 Antibody C LCDR2 = SEQ ID NO: 242 Antibody C LCDR3 = SEQ ID NO: 243Attorney Docket No.057766 / 625782 Antibody C HCVR = SEQ ID NO: 244 Antibody C LCVR = SEQ ID NO: 245 Antibody C HC = SEQ ID NO: 246 or 247 or 248 or 249 Antibody C LC = SEQ ID NO: 250

[0283] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 251 / 252. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.

[0284] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 251; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 252. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 251. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 252. In some embodiments, the anti-CD40 antibody is G28.5. G28.5 HCVR = SEQ ID NO: 251 G28.5 LCVR = SEQ ID NO: 252

[0285] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 259 / 260. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the KabatAttorney Docket No.057766 / 625782 definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.

[0286] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 253, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 254, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 255, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 256, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 257, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 258.

[0287] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 259; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 260. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 259. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 260.

[0288] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 261; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 262. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 261. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 262. In some embodiments, the anti-CD40 antibody is Y12XX-hz28. Y12XX-hz28 (Vh-hzl4;Vk-hz2) HCDR1 = SEQ ID NO: 253Attorney Docket No.057766 / 625782 Y12XX-hz28 (Vh-hzl4;Vk-hz2) HCDR2 = SEQ ID NO: 254 Y12XX-hz28 (Vh-hzl4;Vk-hz2) HCDR3 = SEQ ID NO: 255 Y12XX-hz28 (Vh-hzl4;Vk-hz2) LCDR1 = SEQ ID NO: 256 Y12XX-hz28 (Vh-hzl4;Vk-hz2) LCDR2 = SEQ ID NO: 257 Y12XX-hz28 (Vh-hzl4;Vk-hz2) LCDR3 = SEQ ID NO: 258 Y12XX-hz28 (Vh-hzl4;Vk-hz2) HCVR = SEQ ID NO: 259 Y12XX-hz28 (Vh-hzl4;Vk-hz2) LCVR = SEQ ID NO: 260 Y12XX-hz28 (Vh-hzl4;Vk-hz2) HC = SEQ ID NO: 261 Y12XX-hz28 (Vh-hzl4;Vk-hz2) LC = SEQ ID NO: 262

[0289] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 264 / 265. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.

[0290] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 253, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 263, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 255, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 256, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 257, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 258.

[0291] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 264; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 265. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 264. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ IDAttorney Docket No.057766 / 625782 NO: 265.

[0292] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 266; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 267. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 266. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 267. In some embodiments, the anti-CD40 antibody is Y12XX-hz40. Y12XX-hz40 (Vh-hzl2;Vk-hz3) HCDR1 = SEQ ID NO: 253 Y12XX-hz40 (Vh-hzl2;Vk-hz3) HCDR2 = SEQ ID NO: 263 Y12XX-hz40 (Vh-hzl2;Vk-hz3) HCDR3 = SEQ ID NO: 255 Y12XX-hz40 (Vh-hzl2;Vk-hz3) LCDR1 = SEQ ID NO: 256 Y12XX-hz40 (Vh-hzl2;Vk-hz3) LCDR2 = SEQ ID NO: 257 Y12XX-hz40 (Vh-hzl2;Vk-hz3) LCDR3 = SEQ ID NO: 258 Y12XX-hz40 (Vh-hzl2;Vk-hz3) HCVR = SEQ ID NO: 264 Y12XX-hz40 (Vh-hzl2;Vk-hz3) LCVR = SEQ ID NO: 265 Y12XX-hz40 (Vh-hzl2;Vk-hz3) HC = SEQ ID NO: 266 Y12XX-hz40 (Vh-hzl2;Vk-hz3) LC = SEQ ID NO: 267

[0293] In some embodiments, the present disclosure provides monospecific anti-CD40 antibodies or antigen-binding fragments thereof that specifically bind CD40 (e.g., human CD40). In some embodiments, the anti-CD40 antibody or antigen-binding fragment thereof comprises the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 259 / 265. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Kabat definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the Chothia definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the AbM definition. In some embodiments, the CDRs within the HCVR and / or LCVR are identified according to the IMGT definition.

[0294] In some embodiments, the anti-CD40 antibody or antigen-binding fragment comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 253, an HCDR2Attorney Docket No.057766 / 625782 comprising the amino acid sequence of SEQ ID NO: 254, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 255, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 256, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 257, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 258.

[0295] In some embodiments, the anti-CD40 antibody comprises: an HCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 259; and / or an LCVR having at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 265. In some embodiments, the anti-CD40 antibody comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 259. In some embodiments, the anti-CD40 antibody comprises an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 265.

[0296] In some embodiments, the anti-CD40 antibody comprises: a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 261; and / or a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 267. In some embodiments, the anti-CD40 antibody comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 261. In some embodiments, the anti-CD40 antibody comprises a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 267. In some embodiments, the anti-CD40 antibody is Y12XX-hz42. Y12XX-hz42 (Vh-hzl4;Vk-hz3) HCDR1 = SEQ ID NO: 253 Y12XX-hz42 (Vh-hzl4;Vk-hz3) HCDR2 = SEQ ID NO: 254 Y12XX-hz42 (Vh-hzl4;Vk-hz3) HCDR3 = SEQ ID NO: 255 Y12XX-hz42 (Vh-hzl4;Vk-hz3) LCDR1 = SEQ ID NO: 256 Y12XX-hz42 (Vh-hzl4;Vk-hz3) LCDR2 = SEQ ID NO: 257 Y12XX-hz42 (Vh-hzl4;Vk-hz3) LCDR3 = SEQ ID NO: 258 Y12XX-hz42 (Vh-hzl4;Vk-hz3) HCVR = SEQ ID NO: 259 Y12XX-hz42 (Vh-hzl4;Vk-hz3) LCVR = SEQ ID NO: 265 Y12XX-hz42 (Vh-hzl4;Vk-hz3) HC = SEQ ID NO: 261 Y12XX-hz42 (Vh-hzl4;Vk-hz3) LC = SEQ ID NO: 267Attorney Docket No.057766 / 625782

[0297] In some cases, the antibody comprises a human IgG heavy chain constant region. In some cases, the human IgG heavy chain constant region is isotype IgG1. In some cases, the human IgG heavy chain constant region is isotype IgG4. B. CD40xCD40 Bispecific Antigen-Binding Molecules

[0298] The present disclosure also provides multispecific antigen-binding molecules that specifically bind CD40. In some embodiments, the bispecific antigen-binding molecule comprises two CD40 binding arms comprising distinct HCVRs paired with a common LCVR. In some embodiments, the bispecific antigen-binding molecule comprises two CD40 binding arms comprising distinct HCVRs which are not paired with a common LCVR but instead are paired with distinct LCVRs which can comprise, e.g., without limitation any of the LCVRs, or combination thereof, disclosed herein. In some embodiments, the antigen-binding molecule is a bispecific antigen-binding molecule, e.g., bispecific antibody. Any of the anti-CD40 antibodies disclosed herein can be in bispecific configuration, including a second arm derived from any other anti-CD40 antibody disclosed herein. In some embodiments, the bispecific antigen-binding molecule comprises a first antigen-binding domain (D1) that binds a first epitope of CD40 (e.g., human CD40), and a second antigen-binding domain (D2) that binds a second epitope of CD40 (e.g., human CD40). In some embodiments, D1 and D2 do not compete with one another for binding to CD40 (e.g., human CD40). In some embodiments, D1 and D2 compete with one another for binding to CD40 (e.g., human CD40).

[0299] In some embodiments, the bispecific antigen-binding molecule comprises two different heavy chain immunoglobulin variable regions, wherein at least one heavy chain immunoglobulin variable region comprises an HCDR1-HCDR2-HCDR3 amino acid sequences set contained within an HCVR amino acid sequence selected from the group consisting of SEQ ID NOS: 2, 22, 32, 141, 151, 160, 170, 173, 183, 188, 199, 206, 210, 211, 213, 214, 216, 224, 231, 244, 251, 259, and 264. In some embodiments, the bispecific antigen-binding molecule comprises two different heavy chain immunoglobulin variable regions, wherein at least one heavy chain immunoglobulin variable region comprises an HCDR1-HCDR2-HCDR3 amino acid sequences set contained within an HCVR amino acid sequence selected from the group consisting of SEQ ID NOS: 2, 22, and 32. In some embodiments, each heavy chainAttorney Docket No.057766 / 625782 immunoglobulin variable region comprises an HCDR1-HCDR2-HCDR3 amino acid sequences set contained within an HCVR amino acid sequence selected from the group consisting of SEQ ID NOS: 2, 22, 32, 141, 151, 160, 170, 173, 183, 188, 199, 206, 210, 211, 213, 214, 216, 224, 231, 244, 251, 259, and 264. In some embodiments, each heavy chain immunoglobulin variable region comprises an HCDR1-HCDR2-HCDR3 amino acid sequences set contained within an HCVR amino acid sequence selected from the group consisting of SEQ ID NOS: 2, 22, and 32. In some embodiments, the CDRs within the HCVR are identified according to the Kabat definition. In some embodiments, the CDRs are identified according to the Chothia definition. In some embodiments, the CDRs are identified according to the AbM definition. In some embodiments, the CDRs are identified according to the IMGT definition.

[0300] In some embodiments, at least one heavy chain immunoglobulin variable region comprises an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 2, 22, 32, 141, 151, 160, 170, 173, 183, 188, 199, 206, 210, 211, 213, 214, 216, 224, 231, 244, 251, 259, or 264. In some embodiments, at least one heavy chain immunoglobulin variable region comprises or consists of the amino acid sequence of SEQ ID NO: 2, 22, 32, 141, 151, 160, 170, 173, 183, 188, 199, 206, 210, 211, 213, 214, 216, 224, 231, 244, 251, 259, or 264.

[0301] In some embodiments, one or more of the heavy chain immunoglobulin variable regions comprises a set of HCDR sequences selected from the group consisting of: (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8; (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28; and (c) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38.

[0302] In some embodiments, the bispecific antigen-binding molecule comprises (i) an antigen-binding domain that comprises a heavy chain immunoglobulin variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and an HCDR3 comprising the aminoAttorney Docket No.057766 / 625782 acid sequence of SEQ ID NO: 8; and (ii) an antigen-binding domain that comprises a heavy chain immunoglobulin variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28.

[0303] In some embodiments, the bispecific antigen-binding molecule comprises (i) an antigen-binding domain that comprises a heavy chain immunoglobulin variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8; and (ii) an antigen-binding domain that comprises a heavy chain immunoglobulin variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38.

[0304] In some embodiments, the bispecific antigen-binding molecule comprises (i) an antigen-binding domain that comprises a heavy chain immunoglobulin variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28; and (ii) an antigen-binding domain that comprises a heavy chain immunoglobulin variable region comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38.

[0305] In some embodiments, the bispecific antigen-binding molecule comprises a common light chain variable region. In some embodiments, the light chain variable region comprises an LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the LCVR amino acid sequence of SEQ ID NO: 10. In some embodiments, the light chain variable region comprises an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16.

[0306] In some embodiments, the bispecific antigen-binding molecule comprises a D1 that binds a first epitope of human CD40, wherein the D1 domain comprises a heavy chain immunoglobulin chain comprising: (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and an HCDR3Attorney Docket No.057766 / 625782 comprising the amino acid sequence of SEQ ID NO: 8; or (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28.

[0307] In some embodiments, the D1 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8. In some embodiments, the D1 domain comprises an HCDR1 consisting of the amino acid sequence of SEQ ID NO: 4, an HCDR2 consisting of the amino acid sequence of SEQ ID NO: 6, and an HCDR3 consisting of the amino acid sequence of SEQ ID NO: 8. In some embodiments, the D1 domain comprises an HCVR comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 2. In some embodiments, the D1 domain comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 2

[0308] In some embodiments, the D1 domain comprises a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 46. In some embodiments, the D1 domain comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 46.

[0309] In some embodiments, the D1 domain comprises a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 52. In some embodiments, the D1 domain comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 52.

[0310] In some embodiments, the D1 domain further comprises a light chain immunoglobulin chain comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16.

[0311] In some embodiments, the D1 domain comprises a LCVR comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity toAttorney Docket No.057766 / 625782 SEQ ID NO: 10. In some embodiments, the D1 domain comprises an LCVR comprising the amino acid sequence of SEQ ID NO: 10.

[0312] In some embodiments, the D1 domain comprises a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 20. In some embodiments, the D1 domain comprises a light chain comprising the amino acid sequence of SEQ ID NO: 20.

[0313] In some embodiments, the D1 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28. In some embodiments, the D1 domain comprises an HCDR1 consisting of the amino acid sequence of SEQ ID NO: 24, an HCDR2 consisting of the amino acid sequence of SEQ ID NO: 26, and an HCDR3 consisting of the amino acid sequence of SEQ ID NO: 28. In some embodiments, the D1 domain comprises an HCVR comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 22. In some embodiments, the D1 domain comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 22.

[0314] In some embodiments, the D1 domain comprises a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 42. In some embodiments, the D1 domain comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 42.

[0315] In some embodiments, the D1 domain comprises a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 48. In some embodiments, the D1 domain comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 48.

[0316] In some embodiments, the D1 domain further comprises a light chain immunoglobulin chain comprising an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS (SEQ ID NO: 14), and an LCDR3Attorney Docket No.057766 / 625782 comprising the amino acid sequence of SEQ ID NO: 16.

[0317] In some embodiments, the D1 domain comprises a LCVR comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 10. In some embodiments, the D1 domain comprises an LCVR comprising the amino acid sequence of SEQ ID NO: 10.

[0318] In some embodiments, the D1 domain comprises a light chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 20. In some embodiments, the D1 domain comprises a light chain comprising the amino acid sequence of SEQ ID NO: 20.

[0319] In some embodiments, the bispecific antigen-binding molecule comprises a D2 that binds a second epitope of human CD40, wherein the D2 domain comprises a heavy chain immunoglobulin chain comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38. In some embodiments, the D2 domain comprises an HCDR1 consisting of the amino acid sequence of SEQ ID NO: 34, an HCDR2 consisting of the amino acid sequence of SEQ ID NO: 36, and an HCDR3 consisting of the amino acid sequence of SEQ ID NO: 38. In some embodiments, the D2 domain comprises an HCVR comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 32. In some embodiments, the D2 domain comprises an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 32.

[0320] In some embodiments, the D2 domain comprises a heavy chain comprising an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 44. In some embodiments, the D2 domain comprises a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 44.

[0321] In some embodiments,...

Claims

Attorney Docket No.057766 / 625782 We claim:

1. A method of inserting a nucleic acid encoding a polypeptide of interest into a target genomic locus in a cell or a population of cells in a subject, comprising administering to the subject: (a) a nucleic acid construct comprising a coding sequence for the polypeptide of interest; (b) a nuclease agent or one or more nucleic acids encoding the nuclease agent, wherein the nuclease agent targets a nuclease target site in the target genomic locus; and (c) a CD40 inhibitor, wherein the nuclease agent cleaves the nuclease target site, and the nucleic acid construct is inserted into the target genomic locus.

2. A method of expressing a polypeptide of interest from a target genomic locus in a cell or a population of cells in a subject, comprising administering to the subject: (a) a nucleic acid construct comprising a coding sequence for the polypeptide of interest; and (b) a nuclease agent or one or more nucleic acids encoding the nuclease agent, wherein the nuclease agent targets a nuclease target site in the target genomic locus, (c) a CD40 inhibitor, wherein the nuclease agent cleaves the nuclease target site, the nucleic acid construct is inserted into the target genomic locus to create a modified target genomic locus, and the polypeptide of interest is expressed from the modified target genomic locus.

3. A method of treating an enzyme deficiency in a subject in need thereof, comprising administering to the subject: (a) a nucleic acid construct comprising a coding sequence for a polypeptide of interest, wherein the polypeptide of interest comprises an enzyme to treat the enzyme deficiency; (b) a nuclease agent or one or more nucleic acids encoding the nuclease agent, wherein the nuclease agent targets a nuclease target site in a target genomic locus; and (c) a CD40 inhibitor, wherein the nuclease agent cleaves the nuclease target site, the nucleic acid construct is inserted into the target genomic locus to create a modified target genomic locus, andAttorney Docket No.057766 / 625782 the polypeptide of interest is expressed from the modified target genomic locus, thereby treating the enzyme deficiency.

4. A method of preventing or reducing the onset of a sign or symptom of an enzyme deficiency in a subject in need thereof, comprising administering to the subject: (a) a nucleic acid construct comprising a coding sequence for a polypeptide of interest, wherein the enzyme deficiency is characterized by a loss-of-function of the polypeptide of interest; (b) a nuclease agent or one or more nucleic acids encoding the nuclease agent, wherein the nuclease agent targets a nuclease target site in a target genomic locus; and (c) a CD40 inhibitor, wherein the nuclease agent cleaves the nuclease target site, the nucleic acid construct is inserted into the target genomic locus to create a modified target genomic locus, and the polypeptide of interest is expressed from the modified target genomic locus, thereby preventing or reducing the onset of the sign or symptom of the enzyme deficiency.

5. The method of claim 3 or 4, wherein the subject has a disease of a bleeding disorder characterized by the enzyme deficiency, a disease of an inborn error of metabolism characterized by the enzyme deficiency, or a lysosomal storage disease characterized by the enzyme deficiency, optionally wherein the disease is hemophilia B and the polypeptide of interest is a factor IX protein, the disease is hemophilia A and the polypeptide of interest is a factor VIII protein, or the disease is Pompe disease and the polypeptide of interest is a multidomain therapeutic protein comprising a delivery domain fused to a lysosomal alpha-glucosidase.

6. The method of any preceding claim, wherein the subject does not have preexisting immunity against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or a delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent at the time the CD40 inhibitor is administered.

7. The method of any preceding claim, further comprising a subsequent administration step comprising administering to the subject at one or more subsequent times:Attorney Docket No.057766 / 625782 (a) the nucleic acid construct; (b) the nuclease agent or the one or more nucleic acids encoding the nuclease agent; and optionally (c) the CD40 inhibitor, until a desired level of expression and / or activity of the polypeptide of interest is achieved in the subject.

8. The method of any one of claims 1-6, further comprising a subsequent administration step comprising administering to the subject at one or more subsequent times: (a) the nucleic acid construct; (b) a second nuclease agent or one or more nucleic acids encoding the second nuclease agent, wherein the second nuclease agent targets a second nuclease target site in the target genomic locus, wherein the second nuclease target site is different from the first nuclease target site; and optionally (c) the CD40 inhibitor, until a desired level of expression and / or activity of the polypeptide of interest is achieved in the subject.

9. The method of any one of claims 1-6, further comprising a subsequent administration step comprising administering to the subject at one or more subsequent times: (a) the nucleic acid construct; (b) a second nuclease agent or one or more nucleic acids encoding the second nuclease agent, wherein the second nuclease agent targets a second nuclease target site in a second target genomic locus that is different from the first target genomic locus; and optionally (c) the CD40 inhibitor, until a desired level of expression and / or activity of the polypeptide of interest is achieved in the subject.

10. The method of any one of claims 1-6, further comprising a subsequent administration step comprising administering to the subject at one or more subsequent times:Attorney Docket No.057766 / 625782 (a) a second nucleic acid construct comprising a second coding sequence for the polypeptide of interest, wherein the second coding sequence is different from the first coding sequence; (b) (i) the first nuclease agent or the one or more nucleic acids encoding the first nuclease agent; (ii) a second nuclease agent or one or more nucleic acids encoding the second nuclease agent, wherein the second nuclease agent targets a second nuclease target site in the target genomic locus, wherein the second nuclease target site is different from the first nuclease target site; or (iii) a second nuclease agent or one or more nucleic acids encoding the second nuclease agent, wherein the second nuclease agent targets a second nuclease target site in a second target genomic locus that is different from the first target genomic locus; and optionally (c) the CD40 inhibitor, until a desired level of expression and / or activity of the polypeptide of interest is achieved in the subject.

11. The method of any one of claims 7-10, further comprising the following steps prior to the subsequent administration step: (i) measuring expression and / or activity of the polypeptide of interest in the subject; and (ii) determining the dose of the nucleic acid construct and the nuclease agent or the one or more nucleic acids encoding the nuclease agent for the subsequent administration step in order to achieve the desired level of expression and / or activity of the polypeptide of interest is achieved in the subject.

12. The method of any one of claims 7-10, wherein the dose of the nucleic acid construct for the subsequent administration step is higher than the dose of the nucleic acid construct in the first administration step and / or the dose of the nuclease agent or one or more nucleic acids encoding the nuclease agent for the subsequent step is higher than the dose of the nuclease agent or one or more nucleic acids encoding the nuclease agent in the first administration step,Attorney Docket No.057766 / 625782 optionally wherein the dose of the nucleic acid construct for the subsequent administration step is higher than the dose of the nucleic acid construct in the first administration step.

13. The method of any one of claims 7-10, wherein the dose of the nucleic acid construct for the subsequent administration step is the same as or lower than the dose of the nucleic acid construct in the first administration step and / or the dose of the nuclease agent or one or more nucleic acids encoding the nuclease agent for the subsequent step is the same as or lower than the dose of the nuclease agent or one or more nucleic acids encoding the nuclease agent in the first administration step, optionally wherein the dose of the nucleic acid construct for the subsequent administration step is the same as or lower than the dose of the nucleic acid construct in the first administration step.

14. The method of any one of claims 7-13, wherein the polypeptide of interest is a factor IX protein, and the desired expression level of the factor IX protein in the subject is a serum level of at least about 3 ^g / mL or about 3-5 ^g / mL.

15. The method of any one of claims 7-13, wherein the polypeptide of interest is a multidomain therapeutic protein comprising a delivery domain fused to a lysosomal alpha- glucosidase, and the desired expression level of the multidomain therapeutic protein in the subject is a serum level of at least about 2 ^g / mL or at least about 5 ^g / mL.

16. The method of any one of claims 1-6, further comprising a subsequent administration step comprising administering to the subject at one or more subsequent times: (a) a second nucleic acid construct comprising a coding sequence for a second polypeptide of interest that is different from the first polypeptide of interest; (b) (i) the first nuclease agent or the one or more nucleic acids encoding the first nuclease agent; (ii) a second nuclease agent or one or more nucleic acids encoding the second nuclease agent, wherein the second nuclease agent targets a second nuclease target site in the target genomic locus, wherein the second nuclease target site is different from the first nuclease target site; orAttorney Docket No.057766 / 625782 (iii) a second nuclease agent or one or more nucleic acids encoding the second nuclease agent, wherein the second nuclease agent targets a second nuclease target site in a second target genomic locus that is different from the first target genomic locus; and optionally (c) the CD40 inhibitor, wherein the second nuclease agent cleaves the second nuclease target site, and the second nucleic acid construct is inserted into the second target genomic locus.

17. The method of any one of claims 7-16, wherein the one or more subsequent administration steps is one subsequent administration step.

18. The method of any one of claims 7-16, wherein the one or more subsequent administration steps is two subsequent administration steps or comprises at least two subsequent administration steps.

19. The method of any one of claims 7-18, wherein the CD40 inhibitor is administered in the one or more subsequent administration steps if there is no preexisting CD40 inhibitor in the subject or if preexisting CD40 inhibitor expression and / or activity levels are below a desired threshold level, optionally wherein the method comprises measuring CD40 inhibitor expression and / or activity levels prior to the one or more subsequent administration steps.

20. The method of any preceding claim, wherein the CD40 inhibitor is a CD40 antigen-binding molecule, wherein the CD40 antigen-binding molecule is a monospecific CD40 antigen-binding molecule.

21. The method of any one of claims 1-19, wherein the CD40 inhibitor is a CD40 antigen-binding molecule, wherein the CD40 antigen-binding molecule is a bispecific antigen-binding molecule comprising: (a) a first antigen-binding domain (D1) that binds a first epitope of human CD40; and (b) a second antigen-binding domain (D2) that binds a second epitope of human CD40.

22. The method of claim 21, wherein the bispecific antigen-binding molecule:Attorney Docket No.057766 / 625782 (i) binds human CD40 with a KD of less than 25 nM as measured by surface plasmon resonance at 25°C; (ii) binds human CD40 with a KDof less than 70 nM as measured by surface plasmon resonance at 37°C; (iii) binds human CD40 with a dissociative half-life (t1 / 2) of greater than 75 minutes as measured by surface plasmon resonance at 25°C; (iv) binds a human CD40-expressing cell with an EC50value of about 10 nM or less; (v) inhibits binding of human CD40 monomer to CD40L; (vi) inhibits CD40 ligand (CD40L)-induced activation; and / or (vii) does not significantly agonize CD40 in the absence of CD40L.

23. The method of claim 21 or 22, wherein the D1 domain and the D2 domain each comprise a heavy chain immunoglobulin variable region comprising a set of three heavy chain complementarity determining region sequences HCDR1, HCDR2, and HCDR3 independently selected from the group consisting of: (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8; (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28; and (c) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, and an HCDR3 comprising the amino acid sequence of SEQ ID NO:

38.

24. The method of any one of claims 21-23, wherein the D1 domain and the D2 domain each comprise a light chain immunoglobulin variable region comprising a set of three light chain complementarity determining region sequences LCDR1, LCDR2, and LCDR3, wherein the LCDR1 comprises the amino acid sequence of SEQ ID NO: 12, the LCDR2 comprises the amino acid sequence AAS, and the LCDR3 comprises the amino acid sequence of SEQ ID NO: 16.Attorney Docket No.057766 / 625782 25. The method of any one of claims 21-24, wherein the D1 domain comprises: (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

16.

26. The method of claim 25, wherein the D1 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

16.

27. The method of claim 26, wherein the D1 domain comprises a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:

2.

28. The method of claim 25, wherein the D1 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

16.

29. The method of claim 28, wherein the D1 domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 22.Attorney Docket No.057766 / 625782 30. The method of any one of claims 21-29, wherein the D1 domain comprises a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

10.

31. The method of any one of claims 21-30, wherein the D2 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

16.

32. The method of claim 31, wherein the D2 domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO:

32.

33. The method of claim 31 or 32, wherein the D2 domain comprises an LCVR comprising the amino acid sequence of SEQ ID NO:

10.

34. The method of any one of claims 21-33, wherein the bispecific antigen- binding molecule comprises: a D1 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; and a D2 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

16.

35. The method of claim 34, wherein the D1 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence ofAttorney Docket No.057766 / 625782 SEQ ID NO: 10, and the D2 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 32 and an LCVR comprising the amino acid sequence of SEQ ID NO:

10.

36. The method of any one of claims 21-35, wherein the bispecific antigen- binding molecule comprises: a D1 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; and a D2 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

16.

37. The method of claim 36, wherein the D1 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 22 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and the D2 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 32 and an LCVR comprising the amino acid sequence of SEQ ID NO:

10.

38. The method of any one of claims 21-24, wherein the D1 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

16.

39. The method of claim 38, wherein the D1 domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO:

32.

40. The method of claim 38 or 39, wherein the D1 domain comprises an LCVR comprising the amino acid sequence of SEQ ID NO: 10.Attorney Docket No.057766 / 625782 41. The method of any one of claims 21-24 and 38-40, wherein the D2 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

16.

42. The method of claim 41, wherein the D2 domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO:

2.

43. The method of claim 41, wherein the D2 domain comprises an LCVR comprising the amino acid sequence of SEQ ID NO:

10.

44. The method of any one of claims 21-24 and 38-43, wherein the bispecific antigen-binding molecule comprises: a D1 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; and a D2 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

16.

45. The method of claim 44, wherein the D1 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 32 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and the D2 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO:

10.

46. The method of any one of claims 21-45, wherein the bispecific antigen- binding molecule is a bispecific antibody.Attorney Docket No.057766 / 625782 47. The method of claim 46, wherein the bispecific antibody comprises a human IgG heavy chain constant region, optionally wherein the human IgG heavy chain constant region comprises one or more modifications that increase binding to a neonatal Fc receptor (FcRn) and / or the human IgG heavy chain constant region comprises one or more modifications that decrease binding to an Fc-gamma receptor (FcγR).

48. The method of claim 47, wherein the human IgG heavy chain constant region is isotype IgG4 or IgG1.

49. The method of claim 47 or 48, wherein the human IgG heavy chain constant region comprises one or more modifications that reduce binding to an Fc receptor.

50. The method of any one of claims 21-49, wherein the D1 comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 42 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; or a heavy chain comprising the amino acid sequence of SEQ ID NO: 46 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; or a heavy chain comprising the amino acid sequence of SEQ ID NO: 48 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; or a heavy chain comprising the amino acid sequence of SEQ ID NO: 52 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

51. The method of any one of claims 21-50, wherein the D2 comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 44 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; or a heavy chain comprising the amino acid sequence of SEQ ID NO: 50 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

52. The method of any one of claims 21-51, wherein the bispecific antigen- binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 42 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the aminoAttorney Docket No.057766 / 625782 acid sequence of SEQ ID NO: 44 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

53. The method of any one of claims 21-51, wherein the bispecific antigen- binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 46 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 44 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

54. The method of any one of claims 21-51, wherein the bispecific antigen- binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 48 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 50 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

55. The method of any one of claims 21-51, wherein the bispecific antigen- binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 52 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 50 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

56. The method of any one of claims 21-24 and 38-49, wherein the D1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 283 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

57. The method of any one of claims 21-24, 38-49, and 56, wherein the D2 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 285 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

58. The method of claim 56 or 57, wherein the bispecific antigen-binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising theAttorney Docket No.057766 / 625782 amino acid sequence of SEQ ID NO: 283 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 285 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

59. The method of any one of claims 1-22, wherein the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within an HCVR / LCVR amino acid sequence pair selected from the group consisting of SEQ ID NOS: 2 / 10, 22 / 10, 32 / 10, 183 / 184, 141 / 142, 151 / 268, 231 / 232, 160 / 161, 170 / 171, 173 / 174, 188 / 189, 199 / 200, 183 / 212, 183 / 215, 183 / 217, 211 / 184, 211 / 212, 211 / 215, 211 / 217, 213 / 184, 213 / 212, 213 / 215, 213 / 217, 214 / 184, 214 / 212, 214 / 215, 214 / 217, 216 / 184, 216 / 212, 216 / 215, 216 / 217, 224 / 225, 244 / 245, 251 / 252, 259 / 260, 264 / 265, and 259 / 265; (II) the HCVR and LCVR amino acid sequences contained within an HCVR / LCVR amino acid sequence pair selected from the group consisting of SEQ ID NOS: 2 / 10, 22 / 10, 32 / 10, 183 / 184, 141 / 142, 151 / 268, 231 / 232, 160 / 161, 170 / 171, 173 / 174, 188 / 189, 199 / 200, 183 / 212, 183 / 215, 183 / 217, 211 / 184, 211 / 212, 211 / 215, 211 / 217, 213 / 184, 213 / 212, 213 / 215, 213 / 217, 214 / 184, 214 / 212, 214 / 215, 214 / 217, 216 / 184, 216 / 212, 216 / 215, 216 / 217, 224 / 225, 244 / 245, 251 / 252, 259 / 260, 264 / 265, and 259 / 265; or (III) the heavy chain and light chain amino acid sequences contained within a heavy chain / light chain amino acid sequence pair selected from the group consisting of SEQ ID NOS: 18 / 20, 30 / 20, 40 / 20, 185 / 186, 143 / 144, 152 / 153, 236 / 237, 162 / 163, 172 / 144, 175 / 176, 190 / 192, 191 / 192, 201 / 202, 218 / 153, 229 / 230, 233 / 237, 234 / 237, 235 / 237, 246 / 250, 247 / 250, 248 / 250, 249 / 250, 261 / 262, 266 / 267, and 261 / 267.

60. The method of any one of claims 1-22, wherein the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within an HCVR / LCVR amino acid sequence pair selected from the group consisting of SEQ ID NOS: 2 / 10, 22 / 10, and 32 / 10; (II) (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising theAttorney Docket No.057766 / 625782 amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or (c) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; (III) (a) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 10; (b) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 22 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 10; (c) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 32 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 10; or (IV) (a) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 18 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 20; (b) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 30 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 20; or (c) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 40 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO:

20.

61. The method of any one of claims 1-22, wherein the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 183 / 184; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 177, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 178, an HCDR3 comprising theAttorney Docket No.057766 / 625782 amino acid sequence of SEQ ID NO: 179, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 180, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 181, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 182; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 183 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 184; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 185 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO:

186.

62. The method of any one of claims 1-22, wherein the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 141 / 142; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 135, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 136, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 137, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 138, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 139, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 140; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 141 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 142; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 143 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO:

144.

63. The method of any one of claims 1-22, wherein the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 151 / 268;Attorney Docket No.057766 / 625782 (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 145, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 146, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 147, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 148, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 149, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 150; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 151 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 268; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 152 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO:

153.

64. The method of any one of claims 1-22, wherein the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 231 / 232; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 269, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 220, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 221, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 222, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 223, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 150; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 231 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 232; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 236 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO:

237.

65. The method of any one of claims 1-19, wherein the CD40 inhibitor is a CD40L binding protein.Attorney Docket No.057766 / 625782 66. The method of claim 65, wherein: (I) the CD40L binding protein comprises two Tn3 proteins fused to human serum albumin; or (II) the CD40L binding protein comprises the sequence set forth in SEQ ID NO:

280.

67. The method of claim 65, wherein the CD40L binding protein is an anti- CD40L antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 276 / 277; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 270, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 271, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 272, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 273, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 274, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 275; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 276 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 277; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 278 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO:

279.

68. The method of any preceding claim, wherein the CD40 inhibitor is administered simultaneously with the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent.

69. The method of any one of claims 1-67, wherein the CD40 inhibitor is administered prior to the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent.Attorney Docket No.057766 / 625782 70. The method of any one of claims 1-67, wherein the CD40 inhibitor is administered prior to and after the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent.

71. The method of claim 70, wherein the CD40 inhibitor is administered within about 6 months after the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent, optionally wherein the nucleic acid construct is in a viral vector, and the CD40 inhibitor is administered if the viral vector is still present in the subject.

72. The method of any one of claims 69-71, wherein the CD40 inhibitor is administered about 1 week prior to or within about 1 week prior to the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent.

73. The method of any preceding claim, wherein the nucleic acid construct is administered simultaneously with the nuclease agent or the one or more nucleic acids encoding the nuclease agent.

74. The method of any one of claims 1-72, wherein the nucleic acid construct is administered prior to or after the nuclease agent or the one or more nucleic acids encoding the nuclease agent.

75. The method of any preceding claim, wherein the nucleic acid construct is in a nucleic acid vector, optionally wherein the nucleic acid vector is a viral vector, and optionally wherein the viral vector is administered at a dose of about 3E11 vg / kg to about 5E13 vg / kg.

76. The method of claim 75, wherein the nucleic acid vector is an adeno- associated viral (AAV) vector, optionally wherein the nucleic acid construct is flanked by inverted terminal repeats (ITRs) on each end, optionally wherein the ITR on at least one end comprises, consists essentially of, or consists of SEQ ID NO: 112, and optionally wherein the ITR on each end comprises, consists essentially of, or consists of SEQ ID NO: 112, or optionally wherein the ITR on at least one end comprises, consists essentially of, or consists of SEQ ID NO: 110, andAttorney Docket No.057766 / 625782 optionally wherein the ITR on each end comprises, consists essentially of, or consists of SEQ ID NO:

110.

77. The method of claim 76, wherein the AAV vector is a recombinant AAV8 (rAAV8) vector, optionally wherein the AAV vector is a single-stranded AAV (ssAAV) vector.

78. The method of any preceding claim, wherein the polypeptide of interest is a factor IX protein.

79. The method of claim 78, wherein the factor IX protein coding sequence encodes a factor IX protein comprising SEQ ID NO: 63, optionally wherein the factor IX protein coding sequence comprises or consists of SEQ ID NO: 61, or wherein the factor IX protein coding sequence comprises or consists of SEQ ID NO:

60.

80. The method of claim 78 or 79, wherein the nucleic acid construct is a bidirectional construct, wherein the factor IX protein coding sequence is a first factor IX protein coding sequence, and the bidirectional construct further comprises a reverse complement of a second factor IX protein coding sequence, wherein the first factor IX protein coding sequence and the second factor IX protein coding sequence are different but encode the same factor IX protein sequence.

81. The method of claim 80, wherein the nucleic acid construct comprises from 5’ to 3’: a first splice acceptor, the first factor IX protein coding sequence, a first polyadenylation signal, a reverse complement of a second polyadenylation signal, the reverse complement of the second factor IX protein coding sequence, and a reverse complement of a second splice acceptor, wherein: (i) the first factor IX protein coding sequence comprises SEQ ID NO: 60 and the second factor IX protein coding sequence comprises SEQ ID NO: 61; or (ii) the first factor IX protein coding sequence comprises SEQ ID NO: 61 and the second factor IX protein coding sequence comprises SEQ ID NO: 60, wherein the nucleic acid construct does not comprise a promoter that drives the expression of the factor IX protein, andAttorney Docket No.057766 / 625782 wherein the nucleic acid construct does not comprise homology arms, optionally wherein the nucleic acid construct comprises SEQ ID NO: 64 or 62 or the reverse complement thereof.

82. The method of any one of claims 1-77, wherein the polypeptide of interest is a multidomain therapeutic protein comprising a delivery domain fused to a lysosomal alpha- glucosidase.

83. The method of claim 82, wherein the lysosomal alpha-glucosidase comprises or consists of the sequence set forth in SEQ ID NO: 117, optionally wherein the lysosomal alpha-glucosidase coding sequence comprises or consist of the sequence set forth in SEQ ID NO:

127.

84. The method of claim 82 or 83, wherein the delivery domain is a CD63- binding delivery domain wherein the CD63-binding delivery domain is a single-chain variable fragment (scFv).

85. The method of claim 84, wherein the scFv comprises or consists of the sequence set forth in SEQ ID NO: 119, optionally wherein the scFv coding sequence comprises or consists of the sequence set forth in SEQ ID NO:

129.

86. The method of claim 84 or 85, wherein the multidomain therapeutic protein comprises or consists of the sequence set forth in SEQ ID NO: 120, optionally wherein the coding sequence for the multidomain therapeutic protein comprises or consists of the sequence set forth in SEQ ID NO: 128, optionally wherein the nucleic acid construct comprises the sequence set forth in SEQ ID NO: 134 or 132.

87. The method of any one of claims 84-86, wherein the nucleic acid construct comprises from 5’ to 3’: a splice acceptor, the coding sequence for the multidomain therapeutic protein, and a polyadenylation signal or sequence, wherein the coding sequence for the multidomain therapeutic protein comprises the sequence set forth in SEQ ID NO: 128, optionally wherein the nucleic acid construct comprises the sequence set forth in SEQ ID NO: 134 or 132,Attorney Docket No.057766 / 625782 wherein the polyadenylation signal comprises a BGH polyadenylation signal and a unidirectional SV40 late polyadenylation signal, wherein the nucleic acid construct does not comprise a promoter that drives the expression of the multidomain therapeutic protein, and wherein the nucleic acid construct does not comprise a homology arm.

88. The method of claim 82 or 83, wherein the delivery domain is a TfR- binding delivery domain, wherein the TfR-binding delivery domain comprises a single-chain variable fragment (scFv).

89. The method of claim 88, wherein the scFv comprises or consists of the sequence set forth in SEQ ID NO: 121, optionally wherein the scFv coding sequence comprises or consists of the sequence set forth in SEQ ID NO:

122.

90. The method of claim 88 or 89, wherein the multidomain therapeutic protein comprises or consists of the sequence set forth in SEQ ID NO: 126, optionally wherein the coding sequence for the multidomain therapeutic protein comprises or consists of the sequence set forth in SEQ ID NO: 125, optionally wherein the nucleic acid construct comprises the sequence set forth in SEQ ID NO: 133 or 130.

91. The method of any one of claims 88-90, wherein the nucleic acid construct comprises from 5’ to 3’: a splice acceptor, the coding sequence for the multidomain therapeutic protein, and a polyadenylation signal or sequence, wherein the coding sequence for the multidomain therapeutic protein comprises the sequence set forth in SEQ ID NO: 125, optionally wherein the nucleic acid construct comprises the sequence set forth in SEQ ID NO: 133 or 130, wherein the polyadenylation signal comprises a BGH polyadenylation signal and a unidirectional SV40 late polyadenylation signal, wherein the nucleic acid construct does not comprise a promoter that drives the expression of the multidomain therapeutic protein, and wherein the nucleic acid construct does not comprise a homology arm.Attorney Docket No.057766 / 625782 92. The method of any one of claims 1-77, wherein the polypeptide of interest is a factor VIII protein.

93. The method of any one of claims 1-77, wherein the polypeptide of interest is an antigen-binding protein, optionally wherein the antigen-binding protein is an antibody.

94. The method of any preceding claim, wherein the target genomic locus is an albumin gene, wherein the albumin gene is a human albumin gene, wherein the nuclease target site is in intron 1 of the albumin gene.

95. The method of any preceding claim, wherein the nuclease agent comprises: (a) a zinc finger nuclease (ZFN); (b) a transcription activator-like effector nuclease (TALEN); or (c) (i) a Cas protein or a nucleic acid encoding the Cas protein; and (ii) a guide RNA or one or more DNAs encoding the guide RNA, wherein the guide RNA comprises a DNA-targeting segment that targets a guide RNA target sequence, and wherein the guide RNA binds to the Cas protein and targets the Cas protein to the guide RNA target sequence.

96. The method of any one of claims 1-94, wherein the nuclease agent comprises: (a) a Cas protein or a nucleic acid encoding the Cas protein; and (b) a guide RNA or one or more DNAs encoding the guide RNA, wherein the guide RNA comprises a DNA-targeting segment that targets a guide RNA target sequence, and wherein the guide RNA binds to the Cas protein and targets the Cas protein to the guide RNA target sequence.

97. The method of claim 96, wherein the DNA-targeting segment comprises or consists of SEQ ID NO: 96, optionally wherein the guide RNA comprises SEQ ID NO: 100 or 104.Attorney Docket No.057766 / 625782 98. The method of claim 96 or 97, wherein the method comprises administering the guide RNA in the form of RNA, wherein the guide RNA comprises at least one modification.

99. The method of any one of claims 96-98, wherein the method comprises administering the guide RNA in the form of RNA, the guide RNA comprises SEQ ID NO: 104, and the guide RNA comprises: (i) phosphorothioate bonds between the first four nucleotides at the 5’ end of the guide RNA; (ii) phosphorothioate bonds between the last four nucleotides at the 3’ end of the guide RNA; (iii) 2’-O-methyl-modified nucleotides at the first three nucleotides at the 5’ end of the guide RNA; and (iv) 2’-O-methyl-modified nucleotides at the last three nucleotides at the 3’ end of the guide RNA.

100. The method of any one of claims 96-99, wherein the Cas protein is a Cas9 protein, optionally wherein the Cas protein is derived from a Streptococcus pyogenes Cas9 protein, optionally wherein the Cas protein comprises the sequence set forth in SEQ ID NO:

75.

101. The method of any one of claims 96-100, wherein the method comprises administering the nucleic acid encoding the Cas protein, wherein the nucleic acid comprises an mRNA encoding the Cas protein, wherein the mRNA encoding the Cas protein comprises at least one modification.

102. The method of any one of claims 96-101, wherein the method comprises administering the nucleic acid encoding the Cas protein, wherein the nucleic acid comprises an mRNA encoding the Cas protein, the mRNA encoding the Cas protein comprises the sequence set forth in SEQ ID NO: 65 or 66, and the mRNA encoding the Cas protein is fully substituted with N1-methyl-pseudouridine, comprises a 5’ cap, and comprises a poly(A) tail.

103. The method of any one of claims 96-102, wherein the method comprises administering the guide RNA in the form of RNA, the guide RNA comprises SEQ ID NO: 104, and the guide RNA comprises: (i) phosphorothioate bonds between the first four nucleotides at the 5’ end of the guide RNA; (ii) phosphorothioate bonds between the last four nucleotides at the 3’ end of the guide RNA; (iii) 2’-O-methyl-modified nucleotides at the first three nucleotides atAttorney Docket No.057766 / 625782 the 5’ end of the guide RNA; and (iv) 2’-O-methyl-modified nucleotides at the last three nucleotides at the 3’ end of the guide RNA, and wherein the method comprises administering the nucleic acid encoding the Cas protein, wherein the nucleic acid comprises an mRNA encoding the Cas protein, the mRNA encoding the Cas protein comprises the sequence set forth in SEQ ID NO: 65 or 66, and the mRNA encoding the Cas protein is fully substituted with N1-methyl-pseudouridine, comprises a 5’ cap, and comprises a poly(A) tail.

104. The method of any one of claims 96-103, wherein the Cas protein or the nucleic acid encoding the Cas protein and the guide RNA or the one or more DNAs encoding the guide RNA are associated with a lipid nanoparticle.

105. The method of claim 104, wherein the lipid nanoparticle comprises a cationic lipid, a neutral lipid, a helper lipid, and a stealth lipid, wherein the cationic lipid is Lipid A, the neutral lipid is DSPC, the helper lipid is cholesterol, and the stealth lipid is PEG2k-DMG, optionally wherein the lipid nanoparticle comprises four lipids at the following molar ratios: about 50 mol% Lipid A, about 9 mol% DSPC, about 38 mol% cholesterol, and about 3 mol% PEG2k-DMG.

106. The method of any preceding claim, wherein the cell is a liver cell or a hepatocyte, or the population of cells is a population of liver cells or hepatocytes.

107. The method of any preceding claim, wherein the subject is a human subject.

108. The method of any preceding claim, wherein the subject is a neonatal subject.

109. The method of any preceding claim, further comprising determining whether the subject has immunity against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or a delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent prior to the administering,Attorney Docket No.057766 / 625782 optionally wherein the determining comprises determining the presence of neutralizing antibodies against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or the delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent.

110. The method of any preceding claim, wherein the nucleic acid construct is in an adeno-associated viral (AAV) vector, and wherein the subject does not have preexisting AAV immunity at the time the CD40 inhibitor is administered.

111. The method of any one of claims 1-110, further comprising administering to the subject a plasma cell depleting agent at a time when the subject has preexisting immunity against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or a delivery vehicle for the nucleic acid construct, the nuclease agent when the plasma cell depleting agent is administered, or the one or more nucleic acids encoding the nuclease agent, optionally wherein the subject has preexisting AAV immunity when the plasma cell depleting agent is administered.

112. The method of claim 111, wherein the plasma cell depleting agent is capable of depleting long-lived plasma cells (LLPC).

113. The method of claim 111 or 112, wherein the plasma cell depleting agent is a B cell maturation antigen (BCMA) targeting agent, optionally wherein the BCMA targeting agent is a chimeric antigen receptor against BCMA or an anti-BCMA antibody or a functional fragment thereof, and optionally wherein the anti-BCMA antibody or functional fragment thereof is conjugated to a cytotoxic agent.

114. The method of claim 113, wherein the anti-BCMA antibody is a multispecific antibody or a functional fragment thereof, optionally wherein the multispecific anti-BCMA antibody or functional fragment thereof targets BCMA and CD3.

115. The method of claim 114, wherein the multispecific anti-BCMA antibody or functional fragment thereof is anti-BCMAxCD3 bispecific antibody or functional fragment thereof, optionally wherein the anti-BCMAxCD3 bispecific antibody is selected fromAttorney Docket No.057766 / 625782 linvoseltamab (REGN5458), REGN5459, pacanalotamab (AMG420), teclistamab (JNJ- 64007957), AMG701, alnuctamab (CC-93269), EM801, EM901, elranatamab (PF-06863135), TNB383B (ABBV-383), and TNB384B.

116. The method of claim 115, wherein the anti-BCMAxCD3 bispecific antibody or functional fragment thereof comprises a first antigen-binding domain that specifically binds to BCMA comprising three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 287, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

303.

117. The method of claim 116, wherein the first antigen-binding domain that specifically binds to BCMA comprises a HCDR1 comprising the amino acid sequence of SEQ ID NO: 289, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 291, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 293, a LCDR1 comprising the amino acid sequence of SEQ ID NO: 305, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 307, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:

309.

118. The method of claim 116 or 117, wherein the anti-BCMAxCD3 bispecific antibody or functional fragment thereof comprises a second antigen-binding domain that specifically binds to CD3 comprising three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence selected from the group consisting of SEQ ID NOS: 311 and 319, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

303.

119. The method of claim 118, wherein the second antigen-binding domain that specifically binds to CD3 comprises a HCDR1 comprising the amino acid sequence of SEQ ID NO: 313 or 321, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 315 or 323, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 317 or 325, a LCDR1 comprising the amino acid sequence of SEQ ID NO: 305, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 307, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 309.Attorney Docket No.057766 / 625782 120. The method of any one of claims 116-119, wherein the anti-BCMAxCD3 bispecific antibody or functional fragment thereof comprises: (a) a first antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 289, 291, and 293, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 305, 307, and 309, respectively; and (b) a second antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 313, 315, and 317, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 305, 307, and 309, respectively.

121. The method of claim 120, wherein the anti-BCMAxCD3 bispecific antibody or functional fragment thereof comprises: (a) a first antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 289, 291, and 293, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 305, 307, and 309, respectively; and (b) a second antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 321, 323, and 325, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 305, 307, and 309, respectively.

122. The method of any one of claims 115-121, wherein the anti-BCMAxCD3 bispecific antibody or functional fragment thereof comprises a human IgG heavy chain constant region, optionally wherein the human IgG heavy chain constant region is isotype IgG4 or IgG1, and optionally wherein the human IgG heavy chain constant region comprises one or more modifications that increase binding to a neonatal Fc receptor (FcRn) and / or the human IgG heavy chain constant region comprises one or more modifications that decrease binding to an Fc- gamma receptor (FcγR).

123. The method of any one of claims 111-122, further comprising administering to the subject an effective amount of a B cell depleting agent and / or anAttorney Docket No.057766 / 625782 immunoglobulin depleting agent at a time when the subject has preexisting immunity against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or a delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent when the B cell depleting agent and / or the immunoglobulin depleting agent is administered, optionally wherein the subject has preexisting AAV immunity when the B cell depleting agent and / or the immunoglobulin depleting agent is administered, optionally further comprising administering to the subject an effective amount of a B cell depleting agent and an immunoglobulin depleting agent when the subject has preexisting immunity against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or a delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent when the B cell depleting agent and the immunoglobulin depleting agent are administered, optionally wherein the subject has preexisting AAV immunity when the B cell depleting agent and the immunoglobulin depleting agent are administered.

124. The method of claim 123, wherein the B cell depleting agent is administered before, at the same time as, or after the plasma cell depleting agent.

125. The method of claim 123 or 124, wherein the B cell depleting agent is administered prior to and after the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent.

126. The method of any one of claims 123-125, wherein the immunoglobulin depleting agent is administered prior to and after the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent.

127. The method of any one of claims 123-126, wherein the immunoglobulin depleting agent is administered after an initial dose of the plasma cell depleting agent, or wherein the immunoglobulin depleting agent is administered after an initial dose of the plasma cell depleting agent and after an initial dose of the B cell depleting agent.Attorney Docket No.057766 / 625782 128. The method of any one of claims 123-125, wherein the B cell depleting agent is capable of depleting B cells and plasma cells that express low levels of BCMA.

129. The method of any one of claims 123-128, wherein the B cell depleting agent is an agent that binds to a B cell surface molecule.

130. The method of claim 129, wherein the B cell depleting agent is selected from an anti-CD19 antibody, an anti-CD20 antibody, an anti-CD19 antibody and an anti-CD20 antibody, an anti-CD22 antibody, an anti-CD79 antibody, an anti-CD20xCD3 bispecific antibody, an anti-CD19xCD3 bispecific antibody, an anti-CD22xCD3 bispecific antibody, an anti-CD79xCD3 bispecific antibody, functional fragments of any of said antibodies, and any combinations thereof.

131. The method of any one of claims 123-130, wherein the B cell depleting agent is an anti-CD20 antibody or a functional fragment thereof, wherein the anti-CD20 antibody is a multispecific antibody or a functional fragment thereof, optionally wherein the multispecific anti-CD20 antibody or functional fragment thereof targets CD20 and CD3.

132. The method of claim 131, wherein the multispecific anti-CD20 antibody or functional fragment thereof is anti-CD20xCD3 bispecific antibody or functional fragment thereof.

133. The method of claim 132, wherein the anti-CD20xCD3 bispecific antibody or functional fragment thereof comprises a first antigen-binding domain that specifically binds to CD20 comprising three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 329, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

330.

134. The method of claim 133, wherein the first antigen-binding domain that specifically binds to CD20 comprises a HCDR1 comprising the amino acid sequence of SEQ ID NO: 332, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 333, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 334, a LCDR1 comprising the amino acidAttorney Docket No.057766 / 625782 sequence of SEQ ID NO: 335, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 336, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:

337.

135. The method of claim 133 or 134, wherein the anti-CD20xCD3 bispecific antibody or functional fragment thereof comprises a second antigen-binding domain that specifically binds to CD3 comprising three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 331, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

330.

136. The method of claim 135, wherein the second antigen-binding domain that specifically binds to CD3 comprises a HCDR1 comprising the amino acid sequence of SEQ ID NO: 338, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 339, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 340, a LCDR1 comprising the amino acid sequence of SEQ ID NO: 335, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 336, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:

337.

137. The method of any one of claims 133-136, wherein the anti-CD20xCD3 bispecific antibody or functional fragment thereof comprises: (a) a first antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 332, 333, and 334, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 335, 336, and 337, respectively; and (b) a second antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 338, 339, and 340, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 335, 336, and 337, respectively.

138. The method of any one of claims 132-137, wherein the anti-CD20xCD3 bispecific antibody or functional fragment thereof comprises a human IgG heavy chain constant region, optionally wherein the human IgG heavy chain constant region is isotype IgG4 or IgG1, optionally wherein the human IgG heavy chain constant region comprises one or more modifications that increase binding to a neonatal Fc receptor (FcRn) and / or the human IgG heavyAttorney Docket No.057766 / 625782 chain constant region comprises one or more modifications that decrease binding to an Fc- gamma receptor (FcγR).

139. The method of any one of claims 123-126, wherein the B cell depleting agent is an agent targeting a B cell survival factor.

140. The method of any one of claims 123-126, wherein the B cell depleting agent is a BLyS / BAFF inhibitor, an APRIL inhibitor, a BLyS receptor 3 / BAFF receptor inhibitor, or any combination thereof.

141. The method of any one of claims 123-140, wherein the immunoglobulin depleting agent is capable of accelerating IgG clearance.

142. The method of any one of claims 123-141, wherein the immunoglobulin depleting agent is a neonatal Fc receptor (FcRn) blocker.

143. The method of claim 142, wherein the FcRn blocker is selected from Efgartigimod (ARGX-113), Rozanolixizumab (UCB7665), Batoclimab (RVT-1401), IMVT- 1402, Nipocalimab (M281), Orilanolimab (SYNT001), and any combinations thereof.

144. The method of any one of claims 123-143, wherein the immunoglobulin depleting agent is an immunoglobulin degrading enzyme, and the CD40 inhibitor is resistant to the immunoglobulin degrading enzyme.

145. The method of claim 144, wherein the immunoglobulin degrading enzyme is selected from Imlifidase / IdeS / Fabricator, IdeZ, IdeXork, IceMG, CYR-212, CYR-241, S- 1117, HNSA-5487, and IdeE / KJ103.

146. The method of claim 145, wherein the immunoglobulin degrading enzyme is Imlifidase / IdeS / Fabricator.

147. The method of any one of claims 144-146, wherein the CD40 inhibitor is administered before administration of the immunoglobulin depleting agent to the subject, and the immunoglobulin depleting agent is administered before the administration of the nucleic acidAttorney Docket No.057766 / 625782 construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent to the subject.

148. The method of any one of claims 144-147, wherein the subject does not have preexisting immunity against the immunoglobulin degrading enzyme.

149. The method of any one of claims 144-147, wherein the subject has preexisting immunity against the immunoglobulin degrading enzyme.

150. The method of any one of claims 111-149, wherein the method further comprises plasmapheresis, therapeutic plasma exchange, or immunoadsorption.

151. The method of any one of claims 111-150, wherein the plasma cell depleting agent is administered simultaneously with the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent, prior to the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent, or prior to and after the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent, optionally wherein the plasma cell depleting agent is administered within about 6 months after the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent, optionally wherein the nucleic acid construct is in a viral vector, and the plasma cell depleting agent is administered if the viral vector is still present in the subject, or optionally wherein the plasma cell depleting agent is administered about 1 week prior to or within about 1 week prior to the nucleic acid construct and / or the nuclease agent or the one or more nucleic acids encoding the nuclease agent.

152. The method of any one of claims 111-151, wherein the nucleic acid construct is administered simultaneously with the nuclease agent or the one or more nucleic acids encoding the nuclease agent.

153. The method of any one of claims 111-151, wherein the nucleic acid construct is administered prior to or after the nuclease agent or the one or more nucleic acids encoding the nuclease agent.Attorney Docket No.057766 / 625782 154. The method of any one of claims 111-153, wherein the subject has preexisting immunity against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or a delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent, wherein the plasma cell depleting agent and the CD40 inhibitor are both administered prior to the nucleic acid construct and the nuclease agent or the one or more nucleic acids encoding the nuclease agent, and the plasma cell depleting agent is administered prior to the CD40 inhibitor, optionally wherein the nucleic acid construct is in an AAV vector, the subject has preexisting immunity against the AAV vector, and the plasma cell depleting agent and the CD40 inhibitor are both administered prior to the nucleic acid construct, wherein the plasma cell depleting agent is administered prior to the CD40 inhibitor.

155. The method of any one of claims 111-153, wherein the subject has preexisting immunity against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or a delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent, wherein the plasma cell depleting agent and the CD40 inhibitor are both administered prior to the nucleic acid construct and the nuclease agent or the one or more nucleic acids encoding the nuclease agent, and the plasma cell depleting agent and the CD40 inhibitor are administered simultaneously, optionally wherein the subject has ongoing B cell responses to the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or the delivery vehicle when the plasma cell depleting agent and the CD40 inhibitor are administered, optionally wherein the nucleic acid construct is in an AAV vector, the subject has preexisting immunity against the AAV vector, and the plasma cell depleting agent and the CD40 inhibitor are both administered prior to the nucleic acid construct, wherein the plasma cell depleting agent and the CD40 inhibitor are administered simultaneously, optionally wherein the subject has ongoing B cell responses to the AAV vector when the plasma cell depleting agent and the CD40 inhibitor are administered.Attorney Docket No.057766 / 625782 156. The method of any one of claims 111-153, wherein the subject does not have preexisting immunity against the nucleic acid construct, the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or a delivery vehicle for the nucleic acid construct, the nuclease agent, or the one or more nucleic acids encoding the nuclease agent before administration of the nucleic acid construct and the nuclease agent or the one or more nucleic acids encoding the nuclease agent, the CD40 inhibitor is administered prior to the nucleic acid construct and the nuclease agent or the one or more nucleic acids encoding the nuclease agent, the subject develops immunity against the polypeptide of interest, the nuclease agent, the one or more nucleic acids encoding the nuclease agent, or the delivery vehicle from the administration of the nucleic acid construct and the nuclease agent or the one or more nucleic acids encoding the nuclease agent, and the plasma cell depleting agent is administered after the administration of the nucleic acid construct and the nuclease agent or the one or more nucleic acids encoding the nuclease agent but prior to a second dose of the nucleic acid construct, a second dose of the nuclease agent or the one or more nucleic acids encoding the nuclease agent, a second nucleic acid construct, and / or a second nuclease agent or one or more nucleic acids encoding the second nuclease agent, optionally wherein: (I) the nucleic acid construct is in an AAV vector, the subject does not have preexisting immunity against the AAV vector before administration of a first dose of the nucleic acid construct, the CD40 inhibitor is administered prior to the first dose of the nucleic acid construct, the subject develops immunity against the AAV vector from the first dose of the nucleic acid construct, and the plasma cell depleting agent is administered after the first dose of the nucleic acid construct but prior to a second dose of the nucleic acid construct; or (II) the nucleic acid construct is in an AAV vector, the subject does not have preexisting immunity against the AAV vector before administration of the nucleic acid construct, the CD40 inhibitor is administered prior to the nucleic acid construct, the subject develops immunity against the AAV vector from administration of the nucleic acid construct, and the plasma cell depleting agent is administered after the nucleic acid construct but prior to administration of a second nucleic acid construct in the AAV vector.Attorney Docket No.057766 / 625782 157. A composition or combination comprising a CD40 inhibitor in combination with: (a) a nucleic acid construct comprising a coding sequence for the polypeptide of interest; and (b) a nuclease agent or one or more nucleic acids encoding the nuclease agent, wherein the nuclease agent targets a nuclease target site in a target genomic locus.

158. The composition or combination of claim 157, wherein the CD40 inhibitor is a CD40 antigen-binding molecule, wherein the CD40 antigen-binding molecule is a monospecific antigen-binding molecule.

159. The composition or combination of claim 157, wherein the CD40 inhibitor is a CD40 antigen-binding molecule, wherein the CD40 antigen-binding molecule is a bispecific antigen-binding molecule comprising: (a) a first antigen-binding domain (D1) that binds a first epitope of human CD40; and (b) a second antigen-binding domain (D2) that binds a second epitope of human CD40.

160. The composition or combination of claim 159, wherein the bispecific antigen-binding molecule: (i) binds human CD40 with a KDof less than 25 nM as measured by surface plasmon resonance at 25°C; (ii) binds human CD40 with a KDof less than 70 nM as measured by surface plasmon resonance at 37°C; (iii) binds human CD40 with a dissociative half-life (t1 / 2) of greater than 75 minutes as measured by surface plasmon resonance at 25°C; (iv) binds a human CD40-expressing cell with an EC50value of about 10 nM or less; (v) inhibits binding of human CD40 monomer to CD40L; (vi) inhibits CD40 ligand (CD40L)-induced activation; and / or (vii) does not significantly agonize CD40 in the absence of CD40L.

161. The composition or combination of claim 159 or 160, wherein the D1 domain and the D2 domain each comprise a heavy chain immunoglobulin variable regionAttorney Docket No.057766 / 625782 comprising a set of three heavy chain complementarity determining region sequences HCDR1, HCDR2, and HCDR3 independently selected from the group consisting of: (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8; (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, and an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28; and (c) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, and an HCDR3 comprising the amino acid sequence of SEQ ID NO:

38.

162. The composition or combination of any one of claims 159-161, wherein the D1 domain and the D2 domain each comprise a light chain immunoglobulin variable region comprising a set of three light chain complementarity determining region sequences LCDR1, LCDR2, and LCDR3, wherein the LCDR1 comprises the amino acid sequence of SEQ ID NO: 12, the LCDR2 comprises the amino acid sequence AAS, and the LCDR3 comprises the amino acid sequence of SEQ ID NO:

16.

163. The composition or combination of any one of claims 159-162, wherein the D1 domain comprises: (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16.Attorney Docket No.057766 / 625782 164. The composition or combination of claim 163, wherein the D1 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

16.

165. The composition or combination of claim 164, wherein the D1 domain comprises a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:

2.

166. The composition or combination of claim 163, wherein the D1 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

16.

167. The composition or combination of claim 166, wherein the D1 domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO:

22.

168. The composition or combination of any one of claims 159-167, wherein the D1 domain comprises a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

10.

169. The composition or combination of any one of claims 159-168, wherein the D2 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

16.

170. The composition or combination of claim 169, wherein the D2 domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 32.Attorney Docket No.057766 / 625782 171. The composition or combination of claim 169 or 170, wherein the D2 domain comprises an LCVR comprising the amino acid sequence of SEQ ID NO:

10.

172. The composition or combination of any one of claims 159-171, wherein the bispecific antigen-binding molecule comprises: a D1 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; and a D2 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

16.

173. The composition or combination of claim 172, wherein the D1 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and the D2 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 32 and an LCVR comprising the amino acid sequence of SEQ ID NO:

10.

174. The composition or combination of any one of claims 159-173, wherein the bispecific antigen-binding molecule comprises: a D1 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; and a D2 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence ofAttorney Docket No.057766 / 625782 SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

16.

175. The composition or combination of claim 174, wherein the D1 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 22 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and the D2 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 32 and an LCVR comprising the amino acid sequence of SEQ ID NO:

10.

176. The composition or combination of any one of claims 159-162, wherein the D1 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

16.

177. The composition or combination of claim 176, wherein the D1 domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO:

32.

178. The composition or combination of claim 176 or 177, wherein the D1 domain comprises an LCVR comprising the amino acid sequence of SEQ ID NO:

10.

179. The composition or combination of any one of claims 159-162 and 176- 178, wherein the D2 domain comprises an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

16.

180. The composition or combination of claim 179, wherein the D2 domain comprises an HCVR comprising the amino acid sequence of SEQ ID NO:

2.

181. The composition or combination of claim 179, wherein the D2 domain comprises an LCVR comprising the amino acid sequence of SEQ ID NO: 10.Attorney Docket No.057766 / 625782 182. The composition or combination of any one of claims 159-162 and 176- 181, wherein the bispecific antigen-binding molecule comprises: a D1 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; and a D2 comprising an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence AAS, and an LCDR3 comprising the amino acid sequence of SEQ ID NO:

16.

183. The composition or combination of claim 182, wherein the D1 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 32 and an LCVR comprising the amino acid sequence of SEQ ID NO: 10, and the D2 comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising the amino acid sequence of SEQ ID NO:

10.

184. The composition or combination of any one of claims 159-183, wherein the bispecific antigen-binding molecule is a bispecific antibody.

185. The composition or combination of claim 184, wherein the bispecific antibody comprises a human IgG heavy chain constant region, optionally wherein the human IgG heavy chain constant region comprises one or more modifications that increase binding to a neonatal Fc receptor (FcRn) and / or the human IgG heavy chain constant region comprises one or more modifications that decrease binding to an Fc-gamma receptor (FcγR).

186. The composition or combination of claim 185, wherein the human IgG heavy chain constant region is isotype IgG4 or IgG1.Attorney Docket No.057766 / 625782 187. The composition or combination of claim 185 or 186, wherein the human IgG heavy chain constant region comprises one or more modifications that reduce binding to an Fc receptor.

188. The composition or combination of any one of claims 159-187, wherein the D1 comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 42 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; or a heavy chain comprising the amino acid sequence of SEQ ID NO: 46 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; or a heavy chain comprising the amino acid sequence of SEQ ID NO: 48 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; or a heavy chain comprising the amino acid sequence of SEQ ID NO: 52 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

189. The composition or combination of any one of claims 159-188, wherein the D2 comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO: 44 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; or a heavy chain comprising the amino acid sequence of SEQ ID NO: 50 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

190. The composition or combination of any one of claims 159-189, wherein the bispecific antigen-binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 42 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 44 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

191. The composition or combination of any one of claims 159-189, wherein the bispecific antigen-binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 46 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprisingAttorney Docket No.057766 / 625782 the amino acid sequence of SEQ ID NO: 44 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

192. The composition or combination of any one of claims 159-189, wherein the bispecific antigen-binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 48 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 50 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

193. The composition or combination of any one of claims 159-189, wherein the bispecific antigen-binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 52 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 50 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

194. The composition or combination of any one of claims 159-162 and 176- 187, wherein the D1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 283 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

195. The composition or combination of any one of claims 159-162, 176-187, and 194, wherein the D2 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 285 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

196. The composition or combination of claim 194 or 195, wherein the bispecific antigen-binding molecule is a bispecific antibody comprising (i) a D1 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 283 and a light chain comprising the amino acid sequence of SEQ ID NO: 20; and (ii) a D2 comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 285 and a light chain comprising the amino acid sequence of SEQ ID NO: 20.Attorney Docket No.057766 / 625782 197. The composition or combination of any one of claims 157-160, wherein the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within an HCVR / LCVR amino acid sequence pair selected from the group consisting of SEQ ID NOS: 2 / 10, 22 / 10, 32 / 10, 183 / 184, 141 / 142, 151 / 268, 231 / 232, 160 / 161, 170 / 171, 173 / 174, 188 / 189, 199 / 200, 183 / 212, 183 / 215, 183 / 217, 211 / 184, 211 / 212, 211 / 215, 211 / 217, 213 / 184, 213 / 212, 213 / 215, 213 / 217, 214 / 184, 214 / 212, 214 / 215, 214 / 217, 216 / 184, 216 / 212, 216 / 215, 216 / 217, 224 / 225, 244 / 245, 251 / 252, 259 / 260, 264 / 265, and 259 / 265; (II) the HCVR and LCVR amino acid sequences contained within an HCVR / LCVR amino acid sequence pair selected from the group consisting of SEQ ID NOS: 2 / 10, 22 / 10, 32 / 10, 183 / 184, 141 / 142, 151 / 268, 231 / 232, 160 / 161, 170 / 171, 173 / 174, 188 / 189, 199 / 200, 183 / 212, 183 / 215, 183 / 217, 211 / 184, 211 / 212, 211 / 215, 211 / 217, 213 / 184, 213 / 212, 213 / 215, 213 / 217, 214 / 184, 214 / 212, 214 / 215, 214 / 217, 216 / 184, 216 / 212, 216 / 215, 216 / 217, 224 / 225, 244 / 245, 251 / 252, 259 / 260, 264 / 265, and 259 / 265; or (III) the heavy chain and light chain amino acid sequences contained within a heavy chain / light chain amino acid sequence pair selected from the group consisting of SEQ ID NOS: 18 / 20, 30 / 20, 40 / 20, 185 / 186, 143 / 144, 152 / 153, 236 / 237, 162 / 163, 172 / 144, 175 / 176, 190 / 192, 191 / 192, 201 / 202, 218 / 153, 229 / 230, 233 / 237, 234 / 237, 235 / 237, 246 / 250, 247 / 250, 248 / 250, 249 / 250, 261 / 262, 266 / 267, and 261 / 267.

198. The composition or combination of any one of claims 157-160, wherein the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within an HCVR / LCVR amino acid sequence pair selected from the group consisting of SEQ ID NOS: 2 / 10, 22 / 10, and 32 / 10; (II) (a) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 4, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 6, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 8, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3Attorney Docket No.057766 / 625782 comprising the amino acid sequence of SEQ ID NO: 16; or (b) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 24, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 26, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 28, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; or (c) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 34, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 36, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 38, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 12, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 14, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 16; (III) (a) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 2 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 10; (b) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 22 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 10; (c) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 32 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 10; or (IV) (a) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 18 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 20; (b) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 30 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 20; or (c) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 40 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO:

20.

199. The composition or combination of any one of claims 157-160, wherein the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 183 / 184; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 177, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 178, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 179, an LCDR1 comprising the amino acid sequence ofAttorney Docket No.057766 / 625782 SEQ ID NO: 180, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 181, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 182; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 183 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 184; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 185 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO:

186.

200. The composition or combination of any one of claims 157-160, wherein the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 141 / 142; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 135, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 136, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 137, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 138, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 139, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 140; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 141 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 142; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 143 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO:

144.

201. The composition or combination of any one of claims 157-160, wherein the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising:Attorney Docket No.057766 / 625782 (I) the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 151 / 268; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 145, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 146, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 147, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 148, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 149, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 150; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 151 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 268; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 152 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO:

153.

202. The composition or combination of any one of claims 157-160, wherein the CD40 antigen binding molecule is an anti-CD40 antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 231 / 232; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 269, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 220, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 221, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 222, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 223, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 150; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 231 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 232; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 236 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO: 237.Attorney Docket No.057766 / 625782 203. The composition or combination of claim 157, wherein the CD40 inhibitor is a CD40L binding protein.

204. The composition or combination of claim 203, wherein: (I) the CD40L binding protein comprises two Tn3 proteins fused to human serum albumin; or (II) the CD40L binding protein comprises the sequence set forth in SEQ ID NO:

280.

205. The composition or combination of claim 203, wherein the CD40L binding protein is an anti-CD40L antibody or antigen-binding fragment thereof comprising: (I) the HCDR1-HCDR2-HCDR3-LCDR1-LCDR2-LCDR3 amino acid sequences set contained within the HCVR / LCVR amino acid sequence pair set forth in SEQ ID NOS: 276 / 277; (II) an HCDR1 comprising the amino acid sequence of SEQ ID NO: 270, an HCDR2 comprising the amino acid sequence of SEQ ID NO: 271, an HCDR3 comprising the amino acid sequence of SEQ ID NO: 272, an LCDR1 comprising the amino acid sequence of SEQ ID NO: 273, an LCDR2 comprising the amino acid sequence of SEQ ID NO: 274, and an LCDR3 comprising the amino acid sequence of SEQ ID NO: 275; (III) an HCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 276 and an LCVR comprising or consisting of the amino acid sequence of SEQ ID NO: 277; or (IV) a heavy chain comprising or consisting of the amino acid sequence of SEQ ID NO: 278 and a light chain comprising or consisting of the amino acid sequence of SEQ ID NO:

279.

206. The composition or combination of any one of claims 157-205, wherein the nucleic acid construct is in a nucleic acid vector, optionally wherein the nucleic acid vector is a viral vector.

207. The composition or combination of claim 206, wherein the nucleic acid vector is an adeno-associated viral (AAV) vector,Attorney Docket No.057766 / 625782 optionally wherein the nucleic acid construct is flanked by inverted terminal repeats (ITRs) on each end, optionally wherein the ITR on at least one end comprises, consists essentially of, or consists of SEQ ID NO: 112, and optionally wherein the ITR on each end comprises, consists essentially of, or consists of SEQ ID NO: 112, or optionally wherein the ITR on at least one end comprises, consists essentially of, or consists of SEQ ID NO: 110, and optionally wherein the ITR on each end comprises, consists essentially of, or consists of SEQ ID NO:

110.

208. The composition or combination of claim 207, wherein the AAV vector is a recombinant AAV8 (rAAV8) vector, optionally wherein the AAV vector is a single-stranded AAV (ssAAV) vector.

209. The composition or combination of any one of claims 157-208, wherein the polypeptide of interest is a factor IX protein.

210. The composition or combination of claim 209, wherein the factor IX protein coding sequence encodes a factor IX protein comprising SEQ ID NO: 63, optionally wherein the factor IX protein coding sequence comprises or consists of SEQ ID NO: 61, or wherein the factor IX protein coding sequence comprises or consists of SEQ ID NO:

60.

211. The composition or combination of claim 209 or 210, wherein the nucleic acid construct is a bidirectional construct, wherein the factor IX protein coding sequence is a first factor IX protein coding sequence, and the bidirectional construct further comprises a reverse complement of a second factor IX protein coding sequence, wherein the first factor IX protein coding sequence and the second factor IX protein coding sequence are different but encode the same factor IX protein sequence.

212. The composition or combination of claim 211, wherein the nucleic acid construct comprises from 5’ to 3’: a first splice acceptor, the first factor IX protein coding sequence, a first polyadenylation signal, a reverse complement of a second polyadenylation signal, the reverse complement of the second factor IX protein coding sequence, and a reverse complement of a second splice acceptor,Attorney Docket No.057766 / 625782 wherein: (i) the first factor IX protein coding sequence comprises SEQ ID NO: 60 and the second factor IX protein coding sequence comprises SEQ ID NO: 61; or (ii) the first factor IX protein coding sequence comprises SEQ ID NO: 61 and the second factor IX protein coding sequence comprises SEQ ID NO: 60, wherein the nucleic acid construct does not comprise a promoter that drives the expression of the factor IX protein, and wherein the nucleic acid construct does not comprise homology arms, optionally wherein the nucleic acid construct comprises SEQ ID NO: 64 or 62 or the reverse complement thereof.

213. The composition or combination of any one of claims 157-208, wherein the polypeptide of interest is a multidomain therapeutic protein comprising a delivery domain fused to a lysosomal alpha-glucosidase.

214. The composition or combination of claim 213, wherein the lysosomal alpha-glucosidase comprises or consists of the sequence set forth in SEQ ID NO: 117, optionally wherein the lysosomal alpha-glucosidase coding sequence comprises or consist of the sequence set forth in SEQ ID NO:

127.

215. The composition or combination of claim 213 or 214, wherein the delivery domain is a CD63-binding delivery domain, wherein the CD63-binding delivery domain is a single-chain variable fragment (scFv).

216. The composition or combination of claim 215, wherein the scFv comprises or consists of the sequence set forth in SEQ ID NO: 119, optionally wherein the scFv coding sequence comprises or consists of the sequence set forth in SEQ ID NO:

129.

217. The composition or combination of claim 215 or 216, wherein the multidomain therapeutic protein comprises or consists of the sequence set forth in SEQ ID NO: 120, optionally wherein the coding sequence for the multidomain therapeutic protein comprises or consists of the sequence set forth in SEQ ID NO: 128,Attorney Docket No.057766 / 625782 optionally wherein the nucleic acid construct comprises the sequence set forth in SEQ ID NO: 134 or 132.

218. The composition or combination of any one of claims 215-217, wherein the nucleic acid construct comprises from 5’ to 3’: a splice acceptor, the coding sequence for the multidomain therapeutic protein, and a polyadenylation signal or sequence, wherein the coding sequence for the multidomain therapeutic protein comprises the sequence set forth in SEQ ID NO: 128, optionally wherein the nucleic acid construct comprises the sequence set forth in SEQ ID NO: 134 or 132, wherein the polyadenylation signal comprises a BGH polyadenylation signal and a unidirectional SV40 late polyadenylation signal, wherein the nucleic acid construct does not comprise a promoter that drives the expression of the multidomain therapeutic protein, and wherein the nucleic acid construct does not comprise a homology arm.

219. The composition or combination of claim 213 or 214, wherein the delivery domain is a TfR-binding delivery domain, wherein the TfR-binding delivery domain comprises a single-chain variable fragment (scFv).

220. The composition or combination of claim 219, wherein the scFv comprises or consists of the sequence set forth in SEQ ID NO: 121, optionally wherein the scFv coding sequence comprises or consists of the sequence set forth in SEQ ID NO:

122.

221. The composition or combination of claim 219 or 220, wherein the multidomain therapeutic protein comprises or consists of the sequence set forth in SEQ ID NO: 126, optionally wherein the coding sequence for the multidomain therapeutic protein comprises or consists of the sequence set forth in SEQ ID NO: 125, optionally wherein the nucleic acid construct comprises the sequence set forth in SEQ ID NO: 133 or 130.Attorney Docket No.057766 / 625782 222. The composition or combination of any one of claims 219-221, wherein the nucleic acid construct comprises from 5’ to 3’: a splice acceptor, the coding sequence for the multidomain therapeutic protein, and a polyadenylation signal or sequence, wherein the coding sequence for the multidomain therapeutic protein comprises the sequence set forth in SEQ ID NO: 125, optionally wherein the nucleic acid construct comprises the sequence set forth in SEQ ID NO: 133 or 130, wherein the polyadenylation signal comprises a BGH polyadenylation signal and a unidirectional SV40 late polyadenylation signal, wherein the nucleic acid construct does not comprise a promoter that drives the expression of the multidomain therapeutic protein, and wherein the nucleic acid construct does not comprise a homology arm.

223. The composition or combination of any one of claims 157-208, wherein the polypeptide of interest is a factor VIII protein.

224. The composition or combination of any one of claims 157-208, wherein the polypeptide of interest is an antigen-binding protein, optionally wherein the antigen-binding protein is an antibody.

225. The composition or combination of any one of claims 157-224, wherein the target genomic locus is an albumin gene, optionally wherein the albumin gene is a human albumin gene, wherein the nuclease target site is in intron 1 of the albumin gene.

226. The composition or combination of any one of claims 157-225, wherein the nuclease agent comprises: (a) a zinc finger nuclease (ZFN); (b) a transcription activator-like effector nuclease (TALEN); or (c) (i) a Cas protein or a nucleic acid encoding the Cas protein; and (ii) a guide RNA or one or more DNAs encoding the guide RNA, wherein the guide RNA comprises a DNA-targeting segment that targets a guide RNA target sequence, and wherein the guide RNA binds to the Cas protein and targets the Cas protein to the guide RNA target sequence.Attorney Docket No.057766 / 625782 227. The composition or combination of any one of claims 157-225, wherein the nuclease agent comprises: (a) a Cas protein or a nucleic acid encoding the Cas protein; and (b) a guide RNA or one or more DNAs encoding the guide RNA, wherein the guide RNA comprises a DNA-targeting segment that targets a guide RNA target sequence, and wherein the guide RNA binds to the Cas protein and targets the Cas protein to the guide RNA target sequence.

228. The composition or combination of claim 227, wherein the DNA-targeting segment comprises or consists of SEQ ID NO: 96, optionally wherein the guide RNA comprises SEQ ID NO: 100 or 104.

229. The composition or combination of claim 227 or 228, wherein the composition or combination comprises the guide RNA in the form of RNA, wherein the guide RNA comprises at least one modification.

230. The composition or combination of any one of claims 227-229, wherein the composition or combination comprises the guide RNA in the form of RNA, the guide RNA comprises SEQ ID NO: 104, and the guide RNA comprises: (i) phosphorothioate bonds between the first four nucleotides at the 5’ end of the guide RNA; (ii) phosphorothioate bonds between the last four nucleotides at the 3’ end of the guide RNA; (iii) 2’-O-methyl-modified nucleotides at the first three nucleotides at the 5’ end of the guide RNA; and (iv) 2’-O-methyl-modified nucleotides at the last three nucleotides at the 3’ end of the guide RNA.

231. The composition or combination of any one of claims 227-230, wherein the Cas protein is a Cas9 protein, optionally wherein the Cas protein is derived from a Streptococcus pyogenes Cas9 protein, optionally wherein the Cas protein comprises the sequence set forth in SEQ ID NO:

75.

232. The composition or combination of any one of claims 227-231, wherein the composition or combination comprises the nucleic acid encoding the Cas protein, wherein the nucleic acid comprises an mRNA encoding the Cas protein, wherein the mRNA encoding the Cas protein comprises at least one modification.Attorney Docket No.057766 / 625782 233. The composition or combination of any one of claims 227-232, wherein the composition or combination comprises the nucleic acid encoding the Cas protein, wherein the nucleic acid comprises an mRNA encoding the Cas protein, the mRNA encoding the Cas protein comprises the sequence set forth in SEQ ID NO: 65 or 66, and the mRNA encoding the Cas protein is fully substituted with N1-methyl-pseudouridine, comprises a 5’ cap, and comprises a poly(A) tail.

234. The composition or combination of any one of claims 227-233, wherein the composition or combination comprises the guide RNA in the form of RNA, the guide RNA comprises SEQ ID NO: 104, and the guide RNA comprises: (i) phosphorothioate bonds between the first four nucleotides at the 5’ end of the guide RNA; (ii) phosphorothioate bonds between the last four nucleotides at the 3’ end of the guide RNA; (iii) 2’-O-methyl-modified nucleotides at the first three nucleotides at the 5’ end of the guide RNA; and (iv) 2’-O-methyl-modified nucleotides at the last three nucleotides at the 3’ end of the guide RNA, and wherein the composition or combination comprises the nucleic acid encoding the Cas protein, wherein the nucleic acid comprises an mRNA encoding the Cas protein, the mRNA encoding the Cas protein comprises the sequence set forth in SEQ ID NO: 65 or 66, and the mRNA encoding the Cas protein is fully substituted with N1-methyl-pseudouridine, comprises a 5’ cap, and comprises a poly(A) tail.

235. The composition or combination of any one of claims 227-234, wherein the Cas protein or the nucleic acid encoding the Cas protein and the guide RNA or the one or more DNAs encoding the guide RNA are associated with a lipid nanoparticle.

236. The composition or combination of claim 235, wherein the lipid nanoparticle comprises a cationic lipid, a neutral lipid, a helper lipid, and a stealth lipid, wherein the cationic lipid is Lipid A, the neutral lipid is DSPC, the helper lipid is cholesterol, and the stealth lipid is PEG2k-DMG, optionally wherein the lipid nanoparticle comprises four lipids at the following molar ratios: about 50 mol% Lipid A, about 9 mol% DSPC, about 38 mol% cholesterol, and about 3 mol% PEG2k-DMG.

237. The composition or combination of any one of claims 157-236, wherein the CD40 inhibitor is further in combination with a plasma cell depleting agent.Attorney Docket No.057766 / 625782 238. The composition or combination of claim 237, wherein the plasma cell depleting agent is capable of depleting long-lived plasma cells (LLPC).

239. The composition or combination of claim 237 or 238, wherein the plasma cell depleting agent is a B cell maturation antigen (BCMA) targeting agent, optionally wherein the BCMA targeting agent is a chimeric antigen receptor against BCMA or an anti-BCMA antibody or a functional fragment thereof, and optionally wherein the anti-BCMA antibody or functional fragment thereof is conjugated to a cytotoxic agent.

240. The composition or combination of claim 239, wherein the anti-BCMA antibody is a multispecific antibody or a functional fragment thereof, optionally wherein the multispecific anti-BCMA antibody or functional fragment thereof targets BCMA and CD3.

241. The composition or combination of claim 240, wherein the multispecific anti-BCMA antibody or functional fragment thereof is anti-BCMAxCD3 bispecific antibody or functional fragment thereof, optionally wherein the anti-BCMAxCD3 bispecific antibody is selected from linvoseltamab (REGN5458), REGN5459, pacanalotamab (AMG420), teclistamab (JNJ-64007957), AMG701, alnuctamab (CC-93269), EM801, EM901, elranatamab (PF- 06863135), TNB383B (ABBV-383), and TNB384B.

242. The composition or combination of claim 241, wherein the anti- BCMAxCD3 bispecific antibody or functional fragment thereof comprises a first antigen-binding domain that specifically binds to BCMA comprising three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 287, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

303.

243. The composition or combination of claim 242, wherein the first antigen- binding domain that specifically binds to BCMA comprises a HCDR1 comprising the amino acid sequence of SEQ ID NO: 289, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 291, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 293, a LCDR1 comprisingAttorney Docket No.057766 / 625782 the amino acid sequence of SEQ ID NO: 305, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 307, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:

309.

244. The composition or combination of claim 242 or 243, wherein the anti- BCMAxCD3 bispecific antibody or functional fragment thereof comprises a second antigen- binding domain that specifically binds to CD3 comprising three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence selected from the group consisting of SEQ ID NOS: 311 and 319, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

303.

245. The composition or combination of claim 244, wherein the second antigen-binding domain that specifically binds to CD3 comprises a HCDR1 comprising the amino acid sequence of SEQ ID NO: 313 or 321, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 315 or 323, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 317 or 325, a LCDR1 comprising the amino acid sequence of SEQ ID NO: 305, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 307, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:

309.

246. The composition or combination of any one of claims 242-245, wherein the anti-BCMAxCD3 bispecific antibody or functional fragment thereof comprises: (a) a first antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 289, 291, and 293, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 305, 307, and 309, respectively; and (b) a second antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 313, 315, and 317, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 305, 307, and 309, respectively.

247. The composition or combination of claim 246, wherein the anti- BCMAxCD3 bispecific antibody or functional fragment thereof comprises:Attorney Docket No.057766 / 625782 (a) a first antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 289, 291, and 293, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 305, 307, and 309, respectively; and (b) a second antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 321, 323, and 325, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 305, 307, and 309, respectively.

248. The composition or combination of any one of claims 241-247, wherein the anti-BCMAxCD3 bispecific antibody or functional fragment thereof comprises a human IgG heavy chain constant region, optionally wherein the human IgG heavy chain constant region is isotype IgG4 or IgG1, optionally wherein the human IgG heavy chain constant region comprises one or more modifications that increase binding to a neonatal Fc receptor (FcRn) and / or the human IgG heavy chain constant region comprises one or more modifications that decrease binding to an Fc-gamma receptor (FcγR).

249. The composition or combination of any one of claims 237-248, wherein the plasma cell depleting agent is further in combination with an effective amount of a B cell depleting agent and / or an immunoglobulin depleting agent, optionally wherein the plasma cell depleting agent is further in combination with an effective amount of a B cell depleting agent and an immunoglobulin depleting agent.

250. The composition or combination of claim 249, wherein the B cell depleting agent is capable of depleting B cells and plasma cells that express low levels of BCMA.

251. The composition or combination of claim 249 or 250, wherein the B cell depleting agent is an agent that binds to a B cell surface molecule.

252. The composition or combination of claim 251, wherein the B cell depleting agent is selected from an anti-CD19 antibody, an anti-CD20 antibody, an anti-CD19 antibody and an anti-CD20 antibody, an anti-CD22 antibody, an anti-CD79 antibody, an anti-Attorney Docket No.057766 / 625782 CD20xCD3 bispecific antibody, an anti-CD19xCD3 bispecific antibody, an anti-CD22xCD3 bispecific antibody, an anti-CD79xCD3 bispecific antibody, functional fragments of any of said antibodies, and any combinations thereof.

253. The composition or combination of any one of claims 249-252, wherein the B cell depleting agent is an anti-CD20 antibody or a functional fragment thereof, wherein the anti-CD20 antibody is a multispecific antibody or a functional fragment thereof, optionally wherein the multispecific anti-CD20 antibody or functional fragment thereof targets CD20 and CD3.

254. The composition or combination of claim 253, wherein the multispecific anti-CD20 antibody or functional fragment thereof is anti-CD20xCD3 bispecific antibody or functional fragment thereof.

255. The composition or combination of claim 254, wherein the anti- CD20xCD3 bispecific antibody or functional fragment thereof comprises a first antigen-binding domain that specifically binds to CD20 comprising three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 329, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

330.

256. The composition or combination of claim 255, wherein the first antigen- binding domain that specifically binds to CD20 comprises a HCDR1 comprising the amino acid sequence of SEQ ID NO: 332, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 333, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 334, a LCDR1 comprising the amino acid sequence of SEQ ID NO: 335, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 336, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:

337.

257. The composition or combination of claim 255 or 256, wherein the anti- CD20xCD3 bispecific antibody or functional fragment thereof comprises a second antigen- binding domain that specifically binds to CD3 comprising three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising theAttorney Docket No.057766 / 625782 amino acid sequence of SEQ ID NO: 331, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

330.

258. The composition or combination of claim 257, wherein the second antigen-binding domain that specifically binds to CD3 comprises a HCDR1 comprising the amino acid sequence of SEQ ID NO: 338, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 339, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 340, a LCDR1 comprising the amino acid sequence of SEQ ID NO: 335, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 336, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:

337.

259. The composition or combination of any one of claims 255-258, wherein the anti-CD20xCD3 bispecific antibody or functional fragment thereof comprises: (a) a first antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 332, 333, and 334, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 335, 336, and 337, respectively; and (b) a second antigen-binding domain that comprises HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOS: 338, 339, and 340, respectively, and LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOS: 335, 336, and 337, respectively.

260. The composition or combination of any one of claims 254-259, wherein the anti-CD20xCD3 bispecific antibody or functional fragment thereof comprises a human IgG heavy chain constant region, optionally wherein the human IgG heavy chain constant region is isotype IgG4 or IgG1, optionally wherein the human IgG heavy chain constant region comprises one or more modifications that increase binding to a neonatal Fc receptor (FcRn) and / or the human IgG heavy chain constant region comprises one or more modifications that decrease binding to an Fc-gamma receptor (FcγR).

261. The composition or combination of claim 249 or 250, wherein the B cell depleting agent is an agent targeting a B cell survival factor.Attorney Docket No.057766 / 625782 262. The composition or combination of claim 249 or 250, wherein the B cell depleting agent is a BLyS / BAFF inhibitor, an APRIL inhibitor, a BLyS receptor 3 / BAFF receptor inhibitor, or any combination thereof.

263. The composition or combination of any one of claims 249-262, wherein the immunoglobulin depleting agent is capable of accelerating IgG clearance.

264. The composition or combination of any one of claims 249-263, wherein the immunoglobulin depleting agent is a neonatal Fc receptor (FcRn) blocker.

265. The composition or combination of claim 264, wherein the FcRn blocker is selected from Efgartigimod (ARGX-113), Rozanolixizumab (UCB7665), Batoclimab (RVT- 1401), IMVT-1402, Nipocalimab (M281), Orilanolimab (SYNT001), and any combinations thereof.

266. The composition or combination of any one of claims 157-265, for use in a method of inserting a nucleic acid encoding a polypeptide of interest into a target genomic locus in a cell or a population of cells in a subject.

267. The composition or combination of any one of claims 157-265, for use in a method of expressing a polypeptide of interest from a target genomic locus in a cell or a population of cells in a subject.

268. The composition or combination of any one of claims 157-265, for use in a method of treating an enzyme deficiency in a subject in need thereof.

269. The composition or combination of any one of claims 157-265, for use in a method of preventing or reducing the onset of a sign or symptom of an enzyme deficiency in a subject in need thereof.

270. A kit comprising the composition or combination of any one of claims 157-269.

271. A CD40 inhibitor for use in a method according to any one of claims 1- 156.

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