Compositions and methods for delivering antibody oligonucleotide conjugates for exon skipping

Abstract

The present disclosure provides antibody oligonucleotide conjugates having a 3E10 antibody or an antigen binding fragment thereof conjugated to a single stranded oligonucleotide that may hybridize to an acceptor splice site, a donor splice site, or an exonic splice enhancer element of a pre-mRNA transcript. Compositions including the antibody oligonucleotide conjugates and methods of using the antibody oligonucleotide conjugates are also provided.

Description

COMPOSITIONS AND METHODS FOR DELIVERING ANTIBODYOLIGONUCLEOTIDE CONJUGATES FOR EXON SKIPPINGCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claim priority to U.S. Provisional Patent Application No. 63 / 491,859, filed March 23, 2023, U.S. Provisional Patent Application No. 63 / 515,313, filed July 24, 2023, U.S. Provisional Patent Application No. 63 / 580,898, filed September 6, 2023, U.S. Provisional Patent Application No. 63 / 585,840, filed September 27, 2023, and U.S. Provisional Patent Application No. 63 / 623,890, filed January 23, 2024, the contents of which are hereby incorporated by reference herein, in their entireties, for all purposes.SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted herewith and is hereby incorporated by reference in its entirety. Said .xml copy, created on March 23, 2024, is named 127689-5020-WO-SEQ, and is 978,137 bytes in size.TECHNICAL FIELD

[0003] The present disclosure relates to compositions and methods for treating diseases and disorders with antibody-oligonucleotide conjugates (AOCs).BACKGROUND

[0004] Several diseases result from genetic mutations that disrupt the reading frame of an encoded polypeptide (e.g., nonsense mutations), resulting in expression of truncated polypeptides or, in some cases, no polypeptide at all. Other diseases result from genetic mutations that introduce new splice sites into a pre-mRNA, or activate cryptic splice sites present in the gene, resulting in aberrant splicing and translation of dysfunctional polypeptides. Exon skipping therapies can be used to treat such diseases. Exon skipping methodologies generally use antisense oligonucleotides (ASO) that bind splice sites in pre-mRNA for an exon containing a deleterious mutation, or bind directly to cryptic splice sites, inducing the splicing machinery to skip over the effective exon or cryptic splice site and generate a mature mRNA that lacks the affected exon, or to ignore the cryptic splice site and generate a full-length mature mRNA. Several antisense oligonucleotides are currently undergoing clinical trials for conditions such as spinal muscular atrophy (SMA) and Duchenne muscular dystrophy (DMD), where anti sense-mediated exon skipping can restore theopen reading frame and allow the synthesis of partly or wholly functional proteins instead of non- functional ones.

[0005] Duchenne muscular dystrophy, for example, is caused by the absence of dystrophin protein due to mutations in the dystrophin (DMD) gene. The gene encoding the protein contains 79 exons spread out over more than 2 million nucleotides of DNA. Mutations disrupting the reading frame of the protein cause truncation of the translated dystrophin polypeptide, resulting in Duchenne muscular dystrophy. In September 2016, the US Food and Drug Administration (FDA) conditionally approved the first DMD antisense drug, eteplirsen (Exondys 51, SEQ ID NO:208), which was developed to exclude exon 51 in mature DMD mRNA in patients with deleterious mutations in exon 51. Eteplirsen is an antisense oligonucleotide modified with a phosphorodiamidate morpholino oligomer (morpholino or PMO), an antisense chemistry that has been well-established in terms of its safety and effectiveness.

[0006] However, the effectiveness of polynucleotide-based therapies remains questionable. One possible explanation for the poor efficacy of many nucleic acid-based therapies, including exon-skipping therapies, is poor delivery of the therapeutic nucleic acid into the affected tissues. For instance, naked polynucleotides are readily degraded by a host of extracellular nucleases present in human tissues. Further, naked polynucleotides do not readily cross the cell membrane.

[0007] Conventional approaches to overcoming these obstacles include packaging therapeutic polynucleotides into liposomal-based delivery vehicles or recombinant viral particles. However, these approaches present immunological challenges, because the viral capsids and liposomal vehicles are recognized by the host’s immune system.

[0008] The 3E10 antibody is an ideal molecular delivery vehicle due to its efficiency in penetrating into living cells with specific nuclear localization, absence of toxicity, and successful delivery of therapeutic cargo proteins in vitro and in vivo. 3E10 has not shown any cellular toxicity in vitro or in vivo in studies to date.SUMMARY

[0009] Given the background above, improved methods are needed for delivering therapeutic antisense oligonucleotides in vivo which are amendable to exon skipping. In particular, there is a need to develop antibody-oligonucleotide conjugates (AOCs), which combine the highprecision of siRNA and ASOs, with the deliverability of the 3E10 antibody, taking advantage of both technologies. Advantageously, the present disclosure provides compositions and methods for delivering a conjugate comprising a 3E10 antibody or an antigen binding fragment thereof conjugated to a single stranded oligonucleotide in vivo that are not reliant upon liposomal or viral vector based nucleic acid delivery.

[0010] In some aspects, the disclosure provides a conjugate of Formula (I):A-(L-Pr)qFormula (I), wherein in Formula (I):A is an antibody or antigen-binding fragment thereof comprising a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of SEQ ID NO:58, CDR2 comprising the amino acid sequence of SEQ ID NO:59, CDR3 comprising SEQ ID NO:60; and a light chain variable region (VL) CDR1 comprising the amino acid sequence of SEQ ID NO:61, CDR2 comprising the amino acid sequence of SEQ ID NO:62, CDR3 comprising the amino acid sequence of SEQ ID NO: 63;L is a linker;P is a single stranded oligonucleotide capable of hybridizing to an acceptor splice site, a donor splice site, or an exonic splice enhancer element of a pre-mRNA transcript, wherein the single stranded oligonucleotide induces exon skipping in the pre-mRNA transcript to generate a mRNA transcript encoding a truncated protein; r is an integer from 1 to 4; and q is an integer from 1 to 16.

[0011] In some embodiments, the linker L comprises one or more groups selected from optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted arylene, optionally substituted cycloalkylene, optionally substituted heteroalkylene, optionally substituted heteroarylene, optionally substituted heterocycloalkylene, -NRa-, -N=CRa-, -CRa=N-, -S-, -S(O)-, -S(O)2-, -OP(O)ORa-, -OP(O)ORaO-, -P(O)ORaO-, -O-, -CRb2-, -[(CRb2)1-12O]1-50-, -C(O)-, -C(S)-, -C(=N-OH)-, -C(NRa)-, -C(NH2C1)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NRa-, -NRaC(O)-, -C(O)NRaSO2-, -SO2NRaC(O)-, -OC(O)O-, -OC(O)S-, -SC(O)O-, -OC(O)NRa-, -NRaC(O)O-,-SC(O)NRa-, -NRaC(O)S-, -S(O)tN(Ra)- (where t is 1 or 2), -N(Ra)S(O>- (where t is 1 or 2), and -XAA-; each Rais independently selected at each occurrence from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted alkenyl, optionally substituted fluoroalkenyl, optionally substituted cycloalkenyl, optionally substituted cycloalkenylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; each Rbis independently selected at each occurrence from hydrogen, halide, -OH, -SO3H, -OPO3H2, -PO3H2, -C(0)NRa2, -CO2Ra, -NRa2, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted alkenyl, optionally substituted fluoroalkenyl, optionally substituted cycloalkenyl, optionally substituted cycloalkenylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroaryl alkyl; two independent Rbgroups or an Raand an Rbcan be joined together to form an optionally substituted cycle; and-XAA- is an amino acid sequence comprising 1 to 6 amino acid moieties.

[0012] In some embodiments, each amino acid moiety of -XAA- is independently selected from alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine (Cys), glutamine (Gin), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (He), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), valine (Vai), citrulline (Cit), and homocitrulline (HoCit).

[0013] In some embodiments, the linker L comprises one or more groups selected from optionally substituted C1-C18alkylene, -C=C-, -CRa=CRa-, optionally substituted 6- to 14-membered arylene, optionally substituted C3-C20 cycloalkylene, -[CH2O]1-18-, -[CH2CH2O]1-18-, -[CH2CH2CH2O]1-18-, optionally substituted 5- to 18-membered heteroarylene, optionally substituted 3- to 20-membered heterocycloalkylene, -NRa-, -N=CRa-,-CRa=N-, -S-, -OP(O)ORaO-, -O-, -CRb2-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NRa-, -NRaC(O)-, -OC(O)O-, -OC(O)S-, -SC(O)O-, -OC(O)NRa-, -NRaC(O)O-, -SC(O)NRa-, -NRaC(O)S-, and -XAA-.

[0014] In some embodiments, the linker L comprises one or more groups selected from optionally substituted C1-C16 alkylene, -C=C-, -CRa=CRa-, optionally substituted phenylene, optionally substituted C3-C6cycloalkylene, -[CH2CH2O]1-16-, -[CH2CH2CH2O]1-16-, optionally substituted 5- to 6-membered heteroarylene, optionally substituted 5- to 20-membered heterocycloalkylene, -NRa-, -N=CRa-, -CRa=N-, -S-, -OP(O)ORaO-, -O-, -CRb2-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NRa-, -NRaC(O)-, -OC(O)O-, -OC(O)NRa-, -NRaC(O)O-, and -XAA-.

[0015] In some embodiments, the linker L comprises one or more groups selected from -[C(Rb)2]1-16-, -C=C-. -CR — CR'-, -[CH2CH2O]1-16-, -NRa-, -N=CR;|-, -CRa=N-. -S-, -OP(O)ORaO-, -O-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NRa-, -NRaC(O)-, -OC(O)O-, -OC(O)NRa-,X1, X2, and X3are independently selected at each occurrence from NRa, N, CRb, S, and O.

[0016] In some embodiments, the linker L is of Formula (L-l):Formula (L-l), wherein in Formula (L-l):LAis a connecting moiety through which A is covalently attached to L';L' is a bond or comprises one or more groups selected from optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted arylene, optionally substituted cycloalkylene, optionally substituted heteroalkylene, optionally substituted heteroarylene, optionally substituted heterocycloalkylene, -NRa-, -N=CRa-, -CRa=N-, -S-, -S(O)-, -S(O)2-, -OP(O)ORaO-, -O-, -CRb2-, -[(CRb2)1-12O]1-50-, -C(O)-, -C(S)-, -C(NRa)-, -C(NH2Cl)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NRa-, -NRaC(O)-, -C(O)NRaSO2-, -SO2NRaC(O)-, -OC(O)O-, -OC(O)S-, -SC(O)O-, -OC(O)NRa-, -NRaC(O)O-, -SC(O)NRa-, -NRaC(O)S-, -S(O)tN(Ra)-, -N(Ra)S(O)t-, and -XAA-; andLp is a connecting moiety through which P is covalently attached to L'.

[0017] In some embodiments, the linker L comprises at least one cleavable moiety. In some embodiments, the cleavable moiety comprises an acid-labile moiety, a reducibly-labile moiety, or an enzymatically-labile moiety. In some embodiments, the cleavable moiety comprises one or more groups selected from:wherein: each Rais independently selected at each occurrence from hydrogen, optionally substituted alkyl, and optionally substituted heteroalkyl.

[0018] In some embodiments, the cleavable moiety comprises the reducibly-labile moiety -S-S-.

[0019] In some embodiments, the linker L is of Formula (L-10):Formula (L-10), wherein in Formula (L-10):LAis selected from a bond, -NRa'-, and -S-;L1is a bond or comprises one or more groups selected from optionally substituted C1-C18alkylene, -C=C-, -CR — CRa-, optionally substituted 6- to 14-membered arylene, optionally substituted C3-C20cycloalkylene, -[CH2O]1-18-, -[CH2CH2O]1-18-, -[CH2CH2CH2O]1-18-, optionally substituted 5- to 18-membered heteroarylene, optionally substituted 3- to 20-membered heterocycloalkylene, -NRa-, -N=CRa-, -CRa=N-, -S-, -OP(O)ORaO-, -O-, -CRb2-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NRa-, -NRaC(O)-, -OC(O)O-, -OC(O)S-, -SC(O)O-, -OC(O)NRa-, -NRaC(O)O-, -SC(O)NRa-, -NRaC(O)S-, and -XAA-;Lc is selected from an acid-labile moiety, a reducibly-labile moiety, and an enzymatically- labile moiety;L2is a bond or comprises one or more groups selected from optionally substituted C1-C18 alkylene, -C=C-, -CR — CR1-, optionally substituted 6- to 14-membered arylene, optionally substituted C3-C20 cycloalkylene. -[CH2O]1-18-, -[CH2CH2O]1-18-, -[CH2CH2CH2O]1-18-, optionally substituted 5- to 18-membered heteroarylene, optionally substituted 3- to 20-membered heterocycloalkylene, -NRa-, -N=CRa-, -CRa=N-, -S-, -OP(O)ORaO-, -O-, -CRb2-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NRa-, -NRaC(O)-, -OC(O)O-, -OC(O)S-, -SC(O)O-, -OC(O)NRa-, -NRaC(O)O-, -SC(O)NRa-, -NRaC(O)S-, and -XAA-;LPis selected from a bond, -NRa-, -S-, and -O-; each Rais independently selected at each occurrence from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; each Ra’ is independently selected at each occurrence from hydrogen, optionally substituted alkyl, and optionally substituted heteroalkyl; each Rbis independently selected at each occurrence from hydrogen, halide, -OH, -SO3H, -OPO3H2, -PO3H2, -C(0)NRa2, -CO2Ra, -NRa2, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or two independent Rbgroups are takentogether to form optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocycloalkyl, or optionally substituted heteroaryl; and-XAA- is an amino acid sequence comprising 1 to 4 amino acid moieties.

[0020] In some embodiments, Lc is selected from:wherein: each Rais independently selected at each occurrence from hydrogen, optionally substituted alkyl, and optionally substituted heteroalkyl.

[0021] In some embodiments, Lc is -S-S-.

[0022] In some embodiments, the linker L is of Formula (L-l 1):Formula (L-l 1), wherein in Formula (L-l 1):LAis selected from a bond, -NH-, and -S-;L1is a bond or comprises one or more groups selected from -[C(Rb)2]1-16-, -C=C-, - CRa= CRa-, -[CH2CH2O]1-16-, -NRa-, -N=CRa-, -CRa=N-, -S-, -OP(O)ORaO-, -O-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NRa-, -NRaC(O)-, -OC(O)O-, -XAA-, -OC(O)NRa-, -NRaC(O)O-,L2is a bond or comprises one or more groups selected from -[C(Rb)2]i i6-, -C=C-,-CRa=CRa-, -[CH2CH2O]1-16-, -NRa-, -N=CRa-, -CRa=N-, -S-, -OP(O)ORaO-, -0-, -C(0)-, -C(0)0-, -OC(O)-, -C(O)NRa-, -NRaC(O)-, -OC(O)O-, -XAA-, -OC(O)NRa-, -NRaC(O)O-,Lp is selected from a bond, -NRa-, and -O-; each Ri is independently selected from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or both Ri groups are taken together to form optionally substituted cycloalkyl; each R2is independently selected from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or both R2groups are taken together to form optionally substituted cycloalkyl;each Rais independently selected at each occurrence from hydrogen, optionally substituted C1-C8alkyl, optionally substituted C1-C8fluoroalkyl, optionally substituted C3-C6cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; each Rais independently selected at each occurrence from hydrogen and optionally substituted alkyl; each Rbis independently selected at each occurrence from hydrogen, halide, -OH, -SO3H, -OPO3H2, -PO3H2, -CO2Ra, -NRa2, optionally substituted C1-C8alkyl, optionally substituted C1-C8fluoroalkyl, optionally substituted C3-C6cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; or two independent Rbgroups are taken together to form optionally substituted cycloalkyl; and-XAA- is an amino acid sequence comprising 2 to 4 amino acid moieties.

[0023] In some embodiments, the linker L is of Formula (L-12):Formula (L-12), wherein in Formula (L-12):LAis selected from a bond and -NH-;L1' comprises one or more groups selected from -[C(Rb)2]1-10-, -[CH2CH2O]1-10-, -NRa-, -O-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NRa-, -NRaC(O)-, -OC(O)O-, -XAA-, -OC(O)NRa-,L2’ comprises one or more groups selected from -[C(Rb)2]1-10-, -[CH2CH2O]I-IO-, -NRa-, -O-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NRa-, -NRaC(O)-, -OC(O)O-, -XAA-, -OC(O)NRa-,Lp is selected from a bond and -NRa’-; each Ri is independently selected from hydrogen, optionally substituted C1-C8alkyl, optionally substituted C1-C8fluoroalkyl, optionally substituted C3-C6cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; or both Ri groups are taken together to form optionally substituted C3-C6cycloalkyl; each R2 is independently selected from hydrogen, optionally substituted C1-C8alkyl, optionally substituted C1-C8fluoroalkyl, optionally substituted C3-C6cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; or both R2 groups are taken together to form optionally substituted C3-C6cycloalkyl; each Rais independently selected at each occurrence from hydrogen, optionally substituted C1-C6alkyl, optionally substituted C1-C6fluoroalkyl, and optionally substituted C3-C6cycloalkyl; each Ra' is independently selected at each occurrence from hydrogen and optionally substituted C1-C6alkyl; each Rbis independently selected at each occurrence from hydrogen, halide, -OH, -SO3H, -OPO3H2, -PO3H2, -CO2Ra, -NRa2, optionally substituted C1-C6alkyl, optionally substituted C1-C6fluoroalkyl, and optionally substituted C3-C6cycloalkyl; or two independent Rbgroups attached to the same carbon atom are taken together to form optionally substituted C3-C6cycloalkyl; and-XAA- is an amino acid sequence comprising 2 or 3 amino acid moieties.

[0024] In some embodiments, at least one Ri or R2 is other than hydrogen. In some embodiments, at least one Ri is an optionally substituted C1-C8alkyl. In some embodiments, each Ri is independently an optionally substituted C1-C8alkyl. In some embodiments, at least one R2is an optionally substituted C1-C8alkyl. In some embodiments, each R2 is independently an optionally substituted C1-C8alkyl.

[0025] In some embodiments, the linker L is selected from:

[0027] In some embodiments, the linker is a cleavable linker.

[0028] In some embodiments, the linker L is of Formula (L-20):Formula (L-20), wherein in Formula (L-20):LAis selected from a bond, -NRa'-, and -S-;L3is a bond or comprises one or more groups selected from -[C(Rb)2]1-8-, -NRa-, -C(O)-, -C(S)-, -C(NRa)-, -C(NH2CI)-, -C=C-, - CRa= CRa-, optionally substituted 6- to 14-membered arylene, optionally substituted C3-C20 cycloalkylene, optionally substituted 5- to 18-membered heteroarylene, and optionally substituted 3- to 20-membered heterocycloalkylene;Lx comprises one or more groups selected from optionally substituted C1-C18alkylene, -C=C-, -CRa=CRa-, optionally substituted 6- to 14-membered arylene, optionally substituted C3-C20 cycloalkylene, -[CJ OJ1-18-, -[CJLCI OJ1-18-, -[CH2CH2CH2O]1-18-, optionally substituted 5- to 18-membered heteroarylene, optionally substituted 3- to 20-membered heterocycloalkylene, -NRa-, -S-, -O-, -CRb2-, -C(O)-, -C(S)-, -C(NRa)-, -C(NH2C1)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NRa-, and -NRaC(O)-;L4is a bond or comprises one or more groups selected from -[C(Rb)2]1-8-, -NRa-, -C(O)-, -C(S)-, -C(NRa)-, -C(NH2Cl)-, -C=C-, -CRa=CRa-, optionally substituted 6- to 14-membered arylene, optionally substituted C3-C20 cycloalkylene, optionally substituted 5- to 18-membered heteroarylene, and optionally substituted 3- to 20-membered heterocycloalkylene;Lp is selected from a bond, -NRa-, -S-, and -O-; each Rais independently selected at each occurrence from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; each Rais independently selected at each occurrence from hydrogen, optionally substituted alkyl, and optionally substituted heteroalkyl; and each Rbis independently selected at each occurrence from hydrogen, halide, -OH, -SO3H, -OPO3H2, -PO3H2, -C(O)NRa2, -CO2Ra, -NRa2, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or two independent Rbgroups are taken together to form optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocycloalkyl, or optionally substituted heteroaryl.

[0029] In some embodiments, the linker L is of Formula (L-21):Formula (L-21), wherein in Formula (L-21):LAis selected from a bond and -NH-;LXcomprises one or more groups selected from optionally substituted -[C(Rb)2]1-16-, -C=C-, -CRa=CRa-, -[CH2CH2CH2O]1-16-, -NRa-, -O-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-,LPis selected from a bond and -NRa’-; each Rais independently selected at each occurrence from hydrogen, optionally substituted C1-C8alkyl, optionally substituted C1-C8fluoroalkyl, optionally substituted C3-C6cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; each Rais independently selected at each occurrence from hydrogen and optionally substituted C1-C6alkyl; and each Rbis independently selected at each occurrence from hydrogen, halide, -OH, -SO3H, -OPO3H2, -PO3H2, -CO2Ra, -NRa2, optionally substituted Ci-Cx alkyl, optionally substituted C1-C8fluoroalkyl, optionally substituted C3-C6cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; or two independent Rbgroups are taken together to form optionally substituted cycloalkyl.

[0030] In some embodiments, the linker L is of Formula (L-22a) or Formula (L-22b):Formula (L-22b), wherein in Formulas (L-22a) and (L-22b):LAis selected from a bond and -NH-;Lx comprises one or more groups selected from optionally substituted -[C(Rb)2]1-10-,-C=C-, -CR — CR1-, -[CH2CH2CH2O]1-10-, -NRa-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-,Lp is selected from a bond and -NRa'-; each Rais independently selected at each occurrence from hydrogen, optionally substituted C1-C8alkyl, optionally substituted C1-C8fluoroalkyl, optionally substituted C3-C6cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; each Ra' is independently selected at each occurrence from hydrogen and optionally substituted C1-C6alkyl; and each Rbis independently selected at each occurrence from hydrogen, halide, -OH, -SO3H, -OPO3H2, -PO3H2, -CO2Ra, -NRa2, optionally substituted C1-C8alkyl, optionally substituted C1-C8fluoroalkyl, optionally substituted C3-C6cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl,optionally substituted 5- to 6-membered heteroaryl; or two independent Rbgroups are taken together to form optionally substituted cycloalkyl.

[0031] In some embodiments, the linker L is selected from:

[0032] In some embodiments, the linker is a non-cleavable linker.

[0034] In some embodiments, each amino acid moiety of -XAA- is independently selected from alanine (Ala), arginine (Arg), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), phenylalanine (Phe), tryptophan (Trp), tyrosine (Tyr), valine (Vai), citrulline (Cit), and homocitrulline (HoCit). In some embodiments, each amino acid moiety of -XAA- is independently selected from alanine (Ala), glycine (Gly), lysine (Lys), phenylalanine (Phe), valine (Vai), and citrulline (Cit). In some embodiments, the amino acid sequence -XAA- is selected from -Val-Cit-, -Cit-Val-, -Vai-Ala-, -Ala-Val-, -Phe-Lys-, -Lys-Phe-, -Ala-Ala-, -Val-Val-, -Gly-Gly-, -Ala-Ala-Ala-, -Gly-Gly-Gly-, Gly-Gly-Phe-Gly-(SEQ ID NO: 1032) , -Gly-Phe-Gly-Gly-(SEQ ID NO: 1033),-Gly-Gly-Gly-Phe-(SEQ ID NO: 1034), -Phe-Gly-Gly-Gly-(SEQ ID NO: 1035), and -Gly-Gly-Gly-Gly-(SEQ ID NO: 1036). In some embodiments, the amino acid sequence -XAA- is selected from -Val-Cit-, -Cit-Val-, -Val-Ala-, -Ala-Val-, -Phe-Lys-, -Lys-Phe-, -Ala-Ala-, -Val-Val-, -Gly-Gly-, -Ala-Ala-Ala-, and -Gly-Gly-Gly-.

[0035] In some embodiments, the single stranded oligonucleotide P is a phosphorodiamidate morpholino oligonucleotide or an antisense oligonucleotide. In some embodiments, the single stranded oligonucleotide P is delivered into a muscle cell. In some embodiments, the single stranded oligonucleotide P induces skipping of exon 23 of the DMD gene.

[0036] In some embodiments, the phosphorodiamidate morpholino oligonucleotide comprises the sequence 5'-C6 Amino-GGCCAAACCTCGGCTTACCTGAAAT-3' (SEQ ID NO:408). In some embodiments, the antisense oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 1045, SEQ ID NOs: 158-222, SEQ ID NO:395-405, and SEQ ID NO:410-988. In some embodiments, the antisense oligonucleotide is a peptide nucleic acid (PNA) oligonucleotide. In some embodiments, the peptide nucleic acid (PNA) oligonucleotide comprises the sequence (C)-3’-TAAAGTCCATTCGGCTCCAAACCGG-C6 Amino-5’(N) (SEQ ID NO:409). In some embodiments, the single stranded oligonucleotide P comprises at least from about 10 to about 30 nucleotides in length.

[0037] In some embodiments, the truncated protein modulates muscular dystrophy. In some embodiments, the muscular dystrophy is Duchenne muscular dystrophy or Becker muscular dystrophy.

[0038] In some embodiments, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:9, CDR2 comprises the amino acid sequence of SEQ ID NO: 10, CDR3 comprises SEQ ID NO: 11 ; and the VH CDR1 comprises the amino acid sequence of SEQ ID NO: 15, CDR2 comprises the amino acid sequence of SEQ ID NO:4, CDR3 comprises the amino acid sequence of SEQ ID NO:5.

[0039] In some embodiments, the VL CDR1 comprises the amino acid sequence of SEQ ID NO:29, CDR2 comprises the amino acid sequence of SEQ ID NO: 10, CDR3 comprises SEQ ID NO: 11; and the VH CDR1 comprises the amino acid sequence of SEQ ID NO: 15, CDR2 comprises the amino acid sequence of SEQ ID NO:26, CDR3 comprises the amino acid sequence of SEQ ID NO:5.

[0040] In some embodiments, the antibody or antigen-binding fragment thereof comprises a light chain variable region (VL) comprising an amino acid sequence of SEQ ID NO:21 and a heavy chain variable region (VH) comprising an amino acid sequence of SEQ ID NO: 14.

[0041] In some embodiments, the antibody or antigen-binding fragment thereof comprises a full length light chain (LC) comprising an amino acid sequence of SEQ ID NO:20 and a full length heavy chain (HC) comprising an amino acid sequence of SEQ ID NO: 13.

[0042] In some embodiments, the antibody or antigen-binding fragment thereof comprises: a light chain variable domain (VL) comprising an amino acid sequence that is at least 95% identical to an amino acid sequence selected from the group consisting of 3E10-VL-H1 (SEQ ID NO:85), 3E10-VL-H2 (SEQ ID NO:86), 3E10-VL-H3 (SEQ ID NO:87), 3E10-VL-H4 (SEQ ID NO:88), 3E10-VL-H5 (SEQ ID NO: 89), and 3E10-VL-H6 (SEQ ID NO: 90); and a heavy chain variable domain (VH) comprising an amino acid sequence that is at least 95% identical to an amino acid sequence selected from the group consisting of 3E10-VH-H1 (SEQ ID NO:64), 3E10-VH-H2 (SEQ ID NO:65), 3E1O-VH-H3 (SEQ ID NO:66), 3E10-VH-H4 (SEQ ID NO:67), 3E1O-VH-H5 (SEQ ID NO:68), 3E10-VH-H6 (SEQ ID NO:69), and 3E10-VH-H7 (SEQ ID NO:70).

[0043] In some embodiments, the antibody or antigen-binding fragment thereof comprises: a light chain variable domain (VL) comprising an amino acid sequence selected from the group consisting of 3E10-VL-H1 (SEQ ID NO:85), 3E10-VL-H2 (SEQ ID NO:86), 3E10-VL-H3 (SEQ ID NO:87), 3E10-VL-H4 (SEQ ID NO:88), 3E10-VL-H5 (SEQ ID NO:89), and 3E10-VL-H6 (SEQ ID NO:90); and a heavy chain variable domain (VH) comprising an amino acid sequenceselected from the group consisting of 3E10-VH-H1 (SEQ ID NO:64), 3E10-VH-H2 (SEQ ID NO:65), 3E10-VH-H3 (SEQ ID NO:66), 3E10-VH-H4 (SEQ ID NO:67), 3E10-VH-H5 (SEQ ID NO:68), 3E10-VH-H6 (SEQ ID NO:69), and 3E10-VH-H7 (SEQ ID NO:70).

[0044] In some embodiments, the antibody or antigen-binding fragment thereof comprises a VL / VH pair selected from the group consisting of (a) VL1 (SEQ ID NO:85) and VH1 (SEQ ID NO:64), (b) VL1 (SEQ ID NO:85) and VH2 (SEQ ID NO:65), (c) VL1 (SEQ ID NO:85) and VH3 (SEQ ID NO:66), (d) VL1 (SEQ ID NO:85) and VH4 (SEQ ID NO:67), (e) VL2 (SEQ ID NO:86) and VH1 (SEQ ID NO:64), (f) VL2 (SEQ ID NO:86) and VH2 (SEQ ID NO:65), (g) VL2 (SEQ ID NO:86) and VH3 (SEQ ID NO:66), (h) VL2 (SEQ ID NO:86) and VH4 (SEQ ID NO:67), (i) VL3 (SEQ ID NO:87) and VH1 (SEQ ID NO:64), (j) VL3 (SEQ ID NO:87) and VH2 (SEQ ID NO:65), (k) VL3 (SEQ ID NO:87) and VH3 (SEQ ID NO:66), (1) VL3 (SEQ ID NO:87) and VH4 (SEQ ID NO:67), (m) VL4 (SEQ ID NO:88) and VH1 (SEQ ID NO:64), (n) VL4 (SEQ ID NO:88) and VH2 (SEQ ID NO:65), (o) VL4 (SEQ ID NO:88) and VH3 (SEQ ID NO:66), (p) VL4 (SEQ ID NO:88) and VH4 (SEQ ID NO:67), (q) VL5 (SEQ ID NO:89) and VH5 (SEQ ID NO:68), (r) VL5 (SEQ ID NO:89) and VH6 (SEQ ID NO:69), (s) VL6 (SEQ ID NO:90) and VH5 (SEQ ID NO:68), and (t) VL6 (SEQ ID NO:90) and VH6 (SEQ ID NO:69).

[0045] In some embodiments, the antibody or antigen-binding fragment thereof comprises: a light chain variable domain (VL) comprising 3E10-VL-H6 (SEQ ID NOVO) and a heavy chain variable domain (VH) comprising 3E10-VH-H6 (SEQ ID NO:69).

[0046] In some embodiments, the antibody or antigen-binding fragment thereof comprises: a light chain variable domain (VL) comprising the amino acid sequence (DIQMTQSPSSLSASLGDRATITCRASKTVSTSSYSYMHWYQQKPGQPPKLLIKYASYLE SGVPSRFSGSGSGTDFTLTISSLQPEDAATYYCQHSREFPWTFGGGTKVEIK) (SEQ ID NO: 117) and a heavy chain variable domain (VH) comprising the amino acid sequence (EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGMHWVRQAPGKGLEWVSYISSGSSTI YYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRGLLLDYWGQGTTVTVS S) (SEQ ID NO: 105).

[0047] In some aspects, the present disclosure provides a method for treating a subject in need thereof, the method comprising administering a therapeutically effective amount of a conjugate of the disclosure to the subject.

[0048] In some aspects, the present disclosure provides a method of treating Duchenne muscular dystrophy (DMD) in a subject in need thereof, the method comprising administering a therapeutically effective amount of a conjugate of the disclosure to the subject.

[0049] In another aspect, the disclosure provides a method for delivering a conjugate comprising a 3E10 antibody or an antigen binding fragment thereof conjugated to a single stranded oligonucleotide to a tissue of a subject in vivo, the method including parenterally administering a pharmaceutical composition, as described herein, to the subject. In some embodiments, the antisense oligonucleotide is for treating a disease or disorder including, but not limited to a skeletal muscle disorder, a neurogenetic disease, a cardiovascular disease, a metabolic disease, or a lung disorder for which a known disease-causing mutation.

[0050] In some embodiments of the methods and compositions described herein, the 3E10 antibody or antigen-binding fragment thereof includes (a) a light chain variable region (VL) complementarity determining region (CDR) 1 comprising the amino acid sequence of 3E10-VL- CDR1 (SEQ ID NO: 9), (b) a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2 (SEQ ID NO: 10), (c) a VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3 (SEQ ID NO: 11), (d) a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of 3E10-VH-CDRla (SEQ ID NO: 16), (e) a VH CDR2 comprising the amino acid sequence of 3E10- VH-CDR2 (SEQ ID NO: 4), and (f) a VH CDR3 comprising the amino acid sequence of 3E10- VH-CDR3 (SEQ ID NO: 5).

[0051] In some embodiments of the methods and compositions described herein, the 3E10 antibody or antigen-binding fragment thereof includes (a) a light chain variable region (VL) complementarity determining region (CDR) 1 comprising an amino acid sequence having no more than two amino acid substitutions relative to 3E10-VL-CDR1 (SEQ ID NO: 9), (b) a VL CDR2 comprising an amino acid sequence having no more than two amino acid substitutions relative to 3E10-VL-CDR2 (SEQ ID NO: 10), (c) a VL CDR3 comprising an amino acid sequence having no more than two amino acid substitutions relative to 3E10-VL-CDR3 (SEQ ID NO: 11), (d) a heavy chain variable region (VH) CDR1 comprising an amino acid sequence having no more than two amino acid substitutions relative to 3E10-VH-CDRla (SEQ ID NO: 3), (e) a VH CDR2 comprising an amino acid sequence having no more than two amino acid substitutions relative to 3E10-VH-CDR2 (SEQ ID NO: 4), and (f) a VH CDR3 comprising an amino acid sequence having no more than two amino acid substitutions relative to 3E10-VH-CDR3 (SEQ ID NO: 5).

[0052] In some embodiments of the methods and compositions described herein, the 3E10 antibody or antigen-binding fragment thereof includes (a) a light chain variable region (VL) complementarity determining region (CDR) 1 comprising the amino acid sequence of 3E10-VL- CDRlm (SEQ ID NO: 61), (b) a VL CDR2 comprising the amino acid sequence of 3E10-VL- CDR2m (SEQ ID NO: 62), (c) a VL CDR3 comprising the amino acid sequence of 3E10-VL- CDR3m (SEQ ID NO: 63), (d) a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of 3E10-VH-CDRlm (SEQ ID NO: 58), (e) a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2m (SEQ ID NO: 59), and (f) a VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3m (SEQ ID NO: 60).BRIEF DESCRIPTION OF THE DRAWINGS

[0053] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0054] Figure 1 (SEQ ID NOs: 1-12) illustrates amino acid sequences for the parent 3E10 monoclonal antibody.

[0055] Figures 2A (SEQ ID NOs: 13-25), 2B (SEQ ID NOs:26-30), and 2C (SEQ ID NOs:31-33) illustrate amino acid sequences for the D3 IN variant (Figure 2A), other CDR variants (Figure 2B), and additionally contemplated CDR variants (Figure 2C) of the 3E10 monoclonal antibody, in accordance with some embodiments of the present disclosure.

[0056] Figure 3 (SEQ ID NOs:34-57) illustrates example charge-conserved CDR variants of the 3E10 monoclonal antibody, in accordance with various embodiments of the present disclosure.

[0057] Figure 4 (SEQ ID NOs: 58-63) illustrates example CDR variants containing a combination of amino acid substitutions, charged-conserved amino acid substitutions, and rationally-designed amino acid substitutions of the 3E10 monoclonal antibody, in accordance with various embodiments of the present disclosure.

[0058] Figure 5 (SEQ ID NOs: 103-112) illustrates a sequence alignment of examples of humanized 3E10 heavy chain variable regions, with CDRs underlined as indicated.

[0059] Figure 6 (SEQ ID NOs: 113-121) illustrates a sequence alignment of examples of humanized 3E10 light chain variable regions, with CDRs and putative nuclear localization signals (NLS) underlined as indicated.

[0060] Figures 7A, 7B, 7C, 7D, and 7E (SEQ ID NOs: 122-137) collectively illustrate a sequence alignment of example of humanized di-scFv constructs of the 3E10 monoclonal antibody.

[0061] Figure 8 illustrates 3E10 (V66)-phosphorodiamidate morpholino oligomer (PMO) conjugates using three separate linker chemistries, maleimide, SATA-SAPP, and DBCO.

[0062] Figures 9 illustrates 3E10-phosphorodiamidate morpholino oligomer (PMO) conjugates demonstrating dose-dependent exon skipping in vitro.

[0063] Figure 10 illustrates single dose exon skipping in Duchenne muscular dystrophy (DMD) using a 3E10-phosphorodiamidate morpholino oligomer (PMO) conjugate. Abbreviations shown: Tibialis anterior (T), Gastrocnemius (G), Quadricep (Q), Deltoid (D), and Heart (H).

[0064] Figures 11A and 11B illustrate electrostatic surface potential renderings of a molecular model of a 3E10-scFv construct, revealing a putative Nucleic Acid Binding pocket (NAB1). Figure 11A additionally shows predicted structural and electrostatic potential changes induced by amino acid substitutions at residue HC CDR1 residue 31. Figure 1 IB is an illustration of molecular modeling of 3E10-scFv (Pymol) with NAB1 amino acid residues highlighted by punctate dots.

[0065] Figure 11C illustrates mapping of the putative nucleic acid binding pocket, as identified by the molecular modeling shown in Figures 11 A and 1 IB, onto the amino acid sequence of the 3E10-scFv construct.

[0066] Figure 12 shows the study design for confirmatory single dose exon skipping by 3E10 (V66) in mdx mice.

[0067] Figure 13 shows gel electrophoresis analysis of follow-on single dose IV study performed to confirm DMD functional skipping in mdx mice with 3E10 (V66) non-cleavable linker-PMO conjugate.

[0068] Figure 14A illustrates the results of DMD exon skipping detected across skeletal and heart muscle in mdx mice receiving non-cleavable and disulfide cleavable 3E10 (V66) PMO conjugates.

[0069] Figure 14B illustrates single low-dose administration of 3E10 (V66) - PMO conjugates with cleavable and non-cleavable linkers in selected tissues in mdx mice.

[0070] Figure 15 illustrates nucleotide sequences (SEQ ID NO: 1045, 405, and 1041-1044) for the oligomers designed to cause skipping of exon 23 in dystrophin (DMD) RNA. K denotes lysine residues on PNA; superscript O denotes 2’OMe modifications; superscript F denotes 2’fluoro modifications; superscript L denotes locked nucleic acid (LNA) modifications.

[0071] Figure 16 illustrates amino acid sequences of humanized 3E10 variable heavy (3E10-VH) domains (SEQ ID NOs:64-70), in accordance with various embodiments of the present disclosure.

[0072] Figure 17 illustrates amino acid sequences of mature humanized 3E10 heavy chains (3E10-HC) lacking a signal peptide (SEQ ID NOs:71-77), in accordance with various embodiments of the present disclosure.

[0073] Figure 18 illustrates amino acid sequences of humanized 3E10 heavy chains (3E10-HC) (SEQ ID NOs:78-84), in accordance with various embodiments of the present disclosure.

[0074] Figure 19 illustrates amino acid sequences of humanized 3E10 variable light (3E10-VL) domains (SEQ ID NOs:85-90), in accordance with various embodiments of the present disclosure.

[0075] Figure 20 illustrates amino acid sequences of mature humanized 3E10 light chains (3E10-LC) lacking a signal peptide (SEQ ID NOs:91-96), in accordance with various embodiments of the present disclosure.

[0076] Figure 21 illustrates amino acid sequences of humanized 3E10 light chains (3E10- LC) (SEQ ID NOs:97-98, 100-102, and 1045), in accordance with various embodiments of the present disclosure.

[0077] Figure 22 illustrates western blots of ENT2 protein expression in selected human and mouse healthy tissues.

[0078] Figure 23 illustrates the internalization of a labeled 3E10 (V66) - PMO conjugate in C2C12 muscle myotubes and A427 tumor cells.

[0079] Figure 24 illustrates the experimental design for single and repeat dose studies with 3E10 (V66) - PMO conjugates measuring exon skipping in mdx mice.

[0080] Figures 25A-25C illustrate the durability over time of the 3E10 (V66) - PMO conjugate administration for exon skipping in deltoid and quadriceps (Figure 25A), in tibialis anterior and gastrocnemius (Figure 25B), and in heart and diaphragm (Figure 25C).

[0081] Figure 26 illustrates single dose administration of the 3E10 (V66) - PMO conjugate exhibits dystrophin protein restoration.

[0082] Figures 27A and 27B illustrate dystrophin restoration after administration of the 3E10 (V66) - PMO conjugate in selected tissues on Day 14 (Figure 27A) and on Day 28 (Figure 27B).

[0083] Figure 28A illustrates the process for quantifying attachment sites for Lys-azide conjugation intermediates of 3E10-D31N monoclonal antibody (V66), illustrating peptides KVEPK (SEQ ID NO: 1022) and K*VEPK (SEQ ID NO: 1023) released by proteolysis.

[0084] Figure 28B shows the mass spectroscopy results of mapped Lys-azide conjugation intermediates of 3E10-D31N monoclonal antibody (V66) after proteolysis.

[0085] Figure 28C shows an alignment of chimeric 3E10-D31N variable heavy and variable light chains with the corresponding sequences of a humanized 3E10-D31N antibody (V66), in accordance with various embodiments of the present disclosure. The consensus sequence for the VH is SEQ ID NO: 1024. The 3E10-D31N-VH sequence is SEQ ID NO: 1025. The 3E10- D31N-VH6 sequence is SEQ ID NO:69. The consensus sequence for the VL is SEQ ID NO: 1026. The 3E10-D31N-VL sequence is SEQ ID NO:8. The 3E10-D31N-VL6 sequence is SEQ ID NO:90.

[0086] Figures 29A and 29B collectively illustrate improved cellular internalization of 3E10-D31N monoclonal antibody (V66) oligonucleotide conjugates in A427 cells utilizing trasglutaminase-mediated enzymatic conjugation.

[0087] Figure 30A shows improved exon skipping utilizing transglutaminase-mediated enzymatic conjugation. Figure 30B shows varying the length of the PEG linkers (e.g., PEG4,PEG8, and PEG12) did not significantly impact exon skipping. Figure 30C shows enhanced exon- skipping of cleavable linkers, protease Cathepsin-B and SPDMV (disulfide) compared to non- cleavable linkers with transglutaminase-mediated enzymatic conjugation.

[0088] Figure 31A show a di-methyl-hindered disulfide cleavable linker and protease cleavable linker have greater stability over a single methyl-hindered disulfide linker in mouse serum. Figure 31B show a data table indicating the panel of antibody-oligonucleotide conjugates (AOCs) tested and their oligonucleotide to antibody ratios (OAR or DAR).

[0089] Figure 32 shows a summary table of the in vitro performance of several antibody- oligonucleotides conjugates utilizing lysine and transglutaminase-mediated conjugation.

[0090] Figure 33 shows levels of exon skipping of DMD preRNA at day 7 post-dose in mdx mice that were administered a 3E10 AOC with a noncleavable linker. Doses shown for each muscle group are, from left to right, 2.5 mg / kg, 5 mg / kg, 10 mg / kg, 20 mg / kg, and 30 mg / kg.

[0091] Figure 34 shows quantification of delivered PMO at day 7 post-dose as determined by hybridization ELISA in diaphragm (Di), quadriceps (Q), gastrocnemius (G), tibialis anterior (TA), deltoid (D), and heart (H) muscle tissue samples from mdx mice that were administered a 3E10 AOC with a noncleavable linker. Doses shown for each muscle group are, from left to right, vehicle, 2.5 mg / kg, 5 mg / kg, 10 mg / kg, 20 mg / kg, and 30 mg / kg.

[0092] Figure 35A shows quantification of delivered PMO at day 14 post-dose as determined by hybridization ELISA in diaphragm, quadriceps, and gastrocnemius tissue samples from mdx mice that were administered a 3E10 AOC with a noncleavable linker. Figure 35B shows quantification of delivered PMO at day 14 post-dose as determined by hybridization ELISA in tibialis anterior, deltoid, and heart tissue samples from mdx mice that were administered a 3E10 AOC with a noncleavable linker.

[0093] Figure 36 shows levels over time of exon skipping of DMD preRNA in mdx mice that were administered a 3E10 AOC with a noncleavable linker.

[0094] Figure 37 shows quantification of delivered PMO at day 10 post-dose as determined by hybridization ELISA in diaphragm (Di), quadriceps (Q), gastrocnemius (G), tibialis anterior (TA), deltoid (D), and heart (H) muscle tissue samples from mdx mice that wereadministered a 3E10 AOC with a noncleavable linker. Cohorts shown for each muscle group are, from left to right, vehicle, PMO30, and AOC30.

[0095] Figure 38 shows levels of exon skipping of DMD preRNA at day 10 post-dose in mdx mice that were administered a 3E10 AOC with a noncleavable linker compared to PMO alone (PMO23) or vehicle control.

[0096] Figure 39 shows levels of dystrophin protein restoration at day 28 post-dose in heart, diaphragm, tibialis anterior (TA), and quadriceps (Q) muscle tissue samples from mdx mice that were administered a 3E10 AOC with a noncleavable linker. Doses shown for each muscle group are, from left to right, 18 mg / kg, 30 mg / kg, and 42 mg / kg.

[0097] Figures 40A-40C show day 28 post-dose immunofluorescent detection of dystrophin in the diaphragm of WT mice (Figure 40A), mdx mice (Figure 40B), and mdx mice that were administered a 3E10 AOC with a noncleavable linker (Figure 40C).

[0098] Figure 41 illustrates a 3E10 (V66)-phosphorodiamidate morpholino oligomer (PMO) conjugate having a Cathepsin B cleavable linker.

[0099] Figure 42 shows levels of exon skipping of DMD preRNA at day 14 post-dose in mdx mice that were administered vehicle control, a Tg-noncleavable 3E10 AOC, and a Tg- cleavable 3E10 AOC. The muscle groups shown, from left to right, are, diaphragm (Di), quadriceps (Q), gastrocnemius (G), tibialis anterior (TA), deltoid (D), and heart (H).

[0100] Figure 43 shows levels of exon skipping of DMD preRNA at day 14 post-dose in mdx mice that were administered vehicle control, a Lys-noncleavable 3E10 AOC, and a Tg- noncleavable 3E10 AOC. The muscle groups shown, from left to right, are, diaphragm, quadriceps, gastrocnemius, tibialis anterior, deltoid, and heart.

[0101] Figure 44 shows levels of exon skipping of DMD preRNA at day 28 post-dose in mdx mice that were administered a noncleavable 3E10 AOC and a disulfide cleavable 3E10 AOC. The muscle groups shown, from left to right, are, diaphragm, tibialis anterior (TA), heart, deltoid (Delt), gastrocnemius (Gastroc), and quadriceps (Quad).

[0102] Figure 45 shows day 28 post-dose levels of dystrophin protein restoration in diaphragm, tibialis anterior, quadriceps, and deltoid muscle tissue samples from mdx mice that were administered a noncleavable 3E10 AOC and a disulfide cleavable 3E10 AOC.

[0103] Figure 46 shows day 28 post-dose quantification of dystrophin protein distribution in diaphragm, heart, and tibialis anterior (TA) of mdx mice that were administered a noncleavable 3E10 AOC and a disulfide cleavable 3E10 AOC.

[0104] Figure 47 shows in situ hybridization analysis of PMO distribution in the deltoid muscle of a mouse that was administered a noncleavable 3E10 AOC, at 20x magnification.

[0105] Figure 48 shows in situ hybridization analysis of PMO distribution in the deltoid muscle of a mouse that was administered a noncleavable 3E10 AOC, at 40x and lOOx magnification.

[0106] Figure 49 shows in situ hybridization analysis of PMO distribution in the heart muscle of mice that were administered vehicle control, a cleavable 3E10 AOC, or a noncleavable 3E10 AOC, at 40x magnification.

[0107] Figure 50 shows quantification at day 7 post-dose of delivered PMO as determined by hybridization ELISA in deltoid, diaphragm, heart, and tibialis anterior muscle tissue samples from mice that were administered vehicle, PMO alone, PPMO alone, or a disulfide cleavable 3E10 AOC. Cohorts shown for each muscle group are, from left to right, vehicle, PMO, PPMO, and AOC.

[0108] Figure 51 shows in situ hybridization analysis of PMO distribution in the deltoid muscle of mice that were administered PMO alone, PPMO alone, or a disulfide cleavable 3E10 AOC, at 4x and 40x magnification.

[0109] Figure 52 illustrates various linker designs for generating 3E10 AOCs, i.e., a phosphatase cleavable linker, a glucuronidase cleavable linker, an SPDMV disulfide cleavable linker, and a SPDB disulfide cleavable linker.

[0110] Figures 53A and 53B show levels of exon skipping of DMD preRNA in mdx mice that were administered 3E10 AOCs having (Lys)-DBCO-PEG8-PMO, (Lys)-DBCO-PEG4- SPDMV-PMO, or (Lys)-DBCO-PEG8-PO4-PAB-PMO linkers (Figure 48A), or (Tg)-DBCO- PEG8-PMO, (Tg)-DBCO-PEG4-SPDMV-PMO, or (Tg)-CathB-PEG8-PMO linkers (Figure 48B).

[0111] Figure 54A illustrates the design of a site-specific DAR 4 3E10 AOC transglutaminase conjugate comprising a hindered disulfide SPDMV linker (Tg-SPDP). Figure54B shows mass spectrometry data for the site-specific DAR 4 3E10 AOC transglutaminase conjugate comprising a hindered disulfide SPDMV linker (Tg-SPDP). Figure 54C shows levels of exon skipping of DMD preRNA in mdx mice that were administered a noncleavable 3E10 AOC (Tg-Peg8), or three cleavable 3E10 AOCs (Tg-SPDMV, Tg-CathB, and Tg-SPDP) .DETAILED DESCRIPTIONL _ Introduction

[0112] The present disclosure provides compositions, conjugates, and methods for delivering therapeutic polynucleotides, e g., antisense oligonucleotides, that are amendable to exon skipping in vivo, and that are not reliant upon the conventional viral-based or liposomal-based delivery methodologies associated with difficult and costly production, limited packaging capacity, and adverse immunological events. In some aspects, described in greater detail below, these compositions and methods are based on the covalent attachment of a 3E10 antibody or an antigen binding fragment thereof to a single stranded oligonucleotide, forming a conjugate, increasing the in vivo effectiveness of these complexes. In some embodiments, the methods and compositions find particular use for the treatment of genetic diseases and disorders, including neurogenetic diseases, musculoskeletal disorders, cardiovascular diseases, metabolic diseases, cancers, lung disorders, and other diseases that can be benefitted by exon-skipping therapies. For instance, described herein are compositions comprising a conjugate of (i) a 3E10 antibody or antigen-binding fragment thereof, and (ii) an antisense oligonucleotide, as well as methods for using such compositions for the treatment of diseases and disorders, including neurogenetic diseases, musculoskeletal disorders, cardiovascular diseases, metabolic diseases, cancers, lung disorders, and other diseases that can be benefitted by exon-skipping therapies described herein.

[0113] The studies described herein demonstrate that conjugates comprising a cell- penetrating and nucleic acid-binding antibody (3E10) conjugated via a linker to a therapeutic antisense oligonucleotides can effectively cause exon skipping in vitro and in vivo. For example, Example 2 demonstrates that both stable (non-cleavable) and cleavable 3E10 (V66) conjugates to a phosphorodi ami date morpholino oligomer (PMO) targeting exon 23 of DMD caused dose- dependent exon-skipping in differentiated C2C12 myotubules. Examples 4-6 and 9 demonstrate that V66-PMO conjugates were able to cause exon-skipping in an mdx DMD mouse model. Example 7 demonstrates that there is high ENT2 protein expression in mouse and human muscle,as well as in the heart and diaphragm tissue. Examples 8 and 11 demonstrate that V66-PMO conjugates are internalized in C2C12 muscle myotubes and A427 tumor cells. Example 9 further demonstrates that a V66-PMO conjugate was able to restore dystrophin expression in muscle tissues in mdx mice models of DMD.

[0114] In some embodiments, the advantageous properties of the compositions and methods described herein are based, at least in part, on the substantial targeting and delivery of exon-skipping oligonucleotides to muscle tissues when conjugated to 3E10 antibodies and antigen- binding fragments thereof. For instance, as described in Example 13, and exemplified in Figure 34, high levels of tissue PMO delivery was observed in diaphragm (Di), quadriceps (Q), gastrocnemius (G), tibialis anterior (TA), deltoid (D), and heart (H) muscle tissue at 7 days post- treatment. Moreover, the delivery demonstrated a dose-dependency in all tissues, including the heart. Delivery of the PMO exon skipping oligo to these muscle tissues was approximately 100- fold higher when conjugated to 3E10 than when injected alone without 3E10, as described in Example 14 and exemplified in Figure 37.

[0115] In some embodiments, the advantageous properties of the compositions and methods described herein are based, at least in part, on the persistent localization of exon skipping oligonucleotides to when conjugated to 3E10 antibodies and antigen-binding fragments thereof. For instance, as described in Example 13, and exemplified in Figures 35A-35B, high levels of tissue PMO localization to diaphragm (Di), quadriceps (Q), gastrocnemius (G), tibialis anterior (TA), deltoid (D), and heart (H) muscle tissue persisted 14-days post-treatment. Moreover, the localization followed a dose-dependency.

[0116] In some embodiments, the advantageous properties of the compositions and methods described herein are based, at least in part, on the persistent exon skipping induced by oligonucleotides when conjugated to 3E10 antibodies and antigen-binding fragments thereof. For instance, as described in Example 14, and exemplified in Figure 36, exon skipping in was increased in diaphragm (Di), tibialis anterior (TA), and heart (H) muscle tissue at day 20 post-injection, relative to exon skipping at day 10 post-injection.

[0117] In some embodiments, the advantageous properties of the compositions and methods described herein are based, at least in part, on the even distribution of exon-skipping oligos across muscle tissues when conjugated to 3E10 antibodies and antigen-binding fragmentsthereof. For instance, as described in Example 16, and exemplified in Figures 47-49, PMO oligo was evenly-distributed across deltoid and heart tissues when administered conjugated to 3E10. Moreover, as described in Example 17 and exemplified in Figure 51, significantly more PMO exon-skipping oligonucleotide was delivered to the deltoid, and translocated to the nucleus of muscle cells, when administered conjugated to 3E10 than when administered alone (compared to both PMO and PPMO oligonucleotides).

[0118] Accordingly, in one aspect, the disclosure provides a conjugate of Formula (I):A-(L-Pr)q Formula (I), wherein in Formula (I): A is an antibody, antigen-binding fragment thereof or antigen-binding fragment thereof comprising a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of SEQ ID NO:58, CDR2 comprising the amino acid sequence of SEQ ID NO:59, CDR3 comprising SEQ ID NO:60; and a light chain variable region (VL) CDR1 comprising the amino acid sequence of SEQ ID NO:61, CDR2 comprising the amino acid sequence of SEQ ID NO:62, CDR3 comprising the amino acid sequence of SEQ ID NO:63; L is a linker; P is an oligonucleotide capable of hybridizing to a pre-mRNA transcript, wherein the oligonucleotide induces exon skipping in the pre-mRNA transcript; r is an integer from 1 to 4; and q is an integer from 1 to 16.

[0119] Similarly, in one aspect, the disclosure provides a method for inducing exon skipping in a target tissue in a subject by administering a conjugate of Formula (I). In some embodiments, the target tissue is a muscle tissue. In some embodiments, the target tissue is diaphragm tissue. In some embodiments, the target tissue is quadricep tissue. In some embodiments, the target tissue is gastrocnemius tissue. In some embodiments, the target tissue is tibialis anterior tissue. In some embodiments, the target tissue is deltoid tissue. In some embodiments, the target tissue is heart muscle tissue.

[0120] In some embodiments, the disclosure provides a method for treating a disorder in a subject in need thereof by administering a therapeutically effective amount of a composition comprising a conjugate of Formula (I) to the subject. In some embodiments, the disorder is a muscle disorder. In some embodiments, the muscle disorder is a muscular dystrophy. In some embodiments the muscular dystrophy is Duchenne muscular dystrophy (DMD).II. Definitions

[0121] The terminology used in the present disclosure is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0122] As used herein, the singular forms “a”, “an” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will also be understood that the term “and / or” as used herein refers to and encompasses any and all possible combinations of one or more of the associated listed items. Unless the context requires otherwise, it will be further understood that the terms “includes,” “comprising,” or any variation thereof, when used in this specification, specify the presence of stated features, integers, steps, operations, elements, and / or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups thereof. Furthermore, to the extent that the terms “including,” “includes,” “having,” “has,” “with,” or variants thereof are used in either the detailed description and / or the claims, such terms are intended to be inclusive in a manner similar to the term “comprising.” Additionally, where the terms “comprising,” “including,” “includes,” “having,” “has,” “with,” or variants thereof are used in either the detailed description and / or the claims, alternatives reciting “consisting of’ or “consisting essentially of’ are intended to be encompassed within such disclosures.

[0123] Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein.

[0124] Use of the term “about” is intended to describe values either above or below the stated value in a range of approx. + / - 10%.

[0125] As used herein, the term “antibody” refers to an immunoglobulin molecule that recognizes and specifically binds to a target, such as a protein, polypeptide, peptide, carbohydrate, polynucleotide, lipid, or combinations of the foregoing through at least one antigen recognition site within the variable region of the immunoglobulin molecule. The term “antibody,” as used herein, is used in the broadest sense and encompasses monoclonal antibodies (including full length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), chimeric antibodies, humanized antibodies, human antibodies, fusion proteinscomprising an antigen determination portion of an antibody, and antibody fragments (such as Fab, Fab’, F(ab’)2, Fv fragments, scFv molecules), and any other modified immunoglobulin molecule comprising an antigen recognition site, so long as they exhibit one or more of the desired biological activities. In embodiments, “desired biological activity” of an antibody refers to the ability of the antibody to bind to its target antigen, e.g., a nucleic acid, e.g., DNA. In embodiments, “desired biological activity” can further include antibody binding to its target antigen and resulting in a measurable biological response which can be measured in vitro or in vivo. Such activity can be antagonistic or agonistic. In embodiments, “desired biological activity” of an antibody refers to the ability of the antibody to bind to a target, e.g., nucleic acid molecules. In embodiments, “desired biological activity” of an antibody refers to the ability of the antibody to bind to a cellular receptor, e.g., ENT2. In embodiments, “desired biological activity” of an antibody refers to the ability of the antibody to be internalized by a target cell. “Target antigen,” as used herein, refers to the molecule that is bound specifically by the antigen-binding domain comprising the variable regions of a given antibody. The term “specifically binds” refers to the binding of an antibody to its cognate antigen (e.g., a nucleic acid, e.g., DNA) while not significantly binding to other antigens.

[0126] Depending on the amino acid sequences of the constant domains of their heavy chains, antibodies (immunoglobulins) can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these can be further divided into subclasses or isotypes, e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA . “Isotype,” as used herein, refers to any of the subclasses of immunoglobulins defined by the chemical and antigenic characteristics of their constant regions. The heavy chain constant domains that correspond to the different classes of immunoglobulins are called a, y, s, y, and p. respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known and described generally in, for example, Abbas et al. Cellular and Mol. Immunology, 4thed. (W.B. Saunders, Co., 2000). It should be understood that antibodies disclosed herein can also comprise hybrids of isotypes and / or subclasses.

[0127] Antibodies of the present disclosure are generally isolated or recombinant. “Isolated,” when used to describe the various polypeptides disclosed herein, refers to a polypeptide that has been identified and separated and / or recovered from a cell or cell culture from which it was expressed. Ordinarily, an isolated polypeptide will be prepared by at least one purification step. An “isolated antibody,” refers to an antibody which is substantially free of other antibodies havingdifferent antigenic specificities. As used herein, “recombinant antibody” refers to an antibody that is generated using recombinant nucleic acid techniques in exogenous host cells, and recombinant antibodies can be isolated as well.

[0128] “Native antibodies” are usually heterotetrameric glycoproteins of about 150,000 Daltons, composed of two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide linkages varies among the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has at one end a variable domain (VH) followed by a number of constant domains. Each light chain has a variable domain at one end (VL) and a constant domain at its other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain. Particular amino acid residues are believed to form an interface between the light chain and heavy chain variable domains.

[0129] The term “constant domain” refers to the portion of an immunoglobulin molecule having a more conserved amino acid sequence relative to the other portion of the immunoglobulin, the variable domain, which contains the antigen-binding site. The constant domain contains the CHI, CH2and CH3 domains (collectively, CH) of the heavy chain and the CHL (or CL) domain of the light chain.

[0130] The “variable region” or “variable domain” of an antibody refers to the amino-terminal domains of the heavy or light chain of the antibody. The variable domain of the heavy chain may be referred to as “VH.” The variable domain of the light chain may be referred to as “VL.” These domains are generally the most variable parts of an antibody and contain the antigen-binding sites. The term “variable” refers to the fact that certain portions of the variable domains differ extensively in sequence among antibodies and are used in the binding and specificity of each particular antibody for its particular antigen. However, the variability is not evenly distributed throughout the variable domains of antibodies. It is concentrated in three segments called hypervariable regions (HVRs) or complementary determining regions (CDRs), both in the light- chain and the heavy-chain variable domains, that confer antigen specificity. A “variable heavy domain” pairs with a “variable light domain” to form an antigen-binding domain (ABD) that specifically binds a target antigen. The more highly conserved portions of variable domains arecalled the framework regions (FR). The variable domains of native heavy and light chains each comprise four FR regions, largely adopting a beta-sheet configuration, connected by three CDRs / HVRs, which form loops connecting, and in some cases forming part of, the beta-sheet structure. The CDRs / HVRs in each chain are held together in close proximity by the FR regions and, with the CDRs / HVRs from the other chain, contribute to the formation of the antigen-binding site of antibodies (see Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, Md. (1991)). The constant domains are not involved directly in the binding of an antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody-dependent cellular toxicity.

[0131] The terms “hypervariable region,” “HVR,” “HV,” “complementary determining region,” and “CDR,” used interchangeably herein, refer to the regions of an antibody variable domain which are hypervariable in sequence and / or form structurally defined loops. Generally, antibodies comprise six HVRs or CDRs; three in the VH (Hl, H2, H3; or VH CDR1, VH CDR2, VH CDR3), and three in the VL (LI, L2, L3; or VL CDR1, VL CDR2, VL CDR3).

[0132] The “light chains” of antibodies (immunoglobulins) from any mammalian species can be assigned to one of two clearly distinct types, called kappa (“K”) and lambda (“ ”), based on the amino acid sequences of their constant domains.

[0133] Together, the CDRs of the VH and VL domains form an Fv region. In embodiments, a VH and a VL domain comprise the six CDRs of the ABD. In a “Fab” format, the variable heavy domain (VH; containing VH CDR1, VH CDR2, and VH CDR3) and the variable light domain (VL or VL; containing the VL CDR1, VL CDR2 and VL CDR3), comprise the set of 6 CDRs, with the C-terminus of the VH domain being attached to the N-terminus of the CHI domain of the heavy chain and the C-terminus of the VL domain being attached to the N-terminus of the constant light domain (and thus forming the light chain). In an “scFv” format, the VH and VL domains are covalently attached, generally through the use of a linker (e.g., an “scFv linker”), into a single polypeptide sequence, which can have the N- to C-terminus arrangement of VH-linker-VL or VL- linker-VH. In general, the C-terminus of the scFv domain is attached to the N-terminus of the hinge in the second monomer.

[0134] ‘Fab” or “Fab region,” as used herein, refers to a polypeptide that comprises VH, CHI , VL, and CL immunoglobulin domains, generally on two different polypeptide chains (e.g., VH-CHI on one chain and VL-CL on the other). Fab can refer to this region in isolation, or this region in the context of an antibody of the disclosure. In embodiments, a Fab comprises an Fv region in addition to CHI CL domains.

[0135] Another part of the heavy chain is the hinge region. As used herein, “hinge,” “hinge region,” “antibody hinge region,” or “hinge domain” refers to the flexible polypeptide comprising the amino acids between the first and second constant domains of an antibody. Structurally, the IgG CHl domain ends at EU position 215, and the IgG CH2domain begins at residue EU position 231. Thus, for IgG, the antibody hinge is herein defined to include positions 216 (E216 in IgGl) to 230 (p230 in IgGl), wherein the numbering is according to the EU index as in Kabat. In some cases, a “hinge fragment” is used, which contains fewer amino acids at either or both of the N- and C-termini of the hinge domain.

[0136] “Heavy chain constant region,” as used herein, refers to the CHl-hinge-CH2-CH3 portion of an antibody or fragment thereof, excluding the variable heavy domain. In embodiments, the heavy chain constant region comprises amino acids 118-447 of human IgGl, in EU numbering. As used herein, “heavy chain constant region fragment” refers to a heavy chain constant region that contains fewer amino acids from either or both of the N- and C-termini but still retains the ability to form a dimer with another heavy chain constant region.

[0137] “Fv,” “Fv fragment,” or “Fv region,” as used herein, refers to a polypeptide that comprises VL and VH domains of an antibody binding domain. Fv regions can be formatted as both Fabs and scFvs, where the VL and VH domains are combined (e g., by way of a linker, as discussed herein) to form an scFv.

[0138] “Fc,” “Fc region,” or “Fc domain,” as used herein, refers to a polypeptide comprising CH2 -CH3 domains of an IgG molecule, and, in some cases, inclusive of the hinge. In EU numbering for human IgGl, the CH2-CH3 domain comprises amino acids 231 to 447, and the hinge is 216 to 230. Thus, the definition of “Fc domain” includes both amino acids 231-447 (CH2- CH3) and 216-447 (hinge-CH2-CH3) of IgGl, or fragments thereof. An “Fc fragment” in this context can contain fewer amino acids from either or both of the N- and C-termini but still retains the ability to form a dimer with another Fc domain or Fc fragment as can be detected using standard methods, generally based on size (e g., non-denaturing chromatography, size exclusionchromatography, etc.). In embodiments, the disclosed AOCs comprise human Fc domains. In embodiments, the disclosed AOCs comprise Fc domains from human IgGl, IgG2, or IgG4.

[0139] A “variant Fc domain” contains amino acid modifications as compared to a parental Fc domain. Thus, a “variant human IgGl Fc domain” is one that contains amino acid modifications (generally amino acid substitutions, although in the case of ablation variants, amino acid deletions are included) as compared to the human IgGl Fc domain. In embodiments, variant Fc domains have at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97%, at least about 98%, or at least about 99% identity to the corresponding parental human IgG Fc domain. In embodiments, the percent identity is calculated using the identity algorithms discussed below. In embodiments, the percent identity is calculated using the BLAST algorithm known in the art, using default parameters. In embodiments, variant Fc domains have from 1 to about 20 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20) amino acid modifications as compared to the parental Fc domain. In embodiments, variant Fc domains retain the ability to form a dimer with Ir Fc domain as measured using known techniques as described herein, such as non-denaturing gel electrophoresis.

[0140] For all positions discussed in the present disclosure that relate to antibodies, unless otherwise noted, amino acid position numbering is according to the EU index. The EU index or EU index as in Kabat or EU numbering scheme refers to the numbering of the EU antibody. Kabat et al. collected numerous primary sequences of the variable regions of heavy chains and light chains. Based on the degree of conservation of the sequences, they classified individual primary sequences into the CDR and the framework and made a list thereof. See, SEQUENCES OF IMMUNOLOGICAL INTEREST, 5thedition, NIH publication, No. 91-3242, E.A. Kabat et al.; Edelman et al., 1969, Proc Natl Acad Sci USA 63:78-85, the contents of which are incorporated herein by reference. In embodiments of the present disclosure, amino acid position numbering is according to the IMGT system.

[0141] The terms “full length antibody,” “intact antibody” and “whole antibody” are used herein interchangeably to refer to an antibody in its substantially intact form, not antibody fragments as defined below. The terms particularly refer to an antibody with heavy chains that contain an Fc region.

[0142] An “antibody fragment” comprises a portion of an intact antibody, preferably comprising the antigen-binding region thereof. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.

[0143] A “naked antibody” for the purposes herein is an antibody that is not conjugated to a payload, e.g., an oligonucleotide, cytotoxic moiety, or radiolabel.

[0144] The term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, e g., the individual antibodies comprising the population are identical except for possible mutations, e.g., naturally occurring mutations, that can be present in minor amounts. Thus, the modifier “monoclonal” indicates the character of the antibody as not being a mixture of discrete antibodies. In certain embodiments, such a monoclonal antibody typically includes an antibody comprising a polypeptide sequence that binds a target, wherein the target-binding polypeptide sequence was obtained by a process that includes the selection of a single target binding polypeptide sequence from a plurality of polypeptide sequences. For example, the selection process can be the selection of a unique clone from a plurality of clones, such as a pool of hybridoma clones, phage clones, or recombinant DNA clones. It should be understood that a selected target binding sequence can be further altered, for example, to improve affinity for the target, to humanize the target binding sequence, to improve its production in cell culture, to reduce its immunogenicity in vivo, to create a multispecific antibody, etc., and that an antibody comprising the altered target binding sequence is also a monoclonal antibody of this disclosure. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen. In addition to their specificity, monoclonal antibody preparations are advantageous in that they are typically uncontaminated by other immunoglobulins.

[0145] Antibodies herein specifically include “chimeric” antibodies in which a portion of the heavy and / or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well asfragments of such antibodies, so long as they exhibit one or more of the desired biological activities (see, e.g., U.S. Pat. No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA 81:6851-6855 (1984)). Typically, the variable region of both light and heavy chains corresponds to the variable region of antibodies derived from one species of mammals (e.g., mouse, rat, rabbit, etc.) with the desired specificity, affinity, and / or capability, while the constant regions are homologous to the sequences of antibodies derived from another species of mammals (e.g., human) to avoid eliciting an immune response In that species. Chimeric antibodies include PRIMATTZED® antibodies wherein the antigen-binding region of the antibody is derived from an antibody produced by, e.g., immunizing macaque monkeys with the antigen of interest.

[0146] ‘Humanized” forms of non-human (e.g., murine) antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin. In embodiments, a humanized antibody is a human immunoglobulin (recipient antibody) in which residues from a CDR / HVR of the recipient are replaced by residues from a CDR / HVR of a non-human species (donor antibody) such as mouse, rat, rabbit, or nonhuman primate having the desired specificity, affinity, and / or capacity. In some instances, FR residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies can comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications can be made to further refine antibody performance. In general, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin, and all or substantially all of the FRs are those of a human immunoglobulin sequence. The humanized antibody optionally will also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. See, e.g., Jones et al., Nature 321 :522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593- 596 (1992). See also, e.g., Vaswani and Hamilton, Ann. Allergy, Asthma & Immunol. 1: 105-115 (1998); Harris, Biochem. Soc. Transactions 23: 1035-1038 (1995); Hurle and Gross, Curr. Op. Biotech. 5:428-433 (1994); and U.S. Pat. Nos. 6,982,321 and 7,087,409. Examples of methods used to generate humanized antibodies are described in U.S. Pat. 5,225,539 or 5,639,641, incorporated herein by reference in their entireties.

[0147] As used herein, the term “human antibody” refers to an antibody which possesses an amino acid sequence which corresponds to that of an antibody produced by a human and / or hasbeen made using any technique known in the art. This definition of a human antibody includes intact or full-length antibodies, fragments thereof, and / or antibodies comprising at least one human heavy and / or light chain polypeptide. This definition of a human antibody specifically excludes a humanized antibody comprising non-human antigen-binding residues. Human antibodies can be produced using various techniques known in the art, including phage-display libraries. Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991). Also available for the preparation of human monoclonal antibodies are methods described in Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boemer et al., J. Immunol., 147(l):86-95 (1991). See also van Dijk and van de Winkel, Curr. Opin. Pharmacol., 5: 368-74 (2001). Human antibodies can be prepared by administering the antigen to a transgenic animal that has been modified to produce such antibodies in response to antigenic challenge, but whose endogenous loci have been disabled, e.g., immunized xenomice (see, e.g., U.S. Pat. Nos. 6,075,181 and 6,150,584 regarding XENOMOUSE™ technology). See also, for example, Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006) regarding human antibodies generated via a human B-cell hybridoma technology.

[0148] A “species-dependent antibody” is one which has a stronger binding affinity for an antigen from a first mammalian species than it has for a homologue of that antigen from a second mammalian species. Normally, the species-dependent antibody “binds specifically” to a human antigen (e.g., has a binding affinity (Kd) value of no more than about 1 X 10-7M, preferably no more than about U I0xM and preferably no more than about 1 x 109M) but has a binding affinity for a homologue of the antigen from a second nonhuman mammalian species which is at least about 50 fold, or at least about 500 fold, or at least about 1000 fold, weaker than its binding affinity for the human antigen. The species-dependent antibody can be any of the various types of antibodies as defined above, but preferably is a humanized or human antibody.

[0149] The expression “linear antibodies” refers to the antibodies described in Zapata et al. (1995 Protein Eng, 8(10): 1057-1062). Briefly, these antibodies comprise a pair of tandem Fd segments (VH-CH1-VH-CH1) which, together with complementary light chain polypeptides, form a pair of antigen-binding regions. Linear antibodies can be bispecific or monospecific.

[0150] “Modification,” as used herein, refers to an amino acid substitution, insertion, deletion, and / or any other mutation in a polypeptide sequence.

[0151] ‘Variant protein,” or “protein variant,” or “variant,” as used herein refers to a protein that differs from that of a parent protein by virtue of at least one amino acid modification. The protein variant has at least one amino acid modification compared to the parent protein, yet not so many that the variant protein will not align with the parental protein using an alignment program such as that described below. In general, variant proteins (such as variant Fc domains, etc., described herein, are generally at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% identical to the parent polypeptide, using any alignment program known in the art, such as BLAST.

[0152] Sequence identity between two similar sequences (e.g., antibody variable domains) can be measured by algorithms such as that of Smith, T.F. & Waterman, M.S. (1981) “Comparison Of Biosequences,” Adv. Appl. Math. 2:482 [local homology algorithm]; Needleman, S.B. & Wunsch, CD. (1970) “A General Method Applicable To The Search For Similarities In The Amino Acid Sequence Of Two Proteins,” J. Mol. Biol.48:443 [homology alignment algorithm], Pearson, W.R. & Lipman, D.J. (1988) “Improved Tools For Biological Sequence Comparison,” Proc. Natl. Acad. Sci. (U.S.A.) 85:2444 [search for similarity method]; or Altschul, S.F. et al, (1990) “Basic Local Alignment Search Tool,” J. Mol. Biol. 215:403-10 , the “BLAST” algorithm, see the webpage located at URL blast.ncbi.nlm.nih.gov / Blast.cgi. When using any of the aforementioned algorithms, the default parameters (for Window length, gap penalty, etc.) are used. Unless specifically stated otherwise, sequence identity is determined using the BLAST algorithm, using default parameters.

[0153] In embodiments, a parent polypeptide, for example an Fc parent polypeptide, is a human wild type sequence, such as the heavy constant domain or Fc region from IgGl, IgG2, IgG3 or IgG4, although human sequences with variants can also serve as “parent polypeptides.” In embodiments, antibody sequences described herein have at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%,at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity with a parent polypeptide sequence. Accordingly, “antibody variant” or “variant antibody” as used herein refers to an antibody that differs from a parent antibody by virtue of at least one amino acid modification; “IgG variant” or “variant IgG” as used herein refers to an IgG that differs from a parent IgG (e.g., from a human IgG sequence) by virtue of at least one amino acid modification; “immunoglobulin variant” or “variant immunoglobulin” as used herein refers to an immunoglobulin sequence that differs from that of a parent immunoglobulin sequence by virtue of at least one amino acid modification; and “Fc variant” or “variant Fc” as used herein refers to an Fc that differs from a parent Fc, e.g., an Fc domain of human IgGl, IgG2, IgG3, or IgG4, by virtue of at least one amino acid modification.

[0154] “IgG subclass modification” or “isotype modification,” as used herein, refers to amino acid modifications that convert one amino acid of one IgG isotype to the corresponding amino acid in a different, aligned IgG isotype. For example, because IgGl comprises a tyrosine and IgG2 a phenylalanine at EU position 296, a F296Y substitution in IgG2 is considered an IgG subclass modification.

[0155] ‘Non-naturally occurring modification” as used herein is meant an amino acid modification that is not isotypic. For example, because none of the human IgGs comprise a serine at position 434, the substitution 434S in IgGl, IgG2, IgG3, or IgG4 (or hybrids thereof) is considered a non-naturally occurring modification.

[0156] As used herein, “oligonucleotide” or “polynucleotide,” used interchangeably, refers to a linear polymer of natural or modified nucleoside monomers linked by phosphodiester bonds or analogs thereof. The term “oligonucleotide” usually refers to a shorter polymer, e.g., comprising from about 3 to about 100 monomers, and the term “polynucleotide” usually refers to longer polymers, e.g., comprising from about 100 monomers to many thousands of monomers, e.g., 10,000 monomers, or more. Oligonucleotides and polynucleotides can be natural or synthetic. Oligonucleotides and polynucleotides can include deoxyribonucleosides, ribonucleosides, and / or non-natural analogs thereof. In embodiments, oligonucleotides or polynucleotides are capable of specifically binding to a target genome by way of a regular pattern of monomer-to-monomer interactions, such as Watson-Crick type of base pairing, base stacking, Hoogsteen or reverseHoogsteen types of base pairing, or the like. As used herein, “functional nucleic acid” refers to a nucleic acid having biological functions in vivo or in cells, such as enzymatic functions, catalytic functions, or biologically inhibiting or enhancing functions (e.g., inhibition or enhancement of transcription or translation). In embodiments, examples of functional nucleic acids include, but are not limited to, siRNA, ASO, shRNA, miRNA (including pri-miRNA and pre-miRNA), nucleic acid aptamers (including RNA aptamers and DNA aptamers), ribozymes (including deoxyribozymes), riboswitches, U1 adaptors, molecular beacons, and transcriptional factor- binding regions.

[0157] In certain instances, the term “oligonucleotide” is used in reference to an “antisense oligonucleotide.” For “antisense oligonucleotides,” each subunit consists of: (i) a ribose sugar or a derivative thereof; and (ii) a nucleobase bound thereto, such that the order of the base-pairing moieties forms a base sequence that is complementary to a target sequence in a nucleic acid (typically an RNA) by Watson-Crick base pairing, to form a nucleic acid:oligomer heteroduplex within the target sequence with the proviso that either the subunit, the intersubunit linkage, or both are not naturally occurring. In certain embodiments, the antisense oligonucleotide is a phosphorodiamidate morpholino oligomer (PMO). In other embodiments, the antisense oligonucleotide is a 2'-O-methyl phosphorothioate (2’OMe-PS). In other embodiments, the antisense oligonucleotide is a 2’-fluoro phosphorothioate (2’F-PS). In other embodiments, the antisense oligomer of the disclosure is a peptide nucleic acid (PNA), a locked nucleic acid (LNA), or a bridged nucleic acid (BNA) such as 2'-O,4'-C-ethylene-bridged nucleic acid (ENA).

[0158] Morpholinos as described herein include all stereoisomers and tautomers of the foregoing general structure. The synthesis, structures, and binding characteristics of morpholino oligomers are detailed in U.S. Pat. Nos. 5,698,685; 5,217,866; 5,142,047; 5,034,506; 5,166,315; 5,521,063; 5,506,337; 8,076,476; and 8,299,206; all of which are incorporated herein by reference.

[0159] The terms “complementary” and “complementarity” refer to two or more oligomers (i.e., each comprising a nucleobase sequence) that are related with one another by Watson-Crick base-pairing rules. For example, the nucleobase sequence “T-G-A (5'— >3'),” is complementary to the nucleobase sequence “A-C-T (3'— >5').” Complementarity may be “partial,” in which less than all of the nucleobases of a given nucleobase sequence are matched to the other nucleobase sequence according to base pairing rules. For example, in some embodiments, complementaritybetween a given nucleobase sequence and the other nucleobase sequence may be about 70%, about 75%, about 80%, about 85%, about 90% or about 95%. Or, there may be “complete” or “perfect” (100%) complementarity between a given nucleobase sequence and the other nucleobase sequence to continue the example. The degree of complementarity between nucleobase sequences has significant effects on the efficiency and strength of hybridization between the sequences.

[0160] The terms “nucleobase” (Nu), “base pairing moiety” or “base” are used interchangeably to refer to a purine or pyrimidine base found in naturally occurring, or “native” DNA or RNA (e.g., uracil, thymine, adenine, cytosine, and guanine), as well as analogs of these naturally occurring purines and pyrimidines. These analogs may confer improved properties, such as binding affinity, to the oligomer. Exemplary analogs include hypoxanthine (the base component of inosine); 2,6-diaminopurine; 5-methyl cytosine; C5-propynyl-modified pyrimidines; 10-(9- (aminoethoxy)phenoxazinyl) (G-clamp) and the like.

[0161] The terms “mismatch” or “mismatches” refer to one or more nucleobases (whether contiguous or separate) in an oligomer nucleobase sequence that are not matched to a target pre- mRNA according to base pairing rules. While perfect complementarity is often desired, some embodiments can include one or more but preferably 6, 5, 4, 3, 2, or 1 mismatches with respect to the target pre-mRNA. Variations at any location within the oligomer are included. In certain embodiments, antisense oligomer conjugates of the disclosure include variations in nucleobase sequence near the term variations in the interior, and if present are typically within about 6, 5, 4, 3, 2, or 1 subunits of the 5' and / or 3' terminus.

[0162] As used herein, a “3E10 antibody” refers to an antibody with a set of heavy chain CDRs (VH CDR1, VH CDR2, and VH CDR3), identified according to the Kabat system, comprising amino acid sequences that vary from SEQ ID NOS: 58, 59, and 60 by no more than two amino acids each, respectively, a set of light chain CDRs (VL CDR1, VL CDR2, and VL CRD3) comprising amino acid sequences that vary from SEQ ID NOS: 61, 62, and 63 by no more than two amino acids each, respectively, that binds nucleic acids and is cell-penetrating at least when bound to a nucleic acid, as well as antigen-binding fragments thereof. As described herein, the 3E10 antigen is a polynucleotide. Although generally referred to herein as “3E10” or “3E10 antibodies,” it will be appreciated that fragments, variants, and binding proteins, including antigen- binding fragments and fusion proteins, such as scFv, di-scFv, tr-scFv, and other single chainvariable fragments, and other cell-penetrating, nucleic acid transporting molecules disclosed herein, are encompassed by the phrase and are also expressly provided for use in compositions, conjugates, and methods disclosed herein. Thus, the antibodies and other binding proteins are also referred to herein as cell-penetrating.

[0163] As used herein, the term “cell-penetrating” refers to an antibody or antigen binding fragment thereof that can penetrate a cell, e.g., a mammalian cell, without the aid of an exogeneous transport vehicle, such as a liposome, or a conjugated cell-penetrating peptide. With respect to 3E10 antibodies and antigen binding fragments thereof, the cell-penetrating antibody or antigen binding fragment thereof can penetrate a cell expressing an ENT2 receptor on its cell surface in the presence of nucleic acids, e.g., non-covalently bound and / or conjugated to the 3E10 antibody or antigen binding fragment thereof, resulting in internalization of the 3E10 antibodies and antigen binding fragments thereof. In some embodiments, the cell-penetrating 3E10 antibody or antigen binding fragment thereof is conjugated to a functional molecule, e.g., a chemical agent, polynucleotide, or polypeptide. Although the cell-penetrating molecules are generally referred to herein as “cell-penetrating antibodies,” it will be appreciated that fragments, including antigen-binding fragments, variants, binding proteins and fusion proteins such as scFv, di- scFv, tri-scFv, and other single chain variable fragments, and other cell-penetrating molecules disclosed herein are also expressly provided foruse in compositions, conjugates, and methods disclosed herein. Autoantibodies against double-stranded deoxyribonucleic acid (dsDNA) are frequently identified in the serum of patients with systemic lupus erythematosus (SLE) and are often implicated in disease pathogenesis. Therefore, in embodiments, cell-penetrating antibodies (e.g., cell-penetrating anti-DNA antibodies) can be derived or isolated from patients with SLE or animal models of SLE.

[0164] As used herein, “antibody-oligonucleotide conjugate” or “AOC” refers to an antibody or antigen-binding fragment thereof that is covalently linked or conjugated to a biologically active molecule, for example an oligonucleotide or anti-tumor oligonucleotide, for example, an siRNA molecule, an antisense oligonucleotide.

[0165] As used herein, a “linker” is any chemical moiety that is capable of linking or connecting a molecule, including an oligonucleotide, to a cell-binding agent such as an antibody, such as a 3E10 antibody or a fragment thereof, in a stable, covalent manner. In embodiments, a“linker” is any chemical moiety that is capable of linking or connecting a compound such as an oligonucleotide, a polynucleotide, a DNA damage-inducing agent, a DNA repair inhibitor, an immune modulatory molecule, an alkylating agent, a microtubule inhibitor, an immune checkpoint inhibitor, an angiogenesis inhibitor, an adoptive cell therapy, or a topoisomerase inhibitor, to a cell-binding agent such as a 3E10 antibody or a fragment thereof, in a stable, covalent manner. Linkers can be susceptible to or be substantially resistant to acid-induced cleavage, light-induced cleavage, peptidase-induced cleavage, esterase-induced cleavage, and / or disulfide bond cleavage, at conditions under which the compound and / or the antibody remains active. Suitable linkers are well known in the art and include, for example, disulfide groups, thioether groups, acid labile groups, photolabile groups, peptidase labile groups and esterase labile groups. Linkers also include charged linkers, and hydrophilic forms thereof as described herein and know in the art.

[0166] As used herein, the term “subject” means any individual who is the target of administration. The subject can be any animal (e.g., a mammal. Thus), including, but not limited to, humans, and non-human animals (including, but not limited to, non-human primates, dogs, cats, rodents, horses, cows, pigs, mice, rats, hamsters, rabbits, and the like (e.g., which is to be the recipient of a particular treatment). In embodiments, the subject is a human. In embodiments, methods of the disclosure are useful in treatment a human subject. In embodiments, the human may be referred to as a patient. In embodiments, the human is a female. In embodiments, the human is a male. In embodiments, the human has an age in a range of from about 1 to about 18 months old, from about 18 to about 36 months old, from about 1 to about 5 years old, from about 5 to about 10 years old, from about 10 to about 15 years old, from about 15 to about 20 years old, from about 20 to about 25 years old, from about 25 to about 30 years old, from about 30 to about 35 years old, from about 35 to about 40 years old, from about 40 to about 45 years old, from about 45 to about 50 years old, from about 50 to about 55 years old, from about 55 to about 60 years old, from about 60 to about 65 years old, from about 65 to about 70 years old, from about 70 to about 75 years old, from about 75 to about 80 years old, from about 80 to about 85 years old, from about 85 to about 90 years old, from about 90 to about 95 years old or from about 95 to about 100 years old.

[0167] The terms “subject” and “patient” as used herein include any animal that exhibits a symptom, or is at risk for exhibiting a symptom, which can be treated with an antisense oligomer conjugate of the disclosure, such as a subject (or patient) that has or is at risk for having DMD orBMD, or any of the symptoms associated with these conditions (e.g., muscle fiber loss). Also included are methods of producing dystrophin in a subject (or patient) having a mutation of the dystrophin gene that is amenable to exon 23 skipping.

[0168] The terms “cancer” and “cancerous” refer to or describe the physiological condition in mammals in which a population of cells are characterized by unregulated cell growth. Examples of cancer include, but are not limited to, colorectal cancer, pancreatic cancer, lung cancer, ovarian cancer, liver cancer, breast cancer, brain cancer, kidney cancer, prostate cancer, gastrointestinal cancer, melanoma, cervical cancer, bladder cancer, glioblastoma, head and neck cancer, lymphomas, Hodgkin lymphoma, Non-Hodgkin lymphoma, cutaneous B-cell lymphoma, cutaneous T-cell lymphoma, Waldenstrom macroglobulinemia, chronic lymphocytic leukemia, leukemia, neuroblastoma, Wilms tumor, bone cancer, brain stem tumor, childhood diffuse intrinsic pontine glioma (DIPG), retinoblastoma, rhabdoid tumor, sarcoma, spinal cord tumor, endocrine cancer, esophageal cancer, gastric cancer, hepatobiliary cancer, myeloma, renal cancer, thyroid cancer, uterine cancer, carcinoma, blastoma, papilloma, adenoma, an astrocytic tumor, an oligodendroglial tumor, an oligoastrocytic tumor, an ependymal tumor, a choroid plexus tumor, a neuronal or mixed neuronal-glial tumor, tumor of the pineal region, embryonal tumor, or an otherwise uncategorized neuroepithelial tumors.

[0169] “Tumor” and “neoplasm” refer to any mass of tissue that results from excessive cell growth or proliferation, either benign (noncancerous) or malignant (cancerous), including pre- cancerous lesions.

[0170] The terms “cancer cell,” “tumor cell,” and grammatical equivalents thereof refer to the total population of cells derived from a tumor or a pre-cancerous lesion, including both non- tumorigenic cells, which comprise the bulk of the tumor cell population, and tumorigenic stem cells (cancer stem cells).

[0171] As used herein, the term “pharmaceutically effective amount” means that the amount of the composition used is of sufficient quantity to ameliorate one or more causes or symptoms of a disease or disorder. Such amelioration only requires a reduction or alteration, not necessarily elimination. The precise dosage will vary according to a variety of factors such as subject-dependent variables (e.g., age, immune system health, etc.), the disease or disorder beingtreated, as well as the route of administration and the pharmacokinetics of the agent being administered.

[0172] As used herein, the term “carrier” or “excipient” refers to an organic or inorganic ingredient, natural or synthetic inactive ingredient in a formulation, with which one or more active ingredients are combined. In embodiments, the carrier or excipient is selected to minimize degradation of the active ingredient or to minimize adverse side effects in the subject, as would be well known to one of skill in the art.

[0173] As used herein, the term “treat” refers to the medical management of a patient with the intent to cure, ameliorate, stabilize, or prevent a disease, pathological condition, or disorder. This term includes active treatment, that is, treatment directed specifically toward the improvement of a disease, pathological condition, or disorder, and also includes causal treatment, that is, treatment directed toward removal of the cause of the associated disease, pathological condition, or disorder. In addition, this term includes palliative treatment, that is, treatment designed for the relief of symptoms rather than the curing of the disease, pathological condition, or disorder; preventative treatment, that is, treatment directed to minimizing or partially or completely inhibiting the development of the associated disease, pathological condition, or disorder; and supportive treatment, that is, treatment employed to supplement another specific therapy directed toward the improvement of the associated disease, pathological condition, or disorder.

[0174] As used herein “antibody-oligonucleotide conjugate” or “AOC” refers to an antibody or antigen-binding fragment thereof that is conjugated via a linker to a therapeutic oligonucleotide, for example, an oligonucleotide, an siRNA, or an antisense oligonucleotide (ASO), which may be delivered to specific cells or tissues otherwise not targetable by oligonucleotide delivery. The conjugation of an oligonucleotide with an antibody or antigen- binding fragment thereof may also improve the pharmacokinetic properties of therapeutic oligonucleotides, expanding application of this therapeutic modality.

[0175] A “pharmacologically effective amount,” “pharmacologically effective dose,” “therapeutically effective amount,” or “effective amount” refers to an amount sufficient to produce a desired physiological effect or amount capable of achieving a desired result, particularly for treating or preventing the disorder or disease. An effective amount as used herein would include an amount sufficient to, for example, delay the development of a symptom of the disorder ordisease, alter the course of a symptom of the disorder or disease (e.g., slow the progression of a symptom of the disease), reduce or eliminate one or more symptoms or manifestations of the disorder or disease, and reverse a symptom of a disorder or disease. Therapeutic benefit also includes halting or slowing the progression of the underlying disease or disorder, regardless of whether improvement is realized.

[0176] The terms “effective amount” and “therapeutically effective amount” are used interchangeably herein and refer to an amount of therapeutic compound, such as an antisense oligomer, administered to a mammalian subject, either as a single dose or as part of a series of doses, which is effective to produce a desired therapeutic effect. For an antisense oligomer, this effect is typically brought about by inhibiting translation or natural splice-processing of a selected target sequence, or producing a clinically meaningful amount of dystrophin.

[0177] Effective amounts, toxicity, and therapeutic efficacy can be determined by standard pharmaceutical procedures in cell cultures, tissue samples, tissue homogenates or experimental animals, e.g., for determining the LD50 (the dose lethal to about 50% of the population) and the ED50 (the dose therapeutically effective in about 50% of the population) or the maximum tolerated dose. The dosage can vary depending upon the dosage form employed and the route of administration utilized. The dose ratio between toxic and therapeutic effects is the therapeutic index and can be expressed as the ratio LD50 / ED50. In embodiments, compositions, conjugates, and methods that exhibit large therapeutic indices are preferred. A therapeutically effective dose can be estimated initially from in vitro assays, including, for example, cell culture assays or measurements. Also, a dose can be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 as determined in cell culture, or in an appropriate animal model. Levels of the described compositions in plasma can be measured, for example, by high performance liquid chromatography. The effects of any particular dosage can be monitored by a suitable bioassay. The dosage can be determined by a physician and adjusted, as necessary, to suit observed effects of the treatment.

[0178] In embodiments, the effect will result in a quantifiable change of at least about 10%, at least about 20%, at least about 30%, at least about 50%, at least about 70%, or at least about 90%. In embodiments, the effect will result in a quantifiable change of about 10%, about 20%, about 30%, about 50%, about 70%, or even about 90% or more. Therapeutic benefit also includeshalting or slowing the progression of the underlying disease or disorder, regardless of whether improvement is realized.

[0179] By “enhance” or “enhancing,” or “increase” or “increasing,” or “stimulate” or “stimulating,” refers generally to the ability of one or more antisense oligomer conjugates or pharmaceutical compositions to produce or cause a greater physiological response (i.e., downstream effects) in a cell or a subject, as compared to the response caused by either no antisense oligomer conjugate or a control compound. A greater physiological response may include increased expression of a functional form of a dystrophin protein, or increased dystrophin- related biological activity in muscle tissue, among other responses apparent from the understanding in the art and the description herein. Increased muscle function can also be measured, including increases or improvements in muscle function by about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. The percentage of muscle fibers that express a functional dystrophin can also be measured, including increased dystrophin expression in about 1%, 2%, 5%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of muscle fibers. For instance, it has been shown that around 40% of muscle function improvement can occur if 25-30% of fibers express dystrophin (see, e.g., DelloRusso et al, Proc Natl Acad Sci USA 99: 12979-12984, 2002). An “increased” or “enhanced” amount is typically a “statistically significant” amount, and may include an increase that is 1.1, 1.2, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50 or more times (e g., 500, 1000 times, including all integers and decimal points in between and above 1, e.g., 1.5, 1.6, 1.7, 1.8, etc.) the amount produced by no antisense oligomer conjugate (the absence of an agent) or a control compound.

[0180] As used herein, the terms “function” and “functional” and the like refer to a biological, enzymatic, or therapeutic function.

[0181] A “functional” dystrophin protein refers generally to a dystrophin protein having sufficient biological activity to reduce the progressive degradation of muscle tissue that is otherwise characteristic of muscular dystrophy, typically as compared to the altered or “defective” form of dystrophin protein that is present in certain subjects with Duchenne muscular dystrophy (DMD) or Becker muscular dystrophy (BMD). In certain embodiments, a functional dystrophinprotein may have about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% (including all integers in between) of the in vitro or in vivo biological activity of wild-type dystrophin, as measured according to routine techniques in the art. As one example, dystrophin-related activity in muscle cultures in vitro can be measured according to myotube size, myofibril organization (or disorganization), contractile activity, and spontaneous clustering of acetylcholine receptors (see, e.g., Brown et al., Journal of Cell Science. 112:209-216, 1999). Animal models are also valuable resources for studying the pathogenesis of disease, and provide a means to test dystrophin-related activity. Two of the most widely used animal models for DMD research are the mdx mouse and the golden retriever muscular dystrophy (GRMD) dog, both of which are dystrophin negative (see, e.g., Collins & Morgan, Int J Exp Pathol 84: 165-172, 2003). These and other animal models can be used to measure the functional activity of various dystrophin proteins. Included are truncated forms of dystrophin, such as those forms that are produced following the administration of certain of the exon-skipping antisense oligonucleotides of the present disclosure.

[0182]

[0183] The phrases “parenteral administration” and “administered parenterally” as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.

[0184] The phrase “targeting sequence” refers to a sequence of nucleobases of an oligomer that is complementary to a sequence of nucleotides in a target pre-mRNA. For example, in some embodiments of the disclosure, the sequence of nucleotides in the target pre-mRNA is an exon 23 annealing site in the dystrophin pre-mRNA.

[0185] As used herein, the term “treatment” of a subject (e.g., a mammal, such as a human) or a cell is any type of intervention used in an attempt to alter the natural course of the subject or cell. Treatment includes, but is not limited to, administration of an oligomer or a pharmaceutical composition thereof, and may be performed either prophylactically or subsequent to the initiation of a pathologic event or contact with an etiologic agent. Treatment includes any desirable effect on the symptoms or pathology of a disease or condition associated with the dystrophin protein, asin certain forms of muscular dystrophy, and may include, for example, minimal changes or improvements in one or more measurable markers of the disease or condition being treated. Also included are “prophylactic” treatments, which can be directed to reducing the rate of progression of the disease or condition being treated, delaying the onset of that disease or condition, or reducing the severity of its onset. “Treatment” or “prophylaxis” does not necessarily indicate complete eradication, cure, or prevention of the disease or condition, or associated symptoms thereof.Ill, Antibodv-Oligonucleotide Conjugates (AOCs)

[0186] The present disclosure is directed, in part, to antibody-oligonucleotide conjugates (AOCs) comprising a cell-penetrating antibody, e.g., a 3E10 antibody or antigen-binding fragment thereof, conjugated via a linker to an oligonucleotide, e.g., a therapeutic oligonucleotide. In embodiments, an AOC described herein has the formula A-(L-Pr)q, wherein: A is a 3E10 antibody or antigen-binding fragment thereof, L is a linker, and P is an oligonucleotide moiety as described herein.

[0187] In some aspects, the present disclosure relates to the use of 3E10 antibodies, and derivatives thereof, for delivering antisense oligonucleotides amendable for exon skipping in tissues of a subject, including but not limited to skeletal muscle tissues for treatment of genetic skeletal muscle disorders. As is discussed below, the term antibody is used generally. Antibodies that find use in the present disclosure take on a number of formats as described herein, including traditional antibodies as well as antibody derivatives, fragments, and mimetics, described herein in various embodiments.

[0188] In some aspects, the antibody is conjugated to the biologically active molecule via a linker. In some aspects, the antibody is a 3E10 antibody or antigen-binding fragment thereof, as described herein. In some aspects, the antibody is a humanized 3E10 antibody or antigen-binding fragment thereof, as described herein. Any variety of agents can be transported via conjugation to the 3E10 antibody or antigen-binding fragment thereof, or humanized 3E10 antibody or antigen- binding fragment thereof, herein, such as inorganic and organic molecules, pharmaceutical agents, drugs, peptides, proteins, genetic material, and the like. In some aspects, the antibody- oligonucleotide conjugate (AOC) comprises an oligonucleotide.A . A ntigen-Binding Domains (A BDs)

[0189] As used herein, the term “antigen-binding domain” or “ABD” refers to a domain comprising a three-dimensional structure capable of immunospecifically binding to an epitope of an antigen. Thus, in embodiments, an ABD comprises a hypervariable region, optionally a VH and / or VL domain of an antibody, optionally at least a VH domain. In embodiments, an ABD comprises at least one complementarity determining region (CDR) of an antibody. In embodiments, an ABD comprises at least two CDRs of an antibody. In embodiments, an ABD comprises at least three CDRs of an antibody. In embodiments, an ABD comprises at least four CDRs of an antibody. In embodiments, an ABD comprises at least five CDRs of an antibody. In embodiments, an ABD comprises six CDRs of an antibody.1. 3E10 Antibodies

[0190] In embodiments, the present disclosure relates to the use of 3E10 antibodies and antigen binding fragments thereof, e.g., for delivering therapeutic agents (e.g., oligonucleotides) into a cell within a subject. Although generally referred to herein as “3E10,” “3E10 antibodies,” and the like, it will be appreciated that disclosure herein referring to such antibodies also encompass antigen-binding fragments thereof, e.g., scFv, di-scFv, tr-scFv, regardless of whether it is specifically recited in each instance. Thus, when describing a feature of a 3E10 antibody for use in the various compositions, conjugates, fusion proteins, and methods disclosed herein, that same feature is implicitly disclosed with respect to 3E10 antigen-binding fragments as well.

[0191] In embodiments, a 3E10 antibody comprises VL CDRs of SEQ ID NOs: 61, 62, and 63 and VH CDRs of SEQ ID NOs: 58, 59, and 60. In embodiments, a 3E10 antibody comprises VL CDRs of SEQ ID NOs: 9, 10, and 11 and VH CDRs of SEQ ID NOs: 3, 4, and 5. In embodiments, a 3E10 antibody comprises VL CDRs of SEQ ID NOs: 22, 23, and 24 and VH CDRs of SEQ ID NOs: 15, 17, and 18. In embodiments, a 3E10 antibody comprises VL CDRs of SEQ ID NOs: 9, 10, and 11 and VH CDRs of SEQ ID NOs: 16, 4, and 5. Other examples of 3E10 VL and VH CDR sequences are shown in Figures 1-4.

[0192] 3E10 is known to interact with the ENT2 nucleoside transporter expressed on various cell types, including muscle cells and cancer cells. In fact, ENT2 is overexpressed in most, if not all cancers. Accordingly, an AOC provided herein can widely target cancers based on cell surface expression of ENT2 on cancer cells. Advantageously, an AOC described herein can targetENT2 and extracellular DN A simultaneously. Importantly, 3E10 has been shown to preferentially localize into tumor cell nuclei in vivo, likely due to increased DNA in the local environment released from ischemic and necrotic regions of tumor. Targeting of the 3E10 antibody to extracellular DNA is described in, for example in Weisbart, Sci Reports, 2015, which is herein incorporated by reference. By targeting ENT2 as well as extracellular DNA, an 3E10 AOC described herein presents a platform to target a variety of cancers and deliver therapeutic oligonucleotides to target and kill cancer cells.

[0193] In some aspects of the present disclosure, the antibody or antigen-binding fragment thereof is a murine, chimeric, humanized, or human antibody or antigen-binding fragment thereof.

[0194] In some aspects, an AOC of the present disclosure take on a number of formats as described herein, including traditional antibodies as well as antibody derivatives, fragments, and mimetics, described herein in various embodimentspenetrates into cells and nuclei in an ENT2- dependent manner.

[0195] In embodiments, a second polynucleotide is non-covalently bound to an AOC of the present disclosure, to help facilitate cellular internalization of the AOC. That is, in some embodiments, polynucleotides conjugated to the antibody (cargo polynucleotides) do not interact with the nucleic acid-binding paratope of the antibody, and a second polynucleotide (e.g., carrier nucleic acid) is non-covalently complexed with the paratope to help facilitate internalization. In embodiments, the second polynucleotide is precomplexed with the AOC prior to administering the AOC to a subject. In embodiments, the second polynucleotide is an extracellular polynucleotide that is bound by the AOC at a site of interest in vivo, for example, at a site of tumor ischemia and / or necrosis. In embodiments, the second polynucleotide is DNA. In embodiments, the second polynucleotide is RNA.

[0196] In embodiments, an AOC disclosed herein comprises a VH and VL domain of a 3E10 antibody. In embodiments, the AOC comprises a VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 of a 3E10 antibody.

[0197] In some aspects, the present disclosure provides an antibody-oligonucleotide conjugate having the formula A-(L-Pr)q, wherein: A is a 3E10 antibody or antigen-binding fragment thereof, L is a linker, and P is an oligonucleotide as described herein, wherein the linker L links A to P. In embodiments, L is a cleavable linker and P is an ASO that mediates exonskipping. Tn some aspects of the present disclosure, the amino acid residue corresponding with D31 of the heavy chain CDR1 of the 3E10 antibody or antigen-binding fragment thereof is substituted with N. It is known in the art that mutation of aspartic acid at residue 31 of VH CDR1 to asparagine increases the cationic charge of this residue and enhanced nucleic acid binding and delivery in vivo (3E10-D31N). In embodiments, additional 3E10 antibody variants include mutation of aspartic acid at residue 31 of VH CDR1 to arginine (3E10-D31R), which modeling indicates expands cationic charge, or lysine (3E10-D31K) which modeling indicates changes charge orientation. Thus, in some aspects, the 3E10 antibody or antigen-binding fragment thereof includes a D31R or D3 IK substitution. In embodiments, additional 3E10 antibody variants include R96N, and / or S30D, alone or in combination with D31N, D31R, or D31K. All of the sequences disclosed herein having the residue corresponding to 3E10 D31 or N31, are expressly disclosed with a D31R or D3 IK or N31R or N3 IK substitution.

[0198] In embodiments, the present disclosure provides an antibody-oligonucleotide conjugate (AOC) having the formula A-(L-Pr)q, wherein: A is a 3E10 antibody or antigen-binding fragment thereof, L is a linker, P is an oligonucleotide as described herein, r is an integer from 1 to 4, and q is an integer from 1 to 16, wherein the linker L links A to (P); wherein the 3E10 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of XI YGMX2, where XI is D, E, N, Q, R, or K and X2 is K, R, or H (SEQ ID NO:58). In embodiments, the antibody or antigen-binding fragment thereof comprises (a) a light chain variable region (VL) complementarity determining region (CDR) 1 comprising the amino acid sequence of X1ASX2X3VSTSSYSYX4X5, where XI is K, R, or H, X2 is K, R, or H, X3 is T or S, X4 is M or L, and X5 is K, R, H, or A (SEQ ID NO:61), (b) a VL CDR2 comprising the amino acid sequence of YASYLX1S, where XI is D, E, N, or Q (SEQ ID NO:62), and (c) a VL CDR3 comprising the amino acid sequence of QX1SX2X3FPWT, where XI is K, R, or H, X2 is K, R, or H, and X3 is D or E (SEQ ID NO:63), and (d) a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of X1YGMX2, where XI is D, E, N, Q, R, or K and X2 is K, R, or H (SEQ ID NO: 58), (e) a VH CDR2 comprising the amino acid sequence of YISSX1SSTIYYAX2X3VX4G, where XI is G or S, X2 is D or E, X3 is T or S, and X4 is K, R, or H (SEQ ID NO:59), and (f) a VH CDR3 comprising the amino acid sequence of X1GLLLX2Y, where XI is K, R, or H, and X2 is D or E (SEQ ID NO:60).

[0199] In embodiments, the present disclosure provides an antibody-oligonucleotide conjugate (AOC) having the formula A-(L-Pr)q, wherein: A is a 3E10 antibody or antigen-binding fragment thereof, L is a linker, P is a payload as described herein, r is an integer from 1 to 4, and q is an integer from 1 to 16, wherein the linker L links A to P; wherein the 3E10 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of NYGMH (SEQ ID NO: 15). In embodiments, the antibody or antigen- binding fragment thereof comprises (a) a light chain variable region (VL) complementarity determining region (CDR) 1 comprising the amino acid sequence of RASKSVSTSSYSYMH (SEQ ID NO:9), (b) a VL CDR2 comprising the amino acid sequence of YASYLES (SEQ ID NO: 10), and (c) a VL CDR3 comprising the amino acid sequence of QHSREFPWT (SEQ ID NO: 11), and (d) a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of NYGMH (SEQ ID NO: 15), (e) a VH CDR2 comprising the amino acid sequence of YISSGSSTIYYADTVKG (SEQ ID NO: 4), and (f) a VH CDR3 comprising the amino acid sequence of RGLLLDY (SEQ ID NO: 5). In embodiments, the antibody or antigen-binding fragment thereof comprises a light chain variable region (VL) comprising an amino acid sequence that is identical to SEQ ID NO:21. In embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) comprising an amino acid sequence that is identical to SEQ ID NO: 14. In embodiments, the antibody or antigen-binding fragment thereof comprises a full length light chain (LC) comprising an amino acid sequence that is identical to SEQ ID NO:20. In embodiments, the antibody or antigen-binding fragment thereof comprises a full length heavy chain (HC) comprising an amino acid sequence that is identical to SEQ ID NO: 13.

[0200] In embodiments, the antibody or antigen-binding fragment thereof comprises a light chain variable region (VL) comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO:21. In embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 14. In embodiments, the antibody or antigen-binding fragment thereof comprises a full length light chain (LC) comprising an amino acid sequence that is at least about80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO:20. In embodiments, the antibody or antigen-binding fragment thereof comprises a full length heavy chain (HC) comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 13.

[0201] In some aspects, the 3E10 antibody or antigen-binding fragment thereof can be transported into the cytoplasm and / or nucleus of the cells without the aid of a carrier or conjugate. For example, a monoclonal 3E10 antibody and active fragments thereof that are transported in vivo to the nucleus of mammalian cells without cytotoxic effect are disclosed in U.S. Patent Nos. 4,812,397 and 7,189,396 to Richard Weisbart, the disclosures of which are incorporated by reference herein, in their entireties.

[0202] Amino acid sequences of 3E10 monoclonal antibodies and antigen-binding fragments thereof are known in the art. Example sequences of 3E10 heavy and light chains are provided below herein.

[0203] A murine version of the 3E10 antibody is described in Zack, et al., Immunology and Cell Biology, 72:513-520 (1994), the disclosure of which is incorporated by reference herein, in its entirety.

[0204] Amino acid variants of the 3E10 antibody are also known in the art, for example, as described in Zack, et al., J. Immunol., 157(5):2082-8 (1996). For example, amino acid position 31, in CDR1 of the heavy chain variable region of 3E10, influences nucleic acid binding and the antibody’s ability to penetrate nuclei. Substitution of the ‘wild-type’ (e.g., relative to the original murine antibody) aspartic acid by asparagine (the ‘D3 IN’ mutation) improves nucleic acid binding and nuclei penetration of the antibody, relative to the ‘wild type’ murine antibody. See, for example, Zack, et al., Immunology and Cell Biology, 72:513-520 (1994); Weisbart, et al., J. Autoimmun ., 11, 539-546 (1998); and Weisbart, Int. J. Oncol., 25, 1867-1873 (2004) (which are incorporated by reference herein, in their entireties).

[0205] Sequences for 3E10 antibodies and antigen-binding fragments or variants thereof, with the D3 IN substitution, are disclosed herein. In some aspects, the 3E10 antibodies and antigen- binding fragments thereof disclosed herein include the D31N substitution. In some aspects, otheramino acids are substituted at position 31 in the 3E10 antibodies and antigen-binding fragments thereof disclosed herein. For example, D31R, D31K, or D31R substitutions are incorporated in some aspects of the present disclosure.

[0206] Other 3E10 light chain sequences are known in the art. See, for example, Zack, et al., J. Immunol., 15;154(4):1987-94 (1995); GenBank: L16981.1 - Mouse Ig rearranged L-chain gene, partial cds; GenBank: AAA65681.1 - immunoglobulin light chain, partial [Mus musculus]).

[0207] Traditional antibody structural units typically comprise a tetramer. Each tetramer is typically composed of two identical pairs of polypeptide chains, each pair having one “light” (typically having a molecular weight of about 25 kDa) and one “heavy” chain (typically having a molecular weight of about 50-70 kDa). Human light chains are classified as kappa and lambda light chains. In embodiments, an antibody disclosed herein is an IgA, IgD, IgE, IgG, or IgM antibody, including any subtype or isotype thereof. In embodiments, an antibody disclosed herein is based on the IgG class, which has several. In embodiments, an antibody disclosed herein is based on one of the subclasses of IgG, including, but not limited to IgGl, IgG2, IgG3, and IgG4. In general, IgGl, IgG2 and IgG4 are used more frequently than IgG3. It should be noted that IgGl has different allotypes with polymorphisms at 356 (D or E) and 358 (L or M), and in embodiments, antibodies disclosed herein are based on IgGl having D or E at position 356 and / or L or M at position 358.

[0208] The light chain generally comprises two domains, the variable light domain (containing the light chain CDRs and together with the variable heavy domains forming the Fv region), and a constant light chain region (often referred to as CL or CK). The heavy chain comprises a variable heavy domain and a constant domain, which includes a CHI -optional hinge- Fc domain comprising a CH2-CH3.

[0209] The hypervariable region of an antibody generally encompasses amino acid residues from about amino acid residues 24-34 (LCDR1; “L” denotes light chain), 50-56 (LCDR2) and 89-97 (LCDR3) in the light chain variable region and around about 31-35B (HCDR1; “H” denotes heavy chain), 50-65 (HCDR2), and 95-102 (HCDR3) in the heavy chain variable region; Kabat et al., SEQUENCES OF PROTEINS OF IMMUNOLOGICAL INTEREST. 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991) and / or those residues forming a hypervariable loop (e.g.., residues 26-32 (LCDR1), 50-52 (LCDR2) and 91-96 (LCDR3) in thelight chain variable region and 26-32 (HCDR1), 53-55 (HCDR2) and 96-101 (HCDR3) in the heavy chain variable region; Chothia and Lesk (1987) J. Mol. Biol. 196:901-917. Specific CDRs useful for the compositions, conjugates, and methods described herein are described below.

[0210] As will be appreciated by those in the art, the exact numbering and placement of the CDRs can be different among different numbering systems. However, it should be understood that the disclosure of a variable heavy and / or variable light sequence includes the disclosure of the associated (inherent) CDRs. Accordingly, the disclosure of each variable heavy region is a disclosure of the VH CDRs (e.g. VH CDR1, VH CDR2., and VH CDR3) and the disclosure of each variable light region is a disclosure of the VL CDRs (e.g. VL CDR1, VL CDR2, and vlCDR3). A useful comparison of CDR numbering is described in Lafranc et al., Dev. Comp. Immunol. 27(1): 55-77 (2003)).

[0211] Throughout the present disclosure, the Kabat numbering system is generally used when referring to a residue in the variable domain (approximately, residues 1-107 of the light chain variable region and residues 1-113 of the heavy chain variable region) and the EU numbering system for Fc regions (e.g., Kabat et al., supra (1991)). In some aspects, the present specification uses the IMGT system to define the complementarity determining regions (CDRs) provided herein.

[0212] The present disclosure provides a large number of different CDR sets. In this case, a “full CDR set” comprises the three variable light CDRs, e.g., a VL CDR1, VL CDR2, and VL CDR3, and the three variable heavy CDRs, e.g., a VH CDR1, VH CDR2, and VH CDR3. These can be part of a larger variable light or variable heavy domain, respectfully. In addition, as more fully outlined herein, the variable heavy and variable light domains can be on separate polypeptide chains, when a heavy and light chain is used (for example when Fabs are used), or on a single polypeptide chain in the case of scFv sequences.

[0213] The CDRs contribute to the formation of the antigen-binding, or more specifically, epitope binding site of antibodies. “Epitope” refers to a determinant that interacts with a specific antigen binding site in the variable region of an antibody molecule known as a paratope. Epitopes are groupings of molecules such as nucleic acids, amino acids, or sugar side chains and usually have specific structural characteristics, as well as specific charge characteristics. A single antigen may have more than one epitope. The antibodies described herein bind to nucleic acid epitopes in a partially sequence-independent manner. That is, while the antibodies described herein bind tosome polynucleotide structures and sequences with greater affinity than other nucleic acid structures and sequences, they have some general affinity for polynucleotides.

[0214] The “Fc domain” of the heavy chain includes the -CH2-CH3 domain, and optionally a hinge domain (-H-CH2-CH3). For IgG, the Fc domain comprises immunoglobulin domains CH2and CH3 (Cy2 and Cy3) and the lower hinge region between CHI (Cyl) and CH2(Cy2). Although the boundaries of the Fc region may vary, the human IgG heavy chain Fc region is usually defined to include residues C226 or P230 to its carboxyl-terminus, wherein the numbering is according to the EU index as in Kabat. Accordingly, “CH” domains in the context of IgG are as follows: “CHI” refers to positions 118-215 according to the EU index as in Kabat. “Hinge” refers to positions 216-230 according to the EU index as in Kabat. “CH2” refers to positions 231-340 according to the EU index as in Kabat, and “CH3” refers to positions 341-447 according to the EU index as in Kabat. Thus, the “Fc domain” includes the -CH2-CH3 domain, and optionally a hinge domain (hinge-CH2-CH3). In the embodiments herein, when a scFv is attached to an Fc domain, it is generally the C-terminus of the scFv construct that is attached to all or part of the hinge of the Fc domain; for example, it is generally attached to the sequence EPKS which is the beginning of the hinge. In some embodiments, as is more fully described below, amino acid modifications are made to the Fc region, for example to alter binding to one or more FcyR receptors or to the FcRn receptor, and to enable heterodimer formation and purification, as outlined herein.

[0215] Another part of the heavy chain is the hinge region. By “hinge” or “hinge region” or “antibody hinge region” or “hinge domain” herein is meant the flexible polypeptide comprising the amino acids between the first and second constant domains of an antibody. As noted above herein, the present disclosure refers to different antibody domains of a 3E10 antibody or antigen- binding fragment thereof. Structurally, the IgG CHI domain ends at EU position 215, and the IgG CH2domain begins at residue EU position 231. Thus for IgG the antibody hinge is herein defined to include positions 216 (E216 in IgGl) to 230 (p230 in IgGl), wherein the numbering is according to the EU index as in Kabat. In some cases, a “hinge fragment” is used, which contains fewer amino acids at either or both of the N- and C-termini of the hinge domain.

[0216] A scFv comprises a variable heavy chain, an scFv linker, and a variable light domain. In most of the constructs and sequences outlined herein, the C-terminus of the variableheavy chain is attached to the N-terminus of the scFv linker, the C-terminus of which is attached to the N-terminus of a variable light chain (N-vh-linker-vl-C) although that can be switched (N- vl-linker-vh-C).

[0217] Thus, the present disclosure relates to different antibody domains. These domains include, but are not limited to, the Fc domain, the CHI domain, the CH2domain, the CH3 domain, the hinge domain, the heavy constant domain (CHl-hinge-Fc domain or CHl-hinge-CH2-CH3), the variable heavy (VH) domain, the variable light (VL) domain, the light constant domain, Fab domains and scFv domains.2. Humanized Antibodies

[0218] In certain embodiments, the antibodies of the disclosure comprise a heavy chain variable region from a particular germline heavy chain immunoglobulin gene and / or a light chain variable region from a particular germline light chain immunoglobulin gene. For example, in some embodiments, such an antibody comprises or consists of a murine, chimeric, humanized, or human antibody or antigen-binding fragment thereof comprising heavy or light chain variable regions that are "“the product of'” or "“derived from"” a particular germline sequence, e.g., that of the 3E10 antibody. A human antibody that is "“the product of1” or "“derived from"” a human germline immunoglobulin sequence can be identified as such by comparing the amino acid sequence of the human antibody to the amino acid sequences of human germline immunoglobulins and selecting the human germline immunoglobulin sequence that is closest in sequence (i.e., greatest % identity) to the sequence of the human antibody (using the methods outlined herein). A human antibody that is “the product of’ or “derived from” a particular human germline immunoglobulin sequence may contain amino acid differences as compared to the germline sequence, due to, for example, naturally-occurring somatic mutations or intentional introduction of site-directed mutation. However, a humanized antibody typically is at least 90% identical in amino acids sequence to an amino acid sequence encoded by a human germline immunoglobulin gene and contains amino acid residues that identify the antibody as being derived from human sequences when compared to the germline immunoglobulin amino acid sequences of other species (e.g., murine germline sequences). In some embodiments, a humanized antibody is at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or identical in amino acid sequence to the amino acid sequence encoded by the germline immunoglobulin gene. Typically, a humanized antibody derived from aparticular human germline sequence will display no more than 10-20 amino acid differences from the amino acid sequence encoded by the human germline immunoglobulin gene. In some embodiments, the humanized antibody has no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 amino acid difference from the amino acid sequence encoded by the germline immunoglobulin gene.

[0219] In some embodiments, a parental antibody is affinity matured. Methods for affinity maturation are known in the art. In some embodiments, structure-based methods are employed for humanization and affinity maturation, for example, as described in U.S Patent Publication No. 2006 / 0008883, which is incorporated herein by reference. Selection based methods are also known for humanization and / or affinity maturation of antibody variable regions, including but not limited to methods described in Wu et al., 1999, J. Mol. Biol. 294: 151-162; Baca et al., 1997, J. Biol. Chem. 272(16): 10678-10684; Rosok et al., 1996, J. Biol. Chem. 271(37): 22611-22618; Rader et al., 1998, Proc. Natl. Acad. Sci. USA 95: 8910-8915; Krauss et al., 2003, Protein Engineering 16(10):753-759, all of which are incorporated herein by reference. Yes other known humanization methods include grafting of only parts of the CDRs, including but not limited to methods described in U.S Patent Patent Publication No. 2001 / 0035606; Tan et al., 2002, J. Immunol. 169:1119-1125; De Pascalis et al., 2002, J. Immunol. 169:3076-3084, all of which are incorporated herein by reference.3. Fc variants

[0220] In embodiments, one or more amino acid modifications can be introduced into the Fc region of an antibody provided herein, thereby generating an Fc region variant. The Fc region variant can comprise a human Fc region sequence (e.g., a human IgGl, IgG2, IgG3 or IgG4 Fc region) comprising an amino acid modification (e.g. a substitution) at one or more amino acid positions.

[0221] In embodiments, an Fc region variant possesses some but not all effector functions, which make it a desirable candidate for applications in which the half-life of the antibody in vivo is important yet certain effector functions (such as complement and ADCC) are unnecessary or deleterious. In vitro and / or in vivo cytotoxicity assays can be conducted to confirm the reduction / depletion of CDC and / or ADCC activities. For example, Fc receptor (FcR) binding assays known in the art can be conducted to ensure that the antibody lacks FcyR binding (hencelikely lacking ADCC activity), but retains FcRn binding ability. To assess complement activation, a CDC assay can be performed (see, for example, Gazzano- Santoro et al., J. Immunol. Methods 202: 163 (1996); Cragg, M.S. et al., Blood 101 :1045-1052 (2003); and Cragg, M. S. and M. J. Glennie, Blood 103:2738-2743 (2004)). FcRn binding and in vivo clearance / half life determinations can also be performed using methods known in the art (see, e.g., Petkova, S.B. et al., Int’l. Immunol. 18(12): 1759-1769 (2006)).

[0222] In embodiments, an antibody provided herein can have reduced effector function and thus can comprise a substitution of one or more of Fc region residues 238, 265, 269, 270, 297, 327 and 329 (U.S. Pat. No. 6,737,056). Such Fc mutants include Fc mutants with substitutions at two or more of amino acid positions 265, 269, 270, 297 and 327, including the so-called “DANA” Fc mutant with substitution of residues 265 and 297 to alanine (U.S. Pat. No. 7,332,581).

[0223] In embodiments, an Fc region variant provided herein can have improved or diminished binding to FcRs. See, e.g., U.S. Pat. No. 6,737,056; WO 2004 / 056312, and Shields et al., J. Biol. Chem. 9(2): 6591-6604 (2001), the disclosure of which are incorporated herein by reference, in their entireties.

[0224] In embodiments, an Fc region variant provided herein comprises an Fc region with one or more amino acid substitutions which improve ADCC, e.g., substitutions at positions 298, 333, and / or 334 of the Fc region (EU numbering of residues).

[0225] In embodiments, an Fc region variant provided herein comprises alterations that result in altered (i.e., either improved or diminished) Clq binding and / or Complement Dependent Cytotoxicity (CDC), e.g., as described in U.S. Pat. No. 6,194,551, WO 99 / 51642, and Idusogie et al. J. Immunol. 164: 4178-4184 (2000).

[0226] In embodiments, an Fc region variant provided herein comprises alterations that result in increased half-lives and improved binding to the neonatal Fc receptor (FcRn), which is responsible for the transfer of maternal IgGs to the fetus (Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol. 24:249 (1994)), e.g., as described in US2005 / 0014934A1 (Hinton et al.). Those antibodies comprise an Fc region with one or more substitutions therein which improve binding of the Fc region to FcRn. Such Fc variants include those with substitutions at one or more ofFc region residues: 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362,376, 378, 380, 382, 413, 424 or 434, e.g., substitution of Fc region residue 434 (U.S. Pat. No. 7,371,826).

[0227] In embodiments, an Fc region variant provided herein comprises “knob-in-hole” or “skew” variants, which refer to amino acid engineering that creates stearic influences to favor heterodimeric formation and disfavor homodimeric formation, as described in USSN 61 / 596,846, Ridgway et al, Protein Engineering 9(7):617 (1996); Atwell et al, J. Mol. Biol. 1997 270:26; US Patent No. 8,216,805, all of which are hereby incorporated by reference in their entirety.

[0228] In embodiments, an Fc region variant provided herein comprises alterations described in Duncan & Winter, Nature 322:738-40 (1988); U.S. Pat. No. 5,648,260; U.S. Pat. No. 5,624,821; and WO 94 / 29351.4. Antibody Fragments

[0229] In embodiments, the antibody portion of an AOC described herein comprises an antigen-binding fragment of a 3E10 antibody. In embodiments, the antigen-binding fragment retains the desired biological activity of a 3E10 antibody. In embodiments, the antigen-binding fragment retains at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95% of the desired biological activity of a 3E10 antibody. In embodiments, the antigen-binding fragment retains the ability of the antibody to bind to its target antigen, e.g., a nucleic acid, e.g., DNA. In embodiments, the antigen-binding fragment retains the ability of the antibody to bind to a cellular receptor, e.g., ENT2. In embodiments, the antigen- binding fragment retains the ability of the antibody to be internalized by a target cell.

[0230] In embodiments, the 3E10 antibody or antigen binding fragment thereof comprises a single-chain fragment variable (scFv), a tandem double scFv, an (scFv)2, a minibody, a VHH, an scFv-Fc, a CrossMab, a dual variable domain immunoglobulin (DVD-Ig), a single-chain tandem fragment variable (scTaFv), a diabody, a tandem diabody (TandAb), a Fabsc, a modular IgG-scFv, a Fab, or an F(ab’)2.

[0231] In embodiments, an antigen-binding fragment of a 3E10 antibody or antigen- binding fragment thereof comprises a CrossMab. In the CrossMab format, complementarymutations are introduced in the heavy chain constant region of each arm to generate so-called “holes and knobs,” resulting in preferred association between different arms, forming a heterodimer, rather than a homodimer of two of the same arms. The exact residues that are mutated in the heavy chain constant region of a CrossMab bispecific antibody to form “holes” and “knobs” can vary depending on the specific design and optimization goals of the antibody. For more information on CrossMab antibodies see, for example, Huang, J., et al., Journal of Biological Chemistry, 294(50): 19001-10 (2019), the disclosure of which is incorporated herein by reference in its entirety.

[0232] In embodiments, an antigen-binding fragment of a 3E10 antibody or antigen- binding fragment thereof comprises a divalent, dual-variable domain immunoglobulin (DVD-Ig) of a 3E10 antibody or antigen binding fragment thereof. In the DVD-Ig format, each arm of the antibody contains two VH / VL pairs. In some embodiments, one of the VH / VL pairs comprises 3E10 VH and VL CDRs. For more information on DVD-Ig antibodies see, for example, Polson AG, et al., Journal of immunotherapy. 29(3):241-50 (2006) and U.S. Patent No. 7,612,181, the disclosures of which are incorporated herein by reference in their entireties.

[0233] In embodiments, an antigen-binding fragment of a 3E10 antibody or antigen- binding fragment thereof comprises a single-chain variable fragment (scFv). “Single chain Fv” or “scFv,” as used herein, refers to a VH domain covalently attached to a VL domain through a linker, e g., a scFv linker as discussed herein, to form a continuous protein chain. A scFv domain can be in either arrangement from N- to C-terminus (i.e., VH-linker-VL or VL-linker-VH). In the sequences depicted in the sequence listing and in the figures herein, the order of the VH and VL domain is indicated in the name, e.g., H.X L.Y means the N- to C-terminus arrangement is VH- linker-VL, and L.Y H.X means the N- to C-terminus arrangement is VL-linker-VH.

[0234] In embodiments, an antigen-binding fragment of a 3E10 antibody or antigen- binding fragment thereof comprises a tandem double scFv. A tandem double scFv has two scFv domains linked in a linear fashion. In some embodiments, each scFv domain is derived from a different antibody and provides independent antigen-binding specificity. In some embodiments, one of the scFv domains comprises 3E10 VH and VL CDRs. For more information on tandem double scFvs see, for example, Bossen C, et al., MAbs, 4(2):200-08 (2012), the disclosure of which is incorporated herein by reference in its entirety.

[0235] In embodiments, an antigen-binding fragment of a 3E10 antibody or antigen- binding fragment thereof comprises a dimeric scFv antibody (scFv)2. A dimeric scFv antibody has two scFv domains linked in a dimeric arrangement. In some embodiments, each scFv domain is derived from a different antibody and provides independent antigen-binding specificity. In some embodiments, one of the scFv domains comprises 3E10 VH and VL CDRs. For more information on dimeric scFv antibodies see, for example, Llewellyn C, et al., Journal of immunological methods, 273(l-2):33-44 (2002), the disclosure of which is incorporated herein by reference in its entirety.

[0236] In embodiments, an antigen-binding fragment of a 3E10 antibody or antigen- binding fragment thereof comprises a scFv-Fc. An “scFv-Fc,” as meant herein, is a polypeptide that consists of a heavy and a light chain variable region of an antibody joined by a linker, which is followed by an Fc polypeptide chain of an antibody, optionally the Fc region of a human IgG antibody, such as an IgGl, IgG2, IgG3, or IgG4 antibody.

[0237] In embodiments, an antigen-binding fragment of a 3E10 antibody or antigen- binding fragment thereof comprises a single-chain tandem fragment variable (scTaFv) antibody. A single-chain tandem fragment variable (scTaFv) antibody is a type of bispecific antibody that consists of two variable fragment (VH and VL) domains linked in a tandem arrangement. In some embodiments, one of the variable fragment domains comprises 3E10 VH and VL CDRs. For more information on scTaFv antibodies see, for example, Schramm C, et al., MAbs 5(3):442-49 (2013), the disclosure of which is incorporated herein by reference in its entirety.

[0238] In embodiments, an antigen-binding fragment of a 3E10 antibody or antigen- binding fragment thereof comprises a VHH, also referred to as a nanobody. As used herein, the term “VHH” refers to a variable domain of heavy chain of heavy-chain antibody. A VHH is a molecule that can recognize an antigen through a single domain and is the smallest unit among antibody molecules that have been found to date. In embodiments, a VHH can include one or more variable domains of heavy chain derived from a heavy-chain antibody, and the number of the variable domains of heavy chain included in the VHH is not limited.

[0239] In embodiments, an antigen-binding fragment of a 3E10 antibody or antigen- binding fragment thereof comprises a diabody. As used herein, “diabody” refers to a divalent antibody comprising two polypeptide chains, wherein each polypeptide chain is too short for a pairto form between two domains on the same chain such that each domain is paired with a complementary domain on another polypeptide chain (see, e.g., Holliger et al., 1993, Proc. Natl. Acad. Sci. USA 90: 6444-48 and Poljak et al., 1994, Structure 2: 1121-23). If the two polypeptide chains of the diabody are identical, there will be two identical antigen-binding sites in the diabody resulting from their pairing. In embodiments, one of the antigen binding domains of the diabody comprises 3E10 VH and VL CDRs. For more information on diabodies see, for example, Hoogenboom HR, et al., Trends Biotechnol., 21 (12): 553-57 (2003), the disclosure of which is incorporated herein by reference in its entirety. Polypeptide chains of different sequences can be used to prepare diabodies with two different antigen-binding sites. Similarly, as used herein, “triabodies” and “tetrabodies” refer to antibodies that contain three and four polypeptide chains, respectively, and form three and four antigen-binding sites (which can be the same or different), respectively.

[0240] The term “minibody” is used to refer to an scFv-CH3 fusion protein that self- assembles into a bivalent dimer of 80 kDa (ScFv-CH3)2.

[0241] In embodiments, an antigen-binding fragment of a 3E10 antibody or antigen- binding fragment thereof comprises a tandem diabody (TandAb). A tandem diabody has two antigen-binding domains (VH and VL) linked in a tandem arrangement by a flexible peptide linker. In some embodiments, one of the antigen binding domains comprises 3E10 VH and VL CDRs. For more information on diabodies see, for example, Sidelmann JG, et al., Mol Immunol., 45(9):2597-607 (2008), the disclosure of which is incorporated herein by reference in its entirety.

[0242] In embodiments, an antigen-binding fragment of a 3E10 antibody or antigen- binding fragment thereof comprises a Fabsc. As used herein, a “Fabsc” format antibody molecule typically refers to a bispecific antibody molecule having a Fab fragment, which generally includes a hinge region, which is at the C-terminus of the Fab fragment linked to the N- terminus of a CH2domain, of which the C-terminus is in turn linked to the N-terminus of a scFv fragment.

[0243] In embodiments, an antigen-binding fragment of a 3E10 antibody or antigen- binding fragment thereof comprises a scFab. A scFab, also known as a single-chain fragment antigen binding (Fab), is a type of antibody fragment that combines the variable heavy chain (VH) and variable light chain (VL) domains into a single polypeptide chain, linked by a peptide linker. The domain structure of a Fabsc includes the variable domains of both the heavy chain and lightchain (VH and VL), and a peptide linker that connects the two domains. In addition to the variable domains, a Fabsc also includes the constant domains of the light chain (CL) and the hinge region of the heavy chain. In some embodiments, one of the antigen binding domains comprises 3E10 VH and VL CDRs. For more information on Fabscs see, for example, Kettner, C., et al., Frontiers in Immunology, 8(8):453 (2017), the disclosure of which is incorporated herein by reference in its entirety.

[0244] In embodiments, an antigen-binding fragment of a 3E10 antibody or antigen- binding fragment thereof comprises an IgG-scFv. An IgG-scFv is an antibody in which a scFv is fused to the light chain or heavy chain of an IgG. In some embodiments, the scFv comprises 3E10 VH and VL CDRs. In some embodiments, the IgG comprises 3E10 VH and V LCDRs. In some embodiments, the antibody is an F(ab’)2.5. Bispecific Antibodies

[0245] In embodiments, 3E10 antibodies and antigen-binding fragments thereof can be modified to improve their therapeutic potential. For example, in embodiments, the cell-penetrating anti-DNA antibody is conjugated to another antibody specific for a second therapeutic target in the cytoplasm and / or nucleus of a target cell. For example, in embodiments, the cell-penetrating 3E10 antibody is a bispecific antibody having a first heavy chain and a first light chain from 3E10 and a second heavy chain and a second light chain from a monoclonal antibody that specifically binds a second therapeutic target.

[0246] Bispecific antibodies and other binding proteins having a first heavy chain and a first light chain from 3E10 and a second heavy chain and a second light chain from a monoclonal antibody that specifically binds a second target are discussed in Weisbart, et al., Mol. Cancer Ther., 11(10):2169-73 (2012), and Weisbart, et al., Int. J. Oncology, 25: 1113-8 (2004), and U.S. Patent Application No. 2013 / 0266570, which are specifically incorporated by reference in their entireties. In embodiments, the second target is specific for a target cell-type, tissue, organ, etc. Thus the second heavy chain and second light chain can serve as a targeting moiety that targets the complex to the target cell-type, tissue, organ. In embodiments, the second heavy chain and second light chain target, hematopoietic stem cells, CD34+cells, T cells or any another cell type of interest, e g., by targeting a receptor or ligand expressed on the cell type of interest. In embodiments, the second heavy chain and second light chain target the thymus, spleen, or cancer cells.

[0247] Bispecific antibodies can be used to direct cytotoxic agents or drugs to cells which express a particular antigen. These antibodies possess two binding sites directed at two different antigens or two different epitopes on the same antigen. For example, in embodiments thebispecific can comprise one arm for ENT2 engagement and another arm for a second target. Bispecific antibody design can include a variety of antibody designs with multiple binding arms. Techniques for making bispecific antibodies are common in the art (Millstein et al., 1983, Nature 305:537- 539; Brennan et al., 1985, Science 229:81; Suresh et al, 1986, Methods in Enzymol. 121 : 120; Traunecker et al., 1991, EMBO J. 10:3655-3659; Shalaby et al., 1992, J. Exp. Med. 175:217-225; Kostelny et al., 1992, J. Immunol. 148: 1547-1553; Gruber et al., 1994, J. Immunol. 152:5368; and U.S. Patent 5,731,168). Antibodies with more than two valencies are also contemplated. For example, trispecific antibodies can be prepared (Tutt et al., J. Immunol. 147:60 (1991)). In embodiments the contemplated bispecific antibody disclosed herein can be conjugated as a bispecific antibody-payload conjugate.

[0248] Heteroconjugate antibodies are also within the scope of the present disclosure. Heteroconjugate antibodies are composed of two covalently joined antibodies. Such antibodies have, for example, been proposed to target immune cells to unwanted cells (U.S. Pat. No. 4,676,980). It is contemplated that the antibodies can be prepared in vitro using known methods in synthetic protein chemistry, including those involving crosslinking agents. For example, immunotoxins can be constructed using a disulfide exchange reaction or by forming a thioether bond. Examples of suitable reagents for this purpose include iminothiolate and methyl-4- mercaptobutyrimidate. In embodiments the contemplated herteoconjugate antibody disclosed herein can be conjugated as a heteroconjugate antibody-payload conjugate.6. 3E10 Sequences

[0249] In embodiments, a 3E10 antibody or antigen-binding fragment thereof described herein includes CDR sequences corresponding to the parent 3E10 antibody.

[0250] Accordingly, in embodiments, a 3E10 antibody or antigen-binding fragment thereof comprises (a) a light chain variable region (VL) complementarity determining region (CDR) 1 comprising the amino acid sequence of X1ASX2X3VSTSSYSYX4X5, where XI is K, R, or H, X2 is K, R, or H, X3 is T or S, X4 is M or L, and X5 is K, R, H, or A (SEQ ID NO:61), (b) a VL CDR2 comprising the amino acid sequence of YASYLX1S, where XI is D, E, N, or Q(SEQ ID NO:62), and (c) a VL CDR3 comprising the amino acid sequence of QX1 SX2X3FPWT, where XI is K, R, or H, X2 is K, R, or H, and X3 is D or E (SEQ ID NO:63), and (d) a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of X1YGMX2, where XI is D, E, N, Q, R, or K and X2 is K, R, or H (SEQ ID NO: 58), (e) a VH CDR2 comprising the amino acid sequence of YISSX1SSTIYYAX2X3VX4G, where XI is G or S, X2 is D or E, X3 is T or S, and X4 is K, R, or H (SEQ ID NO: 59), and (f) a VH CDR3 comprising the amino acid sequence of X1GLLLX2Y, where XI is K, R, or H, and X2 is D or E (SEQ ID NO:60).

[0251] In embodiments, a 3E10 antibody or antigen-binding fragment thereof includes a light chain variable region (VL) complementarity determining region (CDR) 1 comprising the amino acid sequence of 3E10-VL-CDR1 (SEQ ID NO:9), a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2 (SEQ ID NO: 10), a VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3 (SEQ ID NO: 11), a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of 3E10-VH-CDR1 (SEQ ID NO:3), a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2 (SEQ ID NO:4), and a VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3 (SEQ ID NO: 5).

[0252] In embodiments, a 3E10 antibody or antigen-binding fragment thereof described herein includes CDR sequences from a variant 3E10 antibody that includes a D31N amino acid substitution in the VH CDR1. Accordingly, in embodiments, the a 3E10 antibody or antigen- binding fragment thereof includes a light chain variable region (VL) complementarity determining region (CDR) 1 comprising the amino acid sequence of 3E10-VL-CDR1 D3 IN (SEQ ID NO:22), a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2 D31N (SEQ ID NO:23), a VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3 D31N (SEQ ID NO:24), a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of 3E10-VH- CDR1 D31N (SEQ ID NO: 15), a VH CDR2 comprising the amino acid sequence of 3E10-VH- CDR2 D31N (SEQ ID NO: 17), and a VH CDR3 comprising the amino acid sequence of 3E10- VH-CDR3 D3 IN (SEQ ID NO: 18).

[0253] In embodiments, a 3E10 antibody or antigen-binding fragment thereof described herein refers to CDR sequences corresponding to the parent 3E10 antibody, optionally including a D3 IN amino acid substitution in the VH CDR1. Accordingly, in embodiments, a 3E10 antibody or antigen-binding fragment thereof includes a light chain variable region (VL) complementaritydetermining region (CDR) 1 comprising the amino acid sequence of 3E10-VL-CDR1 (SEQ ID NO:9), a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2 (SEQ ID NO: 10), a VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3 (SEQ ID NO: 11), a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of 3E10-VH-CDRla (SEQ ID NO: 16), a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2 (SEQ ID NO:4), and a VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3 (SEQ ID NO:5).

[0254] In embodiments, a 3E10 antibody or antigen-binding fragment thereof described herein includes CDR sequences corresponding to the parent 3E10 antibody, with a known amino acid substitution in one or more CDR. Accordingly, in embodiments, a 3E10 antibody or antigen- binding fragment thereof described herein includes one or more amino acid substitution, relative to the CDR sequences of the parent 3E10 or 3E10-D31N variant, selected from a G to S substitution at position 5 of VH CDR2, a T to S substitution at position 14 of VH CDR2, an S to T substitution at position 5 of VL CDR1, an M to L substitution at position 14 of VL CDR1, an H to A substitution at position 15 of VL CDR1, and an E to Q substitution at position 6 of VL CDR2.

[0255] Accordingly, in embodiments, a 3E10 antibody or antigen-binding fragment thereof includes VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2.1 (SEQ ID NO:26) or 3E10-VH-CDR2.2 (SEQ ID NO:27). In embodiments, the 3E10 antibody or antigen- binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 1 and 3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 1 and 3 according to the 3E10- D31N variant. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 1 and 3 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody or relative to the 3E10- D3 IN variant.

[0256] In embodiments, a 3E10 antibody or antigen-binding fragment thereof includes VL CDR1 comprising the amino acid sequence of 3E10-VL-CDR1.1 (SEQ ID NO:28) or 3E10-VL- CDR1.2 (SEQ ID NO:29). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 2 and 3, and VH CDRs 1-3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 2 and 3, and VH CDRs 1-3 according to the 3E10- D3 IN variant. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 2 and 3, and VHCDRs 1-3 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody or relative to the 3E10- D3 IN variant.

[0257] In embodiments, a 3E10 antibody or antigen-binding fragment thereof includes VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2.1 (SEQ ID NO:30). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs1 and 3, and VH CDRs 1-3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3, and VH CDRs 1-3 according to the 3E10- D31N variant. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3, and VH CDRs 1-3 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody or relative to the 3E10- D3 IN variant.

[0258] While some of the amino acid substitutions described above are fairly conservative substitutions — e.g., an S to T substitution at position 5 of VL CDR1 — other substitutions are to amino acids that have vastly different properties — e.g., an M to L substitution at position 14 of VL CDR1, an H to A substitution at position 15 of VL CDR1, and an E to Q substitution at position 6 of VL CDR2. This suggests, without being bound by theory, that at least these positions within the 3E10 CDR framework are tolerant to other amino acid substitutions.

[0259] In embodiments, a 3E10 antibody or antigen-binding fragment thereof includes VHCDR2 comprising the amino acid sequence of 3E10-VH-CDR2.3 (SEQ ID NO:31). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 1 and 3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 1 and 3 according to the 3E10- D31N variant. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 1 and 3 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody or relative to the 3E10- D3 IN variant, e.g., as described herein.

[0260] In embodiments, a 3E10 antibody or antigen-binding fragment thereof includes VL CDR1 comprising the amino acid sequence of 3E10-VL-CDR1.3 (SEQ ID NO:32). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs2 and 3, and VH CDRs 1-3 according to the parent 3E10 antibody. In embodiments, the 3E10antibody or antigen-binding fragment thereof further includes VL CDRs 2 and 3, and VH CDRs 1-3 according to the 3E10- D31N variant. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 2 and 3, and VH CDRs 1-3 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody or relative to the 3E10- D3 IN variant, e g., as described herein.

[0261] In embodiments, a 3E10 antibody or antigen-binding fragment thereof, includes VLCDR2 comprising the amino acid sequence of 3E10-VL-CDR2.2 (SEQ ID NO:33). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3, and VH CDRs 1-3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3, and VH CDRs 1-3 according to the 3E10- D31N variant. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3, and VH CDRs 1-3 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody or relative to the 3E10- D3 IN variant, e g., as described herein.

[0262] In embodiments, a 3E10 antibody or antigen-binding fragment thereof includes VH CDR1 comprising the amino acid sequence of 3E10-VH-CDRl.cl (SEQ ID NO:34), 3E10-VH- CDRl.c2 (SEQ ID NO:35), 3E10-VH-CDRl.c3 (SEQ ID NO:36), 3E10-VH-CDRl.c4 (SEQ ID NO:37), or 3E10-VH-CDRl.c5 (SEQ ID NO:38). In embodiments, the 3E10 antibody or antigen- binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 2 and 3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 2 and 3 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0263] In embodiments, a 3E10 antibody or antigen-binding fragment thereof includes VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2.cl (SEQ ID NO:39), 3E10-VH- CDR2.c2 (SEQ ID NO:40), or 3E10-VH-CDR2.c3 (SEQ ID NO:41). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 1 and 3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 1 and 3 according to the 3E10- D31N variant. In embodiments, the 3E10 antibody or antigen-binding fragment thereof furtherincludes VL CDRs 1 -3, and VH CDRs 1 and 3 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0264] In embodiments, a 3E10 antibody or antigen-binding fragment thereof includes VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3.cl (SEQ ID NO:42), 3E10-VH- CDR3.c2 (SEQ ID NO:43), or 3E10-VH-CDR3.c3 (SEQ ID NO:44). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 1 and 2 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 1 and 2 according to the 3E10- D31N variant. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 -3, and VH CDRs 1 and 2 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0265] In embodiments, a 3E10 antibody or antigen-binding fragment thereof includes VL CDR1 comprising the amino acid sequence of 3E10-VL-CDR1.C1 (SEQ ID NO:45), 3E10-VL- CDR1.C2 (SEQ ID NO:46), 3E10-VL-CDRl.c3 (SEQ ID NO:47), 3E10-VL-CDRl.c4 (SEQ ID NO:48), 3E10-VL-CDR1.C5 (SEQ ID NO:49), or 3E10-VL-CDRl.c6 (SEQ ID NO:50). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs2 and 3, and VH CDRs 1-3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 2 and 3, and VH CDRs 1-3 according to the 3E10- D31N variant. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 2 and 3, and VH CDRs 1-3 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0266] In embodiments, a 3E10 antibody or antigen-binding fragment thereof includes VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2.cl (SEQ ID NO:51). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3, and VH CDRs 1-3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3, and VH CDRs 1- 3 according to the 3E10- D31N variant. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3, and VH CDRs 1-3 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0267] In embodiments, a 3E10 antibody or antigen-binding fragment thereof includes VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3.cl (SEQ ID NO:52), 3E10-VL- CDR3.C2 (SEQ ID NO:53), 3E10-VL-CDR3.c3 (SEQ ID NO:54), 3E10-VL-CDR3.c4 (SEQ ID NO:55), 3E10-VL-CDR3.c5 (SEQ ID NO:56), or 3E10-VL-CDR3.c6 (SEQ ID NO:57). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs1 and 2, and VH CDRs 1-3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 2, and VH CDRs 1- 3 according to the 3E10- D31N variant. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 2, and VH CDRs 1-3 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0268] It is also contemplated that a 3E10 antibody or antigen-binding fragment thereof, as described herein, includes any combination of the 3E10 CDR amino acid substitutions described above.

[0269] Accordingly, in embodiments, a 3E10 antibody or antigen-binding fragment thereof includes VH CDR1 comprising the amino acid sequence of 3E10-VH-CDRlm (SEQ ID NO:58). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 2 and 3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs2 and 3 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0270] In embodiments, a 3E10 antibody or antigen-binding fragment thereof includes VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2m (SEQ ID NO:59). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 1 and 3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 1 and3 according to the 3E10-D31N variant. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 1 and 3 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0271] In embodiments, a 3E10 antibody or antigen-binding fragment thereof includes VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3m (SEQ ID NO:60). Inembodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 1 and 2 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 1 and 2 according to the 3E10-D31N variant. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3, and VH CDRs 1 and 2 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0272] In embodiments, a 3E10 antibody or antigen-binding fragment thereof includes VL CDR1 comprising the amino acid sequence of 3E10-VL-CDRlm (SEQ ID NO:61). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs2 and 3, and VH CDRs 1-3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 2 and 3, and VH CDRs 1-3 according to the 3E10-D31N variant. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 2 and 3, and VH CDRs 1-3 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody, e g., as described herein.

[0273] In embodiments, a 3E10 antibody or antigen-binding fragment thereof includes VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2m (SEQ ID NO:62). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3, and VH CDRs 1-3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3, and VH CDRs 1 - 3 according to the 3E10-D31N variant. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3, and VH CDRs 1-3 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0274] In embodiments, a 3E10 antibody or antigen-binding fragment thereof includes VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3m (SEQ ID NO:63). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 2, and VH CDRs 1-3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 2, and VH CDRs 1- 3 according to the 3E10-D31N variant. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 2, and VH CDRs 1-3 having one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0275] In embodiments, a 3E10 antibody or antigen-binding fragment thereof described herein includes a VL CDR 1 comprising the amino acid sequence of 3E10-VL-CDRlm (SEQ ID NO:61), a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2m (SEQ ID NO:62), a VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3m (SEQ ID NO:63), a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of 3E10-VH-CDRlm (SEQ ID NO:58), a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2m (SEQ ID NO:59), and a VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3m (SEQ ID NO:60).

[0276] In embodiments, a 3E10 antibody or antigen-binding fragment thereof described herein refers to CDR sequences having no more than one amino acid substitution relative to the parent 3E10 antibody optionally including a D31N amino acid substitution in the VH CDR1. Accordingly, in embodiments, a 3E10 antibody or antigen-binding fragment thereof includes a VL CDR 1 comprising an amino acid sequence having no more than one amino acid substitution relative to 3E10-VL-CDR1 (SEQ ID NO:9), a VL CDR2 comprising an amino acid sequence having no more than one amino acid substitution relative to 3E10-VL-CDR2 (SEQ ID NO: 10), a VL CDR3 comprising an amino acid sequence having no more than one amino acid substitution relative to 3E10-VL-CDR3 (SEQ ID NO: 11), a heavy chain variable region (VH) CDR1 comprising an amino acid sequence having no more than one amino acid substitution relative to 3E10-VH-CDRla (SEQ ID NO: 16), a VH CDR2 comprising an amino acid sequence having no more than one amino acid substitution relative to 3E10-VH-CDR2 (SEQ ID NO:4), and a VH CDR3 comprising an amino acid sequence having no more than one amino acid substitution relative to 3E10-VH-CDR3 (SEQ ID NO: 5).

[0277] In embodiments, a 3E10 antibody or antigen-binding fragment thereof described herein refers to CDR sequences having no more than two amino acid substitution relative to the parent 3E10 antibody optionally including a D31N amino acid substitution in the VH CDR1. Accordingly, in embodiments, a 3E10 antibody or antigen-binding fragment thereof includes a VL CDR 1 comprising an amino acid sequence having no more than two amino acid substitutions relative to 3E10-VL-CDR1 (SEQ ID NO:9), a VL CDR2 comprising an amino acid sequence having no more than two amino acid substitutions relative to 3E10-VL-CDR2 (SEQ ID NO: 10), a VL CDR3 comprising an amino acid sequence having no more than two amino acid substitutions relative to 3E10-VL-CDR3 (SEQ ID NO: 11), a heavy chain variable region (VH) CDR1comprising an amino acid sequence having no more than two amino acid substitutions relative to 3E10-VH-CDRla (SEQ ID NO: 16), a VH CDR2 comprising an amino acid sequence having no more than two amino acid substitutions relative to 3E10-VH-CDR2 (SEQ ID NON), and a VH CDR3 comprising an amino acid sequence having no more than two amino acid substitutions relative to 3E10-VH-CDR3 (SEQ ID NO: 5).

[0278] Other variants of a 3E10 antibody or antigen-binding fragment thereof are also known in the art, as disclosed for example, in Zack, et al., J. Immunol., 157(5):2082-8 (1996). For example, amino acid position 31 of the heavy chain variable region of 3E10 has been determined to be influential in the ability of the antibody and fragments thereof to penetrate nuclei and bind to DNA. A D31N mutation in CDR1 penetrates nuclei and binds DNA with much greater efficiency than the original antibody (Zack, et al., Immunology and Cell Biology, 72:513-520 (1994), Weisbart, et al., J. Autoimmun., 11, 539-546 (1998); Weisbart, Int. J. Oncol., 25, 1867-1873 (2004)). In embodiments, the antibody or antigen-binding fragment described herein has the D3 IN substitution.

[0279] Antibody-payload conjugates described herein can be prepared with any 3E10 antibodies or antigen-fragments thereof, or any humanized 3E10 antibodies or antigen-fragments thereof, disclosed in the prior art. See, for example WO 2015 / 106290, 2016 / 033324, WO 2019 / 018426, and WO 2019 / 018428 (each of which is specifically incorporated by reference herein, in its entirety).

[0280] In embodiments, an antibody -payload conjugate provided herein comprises a humanized 3E10 antibody. Generally, a humanized antibody has one or more amino acid residues introduced into it from a source that is non-human. These non-human amino acid residues are often referred to as “import” residues, which are typically taken from an “import” variable domain. Antibody humanization techniques generally involve the use of recombinant DNA technology to manipulate the DNA sequence encoding one or more polypeptide chains of an antibody molecule.

[0281] In embodiments, the disclosure provides humanized antibodies, or antigen-binding fragments thereof, that incorporate any combination of the humanized VL and VH sequences disclosed here, as well as VL and VH sequences having sequence identity thereto, e.g., having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, atleast about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identity to a VH or VL sequence described herein.

[0282] In embodiments, an antibody -pay load conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a heavy chain having the sequence of any one of SEQ ID NOs: l, 13, or 71-84. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 1. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 13. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO:71. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO:72. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO:73. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO:74. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO:75. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO:76. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO:77. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO:78. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO:79. In embodiments, an antibody-payload conjugate provided hereincomprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO:80. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO:81. In embodiments, an antibody -pay load conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 82. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO:83. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 84.

[0283] In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of any one of SEQ ID NOs:2, 14, 64-70, 103-112. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO:2. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 14. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 64. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 65. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO:66. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO:67. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO:68. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, orthree CDRs of a VH having the sequence of SEQ ID NO:69. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO:70. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 103. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 104. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 105. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 106. In embodiments, an antibody -pay load conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 107. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 108. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 109. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 110. In embodiments, an antibody -pay load conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 111. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 112.

[0284] In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a light chain having the sequence of any one of SEQ ID NOs:7, 20, or 91-102. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO:7. Inembodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO:20. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO:91. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO:92. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO:93. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO:94. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO:95. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO:96. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO:97. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO:98. In embodiments, an antibody -pay load conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO:99. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO: 100. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO: 101. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO: 102.

[0285] In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VL having the sequence of any one of SEQ ID NOs:8, 21, 85-90, or 113-121. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 8. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO:21. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO:85. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 86. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO:87. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO:88. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO:89. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NOVO. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 113. In embodiments, an antibody -pay load conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 114. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 115. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 116. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereofcomprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 117. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 118. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 119. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 120. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 121.

[0286] In embodiments, an antibody -pay load conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of any one of SEQ ID NOs: 122-137. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 122. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 123. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 124. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 125. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 126. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 127. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 128. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10antibody or antigen-binding fragment thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 129. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 130. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 131. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 132. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 133. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 134. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 135. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO:136. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 137.

[0287] In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to any one of SEQ ID NOs: l, 13, or 71-84. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 1. In embodiments, an antibody-payload conjugate provided hereincomprises a 3E10 antibody or antigen-binding fragment thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 13. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:71. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:72. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:73. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:74. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:75. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:76. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%identity to SEQ ID NO:77. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%>, at least about 96%, at least about 97%, at least about 98%>, at least about 99%, or 100%> identity to SEQ ID NO:78. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a heavy chain sequence having at least about 75%, at least about 80%>, at least about 85%, at least about 90%, at least about 95%o, at least about 96%, at least about 97%, at least about 98%>, at least about 99%, or 100%) identity to SEQ ID NO:79. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a heavy chain sequence having at least about 75%>, at least about 80%>, at least about 85%>, at least about 90%>, at least about 95%o, at least about 96%, at least about 97%, at least about 98%>, at least about 99%, or 100%> identity to SEQ ID NO:80. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a heavy chain sequence having at least about 75%, at least about 80%>, at least about 85%, at least about 90%, at least about 95%o, at least about 96%, at least about 97%, at least about 98%>, at least about 99%, or 100%) identity to SEQ ID NO:81. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a heavy chain sequence having at least about 75%, at least about 80%>, at least about 85%, at least about 90%, at least about 95%o, at least about 96%, at least about 97%o, at least about 98%>, at least about 99%, or 100%) identity to SEQ ID NO:82. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a heavy chain sequence having at least about 75%, at least about 80%>, at least about 85%, at least about 90%, at least about 95%o, at least about 96%, at least about 97%, at least about 98%>, at least about 99%, or 100%> identity to SEQ ID NO:83. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a heavy chain sequence having at least about 75%, at least about 80%>, at least about 85%, at least about 90%, at least about 95%o, at least about 96%, at least about 97%>, at least about 98%>, at least about 99%, or 100%> identity to SEQ ID NO:84.

[0288] In embodiments, an antibody -pay load conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a light chain sequence having at least about75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to any one of SEQ ID NOs:7, 20, or 91-102. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:7. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:20. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:91. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:92. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:93. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:94. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:95. In embodiments, an antibody -payload conjugate provided hereincomprises a 3E10 antibody or antigen-binding fragment thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:96. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:97. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:98. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:99. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 100. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 101. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 102.

[0289] In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, atleast about 97%, at least about 98%, at least about 99%, or 100% identity to any one of SEQ ID NOs: 2, 14, 64-70, or 103-112. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:2. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 14. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:64. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:65. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:66. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:67. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:68. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:69. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:70. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 103. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 104. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 105. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 106. In embodiments, an antibody -pay load conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 107. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 108. In embodiments, an antibody -pay load conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 109. In embodiments, an antibody -pay load conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 110. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:111. In embodiments, an antibody -pay load conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 112.

[0290] In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to any one of SEQ ID NOs: 8, 21, 85-90, or 113-121. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:8. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:21. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:85. In embodiments, an antibody -payload conjugate provided herein comprises a 3E10 antibody or antigen-bindingfragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 86. In embodiments, an antibody- payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:87. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:88. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:89. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NOVO. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 113. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 114. In embodiments, an antibody -pay load conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 115. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody orantigen-binding fragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 116. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 117. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 118. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:119. In embodiments, an antibody -pay load conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 120. In embodiments, an antibody-payload conjugate provided herein comprises a 3E10 antibody or antigen-binding fragment thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 121.

[0291] In embodiments, an antibody -payload conjugate provided herein comprises a humanized 3E10 antibody, or antigen-binding fragment thereof, comprising a light chain variable domain (3E10-VL) comprising an amino acid sequence that is at least about 97% identical to an amino acid sequence selected from the group consisting of 3E10-VL-hl (SEQ ID NO:85), 3E10- VL-h2 (SEQ ID NO:86), 3E10-VL-h3 (SEQ ID NO:87), 3E10-VL-h4 (SEQ ID NO:88), 3E10- VL-h5 (SEQ ID NO:89), and 3E10-VL-h6 (SEQ ID NOVO) and a heavy chain variable domain (3E10-VH) comprising an amino acid sequence that is at least about 95% identical to an amino acid sequence selected from the group consisting of 3E10-VH-hl (SEQ ID NO:64), 3E10-VH-h2(SEQ ID NO:65), 3E10-VH-h3 (SEQ ID NO:66), 3E10-VH-h4 (SEQ ID NO:67), 3E10-VH-h5 (SEQ ID NO:68), 3E10-VH-h6 (SEQ ID NO:69), and 3E10-VH-h7 (SEQ ID NO:70).

[0292] In embodiments, the sequence of the 3E10-VL is at least about 97% identical to 3E10-VL-hl (SEQ ID NO:85). In embodiments, the sequence of the 3E10-VL is at least about 98% identical to 3E10-VL-hl (SEQ ID NO:85). In embodiments, the sequence of the 3E10-VL is at least about 99% identical to 3E10-VL-hl (SEQ ID NO:85). In embodiments, the sequence of the 3E10-VL is 3E10-VL-hl (SEQ ID NO:85).

[0293] In embodiments, the sequence of the 3E10-VL is at least about 97% identical to 3E10-VL-h2 (SEQ ID NO:86). In embodiments, the sequence of the 3E10-VL is at least about 98% identical to 3E10-VL-h2 (SEQ ID NO:86). In embodiments, the sequence of the 3E10-VL is at least about 99% identical to 3E10-VL-h2 (SEQ ID NO:86). In embodiments, the sequence of the 3E10-VL is 3E10-VL-h2 (SEQ ID NO:86).

[0294] In embodiments, the sequence of the 3E10-VL is at least about 97% identical to 3E10-VL-h3 (SEQ ID NO:87). In embodiments, the sequence of the 3E10-VL is at least about 98% identical to 3E10-VL-h3 (SEQ ID NO:87). In embodiments, the sequence of the 3E10-VL is at least about 99% identical to 3E10-VL-h3 (SEQ ID NO:87). In embodiments, the sequence of the 3E10-VL is 3E10-VL-h3 (SEQ ID NO:87).

[0295] In embodiments, the sequence of the 3E10-VL is at least about 97% identical to 3E10-VL-h4 (SEQ ID NO:88). In embodiments, the sequence of the 3E10-VL is at least about 98% identical to 3E10-VL-h4 (SEQ ID NO:88). In embodiments, the sequence of the 3E10-VL is at least about 99% identical to 3E10-VL-h4 (SEQ ID NO:88). In embodiments, the sequence of the 3E10-VL is 3E10-VL-h4 (SEQ ID NO:88).

[0296] In embodiments, the sequence of the 3E10-VL is at least about 97% identical to 3E10-VL-h5 (SEQ ID NO:89). In embodiments, the sequence of the 3E10-VL is at least about 98% identical to 3E10-VL-h5 (SEQ ID NO:89). In embodiments, the sequence of the 3E10-VL is at least about 99% identical to 3E10-VL-h5 (SEQ ID NO:89). In embodiments, the sequence of the 3E10-VL is 3E10-VL-h5 (SEQ ID NO:89).

[0297] In embodiments, the sequence of the 3E10-VL is at least about 97% identical to 3E10-VL-h6 (SEQ ID NO:90). In embodiments, the sequence of the 3E10-VL is at least about98% identical to 3E10-VL-h6 (SEQ ID NO:90). In embodiments, the sequence of the 3E10-VL is at least about 99% identical to 3E10-VL-h6 (SEQ ID NO:90). In embodiments, the sequence of the 3E10-VL is 3E10-VL-h6 (SEQ ID NO:90).

[0298] In embodiments, the sequence of the 3E10-VH is at least about 95% identical to 3E10-VH-hl (SEQ ID NO:64). In embodiments, the sequence of the 3E10-VH is at least about 96% identical to 3E10-VH-hl (SEQ ID NO:64). In embodiments, the sequence of the 3E10-VH is at least about 97% identical to 3E10-VH-hl (SEQ ID NO:64). In embodiments, the sequence of the 3E10-VH is at least about 98% identical to 3E10-VH-hl (SEQ ID NO:64). In embodiments, the sequence of the 3E10-VH is at least about 99% identical to 3E10-VH-hl (SEQ ID NO:64). In embodiments, the sequence of the 3E10-VH is 3E10-VH-hl (SEQ ID NO:64).

[0299] In embodiments, the sequence of the 3E10-VH is at least about 95% identical to 3E10-VH-h2 (SEQ ID NO:65). In embodiments, the sequence of the 3E10-VH is at least about 96% identical to 3E10-VH-h2 (SEQ ID NO:65). In embodiments, the sequence of the 3E10-VH is at least about 97% identical to 3E10-VH-h2 (SEQ ID NO:65). In embodiments, the sequence of the 3E10-VH is at least about 98% identical to 3E10-VH-h2 (SEQ ID NO:65). In embodiments, the sequence of the 3E10-VH is at least about 99% identical to 3E10-VH-h2 (SEQ ID NO:65). In embodiments, the sequence of the 3E10-VH is 3E10-VH-h2 (SEQ ID NO:65).

[0300] In embodiments, the sequence of the 3E10-VH is at least about 95% identical to 3E10-VH-h3 (SEQ ID NO:66). In embodiments, the sequence of the 3E10-VH is at least about 96% identical to 3E10-VH-h3 (SEQ ID NO:66). In embodiments, the sequence of the 3E10-VH is at least about 97% identical to 3E10-VH-h3 (SEQ ID NO:66). In embodiments, the sequence of the 3E10-VH is at least about 98% identical to 3E10-VH-h3 (SEQ ID NO:66). In embodiments, the sequence of the 3E10-VH is at least about 99% identical to 3E10-VH-h3 (SEQ ID NO:66). In embodiments, the sequence of the 3E10-VH is 3E10-VH-h3 (SEQ ID NO:66).

[0301] In embodiments, the sequence of the 3E10-VH is at least about 95% identical to 3E10-VH-h4 (SEQ ID NO:67). In embodiments, the sequence of the 3E10-VH is at least about 96% identical to 3E10-VH-h4 (SEQ ID NO:67). In embodiments, the sequence of the 3E10-VH is at least about 97% identical to 3E10-VH-h4 (SEQ ID NO:67). In embodiments, the sequence of the 3E10-VH is at least about 98% identical to 3E10-VH-h4 (SEQ ID NO:67). In embodiments,the sequence of the 3E10-VH is at least about 99% identical to 3E10-VH-h4 (SEQ ID NO:67). In embodiments, the sequence of the 3E10-VH is 3E10-VH-h4 (SEQ ID NO:67).

[0302] In embodiments, the sequence of the 3E10-VH is at least about 95% identical to 3E10-VH-h5 (SEQ ID NO:68). In embodiments, the sequence of the 3E10-VH is at least about 96% identical to 3E10-VH-h5 (SEQ ID NO:68). In embodiments, the sequence of the 3E10-VH is at least about 97% identical to 3E10-VH-h5 (SEQ ID NO:68). In embodiments, the sequence of the 3E10-VH is at least about 98% identical to 3E10-VH-h5 (SEQ ID NO:68). In embodiments, the sequence of the 3E10-VH is at least about 99% identical to 3E10-VH-h5 (SEQ ID NO:68). In embodiments, the sequence of the 3E10-VH is 3E10-VH-h5 (SEQ ID NO:68).

[0303] In embodiments, the sequence of the 3E10-VH is at least about 95% identical to 3E10-VH-h6 (SEQ ID NO:69). In embodiments, the sequence of the 3E10-VH is at least about 96% identical to 3E10-VH-h6 (SEQ ID NO:69). In embodiments, the sequence of the 3E10-VH is at least about 97% identical to 3E10-VH-h6 (SEQ ID NO:69). In embodiments, the sequence of the 3E10-VH is at least about 98% identical to 3E10-VH-h6 (SEQ ID NO:69). In embodiments, the sequence of the 3E10-VH is at least about 99% identical to 3E10-VH-h6 (SEQ ID NO:69). In embodiments, the sequence of the 3E10-VH is 3E10-VH-h6 (SEQ ID NO:69).

[0304] In embodiments, the sequence of the 3E10-VH is at least about 95% identical to 3E10-VH-h7 (SEQ ID NO:70). In embodiments, the sequence of the 3E10-VH is at least about 96% identical to 3E10-VH-h7 (SEQ ID NO:70). In embodiments, the sequence of the 3E10-VH is at least about 97% identical to 3E10-VH-h7 (SEQ ID NO:70). In embodiments, the sequence of the 3E10-VH is at least about 98% identical to 3E10-VH-h7 (SEQ ID NO:70). In embodiments, the sequence of the 3E10-VH is at least about 99% identical to 3E10-VH-h7 (SEQ ID NO:70). In embodiments, the sequence of the 3E10-VH is 3E10-VH-h7 (SEQ ID NO:70).

[0305] In embodiments, an antibody -payload conjugate comprising a humanized 3E10 antibody, or antigen-binding fragment thereof, described herein includes a light chain (3E10-LC) comprising an amino acid sequence that is at least about 95% identical to an amino acid sequence selected from the group consisting of 3E10-LC-hlm (SEQ ID NO:91), 3E10-LC-h2m (SEQ ID NO:92), 3E10-LC-h3m (SEQ ID NO:93), 3E10-LC-h4m (SEQ ID NO:94), 3E10-LC-h5m (SEQ ID NO:95), and 3E10-LC-h6m (SEQ ID NO:96) and a heavy chain (3E10-HC) comprising an amino acid sequence that is at least about 95% identical to an amino acid sequence selected fromthe group consisting of 3E10-HC-hlm (SEQ ID NO:71), 3E10-HC-h2m (SEQ ID NO: 72), 3E10- HC-h3m (SEQ ID NO:73), 3E10-HC-h4m (SEQ ID NO:74), 3E10-HC-h5m (SEQ ID NO:75), 3E10-HC-h6m (SEQ ID NO: 76), and 3E10-HC-h7m (SEQ ID NO: 77).

[0306] In embodiments, the sequence of the 3E10-LC is at least about 95% identical to 3E10-LC-hlm (SEQ ID NO:91). In embodiments, the sequence of the 3E10-LC is at least about 96% identical to 3E10-LC-hlm (SEQ ID NO:91). In embodiments, the sequence of the 3E10-LC is at least about 97% identical to 3E10-LC-hlm (SEQ ID NO:91). In embodiments, the sequence of the 3E10-LC is at least about 98% identical to 3E10-LC-hlm (SEQ IDN0:91). In embodiments, the sequence of the 3E10-LC is at least about 99% identical to 3E10-LC-hlm (SEQ ID NO:91). In embodiments, the sequence of the 3E10-LC is 3E10-LC-hlm (SEQ ID NO:91).

[0307] In embodiments, the sequence of the 3E10-LC is at least about 95% identical to 3E10-LC-h2m (SEQ ID NO:92). In embodiments, the sequence of the 3E10-LC is at least about 96% identical to 3E10-LC-h2m (SEQ ID NO:92). In embodiments, the sequence of the 3E10-LC is at least about 97% identical to 3E10-LC-h2m (SEQ ID NO:92). In embodiments, the sequence of the 3E10-LC is at least about 98% identical to 3E10-LC-h2m (SEQ IDNO:92). In embodiments, the sequence of the 3E10-LC is at least about 99% identical to 3E10-LC-h2m (SEQ ID NO:92). In embodiments, the sequence of the 3E10-LC is 3E10-LC-h2m (SEQ ID NO:92).

[0308] In embodiments, the sequence of the 3E10-LC is at least about 95% identical to 3E10-LC-h3m (SEQ ID NO:93). In embodiments, the sequence of the 3E10-LC is at least about 96% identical to 3E10-LC-h3m (SEQ ID NO:93). In embodiments, the sequence of the 3E10-LC is at least about 97% identical to 3E10-LC-h3m (SEQ ID NO:93). In embodiments, the sequence of the 3E10-LC is at least about 98% identical to 3E10-LC-h3m (SEQ ID NO:93). In embodiments, the sequence of the 3E10-LC is at least about 99% identical to 3E10-LC-h3m (SEQ ID NO:93). In embodiments, the sequence of the 3E10-LC is 3E10-LC-h3m (SEQ ID NO:93).

[0309] In embodiments, the sequence of the 3E10-LC is at least about 95% identical to 3E10-LC-h4m (SEQ ID NO:94). In embodiments, the sequence of the 3E10-LC is at least about 96% identical to 3E10-LC-h4m (SEQ ID NO:94). In embodiments, the sequence of the 3E10-LC is at least about 97% identical to 3E10-LC-h4m (SEQ ID NO:94). In embodiments, the sequence of the 3E10-LC is at least about 98% identical to 3E10-LC-h4m (SEQ ID NO:94). In embodiments,the sequence of the 3E10-LC is at least about 99% identical to 3E10-LC-h4m (SEQ ID NO:94). In embodiments, the sequence of the 3E10-LC is 3E10-LC-h4m (SEQ ID NO:94).

[0310] In embodiments, the sequence of the 3E10-LC is at least about 95% identical to 3E10-LC-h5m (SEQ ID NO:95). In embodiments, the sequence of the 3E10-LC is at least about 96% identical to 3E10-LC-h5m (SEQ ID NO:95). In embodiments, the sequence of the 3E10-LC is at least about 97% identical to 3E10-LC-h5m (SEQ ID NO:95). In embodiments, the sequence of the 3E10-LC is at least about 98% identical to 3E10-LC-h5m (SEQ ID NO:95). In embodiments, the sequence of the 3E10-LC is at least about 99% identical to 3E10-LC-h5m (SEQ ID NO:95). In embodiments, the sequence of the 3E10-LC is 3E10-LC-h5m (SEQ ID NO:95).

[0311] In embodiments, the sequence of the 3E10-LC is at least about 95% identical to 3E10-LC-h6m (SEQ ID NO:96). In embodiments, the sequence of the 3E10-LC is at least about 96% identical to 3E10-LC-h6m (SEQ ID NO:96). In embodiments, the sequence of the 3E10-LC is at least about 97% identical to 3E10-LC-h6m (SEQ ID NO:96). In embodiments, the sequence of the 3E10-LC is at least about 98% identical to 3E10-LC-h6m (SEQ IDNO:96). In embodiments, the sequence of the 3E10-LC is at least about 99% identical to 3E10-LC-h6m (SEQ ID NO:96). In embodiments, the sequence of the 3E10-LC is 3E10-LC-h6m (SEQ ID NO:96).

[0312] In embodiments, the sequence of the 3E10-HC is at least about 95% identical to 3E10-HC-hlm (SEQ ID NO:71). In embodiments, the sequence of the 3E10-HC is at least about 96% identical to 3E10-HC-hlm (SEQ ID NO:71). In embodiments, the sequence of the 3E10-HC is at least about 97% identical to 3E10-HC-hlm (SEQ ID NO:71). In embodiments, the sequence of the 3E10-HC is at least about 98% identical to 3E10-HC-hlm (SEQ ID NO:71). In embodiments, the sequence of the 3E10-HC is at least about 99% identical to 3E10-HC-hlm (SEQ ID NO:71). In embodiments, the sequence of the 3E10-HC is 3E10-HC-hlm (SEQ ID NO:71).

[0313] In embodiments, the sequence of the 3E10-HC is at least about 95% identical to 3E10-HC-h2m (SEQ ID NO:72). In embodiments, the sequence of the 3E10-HC is at least about 96% identical to 3E10-HC-h2m (SEQ ID NO:72). In embodiments, the sequence of the 3E10-HC is at least about 97% identical to 3E10-HC-h2m (SEQ ID NO:72). In embodiments, the sequence of the 3E10-HC is at least about 98% identical to 3E10-HC-h2m (SEQ ID NO:72). In embodiments, the sequence of the 3E10-HC is at least about 99% identical to 3E10-HC-h2m (SEQ ID NO:72). In embodiments, the sequence of the 3E10-HC is 3E10-HC-h2m (SEQ ID NO:72).

[0314] In embodiments, the sequence of the 3E10-HC is at least about 95% identical to 3E10-HC-h3m (SEQ ID NO:73). In embodiments, the sequence of the 3E10-HC is at least about 96% identical to 3E10-HC-h3m (SEQ ID NO:73). In embodiments, the sequence of the 3E10-HC is at least about 97% identical to 3E10-HC-h3m (SEQ ID NO:73). In embodiments, the sequence of the 3E10-HC is at least about 98% identical to 3E10-HC-h3m (SEQ ID NO:73). In embodiments, the sequence of the 3E10-HC is at least about 99% identical to 3E10-HC-h3m (SEQ ID NO:73). In embodiments, the sequence of the 3E10-HC is 3E10-HC-h3m (SEQ ID NO:73).

[0315] In embodiments, the sequence of the 3E10-HC is at least about 95% identical to 3E10-HC-h4m (SEQ ID NO:74). In embodiments, the sequence of the 3E10-HC is at least about 96% identical to 3E10-HC-h4m (SEQ ID NO:74). In embodiments, the sequence of the 3E10-HC is at least about 97% identical to 3E10-HC-h4m (SEQ ID NO:74). In embodiments, the sequence of the 3E10-HC is at least about 98% identical to 3E10-HC-h4m (SEQ ID NO:74). In embodiments, the sequence of the 3E10-HC is at least about 99% identical to 3E10-HC-h4m (SEQ ID NO:74). In embodiments, the sequence of the 3E10-HC is 3E10-HC-h4m (SEQ ID NO:74).

[0316] In embodiments, the sequence of the 3E10-HC is at least about 95% identical to 3E10-HC-h5m (SEQ ID NO:75). In embodiments, the sequence of the 3E10-HC is at least about 96% identical to 3E10-HC-h5m (SEQ ID NO:75). In embodiments, the sequence of the 3E10-HC is at least about 97% identical to 3E10-HC-h5m (SEQ ID NO:75). In embodiments, the sequence of the 3E10-HC is at least about 98% identical to 3E10-HC-h5m (SEQ ID NO:75). In embodiments, the sequence of the 3E10-HC is at least about 99% identical to 3E10-HC-h5m (SEQ ID NO:75). In embodiments, the sequence of the 3E10-HC is 3E10-HC-h5m (SEQ ID NO:75).

[0317] In embodiments, the sequence of the 3E10-HC is at least about 95% identical to 3E10-HC-h6m (SEQ ID NO:76). In embodiments, the sequence of the 3E10-HC is at least about 96% identical to 3E10-HC-h6m (SEQ ID NO:76). In embodiments, the sequence of the 3E10-HC is at least about 97% identical to 3E10-HC-h6m (SEQ ID NO:76). In embodiments, the sequence of the 3E10-HC is at least about 98% identical to 3E10-HC-h6m (SEQ ID NO:76). In embodiments, the sequence of the 3E10-HC is at least about 99% identical to 3E10-HC-h6m (SEQ ID NO:76). In embodiments, the sequence of the 3E10-HC is 3E10-HC-h6m (SEQ ID NO:76).

[0318] In embodiments, the sequence of the 3E10-HC is at least about 95% identical to 3E10-HC-h7m (SEQ ID NO:77). In embodiments, the sequence of the 3E10-HC is at least about96% identical to 3E10-HC-h7m (SEQ ID NO:77). In embodiments, the sequence of the 3E10-HC is at least about 97% identical to 3E10-HC-h7m (SEQ ID NO:77). In embodiments, the sequence of the 3E10-HC is at least about 98% identical to 3E10-HC-h7m (SEQ ID NO:77). In embodiments, the sequence of the 3E10-HC is at least about 99% identical to 3E10-HC-h7m (SEQ ID NO:77). In embodiments, the sequence of the 3E10-HC is 3E10-HC-h7m (SEQ ID NO:77).

[0319] In embodiments, an antibody-payload conjugate provided herein comprises a humanized 3E10 antibody, or antigen-binding fragment thereof, comprising a light chain (3E10- LC) comprising an amino acid sequence that is at least about 95% identical to an amino acid sequence selected from the group consisting of 3E10-LC-hl (SEQ ID NO:97), 3E10-LC-h2 (SEQ ID NO:98), 3E10-LC-h3 (SEQ ID NO:99), 3E10-LC-h4 (SEQ ID NO: 100), 3E10-LC-h5 (SEQ ID NO: 101), and 3E10-LC-h6 (SEQ ID NO: 102) and a heavy chain (3E10-HC) comprising an amino acid sequence that is at least about 95% identical to an amino acid sequence selected from the group consisting of 3E10-HC-hl (SEQ ID NO:78), 3E10-HC-h2 (SEQ ID NO:79), 3E10-HC- h3 (SEQ ID NO:80), 3E10-HC-h4 (SEQ ID NO:81), 3E10-HC-h5 (SEQ ID NO:82), 3E10-HC-h6 (SEQ ID NO:83), and 3E10-HC-117 (SEQ ID NO:84).

[0320] In embodiments, the sequence of the 3E10-LC is at least about 95% identical to 3E10-LC-hl (SEQ ID NO:97. In embodiments, the sequence of the 3E10-LC is at least about 96% identical to 3E10-LC-hl (SEQ ID NO:97). In embodiments, the sequence of the 3E10-LC is at least about 97% identical to 3E10-LC-hl (SEQ ID NO:97). In embodiments, the sequence of the 3E10-LC is at least about 98% identical to 3E10-LC-hl (SEQ ID NO:97). In embodiments, the sequence of the 3E10-LC is at least about 99% identical to 3E10-LC-hl (SEQ ID NO:97). In embodiments, the sequence of the 3E10-LC is 3E10-LC-hl (SEQ ID NO:97).

[0321] In embodiments, the sequence of the 3E10-LC is at least about 95% identical to 3E10-LC-h2 (SEQ ID NO:98). In embodiments, the sequence of the 3E10-LC is at least about 96% identical to 3E10-LC-h2 (SEQ ID NO:98). In embodiments, the sequence of the 3E10-LC is at least about 97% identical to 3E10-LC-h2 (SEQ ID NO:98). In embodiments, the sequence of the 3E10-LC is at least about 98% identical to 3E10-LC-h2 (SEQ ID NO:98). In embodiments, the sequence of the 3E10-LC is at least about 99% identical to 3E10-LC-h2 (SEQ ID NO:98). In embodiments, the sequence of the 3E10-LC is 3E10-LC-h2 (SEQ ID NO:98).

[0322] In embodiments, the sequence of the 3E10-LC is at least about 95% identical to 3E10-LC-h3 (SEQ ID NO:99). In embodiments, the sequence of the 3E10-LC is at least about 96% identical to 3E10-LC-h3 (SEQ ID NO:99). In embodiments, the sequence of the 3E10-LC is at least about 97% identical to 3E10-LC-h3 (SEQ ID NO:99). In embodiments, the sequence of the 3E10-LC is at least about 98% identical to 3E10-LC-h3 (SEQ ID NO:99). In embodiments, the sequence of the 3E10-LC is at least about 99% identical to 3E10-LC-h3 (SEQ ID NO:99). In embodiments, the sequence of the 3E10-LC is 3E10-LC-h3 (SEQ ID NO:99).

[0323] In embodiments, the sequence of the 3E10-LC is at least about 95% identical to 3E10-LC-h4 (SEQ ID NO: 100). In embodiments, the sequence of the 3E10-LC is at least about 96% identical to 3E10-LC-h4 (SEQ ID NO: 100). In embodiments, the sequence of the 3E10-LC is at least about 97% identical to 3E10-LC-h4 (SEQ ID NO: 100). In embodiments, the sequence of the 3E10-LC is at least about 98% identical to 3E10-LC-h4 (SEQ ID NO: 100). In embodiments, the sequence of the 3E10-LC is at least about 99% identical to 3E10-LC-h4 (SEQ ID NO: 100). In embodiments, the sequence of the 3E10-LC is 3E10-LC-h4 (SEQ ID NO: 100).

[0324] In embodiments, the sequence of the 3E10-LC is at least about 95% identical to 3E10-LC-h5 (SEQ ID NO: 101). In embodiments, the sequence of the 3E10-LC is at least about 96% identical to 3E10-LC-h5 (SEQ ID NO: 101). In embodiments, the sequence of the 3E10-LC is at least about 97% identical to 3E10-LC-h5 (SEQ ID NO: 101). In embodiments, the sequence of the 3E10-LC is at least about 98% identical to 3E10-LC-h5 (SEQ ID NO: 101). In embodiments, the sequence of the 3E10-LC is at least about 99% identical to 3E10-LC-h5 (SEQ ID NO: 101). In embodiments, the sequence of the 3E10-LC is 3E10-LC-h5 (SEQ ID NO: 101).

[0325] In embodiments, the sequence of the 3E10-LC is at least about 95% identical to 3E10-LC-h6 (SEQ ID NO: 102). In embodiments, the sequence of the 3E10-LC is at least about 96% identical to 3E10-LC-h6 (SEQ ID NO: 102). In embodiments, the sequence of the 3E10-LC is at least about 97% identical to 3E10-LC-h6 (SEQ ID NO: 102). In embodiments, the sequence of the 3E10-LC is at least about 98% identical to 3E10-LC-h6 (SEQ ID NO:102). In embodiments, the sequence of the 3E10-LC is at least about 99% identical to 3E10-LC-h6 (SEQ ID NO: 102). In embodiments, the sequence of the 3E10-LC is 3E10-LC-h6 (SEQ ID NO: 102).

[0326] In embodiments, the sequence of the 3E10-HC is at least about 95% identical to 3E10-HC-hl (SEQ ID NO:78). In embodiments, the sequence of the 3E10-HC is at least about96% identical to 3E10-HC-hl (SEQ ID NO:78). In embodiments, the sequence of the 3E10-HC is at least about 97% identical to 3E10-HC-hl (SEQ ID NO:78). In embodiments, the sequence of the 3E10-HC is at least about 98% identical to 3E10-HC-hl (SEQ ID NO:78). In embodiments, the sequence of the 3E10-HC is at least about 99% identical to 3E10-HC-hl (SEQ ID NO:78). In embodiments, the sequence of the 3E10-HC is 3E10-HC-hl (SEQ ID NO:78).

[0327] In embodiments, the sequence of the 3E10-HC is at least about 95% identical to 3E10-HC-h2 (SEQ ID NO:79). In embodiments, the sequence of the 3E10-HC is at least about 96% identical to 3E10-HC-h2 (SEQ ID NO:79). In embodiments, the sequence of the 3E10-HC is at least about 97% identical to 3E10-HC-h2 (SEQ ID NO:79). In embodiments, the sequence of the 3E10-HC is at least about 98% identical to 3E10-HC-h2 (SEQ ID NO:79). In embodiments, the sequence of the 3E10-HC is at least about 99% identical to 3E10-HC-h2 (SEQ ID NO:79). In embodiments, the sequence of the 3E10-HC is 3E10-HC-h2 (SEQ ID NO:79).

[0328] In embodiments, the sequence of the 3E10-HC is at least about 95% identical to 3E10-HC-h3 (SEQ ID NO:80). In embodiments, the sequence of the 3E10-HC is at least about 96% identical to 3E10-HC-h3 (SEQ ID NO:80). In embodiments, the sequence of the 3E10-HC is at least about 97% identical to 3E10-HC-113 (SEQ ID NO:80). In embodiments, the sequence of the 3E10-HC is at least about 98% identical to 3E10-HC-h3 (SEQ ID NO:80). In embodiments, the sequence of the 3E10-HC is at least about 99% identical to 3E10-HC-h3 (SEQ ID NO:80). In embodiments, the sequence of the 3E10-HC is 3E10-HC-h3 (SEQ ID NO:80).

[0329] In embodiments, the sequence of the 3E10-HC is at least about 95% identical to 3E10-HC-h4 (SEQ ID NO:81). In embodiments, the sequence of the 3E10-HC is at least about 96% identical to 3E10-HC-h4 (SEQ ID NO:81). In embodiments, the sequence of the 3E10-HC is at least about 97% identical to 3E10-HC-h4 (SEQ ID NO:81). In embodiments, the sequence of the 3E10-HC is at least about 98% identical to 3E10-HC-h4 (SEQ ID NO:81). In embodiments, the sequence of the 3E10-HC is at least about 99% identical to 3E10-HC-h4 (SEQ ID NO:81). In embodiments, the sequence of the 3E10-HC is 3E10-HC-h4 (SEQ ID NO:81).

[0330] In embodiments, the sequence of the 3E10-HC is at least about 95% identical to 3E10-HC-h5 (SEQ ID NO:82). In embodiments, the sequence of the 3E10-HC is at least about 96% identical to 3E10-HC-h5 (SEQ ID NO:82). In embodiments, the sequence of the 3E10-HC is at least about 97% identical to 3E10-HC-h5 (SEQ ID NO:82). In embodiments, the sequence ofthe 3E10-HC is at least about 98% identical to 3E10-HC-h5 (SEQ ID NO:82). In embodiments, the sequence of the 3E10-HC is at least about 99% identical to 3E10-HC-h5 (SEQ ID NO:82). In embodiments, the sequence of the 3E10-HC is 3E10-HC-h5 (SEQ ID NO:82).

[0331] In embodiments, the sequence of the 3E10-HC is at least about 95% identical to 3E10-HC-h6 (SEQ ID NO:83). In embodiments, the sequence of the 3E10-HC is at least about 96% identical to 3E10-HC-h6 (SEQ ID NO:83). In embodiments, the sequence of the 3E10-HC is at least about 97% identical to 3E10-HC-116 (SEQ ID NO:83). In embodiments, the sequence of the 3E10-HC is at least about 98% identical to 3E10-HC-h6 (SEQ ID NO:83). In embodiments, the sequence of the 3E10-HC is at least about 99% identical to 3E10-HC-h6 (SEQ ID NO:83). In some aspects, the sequence of the 3E10-HC is 3E10-HC-h6 (SEQ ID NO:83).

[0332] In embodiments, the sequence of the 3E10-HC is at least about 95% identical to 3E10-HC-h7 (SEQ ID NO:84). In embodiments, the sequence of the 3E10-HC is at least about 96% identical to 3E10-HC-h7 (SEQ ID NO:84). In embodiments, the sequence of the 3E10-HC is at least about 97% identical to 3E10-HC-h7 (SEQ ID NO: 84). In embodiments, the sequence of the 3E10-HC is at least about 98% identical to 3E10-HC-h7 (SEQ ID NO:84). In embodiments, the sequence of the 3E10-HC is at least about 99% identical to 3E10-HC-h7 (SEQ ID NO:84). In embodiments, the sequence of the 3E10-HC is 3E10-HC-h7 (SEQ ID NO:84).

[0333] In embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof described herein has CDR sequences corresponding to those in the parent 3E10 antibody, optionally including a D31N amino acid substitution in the VH CDR1. Accordingly, in embodiments, a humanized 3E10 antibody or antigen-binding fragment thereof includes a light chain variable domain (VL) complementarity determining region (CDR) 1 comprising the amino acid sequence of 3E10-VL-CDR1 (SEQ ID NO: 9), a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2 (SEQ ID NO: 10), a VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3 (SEQ ID NO: 11), a heavy chain variable domain (VH) CDR1 comprising the amino acid sequence of 3E10-VH-CDRla (SEQ ID NO: 16), a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2 (SEQ ID NO: 4), and a VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3 (SEQ ID NO: 5).

[0334] In embodiments, an antibody -payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof described herein includes CDR sequences from avariant humanized 3E10 antibody that includes a D31N amino acid substitution in the VH CDR1 (SEQ ID NO: 15).

[0335] In embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof described herein includes a set of complementarity determining regions (CDRs) collectively having no more than seven amino acid substitutions, relative to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11), 3E1O-VH-CDR1 D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NON), and 3E10-VH-CDR3 (SEQ ID NO:5).

[0336] In embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof described herein includes a set of complementarity determining regions (CDRs) collectively having no more than ten amino acid substitutions, relative to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO:9), 3E10- VL-CDR2 (SEQ ID NONO), 3E10-VL-CDR3 (SEQ ID NON 1), 3E10-VH-CDR1 D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NON), and 3E10-VH-CDR3 (SEQ ID NON).

[0337] In embodiments, an antibody -payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof described herein includes a set of complementarity determining regions (CDRs) collectively having no more than nine amino acid substitutions, relative to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NONO), 3E10-VL-CDR3 (SEQ ID NO: 11), 3E1O-VH-CDR1 D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NON), and 3E10-VH-CDR3 (SEQ ID NON).

[0338] In embodiments, an antibody -payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof described herein includes a set of complementarity determining regions (CDRs) collectively having no more than eight amino acid substitutions, relative to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11), 3E1O-VH-CDR1 D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NON), and 3E10-VH-CDR3 (SEQ ID NON).

[0339] In embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof described herein includes a set of complementarity determining regions (CDRs) collectively having no more than seven amino acid substitutions, relative to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9),3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO:11), 3E1O-VH-CDR1 D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NON), and 3E10-VH-CDR3 (SEQ ID NO:5).

[0340] In embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof described herein includes a set of complementarity determining regions (CDRs) collectively having no more than six amino acid substitutions, relative to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NON), 3E10- VL-CDR2 (SEQ ID NONO), 3E10-VL-CDR3 (SEQ ID NO:11), 3E10-VH-CDR1 D31N (SEQ ID NO:15), 3E10-VH-CDR2 (SEQ ID NON), and 3E10-VH-CDR3 (SEQ ID NON).

[0341] In embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof described herein includes a set of complementarity determining regions (CDRs) collectively having no more than five amino acid substitutions, relative to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NON), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11), 3E1O-VH-CDR1 D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NON), and 3E10-VH-CDR3 (SEQ ID NO:5).

[0342] In embodiments, an antibody -payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof described herein includes a set of complementarity determining regions (CDRs) collectively having no more than four amino acid substitutions, relative to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NON), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11), 3E1O-VH-CDR1 D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NON), and 3E10-VH-CDR3 (SEQ ID NON).

[0343] In embodiments, an antibody -payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof described herein includes a set of complementarity determining regions (CDRs) collectively having no more than three amino acid substitutions, relative to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NON), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11), 3E1O-VH-CDR1 D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NON), and 3E10-VH-CDR3 (SEQ ID NON).

[0344] In embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof described herein includes a set of complementarity determining regions (CDRs) collectively having no more than two amino acid substitutions, relative to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NON),3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO:11), 3E1O-VH-CDR1 D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NON), and 3E10-VH-CDR3 (SEQ ID NO:5).

[0345] In embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof described herein includes a set of complementarity determining regions (CDRs) collectively having no more than one amino acid substitution, relative to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NON), 3E10- VL-CDR2 (SEQ ID NONO), 3E10-VL-CDR3 (SEQ ID NO:11), 3E10-VH-CDR1 D31N (SEQ ID NO:15), 3E10-VH-CDR2 (SEQ ID NON), and 3E10-VH-CDR3 (SEQ ID NO:5).

[0346] Accordingly, in embodiments, an antibody-payload conjugate described herein can comprise a humanized 3E10 antibody or antigen-binding fragment thereof includes a light chain variable domain (VL) complementarity determining region (CDR) 1 comprising the amino acid sequence of 3E10-VL-CDR1 (SEQ ID NO: 9), a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2 (SEQ ID NO: 10), a VL CDR3 comprising the amino acid sequence of 3E10- VL-CDR3 (SEQ ID NO: 11), a heavy chain variable domain (VH) CDR1 comprising the amino acid sequence of 3E10-VH-CDR1 D31N (SEQ ID NO: 15), a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2 (SEQ ID NO: 4), and a VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3 (SEQ ID NO: 5).

[0347] In embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof described herein includes a set of complementarity determining regions (CDRs) collectively having no more than 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions, relative to the CDR sequences of 3E10-D31N variant (SEQ ID NOs: 15- 18 and 22-24), selected from, but not limited to, a G to S substitution at position 5 of VH CDR2, a T to S substitution at position 14 of VH CDR2, an S to T substitution at position 5 of VL CDR1, an M to L substitution at position 14 of VL CDR1, an H to A substitution at position 15 of VL CDR1, and an E to Q substitution at position 6 of VL CDR2.

[0348] Accordingly, in embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof includes VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2.1 (SEQ ID NO: 26) or 3E10-VH-CDR2.2 (SEQ ID NO: 27). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3 (SEQ ID NOs:9-l l), and VH CDRs 1 and 3 (SEQ ID NOs:3 and 5) according to theparent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3 (SEQ ID NOs:22-24), and VH CDRs 1 and 3 (SEQ ID NOs: 15 and 18) according to the 3E10-D3 IN variant.

[0349] Similarly, in embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof includes VL CDR1 comprising the amino acid sequence of 3E10-VL-CDR1.1 (SEQ ID NO: 28) or 3E10-VL-CDR1.2 (SEQ ID NO: 29). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 2 and 3 (SEQ ID NOs: 10 and 11), and VH CDRs 1-3 (SEQ ID NOs:3-5) according to the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 2 and 3 (SEQ ID NOs:23 and 24), and VH CRDs 1 -3 (SEQ ID NOs: 15, 17 and 18) according to the 3E10- D31N variant.

[0350] Similarly, in embodiments, an antibody -pay load conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof includes VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2.1 (SEQ ID NO: 30). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3 (SEQ ID NOs:9 and 11), and VH CDRs 1-3 (SEQ ID NOs: 3-5) according to the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3 (SEQ ID NOs:22 and 24), and VH CDRs 1-3 (SEQ ID NOs: 15, 17 and 18)according to the 3E10-D31N variant.

[0351] While some of the amino acid substitutions described above are fairly conservative substitutions — e.g., an S to T substitution at position 5 of VL CDR1 — other substitutions are to amino acids that have vastly different properties — e.g., an M to L substitution at position 14 of VL CDR1, an H to A substitution at position 15 of VL CDR1, and an E to Q substitution at position 6 of VL CDR2. This suggests, without being bound by theory, that at least these positions within the 3E10 CDR framework are tolerant to other amino acid substitutions.

[0352] Accordingly, in embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof includes VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2.3 (SEQ ID NO: 31). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3 (SEQ ID NOs:9-l 1), and VH CDRs 1 and 3 (SEQ ID NOs:3 and 5) according to the parent 3E10 antibody.In embodiments, the 3E10 antibody or antigen-binding fragmentthereof further includes VL CDRs 1-3 (SEQ ID NOs:22-24), and VH CDRs 1 and 3 (SEQ ID NOs: 15 and 18) according to the 3E10- D31N.

[0353] Similarly, in embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof includes VL CDR1 comprising the amino acid sequence of 3E10-VL-CDR1.3 (SEQ ID NO: 32). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 2 and 3 (SEQ ID NOs: 10 and 11), and VH CDRs 1-3 (SEQ ID NOs: 15, 17 and 18) according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 2 and 3 (SEQ ID NOs:23 and 24), and VH CDRs 1-3 (SEQ ID NOs: 15, 17 and 18) according to the 3E10- D3 IN variant.

[0354] Similarly, in embodiments, an antibody -pay load conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof, includes VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2.2 (SEQ ID NO: 33). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3 (SEQ ID NOs:9 and 11), and VH CDRs 1-3 (SEQ ID NOs:3-5) according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3 (SEQ ID NOs:22 and 24), and VH CDRs 1-3 (SEQ ID NOs: 15, 17 and 18) according to the 3E10- D31N variant.

[0355] Accordingly, in embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof includes VH CDR1 comprising the amino acid sequence of 3E10-VH-CDR1.C1 (SEQ ID NO: 34), 3E10-VH-CDRl.c2 (SEQ ID NO: 35), 3E10-VH-CDRl.c3 (SEQ ID NO: 36), 3E10-VH-CDRl.c4 (SEQ ID NO: 37), or 3E10-VH- CDRl.c5 (SEQ ID NO: 38). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3 (SEQ ID NOs:9-l 1), and VH CDRs 2 and 3 (SEQ ID NOs:4 and 5) according to the parent 3E10 antibody.

[0356] Similarly, in embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof includes VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2.cl (SEQ ID NO: 39), 3E10-VH-CDR2.c2 (SEQ ID NO: 40), or 3E10-VH-CDR2.c3 (SEQ ID NO: 41). In embodiments, the humanized 3E10 antibody orantigen-binding fragment thereof further includes VL CDRs 1 -3 (SEQ ID NOs:9-l l), and VH CDRs 1 and 3 (SEQ ID NOs:3 and 5) according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3 (SEQ ID NOs:22-24), and VH CDRs 1 and 3 (SEQ IDNOs: 15 and 18) according to the 3E10-D3 IN variant.

[0357] Similarly, in embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof includes VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3.cl (SEQ ID NO: 42), 3E10-VH-CDR3.c2 (SEQ ID NO: 43), or 3E10-VH-CDR3.c3 (SEQ ID NO: 44). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3 (SEQ ID NOs:9-l l), and VH CDRs 1 and 2 (SEQ ID NOs:3 and 4) according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3 (SEQ ID NOs:22-24), and VH CDRs 1 and 2 (SEQ ID NOs: 15 and 17) according to the 3E10-D3 IN variant.

[0358] Similarly, in embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof includes VL CDR1 comprising the amino acid sequence of 3E10-VL-CDRl.cl (SEQ ID NO: 45), 3E10-VL-CDRl.c2 (SEQ ID NO: 46), 3E10-VL-CDR1.C3 (SEQ ID NO: 47), 3E10-VL-CDRl.c4 (SEQ ID NO: 48), 3E10-VL- CDRl.c5 (SEQ ID NO: 49), or 3E10-VL-CDRl.c6 (SEQ ID NO: 50). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 2 and 3 (SEQ ID NOs:10 and 1 1), and VH CDRs 1-3 (SEQ ID NOs:3-5) according to the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 2 and 3 (SEQ ID NOs:23 and 24), and VH CRDs 1-3 (SEQ ID NOs: 15, 17, 18) according to the 3E10-D31N variant.

[0359] Similarly, in embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof includes VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2.cl (SEQ ID NO: 51). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3 (SEQ ID NOs:9 and 11), and VH CDRs 1-3 (SEQ ID NOs:3-5) according to the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3 (SEQ ID NOs:22 and 24), and VH CDRs 1-3 (SEQ ID NOs:15, 17 and 18) according to the 3E10-D3 IN variant.

[0360] Similarly, in some aspects, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof includes VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3.cl (SEQ ID NO: 52), 3E10-VL-CDR3.c2 (SEQ ID NO: 53), 3E10-VL-CDR3.c3 (SEQ ID NO: 54), 3E10-VL-CDR3.c4 (SEQ ID NO: 55), 3E10-VL- CDR3.c5 (SEQ ID NO: 56), or 3E10-VL-CDR3.c6 (SEQ ID NO: 57). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 2 (SEQ ID NOs:9 and 10), and VH CDRs 1-3 (SEQ ID NOs:3-5) according to the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 2 (SEQ ID NOs:22 and 23), and VH CDRs 1-3 (SEQ ID NOs: 15, 17 and 18) according to the 3E10-D31N variant.

[0361] It is also contemplated that an antibody-payload conjugate comprising a humanized3E10 antibody or antigen-binding fragment thereof, as described herein, includes no more than 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 CDR amino acid substitutions of the CDR amino acid substitutions described above. Further examples of 3E10 variant CDR sequences are described herein (SEQ ID NOs:58-63).

[0362] Accordingly, in embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof includes VH CDR1 comprising the amino acid sequence of 3E10-VH-CDRlm (SEQ ID NO: 58). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3 (SEQ ID NOs:9-l 1), and VH CDRs 2 and 3 (SEQ ID NOs:4 and 5) according to the parent 3E10 antibody.

[0363] Similarly, in embodiments, an antibody -pay load conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof includes VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2m (SEQ ID NO: 59). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3 (SEQ ID NOs:9-l 1), and VH CDRs 1 and 3 (SEQ ID NOs: 3 and 5) according to the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3 (SEQ ID NOs:22-24), and VH CDRs 1 and 3 (SEQ ID NOs: 15 and 18) according to the 3E10-D3 IN variant.

[0364] Similarly, in embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof includes VH CDR3 comprising theamino acid sequence of 3E10-VH-CDR3m (SEQ ID NO: 60). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3 (SEQ ID NOs:9-l 1), and VH CDRs 1 and 2 (SEQ ID NOs:3 and 4) according to the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1-3 (SEQ ID NOs:22-24), and VH CDRs 1 and 2 (SEQ ID NOs: 15 and 17) according to the 3E10-D3 IN variant.

[0365] Similarly, in embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof includes VL CDR1 comprising the amino acid sequence of 3E10-VL-CDRlm (SEQ ID NO: 61). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 2 and 3 (SEQ ID NOs: 10 and 11), and VH CDRs 1-3 (SEQ ID NOs:3-5) according to the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 2 and 3 (SEQ ID NOs:23 and 24), and VH CDRs 1-3 (SEQ ID NOs:15,17, and 18) according to the 3E10-D31N variant.

[0366] Similarly, in embodiments, an antibody -pay load conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof includes VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2m (SEQ ID NO: 62). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3 (SEQ ID NOs:9 and 1 1), and VH CDRs 1 -3 (SEQ ID NOs:3-5) according to the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 3 (SEQ ID NOs: 22 and 24), and VH CDRs 1-3 (SEQ ID NOs: 15, 17 and 18) according to the 3E10-D3 IN variant.

[0367] Similarly, in embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof includes VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3m (SEQ ID NO: 63). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 2 (SEQ ID NOs:9 and 10), and VH CDRs 1-3 (SEQ ID NOs:3-5) according to the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further includes VL CDRs 1 and 2 (SEQ ID NOs:9 and 10), and VH CDRs 1-3 (SEQ ID NOs: 15, 17 and 18) according to the 3E10-D3 IN variant.

[0368] In embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof described herein includes a light chain variable domain (3E10-VL) comprising an amino acid sequence that is at least about 90% identical to an amino acid sequence selected from the group consisting of 3E10-VL-hl (SEQ ID NO:85), 3E10- VL-h2 (SEQ ID NO:86), 3E10-VL-h3 (SEQ ID NO:87), 3E10-VL-h4 (SEQ ID NO:88), 3E10- VL-h5 (SEQ ID NO:89), and 3E10-VL-h6 (SEQ ID NO: 90), where the light chain variable domain (3E10-VL) further comprises one or more amino acid residues selected from proline (Pro) at position 15, threonine (Thr) at position 22, tyrosine (Tyr) at position 49, Thr at position 74, asparagine (Asn) at position 76, alanine (Ala) at position 80, Asn at position 81, Thr at position 83, Asn at position 85, and valine (Vai) at position 104, of the 3E10-VL according to Kabat numbering, and a set of 3E10-VL CDRs collectively having no more than 6 amino acid substitutions relative to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO:9), 3E10-VL-CDR2 (SEQ ID NOTO), 3E10-VL-CDR3 (SEQ ID NO: 11), and where the antibody includes a set of 3E10-VL CDRs collectively having no more than 6 amino acid substitutions relative to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID N0:9), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11).

[0369] In embodiments, an antibody-payload conjugate comprising the humanized 3E10 antibody or antigen-binding fragment thereof includes a set of 3E10-VL CDRs comprising no more than 5 amino acid substitutions relative to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO:9), 3E10-VL-CDR2 (SEQ ID NOTO), 3E10-VL-CDR3 (SEQ ID NO: 11).

[0370] In embodiments, an antibody-payload conjugate comprising the humanized 3E10 antibody or antigen-binding fragment thereof includes a set of 3E10-VL CDRs comprising no more than 4 amino acid substitutions relative to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO:9), 3E10-VL-CDR2 (SEQ ID NOTO), 3E10-VL-CDR3 (SEQ ID NOT 1).

[0371] In embodiments, an antibody-payload conjugate comprising the humanized 3E10 antibody or antigen-binding fragment thereof includes a set of 3E10-VL CDRs comprising no more than 3 amino acid substitutions relative to the set of CDRs having the amino acid sequencesof 3E10-VL-CDR1 (SEQ ID NON), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11).

[0372] In embodiments, an antibody-payload conjugate comprising the humanized 3E10 antibody or antigen-binding fragment thereof includes a set of 3E10-VL CDRs comprising no more than 2 amino acid substitutions relative to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NON), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11).

[0373] In embodiments, an antibody-payload conjugate comprising the humanized 3E10 antibody or antigen-binding fragment thereof includes a set of 3E10-VL CDRs comprising no more than 1 amino acid substitution relative to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NON), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11).

[0374] In embodiments, an antibody-payload conjugate comprising the humanized 3E10 antibody or antigen-binding fragment thereof includes a set of 3E10-VL CDRs comprising a set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NON), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11).

[0375] In one aspect, the present disclosure provides an antibody-payload conjugate comprising a humanized antibody or antigen-binding fragment thereof with a lysine (Lys) residue at position 49 of the 3E10-VL according to Kabat numbering.

[0376] In one aspect, the present disclosure provides an antibody-payload conjugate comprising a humanized antibody or antigen-binding fragment thereof with a glutamic acid (Glu) residue at position 81 of the 3E10-VL according to Kabat numbering.

[0377] In one aspect, the present disclosure provides an antibody-payload conjugate comprising a humanized antibody or antigen-binding fragment thereof with a proline (Pro) residue at position 15 of the 3E10-VL according to Kabat numbering.

[0378] In one aspect, the present disclosure provides an antibody-payload conjugate comprising a humanized antibody or antigen-binding fragment thereof with a valine (Vai) residue at position 104, of the 3E10-VL according to Kabat numbering.

[0379] In embodiments, an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof described herein includes a heavy chain variable domain (3E10-VH) comprising an amino acid sequence that is at least about 90% identical to an amino acid sequence selected from the group consisting of 3E10-VH-hl (SEQ ID NO:64), 3E10- VH-h2 (SEQ ID NO:65), 3E10-VH-h3 (SEQ ID NO:66), 3E10-VH-h4 (SEQ ID NO:67), 3E10- VH-h5 (SEQ ID NO:68), 3E10-VH-h6 (SEQ ID NO:69), and 3E10-VH-h7 (SEQ ID NO:70), where the heavy chain variable domain (3E10-VH) further comprises one or more amino acid residues selected from glutamine (Gin) at position 13, leucine (Leu) at position 18, arginine (Arg) at position 19, glycine (Gly) at position 42, serine (Ser) at position 49, Ser at position 77, tyrosine (Tyr) at position 79, Asn at position 82, Ala at position 84, Vai at position 89, leucine (Leu) at position 108, Vai at position 109, and Ser at position 113, of the 3E10-VH according to Kabat numbering, and where the antibody includes a set of 3E10-VH CDRs collectively having no more than 6 amino acid substitutions relative to the set of CDRs having the amino acid sequences of 3E1O-VH-CDR1 D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO:4), and 3E10-VH- CDR3 (SEQ ID NO:5).

[0380] In embodiments, an antibody-payload conjugate comprising the humanized 3E10 antibody or antigen-binding fragment thereof includes a set of 3E10-VH CDRs comprising no more than 5 amino acid substitutions relative to the set of CDRs having the amino acid sequences of 3E10-VH-CDR1 D31N (SEQ ID NO:15), 3E10-VH-CDR2 (SEQ ID NO:4), and 3E10-VH- CDR3 (SEQ ID NO 5).

[0381] In embodiments, an antibody-payload conjugate comprising the humanized 3E10 antibody or antigen-binding fragment thereof includes a set of 3E10-VH CDRs comprising no more than 4 amino acid substitutions relative to the set of CDRs having the amino acid sequences of 3E10-VH-CDR1 D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO:4), and 3E10-VH- CDR3 (SEQ ID NO:5).

[0382] In embodiments, an antibody-payload conjugate comprising the humanized 3E10 antibody or antigen-binding fragment thereof includes a set of 3E10-VH CDRs comprising no more than 3 amino acid substitutions relative to the set of CDRs having the amino acid sequences of 3E10-VH-CDR1 D31N (SEQ ID NO:15), 3E10-VH-CDR2 (SEQ ID NO:4), and 3E10-VH- CDR3 (SEQ ID NO:5).

[0383] In embodiments, an antibody-payload conjugate comprising the humanized 3E10 antibody or antigen-binding fragment thereof includes set of 3E10-VH CDRs comprising no more than 2 amino acid substitutions relative to the set of CDRs having the amino acid sequences of 3E1O-VH-CDR1 D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NON), and 3E10-VH- CDR3 (SEQ ID NO:5).

[0384] In embodiments, an antibody-payload conjugate comprising the humanized 3E10 antibody or antigen-binding fragment thereof includes a set of 3E10-VH CDRs comprising no more than 1 amino acid substitution relative to the set of CDRs having the amino acid sequences of 3E10-VH-CDR1 D31N (SEQ ID NO:15), 3E10-VH-CDR2 (SEQ ID NON), and 3E10-VH- CDR3 (SEQ ID NO 5)

[0385] In embodiments, an antibody-payload conjugate comprising the humanized 3E10 antibody or antigen-binding fragment thereof includes a set of 3E10-VH CDRs comprising no more than 5, 4, 3, 2, or 1 amino acid substitutions relative to the set of CDRs having the amino acid sequences of 3E10-VH-CDR1 D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NON), and 3E10-VH-CDR3 (SEQ ID NO:5).

[0386] In one aspect, the present disclosure provides an antibody -payload conjugate comprising a humanized antibody or antigen-binding fragment thereof with an arginine (Arg) residue at position 18 of the 3E10-VH according to Kabat numbering.

[0387] In one aspect, the present disclosure provides an antibody-payload conjugate comprising a humanized antibody or antigen-binding fragment thereof with a (Lys) residue at position 19 of the 3E10-VH according to Kabat numbering.

[0388] In one aspect, the present disclosure provides an antibody-payload conjugate comprising a humanized antibody or antigen-binding fragment thereof with an alanine (Ala) residue at position 49 of the 3E10-VH according to Kabat numbering.

[0389] In one aspect, the present disclosure provides an antibody-payload conjugate comprising a humanized antibody or antigen-binding fragment thereof with a glutamine (Gin) residue at position 13, of the 3E10-VH according to Kabat numbering.

[0390] In one aspect, the present disclosure provides an antibody-payload conjugate comprising a humanized antibody or antigen-binding fragment thereof with a leucine (Leu) residue at position 108, of the 3E10-VH according to the Kabat numbering.

[0391] In one aspect, the present disclosure provides an antibody-payload conjugate comprising a humanized antibody or antigen-binding fragment thereof with a valine (Vai) residue at position 109, of the 3E10-VH according to Kabat numbering.

[0392] In one aspect, the present disclosure provides an antibody -payload conjugate comprising a humanized antibody or antigen-binding fragment thereof with a serine (Ser) residue at position 113, of the 3E10-VH according to Kabat numbering.

[0393] In embodiments, the present disclosure provides an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof with a fragment crystallizable (Fc) region.

[0394] In embodiments, the present disclosure provides an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof with an Fc region selected from a human IgGl Fc, a human IgG2a Fc, a human IgG2b Fc, a human IgG3 Fc, and a human IgG4 Fc.

[0395] In embodiments, the present disclosure provides an antibody-payload conjugate comprising humanized 3E10 antibodies or variants thereof, or antigen-binding fragments thereof comprising a heavy chain constant domain (CH).

[0396] In embodiments, an antibody-payload conjugate comprising the humanized 3E10 antibody or antigen-binding fragment thereof comprises an Fc region selected from a human yl CHI, a human γ2 CHI, a human γ3 CHI, and a human γ4 CHI.

[0397] In embodiments, the present disclosure provides an antibody-payload conjugate comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprising a light chain constant domain (CL).

[0398] In one aspect, the present disclosure provides an antibody-payload conjugate comprising a humanized 3E10 antibody or variant comprising an Fc region selected from the group consisting of a human L CL and a human K CL.

[0399] In embodiments, an antibody-payload conjugate comprising the humanized 3E10 antibody or antigen-binding fragment thereof comprising a combination of a light chain variable domain (VL) and a heavy chain variable domain (VH) selected from 3E10-VL-hl (SEQ ID NO:85) and 3E10-VH-hl (SEQ ID NO:64), 3E10-VL-hl (SEQ ID NO:85) and 3E10-VH-h2 (SEQ ID NO:65), 3E10-VL-hl (SEQ ID NO:85) and 3E10-VH-h3 (SEQ ID NO:66), 3E10-VL-hl (SEQ ID NO:85) and 3E10-VH-h4 (SEQ ID NO:67), 3E10-VL-h2 (SEQ ID NO:86) and 3E10-VH-hl (SEQ ID NO:64), 3E10-VL-h2 (SEQ ID NO:86) and 3E10-VH-h2 (SEQ ID NO:65), 3E10-VL- h3 (SEQ ID NO:87) and 3E10-VH-hl (SEQ ID NO:64), 3E10-VL-h5 (SEQ ID NO:89) and 3E10- VH-h5 (SEQ ID NO:68), 3E10-VL-h5 (SEQ ID NO:89) and 3E10-VH-h6 (SEQ ID NO:69), 3E10- VL-h6 (SEQ ID NO:90) and 3E10-VH-h5 (SEQ ID NO:68), and 3E10-VL-h6 (SEQ ID NO:90) and 3E10-VH-h6 (SEQ ID NO:69).

[0400] In embodiments, an antibody-payload conjugate comprising the humanized 3E10 antibody or antigen-binding fragment thereof comprises a combination of a light chain variable domain (VL) of 3E10-VL-h6 (SEQ ID NO:90) and a heavy chain variable domain (VH) of 3E10- VH-h6 (SEQ ID NO:69).

[0401] Antibodies useful in the compositions, conjugates, and methods described herein include whole immunoglobulin (i.e., an intact antibody) of any class, fragments thereof, and synthetic proteins containing at least the antigen-binding variable domain of an antibody. The variable domains differ in sequence among antibodies and are used in the binding and specificity of each particular antibody for its particular antigen. However, the variability is not usually evenly distributed through the variable domains of antibodies. It is typically concentrated in three segments called complementarity determining regions (CDRs) or hypervariable regions both in the light chain and the heavy chain variable domains. The more highly conserved portions of the variable domains are called the framework (FR). The variable domains of native heavy and light chains each comprise four FR regions, largely adopting a beta-sheet configuration, connected by three CDRs, which form loops connecting, and in some cases forming part of, the beta-sheet structure. The CDRs in each chain are held together in close proximity by the FR regions and, with the CDRs from the other chain, contribute to the formation of the antigen-binding site of antibodies. Therefore, the antibodies typically contain at least the CDRs necessary to maintain DNA binding.

[0402] The 3E10 antibody is typically a monoclonal 3E10, or a variant, derivative, fragment, fusion, or humanized form thereof that binds the same or different epitope(s) as 3E10.

[0403] A deposit according to the terms of the Budapest Treaty of a hybridoma cell line producing monoclonal antibody 3E10 was received on September 6, 2000, and accepted by, American Type Culture Collection (ATCC), 10801 University Blvd., Manassas, VA 20110-2209, USA, and given Patent Deposit Number PTA-2439. Thus, the antibody can have the same or different epitope specificity as monoclonal antibody 3E10 produced by ATCC No. PTA 2439 hybridoma. The antibody can have the paratope of monoclonal antibody 3E10. The antibody can be a single chain variable fragment of 3E10, or a variant, e.g., a conservative variant thereof. For example, the antibody can be a single chain variable fragment of 3E10 (3E10 Fv), or a variant thereof.

[0404] Additionally, or alternatively, the heavy chain complementarity determining regions (CDRs) can be defined according to the IMGT system. The complementarity determining regions (CDRs) as identified by the IMGT system include CDR Hl.3 (original sequence): GFTFSDYG (SEQ ID NO:989); CDR Hl.4 (with D31N mutation): GFTFSNYG (SEQ ID NO:990); CDR H2.2: ISSGSSTI (SEQ ID NO:991) and variant ISSSSSTI (SEQ ID NO:992); CDR H3.2: ARRGLLLDY (SEQ ID NO:993).

[0405] Additionally, or alternatively, the light chain complementarity determining regions (CDRs) can be defined according to the IMGT system. The complementarity determining regions (CDRs) as identified by the IMGT system include CDR LI.2 KSVSTSSYSY (SEQ ID NO:994) and variant KTVSTSSYSY (SEQ IDNO:995); CDRL2.2: YAS; CDRL3.2: QHSREFPWT (SEQ ID NO: 996).

[0406] The disclosed compositions, conjugates, and methods typically utilize antibodies that maintain the ability to penetrate cells, and optionally nuclei. The mechanisms of cellular internalization by autoantibodies are diverse. Some are taken into cells through electrostatic interactions or FcR-mediated endocytosis, while others utilize mechanisms based on association with cell surface myosin or calreticulin, followed by endocytosis (Ying-Chyi et al., Eur J Immunol 38, 3178-3190 (2008), Yanase et al., J Clin Invest 100, 25-31 (1997)). The 3E10 antibodies and antigen-binding fragments thereof can transit cellular membranes via an equilibrative nucleoside (ENT) transporter. In embodiments, 3E10 transits cellular membranes via an ENT1, ENT2, ENT3or ENT4 transporter (See, e.g., WO 2015 / 106290 Al and WO 2016 / 033324 Al, each of which is incorporated by reference herein, in its entirety). In embodiments, 3E10 penetrates cells in an Fc- independent mechanism (as evidenced by the ability of 3E10 fragments lacking an Fc to penetrate cells) but involves presence of the nucleoside transporter ENT2 (Weisbart et al., Sci Rep 5: 12022. doi: 10.1038 / srepl2022. (2015), Zack et al., J Immunol 157, 2082-2088 (1996), Hansen et al., J Biol Chem 282, 20790-20793 (2007)). Thus, in embodiments, the antibodies utilized in the disclosed compositions, conjugates, and methods are ones that penetrates cells in an Fc- independent mechanism and involve the presence of the nucleoside transporter ENT2.

[0407] Mutations in 3E10 that interfere with its ability to bind DNA can render the antibody incapable of nuclear penetration. Thus, typically the disclosed variants and humanized forms of the antibody maintain the ability to bind nucleic acids, particularly DNA. In addition, 3E10 scFv has previously been shown capable of penetrating into living cells and nuclei in an ENT2-dependent manner, with efficiency of uptake impaired in ENT2-deficient cells (Hansen, et al., J. Biol. Chem. 282, 20790-20793 (2007)). Thus, in embodiments, the disclosed variants and humanized forms of the antibody maintain the ability penetrate into cell nuclei in an ENT2- dependent manner.

[0408] As discussed in US 2021 / 0054102 and US 2021 / 0137960, some humanized 3E10 variant were found to penetrate cell nuclei more efficiently than the original murine 3E10 (D31N) di-scFv, while others were found to have lost the ability to penetrate nuclei. In particular, variants 10 and 13 penetrated nuclei very well compared to the murine antibody.

[0409] Potential bipartite nuclear localization signals (NLS) in humanized 3E10 VL have been identified and may include part or all of the following sequences:RASKSVSTSSYSYMHWYQQKPGQPPKLLIKY (SEQ ID NO: 138);RASKTVSTSSYSYMHWYQQKPGQPPKLLIKY (SEQ ID NO: 139); or RVTITCRASKSVSTSSYSYMHWYQQKPGKAPKL (SEQ ID NO: 140).

[0410] An example consensus NLS can be, or include, (X)RASKTVSTSSYSYMHWYQQKPGQPPKLL(X)KY (where (X) = any residue, but preferentially is a basic residue (R or K) (SEQ ID NO: 141) or a variant thereof with at least 60, 65, 70, 75, 80, 85, 90, 95, 96, 97, 98, 99 percent sequence identity to SEQ ID NO: 142.

[0411] Thus, in some embodiments, particularly where nuclear importation is important, the disclosed antibodies may include the sequence of any one of SEQ ID NOs: 138-142, or fragments and variants thereof (e.g., at least 70, 75, 80, 85, 90, 95, 96, 97, 98, 99, or 100% amino acid sequence identity with any one of SEQ ID NOs: 138-142) that can translocate into the nucleus of a cell.

[0412] Presence of an NLS indicates that a humanized 3E10 antibody or antigen binding fragment thereof may cross the nuclear envelope via the nuclear import pathway. In some embodiments, the NLS improves importation by interacting with one or more members of the import pathway. Thus, in some embodiments, the NLS can bind to importin- , an importin- β / importin-α heterodimer, or a combination thereof.

[0413] In some embodiments, the disclosed compositions and methods utilize humanized 3E10 antibodies and ENT2-binding fragments thereof that maintain the ability to bind nucleic acids such as DNA, RNA.

[0414] The Examples below illustrate molecular modeling of wild type 3E10 sequences and additional 3E10 variants. Molecular modeling of 3E10 (Pymol) revealed a putative Nucleic Acid Binding pocket (NAB 1) (See, e g., Figures 11 A and 1 IB, and illustrated with underlining the sequences below).WT HEAVY CHAIN scFv SEQUENCEE VQLVESGGGL VKPGGSRKLS CAASGFTFSD YGMHWVRQAP EKGLEWVAYI SSGSSTIYYA DTVKGRFTIS RDNAKNTLFL QMTSLRSEDT AMYYCARRGL LLDYWGQGTT LTVS (SEQ ID NO: 148)LIGHT CHAIN scFv SEQUENCED IVLTQSPASL AVSLGQRATI SCRASKSVST SSYSYMHWYQ QKPGQPPKLL IKYASYLESG VPARFSGSGS GTDFTLNIHP VEEEDAATYY CQHSREFPWT FGGGTKLEIK RADAAPGGGG SGGGGSGGGGS (SEQ ID NO: 149)

[0415] In some embodiments, the disclosed humanized 3E10 antibodies include some or all of the underlined NAB1 sequences. In some embodiments, the humanized 3E10 antibodies include a variant sequence that has an altered ability of bind nucleic acids. In some embodiments, the mutations (e.g., substitutions, insertions, and / or deletions) in the NAB1 improve binding of theantibody to nucleic acids such as DNA, RNA, or a combination thereof. In some embodiments, the mutations are conservative substitutions. In some embodiments, the mutations increase the cationic charge of the NAB1 pocket.

[0416] As discussed and exemplified herein, mutation of aspartic acid at residue 31 of CDR1 to asparagine increased the cationic charge of this residue and enhanced nucleic acid binding and delivery in vivo (3E10-D31N).

[0417] Additional example variants include mutation of aspartic acid at residue 31 of CDR1 to arginine (3E10-D31R), which modeling indicates expands cationic charge, or lysine (3E10-D3 IK) which modeling indicates changes charge orientation. Thus, in some embodiments, the 3E10 binding protein includes a D31R or D3 IK substitution.

[0418] Additional example variants include mutation of arginine (R) 96 to asparagine (N), and / or serine (S) 30 to aspartic acid (D) alone or in combination with D3 IN, D31R, or D3 IK.

[0419] Molecular modeling of 3E10 (Pymol) revealed a putative Nucleic Acid Binding pocket (NAB1) (Figures 11A-1 IB). Mutation of aspartic acid at residue 31 of CDR1 to asparagine increased the cationic charge of this residue and enhanced nucleic acid binding and delivery in vivo (3E10-D31N).

[0420] Mutation of aspartic acid at residue 31 of CDR1 to arginine (3E10-D31R), further expanded the cationic charge while mutation to lysine (3E10-D31K) changed charge orientation (Figure 11A).

[0421] NAB 1 amino acids predicted from molecular modeling have been underlined in the heavy and light chain sequences above. Figure 1 IB is an illustration showing molecular modeling of 3E10-scFv (Pymol) with NAB1 amino acid residues illustrated with punctate dots.

[0422] All of the sequences disclosed herein having the residue corresponding with R96 are expressly disclosed with R96N substitution.

[0423] All of the sequence disclosed herein having the residue corresponding to S30 are expressly disclosed with S30D.

[0424] Any of the substitutions can be included in any combination. The sequence having two or three substitutions at any combination of residues 31, 30, and 96 are expressly provided.

[0425] In particular embodiments, the sequence has 3 IN, 3 IK, or 31R alone or in combination with 30D, and without the R96N substitution. Thus, in some embodiments, the residue corresponding to 96 is not N, and in more specific embodiments remains R.B. Linkers

[0426] In some aspects, the antibody-oligonucleotide conjugates (AOCs) provided herein comprises a linker. The linker (L) described herein can be used to link or conjugate the 3E10 antibody or antigen-binding fragment thereof to an oligonucleotide, e.g., an siRNA or antisense oligonucleotide. The term “linker” as used herein includes, without limitation, any known linker for use in antibody-oligonucleotide-conjugates known in the art. In some aspects, the AOC comprises, one, two, three, four, or more linkers.1. Conjugation Sites and Methods

[0427] In some embodiments, one or more amino acids suitable conjugation of a 3E10 antibody or antigen-binding fragment thereof provided herein are selected from lysine, cysteine, histidine, arginine, aspartic acid, glutamine, serine, threonine and tyrosine. In some embodiments, one or more amino acids suitable for conjugation are introduced by substitution of one or more amino acids in the 3E10 antibody or antigen-binding fragment thereof. In some embodiments, the one or more conjugated amino acids are lysine or arginine, and conjugation is conducted via amine conjugation. In some embodiments, one or more conjugated amino acids are glutamine (Gin) and conjugation is conducted via transglutaminase (TGase) mediated enzymatic conjugation. In some embodiments, one or more conjugated amino acids are cysteine (Cys) and conjugation is conducted via thiol conjugation. For example, NHS-PEG reagent can be used to modify primary amines in a 3E10 antibody or antigen-binding fragment thereof provided herein.

[0428] In some embodiments, a composition comprising an AOC as described herein has an average DAR of at least 4 (an average of at least four drug moieties attached to each antibody). In some embodiments, such a composition has an average DAR of at least 6. In some embodiments, such a composition has an average DAR of at least 8. In some embodiments, such a composition has an average DAR of at least 10. In some embodiments, such a composition has an average DAR of at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or more.

[0429] In some embodiments, a composition comprising an AOC as described herein has an average DAR of no more than 12. In some embodiments, a composition comprising an AOC as described herein has an average DAR of no more than 10. In some embodiments, a composition comprising an AOC as described herein has an average DAR of no more than 8. In some embodiments, a composition comprising an AOC as described herein has an average DAR of no more than 6. In some embodiments, a composition comprising an AOC as described herein has an average DAR of no more than 4.

[0430] In some embodiments, a composition comprising an AOC as described herein has an average DAR of from 2 to 4. In some embodiments, a composition comprising an AOC as described herein has an average DAR of from 2 to 6. In some embodiments, a composition comprising an AOC as described herein has an average DAR of from 2 to 8. In some embodiments, a composition comprising an AOC as described herein has an average DAR of from 2 to 10. In some embodiments, a composition comprising an AOC as described herein has an average DAR of from 2 to 12. In some embodiments, a composition comprising an AOC as described herein has an average DAR of from 3 to 4. In some embodiments, a composition comprising an AOC as described herein has an average DAR of from 3 to 6. In some embodiments, a composition comprising an AOC as described herein has an average DAR of from 3 to 8. In some embodiments, a composition comprising an AOC as described herein has an average DAR of from 3 to 10. In some embodiments, a composition comprising an AOC as described herein has an average DAR of from 3 to 12. In some embodiments, a composition comprising an AOC as described herein has an average DAR of from 4 to 6. In some embodiments, a composition comprising an AOC as described herein has an average DAR of from 4 to 8. In some embodiments, a composition comprising an AOC as described herein has an average DAR of from 4 to 10. In some embodiments, a composition comprising an AOC as described herein has an average DAR of from 4 to 12. In some embodiments, a composition comprising an AOC as described herein has an average DAR of from 6 to 8. In some embodiments, a composition comprising an AOC as described herein has an average DAR of from 6 to 10. In some embodiments, a composition comprising an AOC as described herein has an average DAR of from 6 to 12.[...

Claims

CLAIMS1. A conjugate of Formula (I):A-(L-Pr)qFormula (I), wherein in Formula (I):A is an antibody, antigen-binding fragment thereof or antigen-binding fragment thereof comprising a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of SEQ ID NO:58, CDR2 comprising the amino acid sequence of SEQ ID NO:59, CDR3 comprising SEQ ID NO:60; and a light chain variable region (VL) CDR1 comprising the amino acid sequence of SEQ ID NO:61, CDR2 comprising the amino acid sequence of SEQ ID NO:62, CDR3 comprising the amino acid sequence of SEQ ID NO:63;L is a linker;P is an oligonucleotide capable of hybridizing to a pre-mRNA transcript, wherein the oligonucleotide induces exon skipping in the pre-mRNA transcript; r is an integer from 1 to 4; and q is an integer from 1 to 16.

2. The conjugate of claim 1, wherein the oligonucleotide hybridizes to an acceptor splice site, a donor splice site, or an exonic splice enhancer element of the pre-mRNA transcript.

3. The conjugate of claim 1 or 2, wherein the linker L comprises one or more groups selected from optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted arylene, optionally substituted cycloalkylene, optionally substituted heteroalkylene, optionally substituted heteroarylene, optionally substituted heterocycloalkylene, -NRa-, -N=CRa-, -CRa=N-, -S-, -S(O)-, -S(O)2-, -OP(O)ORa-, -OP(O)ORaO-, -P(O)ORaO-, -O-, -CRb2-, -[(CRb2)1-120]1-50-, -C(O)-, -C(S)-, -C(=N-OH)-, -C(NRa)-, -C(NH2Cl)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NRa-, -NRaC(O)-, -C(O)NRaSO2-, -SO2NRaC(O)-, -OC(O)O-, -OC(O)S-, -SC(O)O-, -OC(O)NRa-, -NRaC(O)O-, -SC(O)NRa-, -NRaC(O)S-, -S(O)tN(Ra)- (where t is 1 or 2), -N(Ra)S(O)t- (where t is 1 or 2), and -XAA-;each Rais independently selected at each occurrence from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted alkenyl, optionally substituted fluoroalkenyl, optionally substituted cycloalkenyl, optionally substituted cycloalkenylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; each Rbis independently selected at each occurrence from hydrogen, halide, -OH, -SO3H, -OPO3H2, -PO3H2, -C(0)NRa2, -CO2Ra, -NRa2, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted alkenyl, optionally substituted fluoroalkenyl, optionally substituted cycloalkenyl, optionally substituted cycloalkenylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; two independent Rbgroups or an Raand an Rbcan be joined together to form an optionally substituted cycle; and-XAA- is an amino acid sequence comprising 1 to 6 amino acid moieties.

4. The conjugate of claim 3, wherein each amino acid moiety of -XAA- is independently selected from alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine (Cys), glutamine (Gin), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), valine (Vai), citrulline (Cit), and homocitrulline (HoCit).

5. The conjugate of any one of claims 1 to 4, wherein the linker L comprises one or more groups selected from optionally substituted C1-C18alkylene, -C=C-, -CR — CRa-, optionally substituted 6- to 14-membered arylene, optionally substituted C3-C20 cycloalkylene, -[CH2O]1-18-, -[CH2CH2O]1-18-, -[CH2CH2CH2O]1-18-, optionally substituted 5- to 18-membered heteroarylene, optionally substituted 3- to 20-membered heterocycloalkylene, -NRa-, -N=CRa-, -CRa=N-, -S-, -OP(O)ORaO-, -O-, -CRb2-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-,-C(O)NRa-, -NRaC(O)-, -OC(O)O-, -OC(O)S-, -SC(O)O-, -OC(O)NRa-, -NRaC(O)O-, -SC(O)NRa-, -NRaC(O)S-, and -XAA-.

6. The conjugate of any one of claims 1 to 5, wherein the linker L comprises one or more groups selected from optionally substituted C1-C16 alkylene, -C=C-, -CR — CRa-, optionally substituted phenylene, optionally substituted C3-C6cycloalkylene, -[CH2CH2O]1-16-, -[CH2CH2CH2O] 1-16-, optionally substituted 5- to 6-membered heteroarylene, optionally substituted 5- to 20-membered heterocycloalkylene, -NRa-, -N=CRa-, -CRa=N-, -S-, -OP(O)ORaO-, -O-, -CRb2-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NRa-, -NRaC(O)-, -OC(O)O-, -OC(O)NRa-, -NRaC(O)O-, and -XAA-.

7. The conjugate of any one of claims 1 to 6, wherein the linker L comprises one or more groups selected from -[C(Rb)2]1-16-, -C=C-, -CRa= CRa-, -[CH2CH2O]1-16-, -NRa-, -N=CRa-, -CRa=N-. -S-, -OP(O)ORaO-, -O-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NRa-, -NRaC(O)-, -OC(O)O-,X1, X2, and X3are independently selected at each occurrence from NRa, N, CRb, S, and O.

8. The conjugate of any one of claims 1 to 7, wherein the linker L is of Formula (L- 1 ):Formula (L-l), wherein in Formula (L-l):LAis a connecting moiety through which A is covalently attached to L';L' is a bond or comprises one or more groups selected from optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted arylene, optionally substituted cycloalkylene, optionally substituted heteroalkylene, optionally substituted heteroarylene, optionally substituted heterocycloalkylene, -NRa-, -N=CRa-, -CRa=N-, -S-, -S(O)-, -S(O)2-, -OP(O)ORaO-, -O-, -CRb2-, -[(CRb2)1-12O]1-50-, -C(O)-, -C(S)-, -C(NRa)-, -C(NH2C1)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NRa-, -NRaC(O)-, -C(O)NRaSO2-, -SO2NRaC(O)-, -OC(O)O-, -OC(O)S-, -SC(O)O-, -OC(O)NRa-, -NRaC(O)O-, -SC(O)NRa-, -NRaC(O)S-, -S(O)tN(Ra)-, -N(Ra)S(O)t-, and -XAA-; andLp is a connecting moiety through which P is covalently attached to L'.

9. The conjugate of any one of claims 1 to 8, wherein the linker L comprises at least one cleavable moiety.

10. The conjugate of claim 9, wherein the cleavable moiety comprises an acid-labile moiety, a reducibly-labile moiety, or an enzymatically-labile moiety.

11. The conjugate of claim 9 or claim 10, wherein the cleavable moiety comprises one or more groups selected from:wherein: each Rais independently selected at each occurrence from hydrogen, optionally substituted alkyl, and optionally substituted heteroalkyl.

12. The conjugate of any one of claims 10 to 11, wherein the cleavable moiety comprises the reducibly-labile moiety -S-S-.

13. The conjugate of any one of claims 1 to 9, wherein the linker L is of Formula (L-10):Formula (L-10), wherein in Formula (L-10):LAis selected from a bond, -NRa'-, and -S-;L1is a bond or comprises one or more groups selected from optionally substituted C1-C18alkylene, -C=C-, -CR — CRa-, optionally substituted 6- to 14-membered arylene, optionally substituted C.3-C20 cycloalkylene, -[CH2O]1-18-, -[CH2CH2O]1-18-, -[CH2CH2CH2O]1-18-, optionally substituted 5- to 18-membered heteroarylene, optionally substituted 3- to 20-membered heterocycloalkylene, -NRa-, -N=CRa-, -CRa=N-, -S-, -OP(O)ORaO-, -O-, -CRb2-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NRa-, -NRaC(O)-, -OC(O)O-, -OC(O)S-, -SC(O)O-, -OC(O)NRa-, -NRaC(O)O-, -SC(O)NRa-, -NRaC(O)S-, and -XAA-;Lc is selected from an acid-labile moiety, a reducibly-labile moiety, and an enzymatically- labile moiety;L2is a bond or comprises one or more groups selected from optionally substituted C1-C18alkylene, -C=C-, -CR — CR1-, optionally substituted 6- to 14-membered arylene, optionally substituted C3-C20 cycloalkylene. -[CH2O]1-18-, -[CH2CH2O]1-18-. -[CH2CH2CH2O]1-18-, optionally substituted 5- to 18-membered heteroarylene, optionally substituted 3- to 20-membered heterocycloalkylene, -NRa-, -N=CRa-, -CRa=N-, -S-, -OP(O)ORaO-, -O-, -CRb2-, -C(O)-. -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NRa-, -NRaC(O)-, -OC(O)O-, -OC(O)S-, -SC(O)O-, -OC(O)NRa-, -NRaC(O)O-, -SC(O)NRa-, -NRaC(O)S-, and -XAA-;LP is selected from a bond, -NRa-, -S-, and -O-; each Rais independently selected at each occurrence from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; each Rais independently selected at each occurrence from hydrogen, optionally substituted alkyl, and optionally substituted heteroalkyl; each Rbis independently selected at each occurrence from hydrogen, halide, -OH, -SO3H, -OPO3H2, -PO3H2, -C(0)NRa2, -CO2Ra, -NRa2, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionallysubstituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or two independent Rbgroups are taken together to form optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocycloalkyl, or optionally substituted heteroaryl; and-XAA- is an amino acid sequence comprising 1 to 4 amino acid moieties.

14. The conjugate of claim 12, wherein Lc is selected from:wherein: each Rais independently selected at each occurrence from hydrogen, optionally substituted alkyl, and optionally substituted heteroalkyl.

15. The conjugate of claim 13 or claim 14, wherein Lc is -S-S-.

16. The conjugate of any one of claims 1 to 8, wherein the linker L is of Formula (L-l 1):Formula (L-l 1), wherein in Formula (L-l 1):LAis selected from a bond, -NH-, and -S-;L1is a bond or comprises one or more groups selected from -[C(Rb)2]1-16-, -C=C-, -CRa=CRa-, -[CH2CH2O]1-16-, -NRa-, -N=CRa-, -CRa=N-, -S-, -OP(O)ORaO-, -O-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NRa-, -NRaC(O)-, -OC(O)O-, -XAA-, -OC(O)NRa-, -NRaC(O)O-,L2is a bond or comprises one or more groups selected from -[C(Rb)2]1-16-, -C=C-,-CRa=CRa-, -[CH2CH2O]1-16-, -NRa-, -N=CRa-, -CRa=N-, -S-, -OP(O)ORaO-, -O-, -C(0)-,-C(O)O-, -OC(O)-, -C(O)NRa-, -NRaC(O)-, -OC(O)O-, -XAA-, -OC(O)NRa-, -NRaC(O)O-,LPis selected from a bond, -NRa-, and -O-; each R1is independently selected from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or both Ri groups are taken together to form optionally substituted cycloalkyl; each R2is independently selected from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or both R2groups are taken together to form optionally substituted cycloalkyl;each Rais independently selected at each occurrence from hydrogen, optionally substituted C1-C8alkyl, optionally substituted C1-C8fluoroalkyl, optionally substituted C3-C6cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; each Rais independently selected at each occurrence from hydrogen and optionally substituted alkyl; each Rbis independently selected at each occurrence from hydrogen, halide, -OH, -SO3H, -OPO3H2, -PO3H2, -CO2Ra, -NRa2, optionally substituted C1-C8alkyl, optionally substituted C1-C8fluoroalkyl, optionally substituted C3-C6cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; or two independent Rbgroups are taken together to form optionally substituted cycloalkyl; and-XAA- is an amino acid sequence comprising 2 to 4 amino acid moieties.

17. The conjugate of any one of claims 1 to 8, wherein the linker L is of Formula (L-12):L2’ comprises one or more groups selected from -[C(Rb)2]1-10-, -[CH2CH2O]1-10-, -NRa-, -O-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NRa-, -NRaC(O)-, -OC(O)O-, -XAA-, -OC(O)NRa-,Lp is selected from a bond and -NRa-; each Ri is independently selected from hydrogen, optionally substituted C1-C8alkyl, optionally substituted C1-C8fluoroalkyl, optionally substituted C3-C6cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; or both Ri groups are taken together to form optionally substituted C3-C6cycloalkyl; each R2 is independently selected from hydrogen, optionally substituted C1-C8alkyl, optionally substituted C1-C8fluoroalkyl, optionally substituted C3-C6cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; or both R2 groups are taken together to form optionally substituted C3-C6cycloalkyl; each Rais independently selected at each occurrence from hydrogen, optionally substituted C1-C6alkyl, optionally substituted C1-C6fluoroalkyl, and optionally substituted C3-C6cycloalkyl; each Rais independently selected at each occurrence from hydrogen and optionally substituted C1-C6alkyl; each Rbis independently selected at each occurrence from hydrogen, halide, -OH, -SO3H, -OPO3H2, -PO3H2, -CO2Ra, -NRa2, optionally substituted C1-C6alkyl, optionally substituted C1-C6fluoroalkyl, and optionally substituted C3-C6cycloalkyl; or two independent Rbgroups attached to the same carbon atom are taken together to form optionally substituted C3-C6cycloalkyl; and-XAA- is an amino acid sequence comprising 2 or 3 amino acid moieties.

18. The conjugate of claim 16 or claim 17, wherein at least one Ri or R2 is other than hydrogen.

19. The conjugate of any one of claims 16 to 18, wherein at least one Ri is an optionally substituted C1-C8alkyl.

20. The conjugate of any one of claims 16 to 18, wherein each Ri is independently an optionally substituted C1-C8alkyl.

21. The conjugate of any one of claims 16 to 20, wherein at least one R2 is an optionally substituted C1-C8alkyl.

22. The conjugate of any one of claims 16 to 20, wherein each R2 is independently an optionally substituted C1-C8alkyl.

23. The conjugate of any one of claims 1 to 11, wherein the linker L is selected from:

24. The conjugate of any one of claims 1 to 17, wherein the linker L is selected from:

25. The conjugate of any one of claims 9 to 24, wherein the linker is a cleavable linker.

26. The conjugate of claim 25, wherein the cleavable linker is a cathepsin-L substrate.

27. The conjugate of claim 26, wherein the linker comprises a dipeptide selected from a -Phe-Gln- dipeptide, a -Val-Gln- dipeptide, a -Leu-Gin- dipeptide, a -Tyr-Met- dipeptide, a -Phe-Arg- dipeptide, a -Phe-Gly- dipeptide, a -Trp-Thr- dipeptide, a -Tyr-Gly- dipeptide, a -Phe-Thr- dipeptide, and a -Val-Gly- dipeptide.

28. The conjugate of claim 26, wherein the linker comprises a -Phe-Gln- dipeptide, a -Val- Gln- dipeptide, a -Leu-Gin- dipeptide, or a -Tyr-Met- dipeptide.

29. The conjugate of claim 26, wherein the linker comprises a -Phe-Gln- dipeptide.

30. The conjugate of claim 26, wherein the linker comprises a -Val-Gln- dipeptide.

31. The conjugate of claim 26, wherein the linker comprises a -Leu-Gin- dipeptide.

32. The conjugate of claim 26, wherein the linker comprises a -Tyr-Met- dipeptide.

33. The conjugate of any one of claims 25-32, wherein the cleavable linker is conjugated to a lysine of the 3E10 antibody or antigen-binding fragment thereof.

34. The conjugate of any one of claims 25-32, wherein the cleavable linker is conjugated to a cysteine of the 3E10 antibody or antigen-binding fragment thereof.

35. The conjugate of any one of claims 25-32, wherein the cleavable linker is conjugated to a histidine of the 3E10 antibody or antigen-binding fragment thereof.

36. The conjugate of any one of claims 25-32, wherein the cleavable linker is conjugated to an arginine of the 3E10 antibody or antigen-binding fragment thereof.

37. The conjugate of any one of claims 25-32, wherein the cleavable linker is conjugated to an aspartic acid of the 3E10 antibody or antigen-binding fragment thereof.

38. The conjugate of any one of claims 25-32, wherein the cleavable linker is conjugated to a glutamine of the 3E10 antibody or antigen-binding fragment thereof.

39. The conjugate of claim 38, wherein the conjugate has a drug to antibody ratio (DAR) of at least 4: 1.

40. The conjugate of claim 38 or 39, wherein the linker is a branched linker attached to at least two copies of the oligonucleotide moiety.

41. The conjugate of any one of claims 25-32, wherein the cleavable linker is conjugated to a serine of the 3E10 antibody or antigen-binding fragment thereof.

42. The conjugate of any one of claims 25-32, wherein the cleavable linker is conjugated to a threonine of the 3E10 antibody or antigen-binding fragment thereof.

43. The conjugate of any one of claims 25-32, wherein the cleavable linker is conjugated to a tyrosine of the 3E10 antibody or antigen-binding fragment thereof.

44. The conjugate of any one of claims 1 to 8, wherein the linker L is of Formula (L-20):Formula (L-20), wherein in Formula (L-20):LAis selected from a bond, -NRa'-, and -S-;L3is a bond or comprises one or more groups selected from -[C(Rb)2]1-8-, -NRa-, -C(O)-, -C(S)-, -C(NRa)-, -C(NH2C1)-, -C=C-, -CRa= CRa-, optionally substituted 6- to 14-membered arylene, optionally substituted C3-C20 cycloalkylene, optionally substituted 5- to 18-membered heteroarylene, and optionally substituted 3- to 20-membered heterocycloalkylene;Lx comprises one or more groups selected from optionally substituted C1-C18alkylene, -C=C-, -CR — CRa-, optionally substituted 6- to 14-membered arylene, optionally substituted C3-C20cycloalkylene, -[CH2O]1-18-, -[CH2CH2O]1-18-, -[CH2CH2CH2O]1-18-, optionally substituted 5- to 18-membered heteroarylene, optionally substituted 3- to 20-memberedheterocycloalkylene, -NRa-, -S-, -O-, -CRb2-, -C(O)-, -C(S)-, -C(NRa)-, -C(NH2C1)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NRa-, and -NRaC(O)-;L4is a bond or comprises one or more groups selected from -[C(Rb)2]i-8-, -NRa-, -C(O)-, -C(S)-, -C(NRa)-, -C(NH2Cl)-, -C=C-, -CR — CRa-, optionally substituted 6- to 14-membered arylene, optionally substituted C3-C20 cycloalkylene, optionally substituted 5- to 18-membered heteroarylene, and optionally substituted 3- to 20-membered heterocycloalkylene;Lp is selected from a bond, -NRa-, -S-, and -O-; each Rais independently selected at each occurrence from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; each Rais independently selected at each occurrence from hydrogen, optionally substituted alkyl, and optionally substituted heteroalkyl; and each Rbis independently selected at each occurrence from hydrogen, halide, -OH, -SO3H, -OPO3H2, -PO3H2, -C(O)NRa2, -CO2Ra, -NRa2, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or two independent Rbgroups are taken together to form optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocycloalkyl, or optionally substituted heteroaryl.

45. The conjugate of any one of claims 1 to 8, wherein the linker L is of Formula (L-21):Formula (L-21), wherein in Formula (L-21):LAis selected from a bond and -NH-;Lx comprises one or more groups selected from optionally substituted -[C(Rb)2]1-16-, -C=C-, -CR — CRa-, -[CH2CH2CH2O]1-16-, -NRa-, -O-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-,Lp is selected from a bond and -NRa-; each Rais independently selected at each occurrence from hydrogen, optionally substituted C1-C8alkyl, optionally substituted C1-C8fluoroalkyl, optionally substituted C3-C6cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; each Ra' is independently selected at each occurrence from hydrogen and optionally substituted C1-C6alkyl; and each Rbis independently selected at each occurrence from hydrogen, halide, -OH, -SO3H, -OPO3H2, -PO3H2, -CO2Ra, -NRa2, optionally substituted C1-C8alkyl, optionally substituted C1-C8fluoroalkyl, optionally substituted C3-C6cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; or two independent Rbgroups are taken together to form optionally substituted cycloalkyl.

46. The conjugate of any one of claims 1 to 8, wherein the linker L is of Formula (L-22a) or Formula (L-22b):wherein in Formulas (L-22a) and (L-22b):LAis selected from a bond and -NH-;Lx comprises one or more groups selected from optionally substituted -[C(Rb)2] 1-10-,-C=C-, -CR:— CRa-, -[CH2CH2CH2O]1-10-, -NRa-, -C(0)-, -C(0)0-, -OC(O)-, -C(O)S-, -SC(O)-,Lp is selected from a bond and -NRa-; each Rais independently selected at each occurrence from hydrogen, optionally substituted C1-C8alkyl, optionally substituted C1-C8fluoroalkyl, optionally substituted C3-C6cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; each Rais independently selected at each occurrence from hydrogen and optionally substituted C1-C6alkyl; and each Rbis independently selected at each occurrence from hydrogen, halide, -OH, -SO3H, -OPO3H2, -PO3H2, -CO2Ra, -NRa2, optionally substituted C1-C8alkyl, optionally substituted C1-C8fluoroalkyl, optionally substituted C3-C6cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; or two independent Rbgroups are taken together to form optionally substituted cycloalkyl.

47. The conjugate of any one of claims 1 to 8, wherein the linker L is selected from:

48. The conjugate of any one of claims 44 to 47, wherein the linker is a non-cleavable linker.

49. The conjugate of claim 48, wherein the non-cleavable linker is conjugated to a lysine of the 3E10 antibody or antigen-binding fragment thereof.

50. The conjugate of claim 48, wherein the non-cleavable linker is conjugated to a cysteine of the 3E10 antibody or antigen-binding fragment thereof.

51. The conjugate of claim 48, wherein the non-cleavable linker is conjugated to a histidine of the 3E10 antibody or antigen-binding fragment thereof.

52. The conjugate of claim 48, wherein the non-cleavable linker is conjugated to an arginine of the 3E10 antibody or antigen-binding fragment thereof.

53. The conjugate of claim 48, wherein the non-cleavable linker is conjugated to an aspartic acid of the 3E10 antibody or antigen-binding fragment thereof.

54. The conjugate of claim 48, wherein the non-cleavable linker is conjugated to a glutamine of the 3E10 antibody or antigen -binding fragment thereof.

55. The conjugate of claim 54, wherein the conjugate has a drug to antibody ratio (DAR) of at least 4: 1.

56. The conjugate of claim 54 or 55, wherein the linker is a branched linker attached to at least two copies of the oligonucleotide moiety.

57. The conjugate of claim 48, wherein the non-cleavable linker is conjugated to a serine of the 3E10 antibody or antigen-binding fragment thereof.

58. The conjugate of claim 48, wherein the non-cleavable linker is conjugated to a threonine of the 3E10 antibody or antigen-binding fragment thereof.

59. The conjugate of claim 48, wherein the non-cleavable linker is conjugated to a tyrosine of the 3E10 antibody or antigen-binding fragment thereof.

61. The conjugate of any one of claims 1 to 17, wherein each amino acid moiety of -XAA- is independently selected from alanine (Ala), arginine (Arg), glycine (Gly), histidine (His), isoleucine (He), leucine (Leu), lysine (Lys), phenylalanine (Phe), tryptophan (Trp), tyrosine (Tyr), valine (Vai), citrulline (Cit), and homocitrulline (HoCit).

62. The conjugate of any one of claims 1 to 17, wherein each amino acid moiety of -XAA- is independently selected from alanine (Ala), glycine (Gly), lysine (Lys), phenylalanine (Phe), valine (Vai), and citrulline (Cit).

63. The conjugate of any one of claims 1 to 16, wherein the amino acid sequence -XAA- is selected from -Val-Cit-, -Cit-Val-, -Vai-Ala-, -Ala-Vai-, -Phe-Lys-, -Lys-Phe-, -Ala-Ala-, -Val-Val-, -Gly-Gly-, -Ala-Ala-Ala-, -Gly-Gly-Gly-, -Gly-Gly-Phe-Gly-(SEQ ID NO: 1032), -Gly-Phe-Gly-Gly-(SEQ ID NO: 1033),-Gly-Gly-Gly-Phe-(SEQ ID NO: 1034), -Phe-Gly-Gly-Gly-(SEQ ID NO: 1035), and -Gly-Gly-Gly-Gly-(SEQ ID NO: 1036).

64. The conjugate of any one of claims 1 to 17, wherein the amino acid sequence -XAA- is selected from -Val-Cit-, -Cit-Val-, -Val-Ala-, -Ala-Vai-, -Phe-Lys-, -Lys-Phe-, -Ala-Ala-, -Val-Val-, -Gly-Gly-, -Ala-Ala-Ala-, and -Gly-Gly-Gly-.

65. The conjugate of any one of claims 1 to 63, wherein the single stranded oligonucleotide P is a phosphorodiamidate morpholino oligonucleotide or an antisense oligonucleotide.

66. The conjugate of any one of claims 1 to 65, wherein the single stranded oligonucleotide P is delivered into a muscle cell.

67. The conjugate of any one of claims 1 to 66, wherein the single stranded oligonucleotide P induces skipping of exon 23 of the DMD gene.

68. The conjugate of claim 65, wherein the phosphorodiamidate morpholino oligonucleotide comprises the sequence 5'-C6 Amino-GGCCAAACCTCGGCTTACCTGAAAT-3' (SEQ ID NO:408).

69. The conjugate of claim 65, wherein the antisense oligonucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 1045, SEQ ID NOs: 158-222, SEQ ID NO:395- 405, and SEQ ID NO:410-988.

70. The conjugate of claim 65, wherein the antisense oligonucleotide is a peptide nucleic acid (PNA) oligonucleotide.

71. The conjugate of claim 70, wherein the peptide nucleic acid (PNA) oligonucleotide comprises the sequence (C)-3’-TAAAGTCCATTCGGCTCCAAACCGG-C6 Amino-5’(N) (SEQ ID NO: 409).

72. The conjugate of any one of claims 1 to 71, wherein the single stranded oligonucleotide P comprises at least from about 10 to about 30 nucleotides in length.

73. The conjugate of claim 1, wherein the truncated protein modulates muscular dystrophy.

74. The conjugate of claim 73, wherein the muscular dystrophy is Duchenne muscular dystrophy or Becker muscular dystrophy.

75. The conjugate of any one of claims 1 to 74, wherein the VL CDR1 comprises the amino acid sequence of SEQ ID NO:9, CDR2 comprises the amino acid sequence of SEQ ID NO: 10, CDR3 comprises SEQ ID NO: 11; and the VH CDR1 comprises the amino acid sequence of SEQ ID NO:15, CDR2 comprises the amino acid sequence of SEQ ID NO:4, CDR3 comprises the amino acid sequence of SEQ ID NO:5.

76. The conjugate of any one of claims 1 to 74, wherein the VL CDR1 comprises the amino acid sequence of SEQ ID NO:29, CDR2 comprises the amino acid sequence of SEQ ID NO: 10, CDR3 comprises SEQ ID NO: 11; and the VH CDR1 comprises the amino acid sequence of SEQ ID NO: 15, CDR2 comprises the amino acid sequence of SEQ ID NO:26, CDR3 comprises the amino acid sequence of SEQ ID NO:5.

77. The conjugate of any one of claims 1 to 74, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable region (VL) comprising an amino acid sequenceof SEQ ID NO:21 and a heavy chain variable region (VH) comprising an amino acid sequence of SEQ ID NO: 14.

78. The conjugate of any one of claims 1 to 74, wherein the antibody or antigen-binding fragment thereof comprises a full length light chain (LC) comprising an amino acid sequence of SEQ ID NO:20 and a full length heavy chain (HC) comprising an amino acid sequence of SEQ ID NO:13.

79. The conjugate of any one of claims 1 to 78, wherein the antibody or antigen-binding fragment thereof comprises: a light chain variable domain (VL) comprising an amino acid sequence that is at least 95% identical to an amino acid sequence selected from the group consisting of 3E10-VL-H1 (SEQ ID NO:85), 3E10-VL-H2 (SEQ ID NO:86), 3E10-VL-H3 (SEQ ID NO:87), 3E10-VL-H4 (SEQ ID NO:88), 3E10-VL-H5 (SEQ ID NO:89), and 3E10-VL-H6 (SEQ ID NO:90); and a heavy chain variable domain (VH) comprising an amino acid sequence that is at least 95% identical to an amino acid sequence selected from the group consisting of 3E10-VH-H1 (SEQ ID NO:64), 3E10-VH-H2 (SEQ ID NO:65), 3E10-VH-H3 (SEQ ID NO:66), 3E10-VH-H4 (SEQ ID NO:67), 3E1O-VH-H5 (SEQ ID NO:68), 3E10-VH-H6 (SEQ ID NO:69), and 3E10- VH-H7 (SEQ ID NO:70).

80. The conjugate of any one of claims 1 to 78, wherein the antibody or antigen-binding fragment thereof comprises: a light chain variable domain (VL) comprising an amino acid sequence selected from the group consisting of 3E10-VL-H1 (SEQ ID NO:85), 3E10-VL-H2 (SEQ ID NO:86), 3E10-VL- H3 (SEQ ID NO:87), 3E10-VL-H4 (SEQ ID NO:88), 3E10-VL-H5 (SEQ ID NO:89), and 3E10- VL-H6 (SEQ ID NOVO); and a heavy chain variable domain (VH) comprising an amino acid sequence selected from the group consisting of 3E10-VH-H1 (SEQ ID NO:64), 3E10-VH-H2 (SEQ ID NO:65), 3E10-VH- H3 (SEQ ID NO:66), 3E10-VH-H4 (SEQ ID NO:67), 3E10-VH-H5 (SEQ ID NO:68), 3E10- VH-H6 (SEQ ID NO:69), and 3E10-VH-H7 (SEQ ID NO:70).

81. The conjugate of any one of claims 1 to 78, wherein the antibody or antigen-binding fragment thereof comprises a VL / VH pair selected from the group consisting of:(a) VL-hl (SEQ ID NO:85) and VH-hl (SEQ ID NO:64),(b) VL-hl (SEQ ID NO:85) and VH-h2 (SEQ ID NO:65),(c) VL-hl (SEQ ID NO:85) and VH-h3 (SEQ ID NO:66),(d) VL-hl (SEQ ID NO:85) and VH-h4 (SEQ ID NO:67),(e) VL-h2 (SEQ ID NO:86) and VH-hl (SEQ ID NO:64),(f) VL-h2 (SEQ ID NO:86) and VH-h2 (SEQ ID NO:65),(g) VL-h2 (SEQ ID NO:86) and VH-h3 (SEQ ID NO:66),(h) VL-h2 (SEQ ID NO:86) and VH-h4 (SEQ ID NO:67),(i) VL-h3 (SEQ ID NO: 87) and VH-hl (SEQ ID NO: 64),(j) VL-h3 (SEQ ID NO: 87) and VH-h2 (SEQ ID NO: 65),(k) VL-h3 (SEQ ID NO:87) and VH-h3 (SEQ ID NO:66),(l) VL-h3 (SEQ ID NO: 87) and VH-h4 (SEQ ID NO: 67),(m) VL-h4 (SEQ ID NO: 88) and VH-hl (SEQ ID NO: 64),(n) VL-h4 (SEQ ID NO:88) and VH-h2 (SEQ ID NO:65),(o) VL-h4 (SEQ ID NO:88) and VH-h3 (SEQ ID NO:66),(p) VL-h4 (SEQ ID NO:88) and VH-h4 (SEQ ID NO:67),(q) VL-h5 (SEQ ID NO:89) and VH-h5 (SEQ ID NO:68),(r) VL-h5 (SEQ ID NO:89) and VH-h6 (SEQ ID NO:69),(s) VL-h6 (SEQ ID NO:90) and VH-h5 (SEQ ID NO:68), and(t) VL-h6 (SEQ ID NO:90) and VH-h6 (SEQ ID NO:69).

82. The conjugate of any one of claims 1 to 78, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable domain (VL) comprising an amino acid sequence that is at least 95% identical to 3E10-VL-H6 (SEQ ID NO:90) and a heavy chain variable domain (VH) comprising an amino acid sequence that is at least 95% identical to 3E10-VH-H6 (SEQ ID NO:69).

83. The conjugate of any one of claims 1 to 78, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable domain (VL) comprising the amino acid sequence of 3E10-VL-H6 (SEQ ID NO:90) and a heavy chain variable domain (VH) comprising the amino acid sequence of 3E10-VH-H6 (SEQ ID NO:69).

84. The conjugate of any one of claims 1 to 78, wherein the antibody or antigen-binding fragment thereof comprises: a light chain variable domain (VL) comprising an amino acid sequence that is at least 95% identical to SEQ ID NO: 105 and a heavy chain variable domain (VH) comprising an amino acid sequence that is at least 95% identical to SEQ ID NO: 117.

85. The conjugate of any one of claims 1 to 78, wherein the antibody or antigen-binding fragment thereof comprises: a light chain variable domain (VL) comprising the amino acid sequence of SEQ ID NO: 105 and a heavy chain variable domain (VH) comprising the amino acid sequence of SEQ ID NO: 117.

86. A method for treating a genetic disease or disorder in a subj ect in need thereof, the method comprising administering a therapeutically effective amount of a conjugate according to any one of claims 1 to 85 to the subject.

87. The method of claim 86, wherein the disease or disorder is a neurogenetic disease, a musculoskeletal disorder, a cardiovascular disease, a metabolic disease, a cancer, a lung disorder, or a disease that can be benefitted by exon-skipping therapies.

88. The method of claim 87, wherein the neurogenetic disease is CDG, FD, PBD1A, MRD23, EPM5, FHM1, AHC, AGS6, EIEEF4, VWM, ICCA, BFIE, EKD1, ALGS, TSC, USH, PTHS, SMS, GEFSP9, HGPS, FTDP-17, PHMDS, SCZD15, NF2, or PARK8.

89. The method of claim 87, wherein the musculoskeletal disorder is DMD, DM1, DM2, SMA, or IBMPFDl.

90. The method of claim 87, wherein the cardiovascular disease is FH.

91. The method of claim 87, wherein the metabolic disease is ACADMD, PKU, or NPC1.

92. The method of claim 87, wherein the cancer is a RAS-associated cancer or MLD.

93. The method of claim 87, wherein the lung disorder is CF.

94. The method of claim 87, wherein the disease that can be benefitted by exon-skipping therapies is IBS or DEB.

95. A method of treating Duchenne muscular dystrophy (DMD) in a subject in need thereof, the method comprising administering a therapeutically effective amount of a conjugate according to any one of claims 1 to 85 to the subject.

96. A conjugate according to any one of claims 1 to 85 for use in the treatment of a genetic disease or disorder.

97. The conjugate of claim 96, wherein the disease or disorder is a neurogenetic disease, a musculoskeletal disorder, a cardiovascular disease, a metabolic disease, a cancer, a lung disorder, or a disease that can be benefitted by exon-skipping therapies.

98. The conjugate of claim 96, wherein the neurogenetic disease is CDG, FD, PBD1A, MRD23, EPM5, FHM1, AHC, AGS6, EIEEF4, VWM, ICCA, BFIE, EKD1, ALGS, TSC, USH, PTHS, SMS, GEFSP9, HGPS, FTDP-17, PHMDS, SCZD15, NF2, or PARK8.

99. The conjugate of claim 96, wherein the musculoskeletal disorder is DMD, DM1, DM2, SMA, or IBMPFDl.

100. The conjugate of claim 96, wherein the cardiovascular disease is FH.

101. The conjugate of claim 96, wherein the metabolic disease is ACADMD, PKU, or NPC1.

102. The conjugate of claim 96, wherein the cancer is a RAS-associated cancer or MLD.

103. The conjugate of claim 96, wherein the lung disorder is CF.

104. The conjugate of claim 96, wherein the disease that can be benefitted by exon-skipping therapies is IBS or DEB.

105. A conjugate according to any one of claims 1 to 85 for use in the treatment of Duchenne muscular dystrophy (DMD).

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