Antigen binding molecules to CD3 and CD19

Abstract

Disclosed herein are anti-CD3 antigen binding molecules, anti-CD19 antigen binding molecules, anti-CD19 and anti-CD3 bi-specific antigen binding molecules, and methods for the use thereof.

Description

ANTIGEN BINDING MOLECULES TO CD3 AND CD19CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U. S. Provisional Application No. 63 / 793,511, filed on April 23, 2025, and U. S. Provisional Application No. 63 / 692,069, filed on September 7, 2024, the disclosures of each of which are hereby incorporated by reference in their entireties.SEQUENCE LISTING

[0002] The contents of the electronic sequence listing(121168000407_Sequence_Listing_ST26.xml; Size: 3,392,961 bytes; and Date of Creation: August 25, 2025) is herein incorporated by reference in its entirety.TECHNICAL FIELD

[0003] Disclosed herein are antigen binding molecules binding CD3 and CD 19, and methods for the use thereof.BACKGROUND

[0004] CD3 (Cluster of Differentiation 3) is a T-cell co-receptor that is present on both cytotoxic and helper T-cells. CD3 is involved in activating T-cells making CD3 a potential target to stimulate or block T cell activation. CD19 (Cluster of Differentiation 19) is a signaling glycoprotein that is present on all B-cells, which makes CD 19 an important marker and potential target for therapies aimed to modulate for B-cells.SUMMARY

[0005] In a first aspect, the disclosure is directed to a bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprising an anti-CD19 antigen binding domain and an anti-CD3 antigen binding domain.

[0006] In some variations, the anti-CD19 antigen binding domains comprise a first binding subunit comprising the CDR sequences of SEQ ID NOs: 1, 2, and 3, and a second binding subunit comprising the CDR sequences of SEQ ID NOs: 4, 5, and 6. The anti-CD3 antigen binding domains comprise a first binding subunit comprising the CDR sequences of SEQ ID NO: 9, SEQ ID NO: 10 or 11, and SEQ ID NO: 12, 13, 14, 15, or 16, and a second binding subunit comprising the CDR sequences of SEQ ID NOs: 17, 18, and 19.- 1 - 1211680004074923-9899- 2484 1P190500. W0.01

[0007] In some variations, the bi-specific anti-CD19 and anti-CD3 antigen binding molecules include a first monomer, second monomer, and light chain. The first monomer comprises a scFv-domain linker-CH2-CH3 domain in which the scFv comprises an antigen binding domain that binds CD3, and the CH2-CH3 is a first variant Fc domain. The second monomer comprises a VH-CHl-hinge-CH2-CH3, in which the CH2-CH3 is a second variant Fc domain complementary to the first variant Fc domain. The first variant Fc domain and second variant Fc domain form a heterodimeric backbone. The light chain together with the second monomer forms a Fab domain that binds CD19. In some variations, the Fab domain is a Fv domain.

[0008] In some variations, the bi-specific anti-CD19 and anti-CD3 antigen binding molecules include first monomer, a second monomer, a first light chain, and a second light chain. The first monomer comprises a VH-CH1 -first domain linker-scFv-second domain linker-CH2-CH3 in which the the scFv comprises an antigen binding domain that binds CD3, and the CH2-CH3 is a first variant Fc domain. The second monomer comprises a VH-CHl-hinge-CH2-CH3, in which the CH2-CH3 is a second variant Fc domain complementary to the first variant Fc domain. The first variant Fc domain and second variant Fc domain form a heterodimeric backbone. The first light chain together with the first monomer forms a Fab domain that binds CD 19. The second light chain together with the second monomer forms a Fab domain that binds CD19. In some variations, the Fab domain is a Fv domain.

[0009] In a second aspect, the disclosure is directed to an anti-CD19 antigen binding molecules comprising an antigen binding domain. The antigen binding domain comprises a first binding subunit comprising the CDR sequences of SEQ ID NOs: 1, 2, and 3, and a second binding subunit comprising the CDR sequences of SEQ ID NOs: 4. 5, and 6.

[0010] In some variations, the anti-CD19 antigen binding molecules, comprising an antigen binding domain. The antigen binding domain comprises a first binding subunit, a second binding subunit, and a framework region comprising at least 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 20. 21. 22. 23. 24, 25, 26, and 27.

[0011] In some variations, the anti-CD19 antigen binding molecules, comprising an antigen binding domain. The antigen binding domain comprises an amino acid sequence comprising at least 95% identity to SEQ ID NO: 20 N-terminal to a first binding subunit CDR1, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 21 between the first binding subunit CDR1 and a first binding subunit CDR2, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 22 between the first binding subunit CDR2 and a first binding subunit CDR3, an amino acid sequence comprising at least 95% identity7to SEQ ID NO: 23 C-terminal to the first binding subunit CDR3, an amino acid sequence comprising at least 95%P190500. W0.01 identity to SEQ ID NO: 24 N-terminal to a second binding subunit CDR1, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 25 between the second binding subunit CDR1 and a second binding subunit CDR2, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 26 between the second binding subunit CDR2 and a second binding subunit CDR3, and an amino acid sequence comprising at least 95% identity to SEQ ID NO: 27 C-terminal to the second binding subunit CDR3.

[0012] In a third aspect, the disclosure is directed to an anti-CD3 antigen binding molecules comprising an antigen binding domain. The antigen binding domain comprises a first binding subunit comprising the CDR sequences of SEQ ID NO: 9, SEQ ID NO: 10 or 11, and SEQ ID NO: 12, 13, 14, 15, or 16, and a second binding subunit comprising the CDR sequences of SEQ ID NOs: 17, 18, and 19.

[0013] In some variations, the anti-CD3 antigen binding molecules, comprising a first binding subunit and a second binding subunit, and a framework region comprising at least 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 28, 29, 30, 31, 32, 33, 34, and 35.

[0014] In some variations, the anti-CD3 antigen binding molecules, comprising an amino acid sequence comprising at least 95% identity to SEQ ID NO: 28 N-terminal to a first binding subunit CDR1. an amino acid sequence comprising at least 95% identity to SEQ ID NO: 29 between the first binding subunit CDR1 and a first binding subunit CDR2, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 30 between the first domain CDR2 and a first domain CDR3, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 31 C-terminal to the first domain CDR3, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 32 N-terminal to a second domain CDR1, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 33 between the second domain CDR1 and a second domain CDR2, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 34 between the second domain CDR2 and a second domain CDR3, and an amino acid sequence comprising at least 95% identity to SEQ ID NO: 35 C-terminal to the second domain CDR3.BRIEF DESCRIPTION OF THE DRAWINGS

[0015] The summary, as well as the following detailed description, is further understood when read in conjunction with the appended drawings. For the purpose of illustrating the disclosed compositions and methods, the drawings show example embodiments of the compositions and methods; however, the compositions and methods are not limited to the specific embodiments disclosed. In the drawings:P190500. W0.01

[0016] FIG. 1 depicts sequences for human, mouse, and cynomolgus CD3. Such CD3 sequences are useful for the development of cross-reactive CD3 antigen binding domains for ease of clinical development.

[0017] FIG. 2 depicts sequences for human, mouse, and cynomolgus CD 19. Such CD 19 sequences are useful for the development of cross-reactive CD 19 antigen binding domains for ease of clinical development.

[0018] FIG. 3 A - FIG.3F depict useful pairs of heterodimerization variant sets (including skew and pl variants). There are variants for which there are no corresponding “monomer 2"’ variants (FIG.3F). Such variants are pl variants that can be used alone on either monomer of a bispecific antibody (e.g., CD19 x CD3 bsAb), or included, for example, on the non-scFv side of a format that utilizes an scFv as a component and an appropriate charged scFv linker can be used on the second monomer that utilizes an scFv as the CD3 binding domain. Suitable charged linkers are shown in FIG. 6. Skew variants and pl variants are described, for example, in U. S. Patent No. 11,299,554. U. S. Patent No. 9,850.320 U. S. Patent Publication No.2023 / 0048390, PCT Publication WO2014 / 145806, and U. S. Patent No. 9,650,446, each of which is hereby incorporated by reference in its entirety.

[0019] FIG. 4 depicts a list of isosteric variant antibody constant regions and their respective substitutions, pl (-) indicates lower pl variants, while pl_(+) indicates higher pl variants. These variants can be optionally and independently combined with other variants, including heterodimerization variants, outlined herein, according to an illustrative embodiment.

[0020] FIG. 5A and FIG. 5B depict useful ablation variants that ablate FcyR binding (also referred to as “knockouts” or “KO” variants). In some embodiments, such ablation variants are included in the Fc domain of both monomers of the subject antibody described herein. In other embodiments, the ablation variants are only included on only one variant Fc domain.

[0021] FIG. 6 depicts a number of charged scFv linkers that find use in increasing or decreasing the pl of the subject heterodimeric bispecific antibodies that utilize one or more scFv as a component, as described herein (e.g., CD19 x CD3 bispecific antibodies ( bsAbs)). The (+H) positive linker finds particular use herein, particularly with anti-CD3 VL and VH sequences shown herein. A single prior art scFv linker with a single charge is referenced as “Whitlow,” from Whitlow et al., Protein Engineering 6(8): 989-995 (1993). It should be noted that this linker was used for reducing aggregation and enhancing proteolytic stability in scFvs. Such charged scFv linkers can be used in any of the subject antibody formats disclosed herein that include scFvs (e.g., 1 + 1 Fab-scFv-Fc and 2 + 1 Fab2-scFv-Fc formats).

[0022] FIG. 7A and FIG. 7B depict a number of example domain linkers. In some embodiments, these linkers find use linking a single-chain Fv to an Fc chain. In someP190500. W0.01 embodiments, these linkers may be combined in any orientation. For example, a GGGGS linker may be combined with a '‘lower half hinge” linker at the N-terminus or at the C-terminus.

[0023] FIG. 8 shows a particularly useful bispecific antibody backbone for the CD 19 x CD3 bispecific antibodies (bsAbs) of the disclosure. Although the backbone is described here in the context of the 1 + 1 Fab-scFv-Fc format, they can be adapted for use in other bispecific antibody formats.

[0024] FIG. 9 depicts various heterodimeric skew variant amino acid substitutions that can be used with the heterodimeric antibodies described herein.

[0025] FIG. 10 - FIG. 10F show the sequences of several heterodimeric CD 19 x CD3 bsAb backbones based on human IgGl, without the variable domains. Heterodimeric Fc backbone 1 is based on human IgGl (356E / 358M allotype), and includes the L368D / K370S skew variants and the Q295E / N384D / Q418E / N421D pl variants on a first heterodimeric Fc chain, the S364K / E357Q skew variants on a second heterodimeric Fc chain, and the E233P / L234V / L235A / G236del / S267K ablation variants on both chains. Heterodimeric Fc backbone 2 is based on human IgGl (356E / 358M allotype), and includes the L368D / K370S skew variants and the Q295E / N384D / Q418E / N421D pl variants on a first heterodimeric Fc chain, the S364K skew variant on a second heterodimeric Fc chain, and the E233P / L234V / L235A / G236del / S267K ablation variants on both chains. Heterodimeric Fc backbone 3 is based on human IgGl (356E / 358M allotype), and includes the L368E / K370S skew variants and the Q295E / N384D / Q418E / N421D pl variants on a first heterodimeric Fc chain, the S364K skew variant on a second heterodimeric Fc chain, and the E233P / L234V / L235A / G236del / S267K ablation variants on both chains. Heterodimeric Fc backbone 4 is based on human IgGl (356E / 358M allotype), and includes the K360E / Q362E / T411E skew variants and the Q295E / N384D / Q418E / N421D pl variants on afirst heterodimeric Fc chain, the D401K skew variant on a second heterodimeric Fc chain, and the E233P / L234V / L235A / G236del / S267K ablation variants on both chains. Heterodimeric Fc backbone 5 is based on human IgGl (356D / 358L allotype), and includes the L368D / K370S skew variants and the Q295E / N384D / Q418E / N421D pl variants on a first heterodimeric Fc chain, the S364K / E357Q skew variants on a second heterodimeric Fc chain, and the E233P / L234V / L235A / G236del / S267K ablation variants on both chains. Heterodimeric Fc backbone 6 is based on human IgGl (356E / 358M allotype), and includes the L368D / K.370S skew variants and the Q295E / N384D / Q418E / N421D pl variants on a first heterodimeric Fc chain, the S364K / E357Q skew variants on a second heterodimeric Fc chain, and the E233P / L234V / L235A / G236del / S267K ablation variants and N297A variant that removes glycosylation on both chains. Heterodimeric Fc backbone 7 is based on human IgGlP190500. W0.01 (356E / 358M allotype), and includes the L368D / K370S skew variants and the Q295E / N384D / Q418E / N421D pl variants on a first heterodimeric Fc chain, the S364K / E357Q skew variants on a second heterodimeric Fc chain, and the E233P / L234V / L235A / G236del / S267K ablation variants and N297S variant that removes glycosylation on both chains. Eleterodimeric Fc backbone 8 is based on human IgG4. and includes the L368D / K370S skew variants and the Q295E / N384D / Q418E / N421D pl variants on a first heterodimeric Fc chain, the S364K / E357Q skew variants on a second heterodimeric Fc chain, and the S228P (according to EU numbering, S241P in Kabat) variant that ablates Fab arm exchange (as is known in the art) on both chains. Eleterodimeric Fc backbone 9 is based on human IgG2, and includes the L368D / K370S skew variants and the Q295E / N384D / Q418E / N421D pl variants on a first heterodimeric Fc chain, the S364K / E357Q skew variants on a second heterodimeric Fc chain. Heterodimeric Fc backbone 10 is based on human IgG2, and includes the L368D / K370S skew variants and the Q295E / N384D / Q418E / N421D pl variants on a first heterodimeric Fc chain, the S364K / E357Q skew variants on a second heterodimeric Fc chain, and the S267K ablation variant on both chains. Heterodimeric Fc backbone 11 is based on human IgGl (356E / 358M allotype), and includes the L368D / K370S skew variants and the Q295E / N384D / Q418E / N421D pl variants on a first heterodimeric Fc chain, the S364K / E357Q skew variants on a second heterodimeric Fc chain, and the E233P / L234V / L235A / G236del / S267K ablation vanants and M428L / N434S Xtend variants on both chains. Heterodimeric Fc backbone 12 is based on human IgGl (356E / 358M allotype), and includes the L368D / K370S skew variants on a first heterodimeric Fc chain, the S364K / E357Q skew variants and P217R / P229R / N276K pl variants on a second heterodimeric Fc chain, and the E233P / L234V / L235A / G236del / S267K ablation variants on both chains. Heterodimeric Fc backbone 13 is based on human IgGl (356E / 358M allotype), and includes the T366W skew variant on a first heterodimeric Fc chain, the T366S / L36 A / Y407V skew variants and H435R / Y436F purification variants on a second heterodimeric Fc chain, and the L234A / L235A / D265S ablation variants on both chains. Heterodimeric Fc backbone 14 is based on human IgGl (356E / 358M allotype), and includes the T366W skew variant on a first heterodimeric Fc chain, the T366S / L368A / Y407V skew variants and H435R / Y436F purification variants on a second heterodimeric Fc chain, and the L234A / L235A / D265S ablation variants and M252Y / S254T / T256E half-life extension variants on both chains. Heterodimeric Fc backbone 15 is based on human IgGl (356D / 358L allotype), and includes the L368D / K370S skew variants and the Q295E / N384D / Q418E / N421D pl variants on a first heterodimeric Fc chain, the S364K / E357Q skew variants on a second heterodimeric Fc chain, and the E233P / L234V / L235A / G236del / S267K ablation variants and M428L / N434S Xtend variants onP190500. W0.01 both chains. Heterodimeric Fc backbone 16 is based on human IgGl (356E / 358M allotype), and includes the L368D / K370S skew variants and the Q295E / N384D / Q418E / N421D pl variants on a first heterodimeric Fc chain, the S364K / E357Q skew variants on a second heterodimeric Fc chain, and the E233P / L234V / L235A / G236del / S267K ablation variants and M428L / N434A Xtend variants on both chains. Heterodimeric Fc backbone 17 is based on human IgGl (356D / 358L allotype), and includes the L368D / K370S skew variants and the Q295E / N384D / Q418E / N421D pl variants on a first heterodimeric Fc chain, the S364K / E357Q skew variants on a second heterodimeric Fc chain, and the E233P / L234V / L235A / G236del / S267K ablation variants and M428L / N434A Xtend variants on both chains. Included within each of these backbones are sequences that are 90, 95, 98 and 99% identical (as defined herein) to the recited sequences, and / or contain from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 additional amino acid substitutions (as compared to the “parent” of the Figure, which, as will be appreciated by those in the art, already contain a number of amino acid modifications as compared to the parental human IgGl (or IgG2 or IgG4, depending on the backbone). That is, the recited backbones may contain additional amino acid modifications (generally amino acid substitutions) in addition or as an alternative to the skew, pl and ablation variants contained within the backbones of this Figure. Additionally, the backbones depicted herein may include deletion of the C-terminal glycine (G446_) and / or lysine (K447_)- The C-terminal glycine and / or lysine deletion may be intentionally engineered to reduce heterogeneity or in the context of certain bispecific formats, such as the mAb-scFv format. Additionally, C-terminal glycine and / or lysine deletion may occur naturally for example during production and storage. As used herein, a “heterodimeric Fc chain" is also referred to as a “variant Fc domain.”

[0026] FIG. 11 depicts sequences for “CHI” that find use in embodiments of CD 19 x CD3 bsAbs.

[0027] FIG. 12 depicts sequences for “hinge” that find use in embodiments of CD 19 x CD3 bsAbs.

[0028] FIG. 13 depicts the constant domain of the cognate light chains that find use in the subject CD19 x CD3 bsAbs that utilize a Fab binding domain.

[0029] FIG. 14 depicts the variable heavy and variable light chain sequences for CD 19-OG (VH=SEQ ID NO: 151 / VL=SEQ ID NO: 152), an example CD19 binding domain. CDRs are underlined and slashes indicate the border(s) between the variable regions and constant domain. As noted herein and is true for every sequence herein containing CDRs, the exact identification of the CDR locations may be slightly different depending on the numbering used as is shown in Table 1, and thus included herein are not only the CDRs that are underlined but also CDRs included within the VH and VL domains using other numbering systems.P190500. W0.01 Furthermore, as for all the sequences in the Figures, these VH and VL sequences can be used either in a scFv format or in a Fab format.

[0030] FIG. 15 depicts the variable heavy and variable light chain sequences for CD 19-NEW (VH=SEQ ID NO: 7 / VL=SEQ ID NO: 8), an example CD19 binding domain that is a reengineered variant of CD19-OG. CDRs are underlined and slashes indicate the border(s) between the variable regions and constant domain. As noted herein and is true for every sequence herein containing CDRs, the exact identification of the CDR locations may be slightly different depending on the numbering used as is shown in Table 1, and thus included herein are not only the CDRs that are underlined but also CDRs included within the VH and VL domains using other numbering systems. Furthermore, as for all the sequences in the Figures, these VH and VL sequences can be used either in a scFv format or in a Fab format.

[0031] FIG. 16A - FIG. 16F depict the variable heavy and variable light chain sequences for illustrative CD3 binding domains which find use in the CD19 x CD3 bsAbs of the disclosure. The CDRs are underlined, the scFv linker is double underlined (in the sequences, the scFv linker is a positively charged scFv (GKPGS)4 linker (SEQ ID NO: 36), although as will be appreciated by those in the art, this linker can be replaced by other linkers, including uncharged or negatively charged linkers, some of which are depicted in FIG. 6), and the slashes indicate the border(s) of the variable domains. In addition, the naming convention illustrates the orientation of the scFv from N- to C-terminus. As noted herein and is true for every sequence herein containing CDRs, the exact identification of the CDR locations may be slightly different depending on the numbering used as is shown in Table 1, and thus included herein are not only the CDRs that are underlined but also CDRs included within the VH and VL domains using other numbering systems. Furthermore, as for all the sequences in the Figures, these VH and VL sequences can be used either in a scFv format or in a Fab format.

[0032] FIG. 17A - FIG. 17J depict the variable heavy’ and variable light chain sequences for example CD3 binding domains re-engineered to improve stability' while retaining an affinity ladder. The CDRs are underlined, the scFv linker is double underlined (in the sequences, the scFv linker is a positively charged scFv (GKPGS)4 linker (SEQ ID NO: 36), although as will be appreciated by those in the art, this linker can be replaced by other linkers, including uncharged or negatively charged linkers, some of which are depicted in FIG. 6), and the slashes indicate the border(s) of the variable domains. In addition, the naming convention illustrates the onentation of the scFv from N- to C-terminus. As noted herein and is true for every sequence herein containing CDRs, the exact identification of the CDR locations may be slightly different depending on the numbering used as is shown in Table 1, and thus included herein are not only the CDRs that are underlined but also CDRs included within the VH and VLP190500. W0.01 domains using other numbering systems. Furthermore, as for all the sequences in the Figures, these VH and VL sequences can be used either in a scFv format or in a Fab format.

[0033] FIG. ISA - FIG. 18N depict bispecific formats of the present disclosure. FIG.18A depicts the " I + 1 Fab-scFv-Fc'’ format, with a first Fab arm binding a first antigen and a second scFv arm binding second antigen. The 1 + 1 Fab-scFv-Fc format comprises a first monomer comprising a first heavy chain variable region (VH1) covalently attached to the N-terminus of a first heterodimeric Fc backbone (optionally via a linker), a second monomer comprising a single-chain Fv covalently attached to the N-terminus of a second corresponding heterodimeric Fc backbone (optionally via a linker), and a third monomer comprising a light chain variable region covalently to a light chain constant domain, wherein the light chain variable region is complementary to the VH1. FIG. 18B depicts the “2 + 1 Fab2-scFv-Fc” format, with a first Fab arm and a second Fab-scFv arm, wherein the Fab binds a first antigen and the scFv binds second antigen. The 2 + 1 Fab2-scFv-Fc format comprises a first monomer comprising a first heavy chain variable region (VH1) covalently attached to the N-terminus of a first heterodimeric Fc backbone (optionally via a linker), a second monomer comprising the VH1 covalently attached (optionally via a linker) to a single-chain Fv covalently attached (optionally via a linker) to the N-terminus of a second corresponding heterodimeric Fc backbone, and a third monomer comprising a light chain variable region covalently to a light chain constant domain, wherein the light chain variable region is complementary to the VH1. FIG. 18C depicts the “1 + 1 Common Light Chain” or ‘ + 1 CLC” format, with a first Fc comprising a first Fab arm binding a first antigen and a second Fc comprising a second Fab arm binding second antigen. The 1 + 1 CLC format comprises a first monomer comprising VHl-CHl-hinge-CH2-CH3, a second monomer comprising VH2-CHl-hinge-CH2-CH3. and a third monomer comprising VL-CL. The VL pairs with the VH1 to form a binding domain with a first antigen binding specificity: and the VL pairs with the VH2 to form a binding domain with a second antigen binding specificity. FIG. 18D depicts the "2 + 1 Common Light Chain” or “2 + 1 CLC” format, with a first Fc comprising 2 Fab arms each binding a first antigen and a second Fc comprising 1 Fab arm binding a second antigen. The 2 + 1 CLC format comprises a first monomer comprising VHl-CHl-hinge-VHl-CHl-hinge-CH2-CH3, a second monomer comprising VH2-CHl-hinge-CH2-CH3, and a third monomer comprising VL-CL. The VL pairs with the first and second VH1 to form binding domains with a first antigen binding specificity; and the VL pairs with the VH2 to form a binding domain with a second antigen binding specificity. FIG. 18E depicts the “2 + 1 mAb-scFv” format, with a first Fc comprising an N-terminal Fab arm binding a first antigen and a second Fc comprising an N-terminal Fab arm binding the first antigen and a C-terminal scFv binding a second antigen. The 2 + 1 mAb-scFv format comprises a first monomerP190500. W0.01 comprising VHl-CHl-hinge-CH2-CH3. a second monomer comprising VHl-CHl-hinge-CH2- CH3-scFv, and a third monomer comprising VL-CL. The VL pairs with the first and second VH1 to form binding domains with binding specificity for the first antigen. FIG. 18F depicts the “2 + 1 mAb-scFv” format which comprises a first monomer comprising fromN-terminal to C-terminal VHl-CHl-linker-VH2-CHl-hinge-CH2-CH3 wherein CH2-CH3 is a first heterodimeric Fc domain; a second monomer comprising from N-terminal to C-terminal scFv-linker-CH2-CH3 wherein CH2-CH3 is a second heterodimeric Fc domain complementary to the first heterodimeric Fc domain and wherein the scFv has a first antigen specificity; and a third monomer that is a common light chain comprising from N-terminal to C-terminal VL-CL wherein the VL pairs with VH1 and VH2 of the first monomer to form two antigen binding domains each having a specificity for a second antigen binding domain. Additional bispecific formats include dual scFv (FIG. 18G), one-arm scFv-mAb (FIG. 18H), scFv-mAb (FIG. 181), bispecific mAb (FIG. 18J), one-arm central-scFv (FIG. 18K), mAb-Fv (FIG. 18L), central-Fv (FIG. 18M), and trident (FIG. 18N).

[0034] FIG. 19A - FIG. 19C depict the sequences for illustrative CD19 x CD3 bsAbs in the 1 + 1 Fab-scFv-Fc format. It should be noted that any other suitable backbones including those depicted in FIG. 10A - FIG. 10F may be used. CDRs are underlined and slashes indicate the border(s) between the variable regions and other domains. It should be noted that the CD 19 x CD3 bsAbs can utilize variable region, Fc region, and constant domain sequences that are 90, 95, 98 and 99% identical (as defined herein), and / or contain from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid substitutions. In addition, each sequence outlined herein can include or exclude the M428L / N434S, M428L / N434A, or M252Y / S254T / T256E variants in one or preferably both Fc domains, which including the variants results in longer half-life in serum.

[0035] FIG. 20A - FIG. 20 AD depict the sequences for illustrative CD 19 x CD3 bsAbs in the 2 + 1 Fab2-scFv-Fc format. It should be noted that any other suitable backbones including those depicted in FIG. 1 OA - FIG. 1 OF may be used. CDRs are underlined and slashes indicate the border(s) between the variable regions and other domains. It should be noted that the CD19 x CD3 bsAbs can utilize variable region, Fc region, and constant domain sequences that are 90, 95, 98 and 99% identical (as defined herein), and / or contain from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid substitutions. In addition, each sequence outlined herein can include or exclude the M428L / N434S. M428L / N434A, or M252Y / S254T / T256E variants in one or preferably both Fc domains, which including the variants results in longer half-life in serum.

[0036] FIG. 21 depicts in vitro depletion of CD19+Ramos cells by CD19 x CD3 bsAbs (in 1 + 1 or 2 + 1 formats with either high or intermediate affinity CD3 binding domains).P190500. W0.01

[0037] FIG. 22 depicts in vivo depletion of peripheral B cells in cynomolgus monkeys dosed with CD 19 x CD3 bsAbs (in 1 + 1 or 2 + 1 formats with either high or intermediate affinity CD3 binding domains).

[0038] FIG. 23 depicts in vivo depletion of peripheral B cells in cynomolgus monkeys dosed with CD 19 x CD3 bsAbs (in 1 + 1 or 2 + 1 formats with either high or intermediate affinity CD3 binding domains) with logio y-axis.

[0039] FIG. 24 depicts peripheral T cell activation in cynomolgus monkeys dosed with CD19 x CD3 bsAbs (in 1 + 1 or 2 + 1 formats with either high or intermediate affinity7CD3 binding domains).

[0040] FIG. 25A depicts in vivo depletion of bone marrow B cell and FIG. 25B depicts lymph node B cell in cynomolgus monkeys dosed with CD 19 x CD3 bsAbs (in 1 + 1 or 2 + 1 formats with either high or intermediate affinity7CD3 binding domains).

[0041] FIG. 26 depicts serum concentration of CD 19 x CD3 bsAbs (in 1 + 1 or 2 + 1 formats with either high or intermediate affinity CD3 binding domains) over time in cynomolgus monkeys.

[0042] FIG. 27A depicts B cell depletion by XENP26982 and FIG. 27B serum concentration of XENP26982 in cynomolgus monkeys over time.

[0043] FIG. 28A and FIG.28B depict comparison of CD19-OG and re-engineered CD19-NEW binding domains.

[0044] FIG. 29 depicts improvements in re-engineered CD19-NEW binding domain relative to CD19-OG.

[0045] FIG. 30 depicts affinity of original CD3 binding domains and CD3 binding domains re-engineered with disulfide bridges and deamidation fix.

[0046] FIG. 31 depicts induction of CD19+Ramos cell kill by CD 19 x CD3 bsAbs with CD19-OG and CD19-NEW binding domains (in 1 + 1 and 2 + 1 formats with varied CD3 affinity).

[0047] FIG. 32A - FIG. 32D depict induction of T cell activation by CD 19 x CD3 bsAbs having original and re-engineered CD3 binding domains in the presence of human PBMC (FIG. 32A), cyno PBMC (FIG. 32B), and CD19+ Ramos cell kill (FIG. 32C).

[0048] FIG. 33 depicts induction of CD 19+ Ramos cell kill, T cell activation, and IFNy release by CD 19 x CD3 bsAbs with CD19-OG and CD19-NEW binding domains and original and re-engineered CD3 binding domains (in 1 + 1 and 2 + 1 formats).

[0049] FIG. 34 depicts induction of human B cell kill by CD19 x CD3 bsAbs with CD19-NEW and the stability and affinity -enhanced CD3 binding domains.P190500. W0.01

[0050] FIG. 35 depicts the average pharmacokinetics at various relative doses of various CD 19 x CD3 bsAbs (along with ADA status), non-compartmental analysis.

[0051] FIG. 36 depicts depletion of cyno B cells in peripheral blood over time after dosing with various CD19 x CD3 bsAbs at indicated relative doses. Certain of the points denoted 0.01 are 0 B cells per pL as shown in Fig. 41.

[0052] FIG. 37 depicts depletion of cyno B cells among lymphocytes over time after dosing with various CD 19 x CD3 bsAbs at indicated relative doses.

[0053] FIG. 38 depicts depletion of cyno B cells in lymph nodes over time after dosing with various CD 19 x CD3 bsAbs at indicated relative doses.

[0054] FIG. 39 depicts depletion of cyno B cells among bone marrow lymphocytes over time after dosing with various CD19 x CD3 bsAbs at indicated relative doses.

[0055] FIG. 40 depicts depletion of B cell over time in all cynomolgus monkeys dosed with XENP49657 at IX (top graph) or 3X dose level (bottom graph).

[0056] FIG. 41 depicts B cell depletion in periphery, bone marrow, and lymph nodes of cynomolgus monkeys dosed with IX or 3X dose XENP49657. * indicates point is peripheral B cells <1 B cell per pL and + indicates data point is 0 B cells per pL. B cells were gated as CD45+CD2- / 1O wCD20+CD4-CD8a-CD 159a-

[0057] FIG. 42 depicts plasma deletion in the bone marrow of cynomolgus monkeys dosed with IX (top graph) or 3X dose (bottom graph) XENP49657. Plasma cells defined as SSlow / CD45+CD2- / lowCD20- / lowCD4-CD8a-CD159a-CD27-CD38high.

[0058] FIG. 43 depicts reduction in serum immunoglobulin in cynomolgus monkeys dosed with IX or 3X XENP49657 that were not impacted by ADA.

[0059] FIG. 44 depicts affinity for human and cyno CD 19 (apparent dissociation constant to account for bivalent binding, except in the case of 49653 which has monovalent CD19 binding), human and cyno CD3, and melting temperatures of CD19 x CD3 bsAbs having the old CD3 and new stability -enhanced CD3 binding domains.

[0060] FIG. 45 depicts depletion of bone marrow plasma cells over time in cynomolgus monkeys dosed with IX or 3X dose XENP49657.

[0061] FIG. 46 depicts pharmacokinetics of IX and 3X intravenous dose XENP49657 in cynomolgus monkeys, half life -15 days.

[0062] FIG. 47 depicts pharmacokinetics of 3X subcutaneous dose XENP49657 in cynomolgus monkeys, half-life -15 days.

[0063] FIG. 48 depicts depletion of B cell over time in cynomolgus monkeys dosed subcutaneously with XENP49657 3X dose level.P190500. W0.01

[0064] FIG. 49 depicts deep B-depletion sustained for at least 42 days in cynomolgus monkeys dosed intravenously with XENP49657 at 3X dose level.

[0065] FIG. 50 depicts depletion of CD 19+ plasma cells in bone marrow in cynomolgus monkeys dosed with CD 19 x CD3 bsAbs.

[0066] FIG. 51 depicts melting temperature of bispecific antibodies in the 2 + 1 Fab2-scFv-Fc format having original CD3 binding domains and CD3 binding domains re-engineered with disulfide bridges (as denoted by S-S).

[0067] FIG. 52 depicts affinity of original CD3 binding domains and CD3 binding domains re-engineered with disulfide bridges (as denoted by S-S) and deamidation fix (in the context of 2 + 1 Fab2-scFv-Fc bispecific antibodies).

[0068] FIG. 53 depicts induction of B cell killing (top panel), IFNy secretion (middle panel), and IL-6 secretion (bottom panel) by XENP49657 in human PBMCs from healthy donors (average of 4 donors) vs. donors with rheumatoid arthritis (average of 7 donors).

[0069] FIG. 54 depicts induction of A) B cell killing, B) IFNy secretion, and C) IL-6 secretion by XENP49657 in human PBMCs from healthy donors (average of 4 donors) vs. donors with systemic lupus erythematosus (average of 8 donors). depicts induction of A) B cell killing, B) IFNy secretion, and C) IL-6 secretion by XENP49657 in human PBMCs from healthy donors (average of 4 donors) vs. donors with systemic lupus erythematosus (average of 8 donors).DETAILED DESCRIPTIONI. Overview

[0070] Disclosed herein are anti-CD3 and anti-CD19 bispecific antigen binding molecules comprising an anti-CD19 antigen binding domain and an anti-CD3 antigen binding domain, such that the binding molecules bind to the extracellular domains of human CD19 and CD3.

[0071] In some variations, the anti-CD19 antigen binding domain, anti-CD3 antigen binding domain, or both have improved stability while maintaining binding affinity. For example, anti-CD3 antigen binding domain variants of the bispecific antigen binding molecules are engineered to fix deamidation and incorporate disulfide bridges, while maintaining affinity and improved stability of the anti-CD3 antigen binding domain. The anti-CD3 antigen binding domain can have binding affinity similar to a non-engineered anti-CD3 antigen binding molecule. Likewise, the anti-CD19 antigen binding domain of the bispecific antigen binding molecules can be, for example, re-humanized and demonstrate enhanced stability7, such as a +7°C improved Tm(FIG. 29). The herein disclosed anti-CD3 and anti-CD19 bispecific antigen bindingP190500. W0.01 molecules exhibit comparable CD 19+ Ramos cell killing and T-cell activation compared to the reference bispecific molecules that lack the sequence changes that mediate the improvements described herein.

[0072] The disclosed compositions and methods may be understood more readily by¬ reference to the following detailed description taken in connection with the accompanying figures, which form a part of this disclosure. It is to be understood that the disclosed compositions and methods are not limited to the specific compositions and methods described and / or shown herein, and that the terminology used herein is for the purpose of describing particular embodiments by way of example only and is not intended to be limiting of the claimed compositions and methods.

[0073] Unless specifically stated otherwise, any description as to a possible mechanism or mode of action or reason for improvement is meant to be illustrative only, and the disclosed compositions and methods are not to be constrained by the correctness or incorrectness of any such suggested mechanism or mode of action or reason for improvement.

[0074] Throughout this text, the descriptions refer to compositions and methods of using said compositions. Where the disclosure describes or claims a feature or embodiment associated with a composition, such a feature or embodiment is equally applicable to the methods of using said composition. Likewise, where the disclosure describes or claims a feature or embodiment associated with a method of using a composition, such a feature or embodiment is equally applicable to the composition.

[0075] Where a range of numerical values is recited or established herein, the range includes the endpoints thereof and all the individual integers and fractions within the range, and also includes each of the narrower ranges therein formed by all the various possible combinations of those endpoints and internal integers and fractions to form subgroups of the larger group of values within the stated range to the same extent as if each of those narrower ranges was explicitly recited. Where a range of numerical values is stated herein as being greater than a stated value, the range is nevertheless finite and is bounded on its upper end by a value that is operable within the context of the herein disclosure. Where a range of numerical values is stated herein as being less than a stated value, the range is nevertheless bounded on its lower end by a non-zero value. It is not intended that the scope of the compositions and methods be limited to the specific values recited when defining a range. All ranges are inclusive and combinable.

[0076] When values are expressed as approximations, by use of the antecedent "about.” it will be understood that the particular value forms another embodiment. Reference to a particular numerical value includes at least that particular value, unless the context clearly dictates otherwise.P190500. W0.01

[0077] It is to be appreciated that certain features of the disclosed compositions and methods which are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the disclosed compositions and methods that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any subcombination.

[0078] The disclosures of each patent, patent application, and publication cited or described in this document are hereby incorporated herein by reference, in its entirety'.III. Definitions

[0079] As used herein, the singular forms “a,” “an,” and “the” include the plural.

[0080] When a list is presented, unless stated otherwise, it is to be understood that each individual element of that list, and every' combination of that list, is a separate embodiment. For example, a list of embodiments presented as “A. B, or C” is to be interpreted as including the embodiments: “A;” “B;” “C;” “A or B;” “A or C;” “B or C;” or “A. B, or C.”

[0081] Various terms relating to aspects of the description are used throughout the specification and claims. Such terms are to be given their ordinary' meaning in the art unless otherwise indicated. Other specifically defined terms are to be construed in a manner consistent with the definitions provided herein.

[0082] The term “comprising” is intended to include examples encompassed by the terms “consisting essentially' of’ and “consisting of’; similarly, the term “consisting essentially of’ is intended to include examples encompassed by the term “consisting of.”

[0083] “Antigen binding molecule” refers to a protein or peptide that specifically binds to an antigenic determinant. Examples of antigen binding molecules are immunoglobulin and derivatives or fragments thereof, e.g.. Fab, scFv, 1+1 Fab-scFv-Fc, 2+1 Fab2-scFv-Fc, 1+1 Common Light Chain, 2+1 Common Light Chain, 2+1 mAb-scFv, 2+1 stackFab2-scFv-Fc, Dual scFv, One-arm scFv-mAb, scFv-mAb, Bispecific mAb, One-arm central-scFv, mAb-Fv, central-Fv, and Trident. Additional examples of antigen binding molecules are receptors and ligands.

[0084] ‘ ’Binding subunit” as used herein refers to a protein subunit that comprises three “Complementarity Determining Regions” (CDRs). More than one binding subunit can be combined to form an antigen binding domain, which has specificity to an antigenic determinant. Multiple binding units may be linked directly or indirectly. A binding subunit can be a variable heavy domain or a variable light domain; that is, a variable heavy binding subunit and a variable light binding subunit.

[0085] “Antigen binding domain” or “ABD” herein is meant the part of an antigen binding molecule which confers its binding specificity to an antigenic determinant. Herein, theP190500. W0.01 "antigen binding domain” refers to a set of six “Complementarity Determining Regions” (CDRs) that, when present as part of a polypeptide sequence, specifically binds a target antigen as discussed herein. The antigen binding domain can include a first binding subunit comprising three CDRs, and a second binding subunit comprising three CDRs. In some variations, the first binding subunit can correspond to a first set of variable heavy CDRs (vhCDRs or VHCDRs), while the second binding subunit can correspond to a second set of variable light CDRs (vlCDRs or VLCDRs). For example, as is known in the art, CDRs can be present as a first set of variable heavy CDRs and a second set of variable light CDRs, each comprising three CDRs: vhCDRl, vhCDR2. vhCDR3 for the heavy chain and vlCDRl. vlCDR2 and vlCDR3 for the light chain. The CDRs can be present in the variable heavy and variable light domains, respectively. In an antigen binding domain, when a first binding subunit is a variable heavy domain and a second binding subunit is a variable light domain, together the antigen binding domain can form an Fv region. In a “Fab'’ format, the set of 6 CDRs are contributed by two different polypeptide sequences, the variable heavy domain (vh or VH; containing the vhCDRl, vhCDR2 and vhCDR3) and the variable light domain (vl or VL; containing the vlCDRl, vlCDR2 and vlCDR3), 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 a scFv format, the vh and vl domains are covalently attached, generally through the use of a linker (a “scFv linker”) as outlined herein, into a single polypeptide sequence, which can be either (starting from the N-terminus) vh-linker-vl or vl-linker-vh (including optional domain linkers on each side. In general, the C-terminus of the scFv domain is attached to the N-terminus of the hinge in the second monomer.TABLE 1: CDR PositionsKabat+ IMGT Kabat AbM Chothia Contact Xencor ChothiavhCDRl 26-35 27-38 31-35 26-35 26-32 30-35 27-35 vhCDR2 50-65 56-65 50-65 50-58 52-56 47-58 54-61 vhCDR3 95-102 105-117 95-102 95-102 95-102 93-101 103-116 vlCDRl 24-34 27-38 24-34 24-34 24-34 30-36 27-38 V1CDR2 50-56 56-65 50-56 50-56 50-56 46-55 56-62 V1CDR3 89-97 105-117 89-97 89-97 89-97 89-96 97-105P190500. W0.01

[0086] The term “antibody,” and like terms are a category of antigen binding molecule. In a broad sense antibody refers to immunoglobulin molecules including, monoclonal antibodies, antibody fragments, bispecific or multispecific antibodies, dimeric, tetrameric or multimeric antibodies, and single chain antibodies. Antibody formats include, but are not limited to Fab, scFv, 1+1 Fab-scFv-Fc, 2+1 Fab2— scFv-Fc. 1+1 Common Light Chain, 2+1 Common Light Chain, 2+1 mAb-scFv, 2+1 stackFab2-scFv-Fc, Dual scFv, One-arm scFv-mAb, scFv-mAb, Bispecific mAb, One-arm central-scFv, mAb-Fv, central-Fv, and Trident. Antibodies of the invention are generally derived or use components from human IgG, which has four isot pes IgGL IgG2, IgG3 and IgG4. Of particular use herein are heavy chain constant regions from IgGL IgG2 and IgG4, with IgGl finding particular use in many embodiments. Antibody light chains of any vertebrate species can be assigned to one of two clearly distinct types, namely kappa (K) and lambda (X), based on the amino acid sequences of their constant domains.

[0087] “Bispecific” and “bi-specific” refer to the characteristic of a molecule that binds two different antigens or epitopes at the same time. A “bispecific antigen binding molecule,” or a “bispecific antibody” refers to a protein comprising at least two antigen-binding domains that are capable of simultaneously binding two different antigens (targets). Bispecific antibodies (also referred to as “bsAbs”) can be heterodimeric antibodies. Bispecific antigen binding molecules can, for example, be used to form a link between T cells and target cells, leading to the activation of T cells and generation of an immunological synapse. As shown herein, bispecific antigen binding constructs can be bivalent and bispecific (e.g. containing two antigen binding domains, each one binding a different target), or trivalent and bispecific (e.g. where the construct includes three antigen binding domains, two of which bind to one target (e.g. CD19) and one ABD that binds a separate target (e.g. CD3).

[0088] By “Fc” or “Fc region” or “Fc domain” as used herein is meant the polypeptide comprising the 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) or 216-447 (hinge-CH2-CH3), or fragments thereof. An “Fc fragment” in this context may 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 exclusion chromatography, etc.) Human IgG Fc domains are of particular use in the methods provided herein, and can be the Fc domain from human IgGl, IgG2 or IgG4. Two Fc domains can self-assemble to form the full constant region or backbone.P190500. W0.01

[0089] By “heavy chain constant region’7herein is meant the CHl-hinge-CH2-CH3 portion of an antibody (or fragments thereof), excluding the variable heavy domain; in EU numbering of human IgGl this is amino acids 118-447. By “heavy chain constant region fragment’’ herein is meant 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, and more particularly the ability to form a dimeric Fc domain.

[0090] “Heterodimeric antibody” or “heterodimeric domain,” means that the protein has at least two distinct sequences that associate to form a domain. For example, at least distinct two Fc sequences can assemble into a heterodimeric Fc domain, which can further comprise at least two Fv regions.

[0091] “Antibody fragment” refers to a portion of an immunoglobulin molecule that retains the specific antigen binding properties of the parental full length antibody (i. e. antigenbinding fragment thereof). By way of example and not limitation, antibody fragments comprise two binding domains, wherein each binding domain comprises a CDR1, a CDR2, and a CDR3. In some embodiments, the first binding domain can be a heavy chain comprising heavy chain complementarity determining regions (HCDR) 1, 2, and 3, and the second binding domain can be a light chain comprising complementarity determining regions (LCDR) 1, 2, and 3. Other example antibody fragments comprise a heavy chain variable region (VH) and a light chain variable region (VL). By “Fab” or “Fab region” as used herein is meant the polypeptide that comprises the VH, CHI, VL, and CL immunoglobulin domains. By “Fv” or “Fv fragment” or “Fv region” as used herein is meant a polypeptide that comprises the VL and VH domains of a single antibody. For example, antibody fragments include without limitation: a Fab fragment, a monovalent fragment consisting of the VL, VH. constant light (CL), and constant heavy 1 (CHI) domains; a F(ab)2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; and a Fv fragment consisting of the VL and VH domains of a single arm of an antibody. Further, VH and VL domains can be engineered and linked together via a synthetic linker to form various types of single chain antibody designs where the VH / VL domains pair intramolecularly, or intermolecularly in those cases when the VH and VL domains are expressed by separate single chain antibody constructs, to form a monovalent antigen binding site, such as single chain Fv (scFv) or diabody. These antibody fragments are obtained using techniques well known to those of skill in the art, and the fragments are screened for utility in the same manner as are full length antibodies.

[0092] An antigen binding molecule, antigen binding region, or antibody binding domain can have up to four “framework” regions (FRs) flanking CDRs in each binding subunit. In some variations, FRs alternate with three CDRs, in the sequence FR1, CDR1, FR2, CDR2,P190500. W0.01 FR3, CDR3, FR4 (from amino to carboxy termini). If the binding subunit is a heavy chain variable region (VH), the three CDRs are identified as HCDR1, HCDR2, HCDR3 and the FRs are VHFR1, VHFR2, VHFR3 and VHFR4, such that the variable heavy domain comprises, from N- to C-terminal, VHFRI-VHCDR1-VHFR2-VHCDR2-VHFR3-VHCDR3-VHFR4. If the binding subunit is a light chain variable region (VL), the three CDRs are identified as LCDR1. LCDR2, LCDR3, the FRs are VLFR1, VLFR2, VLFR3 and VLFR4, such that the variable heavy domain comprises, from N- to C-terminal, VLFR1-VLCDR1-VLFR2-VLCDR2-VLFR3-VLCDR3-VLFR4. The location and size of the CDRs are defined based on rules which identify regions of sequence variability within the immunoglobulin variable regions (Wu and Kabat J Exp Med 132:211-50, 1970; Kabat et al. Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Sendee, National Institutes of Health, Bethesda, Md., 1991).

[0093] “Humanized antibody” refers to an antibody in which the antigen binding sites are derived from non-human species and the framework regions are derived from human immunoglobulin sequences. Humanized antibodies may include substitutions in the framework regions so that the framework may not be an exact copy of expressed human immunoglobulin or germline gene sequences. If the antibody contains a constant region, the constant region is also derived from sequences of human origin. Methods of antibody humanization include, for example, string content optimization as described in U. S. Patent No. 7,657.380.

[0094] “Human antibody” refers to an antibody having heavy and light chain variable regions in which both the framework and the antigen binding sites are derived from sequences of human origin. If the antibody contains a constant region, the constant region also is derived from sequences of human origin. A human antibody comprises heavy or light chain variable regions that are “derived from” sequences of human origin if the variable regions of the antibody are obtained from a system that uses human germline immunoglobulin or rearranged immunoglobulin genes. Such systems include human immunoglobulin gene libraries displayed on phage, and transgenic non-human animals such as mice or rats carrying human immunoglobulin loci. “Human antibody” may contain amino acid differences when compared to the human germline or rearranged immunoglobulin sequences due to, for example, naturally occurring somatic mutations or intentional introduction of substitutions in the fra ework or antigen binding sites. Typically, a “human antibody” is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical in amino acid sequence to an amino acid sequence encoded by a human germline or rearranged immunoglobulin gene.

[0095] The terms “bind,” “binding,” and the like, refer to the ability7of the disclosed antigen binding molecules to preferentially bind to its target (e.g. CD3 in the case of anti-CD3P190500. W0.01 antigen binding molecules or CD 19 in the case of anti-CD19 antigen binding molecules) without preferentially binding other molecules in a sample containing a mixed population of molecules. Antigen binding molecules that immunospecifically bind, for example, CD3 are substantially free of other antigen binding molecules having different antigenic specificities (e g., an anti-CD3 antigen binding molecule is substantially free of antigen binding molecules that specifically bind antigens other than CD3). Antigen binding molecules that immunospecifically bind, for example, CD3 and CD 19, however, can have cross-reactivity to other antigens, such as orthologs of human CD3 and CD 19, including Macaca fascicularis (cynomolgus monkey) CD3 and CD 19, respectively. The antigen binding molecules disclosed herein are able to immunospecifically bind both naturally-produced human CD3 and CD 19 and to CD3 and CD 19 which is recombinantly produced in mammalian or prokaryotic cells.

[0096] By “target antigen” as used herein is meant the molecule that is bound specifically by the antigen binding domain comprising the variable regions of a given antibody. As discussed herein, in the target antigens are CD 19 and CD3.

[0097] The strength, or affinity, of specific binding can be expressed in terms of dissociation constant (KD) of the interaction, wherein a smaller KD represents greater affinity and a larger KD represents lower affinity. Binding properties can be determined by methods well known in the art such as bio-layer interferometry and surface plasmon resonance based methods. One such method entails measuring the rates of antigen-binding site / antigen or receptor / ligand complex association and dissociation, wherein rates depend on the concentration of the complex partners, the affinity of the interaction, and geometric parameters that equally influence the rate in both directions. Thus, both the association rate (ka) and the dissociation rate (kd) can be determined, and the ratio of kd / ka is equal to the dissociation constant KD (See Nature 361:186-187 (1993) and Davies et al. (1990) Annual Rev Biochem 59:439-473).

[0098] Spes 2i fie binding for a particular molecule or an epitope can be exhibited, for example, by an antigen binding molecule having a KD for an antigen or epitope of at least about 10 ~4M, at least about 105M, at least about 10 "6M, at least about 10 ~7M, at least about 10sM, at least about 10 M, alternatively at least about 10 ”!0M, at least about 10 ”nM, at least about 10 ~i2M, or lesser. Typically, an antigen binding moleculethat specifically binds an antigen will have a KD that is 20-, 50-, 100-, 500-, 1000-, 5,000-, 10,000- or more times greater for a control molecule relative to the antigen or epitope.

[0099] Also, specific binding for a particular molecule or an epitope can be exhibited, for example, by an antigen binding molecule having a ka or association rate for an antigen or epitope of at least 20-, 50-, 100-, 500-, 1000-, 5,000-, 10,000- or more times greater for the epitope relative to a control.P190500. W0.01

[0100] By “epitope’7is herein meant a determinant that interacts with a specific antigen binding domain, for example variable region of an antibody molecule, known as a paratope. Epitopes are groupings of molecules such as amino acids or sugar side chains and usually have specific structural characteristics, as well as specific charge characteristics. A single molecule may have more than one epitope. The epitope may comprise amino acid residues directly involved in the binding (also called immunodominant component of the epitope) and other amino acid residues, which are not directly involved in the binding, such as amino acid residues which are effectively blocked by the specifically antigen binding peptide; in other words, the amino acid residue is within the footprint of the specifically antigen binding peptide. Epitopes may be either conformational or linear. A conformational epitope is produced by spatially juxtaposed amino acids from different segments of the linear polypeptide chain. A linear epitope is one produced by adjacent amino acid residues in a polypeptide chain.Conformational and nonconformational epitopes may be distinguished in that the binding to the former but not the latter is lost in the presence of denaturing solvents. An epitope typically includes at least 3, and more usually, at least 5 or 8-10 amino acids in a unique spatial conformation. Antigen binding molecules that recognize the same epitope can be verified in a simple immunoassay showing the ability of one antigen binding molecule to block the binding of another antigen binding molecule to a target antigen, for example “binning.” As outlined below, the invention not only includes the enumerated antigen binding molecules and antigen binding domains herein, but those that compete for binding with the epitopes bound by the enumerated antigen binding molecules or antigen binding domains.

[0101] The term “CD3” refers to cluster of differentiation 3 protein complex or CD3 chains that form the CD3 complex, such as CD3 gamma, CD3 delta, CD3 zeta, and CD3 epsilon. In particular variations, CD3 refers to the human CD3-epsilon polypeptide of SEQ ID NO: 37. Example CD3 sequences to which the herein disclosed CD3 antigen binding molecules can bind include, for example, SEQ ID NOs: 37-42, which include human, mouse, and cynomolgus CD3 sequences (FIG. 1).

[0102] The term “CD19” refers to Cluster of Differentiation 19, which includes a transmembrane domain, a cytoplasmic C-terminus, and extracellular N-terminus. The herein disclosed CD19 antigen binding molecules can bind the extracellular N-terminus of CD19, which has two C2-type Ig-like domains divided by a smaller potential disulfide linked non-Ig-hke domain. Example CD 19 sequences to which the herein disclosed CD 19 antigen binding molecules can bind include, for example, SEQ ID NOs: 43-48, which include human, mouse, and cynomolgus CD 19 sequences (FIG. 1).P190500. W0.01

[0103] “Variant” refers to a polypeptide or a polynucleotide that differs from a reference polypeptide or a reference polynucleotide by one or more modifications for example, substitutions, insertions, or deletions.

[0104] By “position” as used herein is meant a location in the sequence of a protein. Positions may be numbered sequentially, or according to an established format, for example the EU index for antibody numbering.

[0105] “Parent polypeptide” as used herein is meant a starting polypeptide that is subsequently modified to generate a variant. The parent polypeptide may be a naturally occurring polypeptide, or a variant or engineered version of a naturally occurring polypeptide. Parent polypeptide may refer to the polypeptide itself, compositions that comprise the parent polypeptide, or the amino acid sequence that encodes it.

[0106] The term “percent identity ” as used herein refers to the percent of amino acids that are the same between two or more polypeptides. Sequence identity between two polypeptides can be determined by aligning the amino acid sequences of the polypeptides and scoring the number of positions in the aligned polypeptides that contain the same amino acid residue and comparing that to the number of positions in the aligned polypeptides that differ. Polypeptides can differ at a position, for example, by containing a different amino acid (i.e. substitution or mutation) or by lacking an amino acid (i.e. amino acid insertion or amino acid deletion in one or both of the polypeptides). Sequences with at least 95% sequence identity to a given sequence also include, for example, sequences with 96%, 97%, 98%, and 99% sequence identity. Sequence identity between two similar sequences can be calculated by algorithms such as 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 [homology7alignment 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 (blast.ncbi.nlm.nih.gov / Blast_cgi).

[0107] “Treat,” “treatment,” and like terms refer to both therapeutic treatment and prophylactic or preventative measures, and includes reducing the severity and / or frequency of symptoms, eliminating symptoms and / or the underlying cause of the symptoms, reducing the frequency or likelihood of symptoms and / or their underlying cause, and improving or remediating damage caused, directly7or indirectly, by, for example, an autoimmune disease. Treatment also includes prolonging survival as compared to the expected survival of a subject not receiving treatment. Subjects to be treated include those that have the condition or disorderP190500. W0.01 as well as those prone to have the condition or disorder or those in which the condition or disorder is to be prevented.

[0108] The term “subject” as used herein is intended to mean any animal, in particular, mammals. Any type of mammal can be treated using the disclosed methods. Thus, the methods are applicable to human and nonhuman animals, although preferably used with mice, monkeys, and humans, and most preferably with humans. “Subject” and “patient” are used interchangeably herein.

[0109] As used herein, “administering to said patient” and similar terms indicate a procedure by which a herein disclosed antigen binding molecule is injected into a patient such that target cells, tissues, or segments of the body of the subject are contacted with a herein disclosed antigen binding molecule. Suitable routes of administration include, for example, intravenous administration and subcutaneous administration.

[0110] As used herein, “cancer” and “cancerous” refers to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth, proliferation, or metastasis, and includes both solid cancers and non-solid cancers. Examples of cancer include but are not limited to carcinoma, lymphoma, blastoma, sarcoma (including liposarcoma). neuroendocrine tumors, mesothelioma, schwanoma, meningioma, adenocarcinoma, melanoma, and leukemia or lymphoid malignancies.

[0111] As used herein, the phrase “therapeutically effective amount” refers to an amount of a herein disclosed antigen binding molecule effective to achieve a particular biological or therapeutic result such as, but not limited to, biological or therapeutic results disclosed, described, or exemplified herein. The therapeutically effective amount may vary according to factors such as the disease state, age. sex, and weight of the individual, and the ability of the composition to cause a desired response in a subject.

[0112] “Autoimmune diseases” and like terms as used herein include allogenic islet graft rejection, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, antineutrophil cytoplasmic autoantibodies (ANCA), autoimmune diseases of the adrenal gland, anti-MDA5 syndrome, antisynthetase syndrome, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune autonomic ganglionopathy (AAG), autoimmune encephalitis (anti-NMDAR, anti-LGIl), autoimmune myocarditis, autoimmune neutropenia, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, autoimmune urticaria, Behcet's disease, bullous pemphigoid, cardiomyopathy, Castleman's syndrome, celiac sprucedermatitis, chronic fatigue immune disfunction syndrome, chronic inflammatory demyelinating polyneuropathy, Churg-Strauss syndrome, cicatrical pemphigoid, cold agglutinin disease, CREST syndrome, cold agglutinin disease, Crohn's disease, dermatomyositis, discoid lupus,P190500. W0.01 Evans syndrome, essential mixed cryoglobulinemia, factor VIII deficiency, fibromyalgia-fibromyositis, glomerulonephritis, Grave's disease, Guillain-Barre, Goodpasture's syndrome, graft-versus-host disease (GVHD), Hashimoto's thyroiditis, hemophilia A, idiopathic pulmonary7fibrosis, idiopathic thrombocytopenia purpura (ITP), IgA neuropathy, IgM polyneuropathies. IgA vasculitis (Henoch-Schonlein purpura), IgG4-related disease, immune mediated thrombocytopenia, immune thrombocytopenic purpura (ITP) juvenile arthritis, Kawasaki's disease, lichen planus, lupus erthematosis, Meniere's disease, mixed connective tissue disease, multiple sclerosis, type 1 diabetes mellitus, myasthenia gravis, Neuromyelitis optica spectrum disorder (NMOSD), Overlap myositis, pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, polychrondritis, polyglandular syndromes, polymyalgia rheumatica, polymyositis and dermatomyositis, primary agammaglobinulinemia, primary biliary cirrhosis, primary biliary cholangitis (CD19+plasma-cell dominant), psoriasis, psoriatic arthritis, Reynauld's phenomenon, Reiter's syndrome, rheumatoid arthritis, sarcoidosis, scleroderma, Sjogren's syndrome, solid organ transplant rejection, stiff-man syndrome, systemic lupus erythematosus, systemic sclerosis (diffuse cutaneous subtype), type 1 diabetes mellitus (early or new-onset), takayasu arteritis, temporal arteristis / giant cell arteritis, thrombotic thrombocytopenia purpura, ulcerative colitis, uveitis, vasculitides such as dermatitis herpetiformis vasculitis, vitiligo, and Wegner's granulomatosis.■‘Inflammatory disorders” and like terms as used herein include acute respiratory distress syndrome (ARDS), acute septic arthritis, adjuvant arthritis (Prakken et al., Springer Semin Immunopathol., 2003 August; 25(l):47-63, incorporated entirely by reference), juvenile idiopathic arthritis (de Kleer et al., Arthritis Rheum. 2003 July; 47(7):200I-10, incorporated entirely by reference), allergic encephalomyelitis, allergic rhinitis, allergic vasculitis, allergy, asthma, atherosclerosis, chronic inflammation due to chronic bacterial or viral infections, chronic obstructive pulmonary disease (COPD), coronary artery disease, encephalitis, inflammatory bowel disease, inflammatory osteolysis, inflammation associated with acute and delayed hypersensitivity reactions, inflammation associated with tumors, peripheral nerve injury or demyelinating diseases, inflammation associated with tissue trauma such as bums and ischemia, inflammation due to meningitis, multiple organ injury syndrome, pulmonary fibrosis, sepsis and septic shock, Stevens-Johnson syndrome, undifferentiated arthropy, and undifferentiated spondyloarthropathy.Bi-Specific CD3 x CD19 Antigen Binding MoleculesP190500. W0.01

[0113] Disclosed herein are bi-specific antigen binding molecules that bind CD3 and CD 19. In further variations, antigen binding molecules that specifically bind to CD3 and antigen binding molecules that immunospecifically bind to CD 19 are also provided.

[0114] As described herein, the bi-specific CD3 x CD 19 antigen binding molecules combine the antigen binding domains of an anti-CD3 antigen binding molecule and the antigen binding domains of an anti-CD19 antigen binding molecule. Anti-CD3 and anti-CD19 antigen binding molecules are disclosed separately herein, and can be combined in the bi-specific CD3 x CD 19 antigen binding molecule as described herein.CD3 Antigen Binding Molecules

[0115] Disclosed herein are anti-CD3 antigen binding molecules with improved stability. Sequence changes that mediate improved stability include, but are not limited to, changes in the framework region and changes in the CDRs. Improvements in stability include increase thermostability of the antigen binding molecules. Improving stability' of the antigen binding molecule is important if antigen binding molecules are, for example, to be used in high concentrations such as in a subcutaneous formulation.CDRs

[0116] Disclosed herein are anti-CD3 antigen binding molecules. In one variation, the anti-CD3 antigen binding molecule comprises an antigen binding domain comprising a first binding subunit comprising the CDR sequences of SEQ ID NO: 9, SEQ ID NO: 10 or 11, and SEQ ID NO: 12, 13, 14, 15, or 16, and a second binding subunit comprising the CDR sequences of SEQ ID NOs: 17, 18, and 19. In some variations, the first binding subunit comprises the CDR sequences of SEQ ID NOs: 9, 10, and 12 and the second binding subunit comprises the CDR sequences of SEQ ID NOs: 17, 18, and 19. In some variations, the first binding subunit comprises the CDR sequences of SEQ ID NOs: 9, 10, and 13 and the second binding subunit comprises the CDR sequences of SEQ ID NOs: 17, 18, and 19. In some variations, the first binding subunit comprises the CDR sequences of SEQ ID NOs: 9, 11, and 12 and the second binding subunit comprises the CDR sequences of SEQ ID NOs: 17, 18, and 19. In some variations, the first binding subunit comprises the CDR sequences of SEQ ID NOs: 9, 11, and 13 and the second binding subunit comprises the CDR sequences of SEQ ID NOs: 17, 18, and 19. In some variations, the first binding subunit comprises the CDR sequences of SEQ ID NOs: 9, 10, and 14 and the second binding subunit comprises the CDR sequences of SEQ ID NOs: 17, 18, and 19. In some variations, the first binding subunit comprises the CDR sequences of SEQ ID NOs: 9, 10, and 15 and the second binding subunit comprises the CDR sequences of SEQ ID NOs: 17, 18, and 19. In some variations, the first binding subunit comprises the CDR sequencesP190500. W0.01 of SEQ ID NOs: 9. 11. and 14 and the second binding subunit comprises the CDR sequences of SEQ ID NOs: 17, 18, and 19. In some variations, the first binding subunit comprises the CDR sequences of SEQ ID NOs: 9, 11, and 15 and the second binding subunit comprises the CDR sequences of SEQ ID NOs: 17, 18, and 19. In some variations, the first binding subunit comprises the CDR sequences of SEQ ID NOs: 9. 10, and 16 and the second binding subunit comprises the CDR sequences of SEQ ID NOs: 17, 18, and 19.Linkers

[0117] The anti-CD3 antigen binding molecules can comprise a linker between the first binding subunit and the second binding subunit. The linker can comprise at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 36 between the first binding subunit and the second binding subunit. The linker can comprise the amino acid sequence of SEQ ID NO: 36 between the first binding subunit and the second binding subunit. The linker can comprise at least 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, or 95. At least 95% sequence identity includes, for example, 96%, 97%, 98%, and 99% sequence identity. The linker can comprise the amino acid sequence of any one of SEQ ID NOs: 49, 50, 51, 52, 53, 54, 55. 56.57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, or 95.Variable Regions

[0118] In some variations of the anti-CD3 antigen binding molecule, the first binding subunit is a heavy chain variable region (VH) and the second binding subunit is a light chain variable region (VL). In some variations, the VH comprises a VH complementarity determining region (HCDR)1. HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 9. 10. and 12. respectively, and the VL comprises the VL LCDR1, LCDR2. and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 17, 18, and 19, respectively. In some variations, the VH comprises a VH HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 9, 10, and 13, respectively, and the VL comprises the VL LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 17, 18, and 19, respectively. In some variations, the VH comprises a VH HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 9, 11, and 12, respectively, and the VL comprises the VL LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 17, 18, and 19, respectively. In some variations, the VH comprises a VH HCDR1,P190500. W0.01 HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 9, 11. and 13, respectively, and the VL comprises the VL LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 17, 18, and 19, respectively. In some variations, the VH comprises a VH HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 9. 10. and 14, respectively, and the VL comprises the VL LCDR1, LCDR2. and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 17, 18, and 19, respectively. In some variations, the VH comprises a VH HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 9, 10, and 15, respectively, and the VL comprises the VL LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 17, 18, and 19, respectively. In some variations, the VH comprises a VH HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 9, 11, and 14, respectively, and the VL comprises the VL LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 17, 18, and 19, respectively. In some variations, the VH comprises a VH HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 9, 11. and 15, respectively, and the VL comprises the VL LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 17, 18, and 19, respectively. In some variations, the VH comprises a VH HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 9. 10. and 16, respectively, and the VL comprises the VL LCDR1, LCDR2. and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 17, 18, and 19, respectively.

[0119] The anti-CD3 antigen binding molecules can comprise a heavy chain variable region comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 179, 181, 182, 183. 184, 185, 186, 187, 188. or 189, and a VL comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 179 and a VL comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 181 and a VL comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 182 and a VL comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 183 and a VL comprising at least 95% sequence identity7to the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising at leastP190500. W0.01 95% sequence identity to the amino acid sequence of SEQ ID NO: 184 and a VL comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 185 and a VL comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 186 and a VL comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 187 and a VL comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising at least 95% sequence identity7to the amino acid sequence of SEQ ID NO: 188 and a VL comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 189 and a VL comprising at least 95% sequence identity7to the amino acid sequence of SEQ ID NO: 180. At least 95% sequence identity includes, for example, 96%, 97%, 98%, and 99% sequence identity.

[0120] The anti-CD3 antigen binding molecules can comprise a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 179, 181, 182, 183, 184, 185, 186, 187, 188, or 189, and aVL comprising the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 179 and a VL comprising the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 181 and a VL comprising the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 182 and a VL comprising the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 183 and a VL comprising the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 184 and a VL comprising the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 185 and a VL comprising the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising the amino acid sequence of SEQP190500. W0.01 ID NO: 186 and a VL comprising the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 187 and a VL comprising the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 188 and a VL comprising the amino acid sequence of SEQ ID NO: 180. In some variations, the anti-CD3 antigen binding molecule comprises a VH comprising the amino acid sequence of SEQ ID NO: 189 and a VL comprising the amino acid sequence of SEQ ID NO: 180.

[0121] The anti-CD3 antigen binding molecule can comprise a VH of any one of SEQ ID NOs: 250-2644. The VH of any one of SEQ ID NOs: 250-2644 can comprise a G44C substitution. The VH of any one of SEQ ID NOs: 250-2644 can comprise aN97Q, G96T, G96E, D100G, or any combination thereof, such as G96T / D100G or G96E / D100G. In some embodiments, the substitution is N97Q. In some embodiments, the substitution is D100G. In some embodiments, the substitutions are N97Q and D100G. The anti-CD3 antigen binding molecule can comprise a VL of any one of SEQ ID NOs: 2645-3380. The VL of any one of SEQ ID NOs: 2645-3380 can comprise a G100C substitution. The herein disclosed anti-CD3 antigen binding molecules can comprise any combination of disclosed VH or VL. For example, the anti-CD3 antigen binding molecule can comprise a VH and a VL, wherein the VH comprises any one of SEQ ID NOs: 250-2644 or a variant thereof comprising G44C, N97Q, G96T, G96E, DI 00G, or a combination thereof, and the VL comprises any one of SEQ ID NOs: 2645-3380 or a variant thereof comprising G100C.

[0122] The herein disclosed anti-CD3 antigen binding molecules can comprise a single-chain variable fragment (scFv) comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, or 209. In some variations, the scFV comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 190. In some variations, the scFV comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 191. In some variations, the scFV comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 192. In some variations, the scFV comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 193. In some variations, the scFV comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 194. In some variations, the scFV comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 195. In some variations, the scFV comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 196. In some variations, the scFV comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 197. In some variations, the scFV comprisesP190500. W0.01 at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 198. In some variations, the scFV comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 199. In some variations, the scFV comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 200. In some variations, the scFV comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 201. In some variations, the scFV comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 202. In some variations, the scFV comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 203. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 204. In some variations, the scFV comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 205. In some variations, the scFV comprises at least 95% sequence identity7to the amino acid sequence of SEQ ID NO: 206. In some variations, the scFV comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 207. In some variations, the scFV comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 208. In some variations, the scFV comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 209. At least 95% sequence identity includes, for example, 96%, 97%, 98%, and 99% sequence identity.

[0123] The herein disclosed anti-CD3 antigen binding molecules can comprise a single-chain variable fragment (scFv) comprising the amino acid sequence of SEQ ID NO: 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, or 209. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 190. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 191. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 192. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 193. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 194. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 195. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 196. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 197. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 198. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 199. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 200. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 201. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 202. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 203. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 204. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 205. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 206. In some variations, the scFV comprises the amino acid sequenceP190500. W0.01 of SEQ ID NO: 207. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 208. In some variations, the scFV comprises the amino acid sequence of SEQ ID NO: 209.Framework Regions

[0124] The anti-CD3 antigen binding molecule can comprise one or more framework regions between CDRs. The framework region described herein can be at specific positions relative to the CDRs of the first binding subunit and the CDRs of the second binding subunit in an antigen binding domain.

[0125] Disclosed herein are anti-CD3 antigen binding molecules comprising an antigen binding domain. The antigen binding domain comprises a first binding subunit, a second binding subunit, and a framework region comprising at least 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 28, 29, 30, 31, 32, 33, 34, and 35. In some variations, the framework region comprises at least 95% identity to the amino acid sequence of SEQ ID NOs: 28. In some variations, the framework region comprises the amino acid sequence of SEQ ID NOs: 28. In some variations, the framework region comprises at least 95% identity to the amino acid sequence of SEQ ID NOs: 29. In some variations, the framework region comprises the amino acid sequence of SEQ ID NOs: 29. In some variations, the framework region comprises at least 95% identity to the amino acid sequence of SEQ ID NOs: 30. In some variations, the framework region comprises the amino acid sequence of SEQ ID NOs: 30. In some variations, the framework region comprises at least 95% identity to the amino acid sequence of SEQ ID NOs: 31. In some variations, the framework region comprises the amino acid sequence of SEQ ID NOs: 31. In some variations, the framework region comprises at least 95% identity to the amino acid sequence of SEQ ID NOs: 32. In some variations, the framework region comprises the amino acid sequence of SEQ ID NOs: 32. In some variations, the framework region comprises at least 95% identity to the amino acid sequence of SEQ ID NOs: 33. In some variations, the framework region comprises the amino acid sequence of SEQ ID NOs: 33. In some variations, the framework region comprises at least 95% identity to the amino acid sequence of SEQ ID NOs: 34. In some variations, the framework region comprises the amino acid sequence of SEQ ID NOs: 34. In some variations, the framework region comprises at least 95% identity to the amino acid sequence of SEQ ID NOs: 35. In some variations, the framework region comprises the amino acid sequence of SEQ ID NOs: 35. At least 95% sequence identity includes, for example, 96%, 97%, 98%, and 99% sequence identity.

[0126] The anti-CD3 antigen binding molecules can comprise a framework region 1 (FR1) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 28, a framework region 2 (FR2) comprising at least 95% identity to the amino acid sequence of SEQP190500. W0.01 ID NO: 29. a framework region 3 (FR3) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 30, a framework region 4 (FR4) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 31, a framework region 5 (FR5) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 32, a framework region 6 (FR6) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 33, a framework region 7 (FR7) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 34, and a framework region 8 (FR8) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 35. At least 95% sequence identity includes, for example, 96%, 97%, 98%, and 99% sequence identity.

[0127] The anti-CD3 antigen binding molecules can comprise FR1 comprising the amino acid sequence of SEQ ID NO: 28, a FR2 comprising the amino acid sequence of SEQ ID NO: 29, a FR3 comprising the amino acid sequence of SEQ ID NO: 30, a FR4 comprising the amino acid sequence of SEQ ID NO: 31, a FR5 comprising the amino acid sequence of SEQ ID NO: 32, a FR6 comprising the amino acid sequence of SEQ ID NO: 33. a FR7 comprising the amino acid sequence of SEQ ID NO: 34, and a FR8 comprising the amino acid sequence of SEQ ID NO: 35.

[0128] Disclosed herein are anti-CD3 antigen binding molecules comprising an amino acid sequence comprising at least 95% identity to SEQ ID NO: 28 N-terminal to a first binding subunit CDR1, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 29 between the first binding subunit CDR1 and a first binding subunit CDR2, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 30 between the first binding subunit CDR2 and a first binding subunit CDR3, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 31 C-terminal to the first binding subunit CDR3. an amino acid sequence comprising at least 95% identity to SEQ ID NO: 32 N-terminal to a second binding subunit CDR1, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 33 between the second binding subunit CDR1 and a second binding subunit CDR2, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 34 between the second binding subunit CDR2 and a second binding subunit CDR3; and an amino acid sequence comprising at least 95% identity to SEQ ID NO: 35 C-terminal to the second binding subunit CDR3. At least 95% sequence identity includes, for example, 96%, 97%, 98%, and 99% sequence identity. In some variations, the first binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 9, the first binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 10 or 11, the first binding subunit CDR3 comprises the amino acid sequence of SEQ ID NO: 12, 13, 14, 15, or 16, the second binding subunit CDR1 comprising the amino acid sequence of SEQ ID NO: 17, the second binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO:P190500. W0.01 18, and the second binding subunit CDR3 comprises the amino acid sequence of SEQ ID NO: 19.

[0129] In some variations, the first binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 9, 10. and 12, respectively, and the second binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acids sequences of SEQ ID NOs: 17, 18, and 19, respectively. In some variations, the first binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 9, 10, and 13, respectively, and the second binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acids sequences of SEQ ID NOs: 17, 18, and 19, respectively. In some variations, the first binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 9, 11, and 12, respectively, and the second binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acids sequences of SEQ ID NOs: 17, 18, and 19, respectively. In some variations, the first binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 9, 11, and 13, respectively, and the second binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acids sequences of SEQ ID NOs: 17, 18, and 19, respectively. In some variations, the first binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 9. 10, and 14. respectively, and the second binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acids sequences of SEQ ID NOs: 17, 18, and 19, respectively. In some variations, the first binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 9, 10, and 15, respectively, and the second binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acids sequences of SEQ ID NOs: 17. 18. and 19, respectively. In some variations, the first binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 9, 11, and 14, respectively, and the second binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acids sequences of SEQ ID NOs: 17, 18, and 19, respectively. In some variations, the first binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 9, 11, and 15, respectively, and the second binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acids sequences of SEQ ID NOs: 17, 18, and 19, respectively. In some variations, the first binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acid sequences of SEQ ID NOs: 9, 10, and 16, respectively, and the second binding subunit comprises a CDR1, CDR2, and CDR3 comprising the amino acids sequences of SEQ ID NOs: 17, 18, and 19, respectively.StabilityP190500. W0.01

[0130] An antigen binding molecule, such as an anti-CD3 antigen binding molecule can include, for example, a non-canonical deamidation motif GNF at positions 96-98 (Kabat numbering) in the first binding subunit CDR3. Consequentially, variants are made to remove this deamidation liability as well as to engineer stabilizing disulfide bridges. N97 is important for binding, but variants at N97 abrogated binding affinity (with the exception of N97Q which retains some binding). Variants at G96 is also important to binding, and G96E or G96T was found to improve binding affinity (albeit with decrease in stability’ by ~2-4°C).

[0131] The disulfide bridges can increase the thermodynamic stability of the antigen binding molecule. In some variations, the disulfide bridges can increase the denaturation temperature by at least 2 °C). In some variations, the disulfide bridges can increase the denaturation temperature by at least 3 °C). In some variations, the disulfide bridges can increase the denaturation temperature by at least 4 °C).

[0132] D100G, for example, improves affinity. Further variants are engineered to fix the deamidation and incorporate disulfide bridges while maintaining an affinity ladder and improved stability. Sequences for these variants are depicted in, for example, FIG. 17A - FIG.17J, and affinity (in the context of 2 + 1 Fab2-scFv-Fc molecules of FIG. 20A - FIG. 20AD) are depicted in FIG. 30. The re-engineered anti-CD3 antigen binding molecules can have binding affinity similar to a non-engineered anti-CD3 antigen binding molecule, albeit with removal of the deamidation motif and introduction of stabilizing disulfide bridges.CD 19 Antigen Binding Molecules

[0133] The anti-CD19 antigen binding molecules, for example, are re-humanized and demonstrate enhanced stability.CDRs

[0134] Disclosed herein are anti-CD19 antigen binding molecules comprising a first binding subunit comprising the CDR sequences of SEQ ID NOs: 1. 2, and 3, and a second binding subunit comprising the CDR sequences of SEQ ID NOs: 4, 5, and 6.Linkers

[0135] The anti-CD19 antigen binding molecules can comprise a linker between the first binding subunit and the second binding subunit. The linker can comprise at least 95% sequence identity7to the amino acid sequence of SEQ ID NO: 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93. 94. or 95. At least 95% sequence identity includes, forP190500. W0.01 example. 96%, 97%. 98%. and 99% sequence identity. The linker can comprise the amino acid sequence of SEQ ID NO: 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, or 95.Variable Regions

[0136] The herein disclosed anti-CD19 antigen binding molecules can comprise a heavy chain variable region (VH) and a light chain variable region (VL). In some variations, the disclosed first binding subunit is a VH, and the disclosed second binding subunit is a VL. In some variations, the VH comprises a heavy chain HCDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 3, and the VL comprises a light chain LCDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 6.

[0137] The anti-CD19 antigen binding molecules can comprise a heavy chain variable region comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 7, and a light chain variable region comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 8. At least 95% sequence identity includes, for example, 96%, 97%, 98%. and 99% sequence identity.

[0138] The anti-CD19 antigen binding molecules can comprise a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8.Framework Regions

[0139] Disclosed herein are anti-CD19 antigen binding molecules comprising an antigen binding domain. The antigen binding domain comprises a first binding subunit, a second binding subunit, and one or more framework regions. In one variation, a framework region comprising at least 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 20, 21, 22, 23, 24, 25, 26, and 27. In some variations, the framework region comprises at least 95% identity to the amino acid sequence of SEQ ID NOs: 20. In some variations, the framework region comprises the amino acid sequence of SEQ ID NOs: 20. In some variations, the framework region comprises at least 95% identity to the amino acid sequence of SEQ ID NOs: 21. In some variations, the framework region comprises the aminoP190500. W0.01 acid sequence of SEQ ID NOs: 21. In some variations, the framework region comprises at least 95% identity to the amino acid sequence of SEQ ID NOs: 22. In some variations, the framework region comprises the amino acid sequence of SEQ ID NOs: 22. In some variations, the framework region comprises at least 95% identity to the amino acid sequence of SEQ ID NOs: 23. In some variations, the framework region comprises the amino acid sequence of SEQ ID NOs: 23. In some variations, the framework region comprises at least 95% identity to the amino acid sequence of SEQ ID NOs: 24. In some variations, the framework region comprises the amino acid sequence of SEQ ID NOs: 24. In some variations, the framework region comprises at least 95% identity to the amino acid sequence of SEQ ID NOs: 25. In some variations, the framework region comprises the amino acid sequence of SEQ ID NOs: 25. In some variations, the framework region comprises at least 95% identity to the amino acid sequence of SEQ ID NOs: 26. In some variations, the framework region comprises the amino acid sequence of SEQ ID NOs: 26. In some variations, the framework region comprises at least 95% identity to the amino acid sequence of SEQ ID NOs: 27. In some variations, the framework region comprises the amino acid sequence of SEQ ID NOs: 27. At least 95% sequence identity includes, for example, 96%, 97%, 98%, and 99% sequence identity.

[0140] The anti-CD19 antigen binding molecules can comprise a framework region 1 (FR1) comprising at least 95% identity to an amino acid sequence of SEQ ID NO: 20, a framework region 2 (FR2) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 21, a framework region 3 (FR3) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 22, a framework region 4 (FR4) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 23, a framework region 5 (FR5) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 24, a framework region 6 (FR6) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 25, a framework region 7 (FR7) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 26, and a framework region 8 (FR8) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 27. At least 95% sequence identity includes, for example. 96%. 97%. 98%. and 99% sequence identity.

[0141] The anti-CD19 antigen binding molecules can comprise a framework region 1 (FR1) comprising an amino acid sequence of SEQ ID NO: 20, a framework region 2 (FR2) comprising the amino acid sequence of SEQ ID NO: 21, a framework region 3 (FR3) comprising the amino acid sequence of SEQ ID NO: 22, a framework region 4 (FR4) comprising the amino acid sequence of SEQ ID NO: 23, a framework region 5 (FR5) comprising the amino acid sequence of SEQ ID NO: 24, a framework region 6 (FR6) comprising the amino acid sequence of SEQ ID NO: 25. a framework region 7 (FR7) comprising the amino acid sequence of SEQ IDP190500. W0.01 NO: 26, and a framework region 8 (FR8) comprising the amino acid sequence of SEQ ID NO: 27.

[0142] Disclosed herein are anti-CD19 antigen binding molecules comprising an amino acid sequence comprising at least 95% identity to SEQ ID NO: 20 N-terminal to a first binding subunit CDR1. an amino acid sequence comprising at least 95% identity to SEQ ID NO: 21 between the first binding subunit CDR1 and a first binding subunit CDR2, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 22 between the first binding subunit CDR2 and a first binding subunit CDR3, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 23 C-terminal to the first binding subunit CDR3, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 24 N-terminal to a second binding subunit CDR1, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 25 between the second binding subunit CDR1 and a second binding subunit CDR2, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 26 between the second binding subunit CDR2 and a second binding subunit CDR3, and an amino acid sequence comprising at least 95% identity to SEQ ID NO: 27 C-terminal to the second binding subunit CDR3. At least 95% sequence identity includes, for example, 96%, 97%, 98%, and 99% sequence identity.

[0143] In some variations, the first binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 1, the first binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 2, the first binding subunit CDR3 comprises the amino acid sequence of SEQ ID NO: 3, the second binding subunit CDR1 comprising the amino acid sequence of SEQ ID NO: 4, the second binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 5, and the second binding subunit CDR3 comprises the amino acid sequence of SEQ ID NO: 6.

[0144] The anti-CD19 antigen binding molecules can comprise an amino acid sequence comprising SEQ ID NO: 20 N-terminal to a first binding subunit CDR1, an amino acid sequence comprising SEQ ID NO: 21 between the first binding subunit CDR1 and a first binding subunit CDR2, an amino acid sequence comprising SEQ ID NO: 22 between the first binding subunit CDR2 and a first binding subunit CDR3, an amino acid sequence comprising SEQ ID NO: 23 C-terminal to the first binding subunit CDR3, an amino acid sequence comprising SEQ ID NO: 24 N-terminal to a second binding subunit CDR1, an amino acid sequence comprising SEQ ID NO: 25 between the second binding subunit CDR1 and a second binding subunit CDR2, an amino acid sequence comprising SEQ ID NO: 26 between the second binding subunit CDR2 and a second binding subunit CDR3, and an amino acid sequence comprising SEQ ID NO: 27 C-terminal to the second binding subunit CDR3.

[0145] In some variations, the first binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 1, the first binding subunit CDR2 comprises the amino acid sequenceP190500. W0.01 of SEQ ID NO: 2, the first binding subunit CDR3 comprises the amino acid sequence of SEQ ID NO: 3, the second binding subunit CDR1 comprising the amino acid sequence of SEQ ID NO: 4, the second binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 5, and the second binding subunit CDR3 comprises the amino acid sequence of SEQ ID NO: 6.Stability

[0146] Germline identities and 9mers are increased across both the VH and VL, while maintaining binding affinity7(FIG. 29). Further, the re-humanized anti-CD19 antigen binding molecule has +7°C improved Tm(FIG. 29). For example, FIG. 29 shows that the anti-CD19 antigen binding molecule has a Tmof 78.5 °C compared to a reference anti-CD19 molecule with a Tmof 71.5 °C.

[0147] An A100Q substitution in the VL FR4 alone improves stability7by I °C and removed a predicted MHC -binding peptide (data not shown). Improving stability of the antigen binding molecule is important if antigen binding molecules are. for example, to be used in high concentrations such as in a subcutaneous formulation. Additionally, a VL CDR2 methionine that may be liable to oxidation is substituted with alanine (FIG. 28B). The M51A substitution does not damage binding affinity7. The re-engineered anti-CD19 antigen binding molecule can contain, for example, 33 amino acid substitutions relative to the parental sequence.Bi-Specific Antigen Binding Molecules

[0148] Disclosed herein are anti-CD3 and anti-CD19 bispecific antigen binding molecules comprising an anti-CD19 antigen binding domain with improved stability and an anti-CD3 antigen binding domain with improved stability. Sequence changes that mediate improved stability include, but are not limited to, changes in the framework region. Improvements in stability include increase thermostability7of the antigen binding molecules. As described above, improving stability of the antigen binding molecule is important if antigen binding molecules are, for example, to be used in high concentrations such as in a subcutaneous formulation.CDRs

[0149] Disclosed herein are bispecific anti-CD3 and anti-CD19 antigen binding molecules. In some embodiments, the bispecific anti-CD3 and anti-CD19 antigen binding molecules comprise any of the herein disclosed anti-CD3 antigen binding molecules and means for binding CD 19. In some embodiments, the bispecific anti-CD3 and anti-CD19 antigen binding molecules comprise any of the herein disclosed anti-CD19 antigen binding molecules and means for binding CD3.P190500. W0.01

[0150] The means for binding CD 19 and means for binding CD3 can correspond to Table 2:Table 2: linking the Group I means clauseMeans for Element Corresponding Element Disclosure and / or materials and / or acts disclosed in the Specificationmeans for binding CD 19 Species of CD 19 antibodies disclosed in WO2023062188,WO2021055528, WO2020221792, WO2019075366, WO2012146394. WO2012055961, WO2011051307, W02010052013, W02010052014, W02009070642, W02007068354. W02005052004, W02004106380, W02004106381, WO1999054440, WO2024227055, WO2022254337. 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WO2023097219, W02023044402, WO2020247929, WO2018208864, WO2020247932,WO2018220584. WO2018222935, WO2018237341,P190500. W0.01 W02019023097, WO2023056391, W02019033050, WO2023180346, W02023006809, WO2019034580, WO2019045856. W02019057099, WO2019057122, WO2019057124, WO2019068633, W02020204305, WO2019078697. WO2019086534, WO2019095641, WO2021195513, W02019104075, W02024079010, W02024079015. WO2019152705, WO2019143636, WO2019167962, WO2019175806, WO2019184549, W02022097061. W02022097060, WO2020236792, WO2019195535, WO2019224715, W02020156405, WO2019228406. WO2023031397, WO2022167474, WO2022069712, WO2021110935, WO2019234241, WO2021119551. WO2019237081, WO2019241216, W02020001344, W02020041537, W02020061482, W02020088403. W02020088608, W02020088437, W02020088605, WO2018224443, W02025131071, W02020114478. WO2024184287, WO2020127619, WO2024012457, WO2020168555, W02020209559, WO2020232165. WO2020237173, W02020252907, W02020264200, W02021003189, W02021009263, W02021022304. 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WO2022105787, WO2022116480, WO2022117050, WO2022129560, WO2022135468, WO2023036326. WO2022135536, W02022150803, WO2022165171, WO2022166940, WO2022171192, W02022170740. WO2022170619, W02025134050, WO2025134049, WO2022175255, WO2022192895, WO2022194264. WO2022197907, W02022200443, WO2022229434, WO2022234102, WO2025179012,W02024100451. WO2024102723, WO2022240637,P190500. W0.01 WO2022246244, WO2022262868, WO2022262733, WO2022266660, WO2022263507, WO2023273595, WO2023273914. W02025083076, WO2025082594, WO2023285560, W02023001154, W02023000791, W02023002392. W02023002390, W02023011338, W02023015170, WO2023015169, W02023014809, WO2023236099. W02023029089, W02023049150, WO2023057571, W02023060157, WO2023061419, WO2023061417. WO2023222027, WO2025140489, WO2023217289, W02023078450, WO2023078446, WO2023079425. W02025051141, WO2023103335, WO2023104938, WO2023110788, WO2023125729, WO2024131751. WO2023125611, WO2023131328, WO2023213998, WO2023138551, WO2023141856, WO2023164698. WO2023169419, WO2023183766, WO2023178357, WO2023173258, WO2023174925, WO2023179716. WO2023178645, WO2023183477, WO2023186112, WO2023192850, WO2023187130, WO2023196954. W02023060169, WO2023199235, W02023208014, WO2023206985, WO2023219120, WO2023221618. WO2023222024, WO2023230626, WO2024251242, WO2023250272, W02024002308, WO2024041435. W02024144400, WO2024067222, WO2024074145, WO2024081918, W02024092038, W02025002014. WO2024087521, WO2024102948, W02024108032, WO2024118771, W02024130032, WO2024129897. WO2024131861, WO2024131874, WO2024152963, WO2024156672, WO2024160269, WO2024165403, WO2024229238, WO2024173852, WO2024178202, WO2024178414, WO2024184812, WO2024184811, WO2024184810, WO2024191322, WO2024191168, WO2024188319, WO2024188965, WO2024188966, WO2024188355, WO2024199294, WO2024199454, W02024214007, WO2024213782, WO2024220397, WO2024235317, WO2024241206, WO2024254757. W02025006794, WO2025024646, W02025051309, W02025056013, WO2025064885, W02025064890. WO2025103425, W02025061171, WO2025064369, WO2025077737, W02025080533, W02025080534. WQ2025087681, WO2025092742, WO2025092742, WO2025091264, WO2025113615, WO2025122927. WO2025124266, WO2025124523, WO2025131075, WO2025131077, WO2025132907, W02025140010. WO2025157132, WO2025162453, WO2025175056, incorporated herein by reference to the extent the species' therein describe binding to CD3.Table 3 below links Group 2 means clauses to the relevant portions of the specification Means for Element Corresponding Element DisclosureP190500. W0.01 means for binding CD 19 Species of CD19 antibodies disclosed in paragraph

[0006] ,

[0009] -

[0011] ,means for binding CD3 Species of CD3 antibodies disclosed in paragraph

[0006] ,

[0012] -

[0014] ,

[0151] The herein disclosed bispecific anti-CD3 and anti-CD19 antigen binding molecules can comprise any of the herein disclosed anti-CD3 antigen binding molecules in combination with any of the herein disclosed anti-CD19 antigen binding molecules. For example, the bi-specific anti-CD19 and anti-CD3 antigen binding molecule can comprise an anti-CD19 antigen binding domain comprising a first binding subunit comprising the CDR sequences of SEQ ID NOs: 1, 2, and 3, and a second binding subunit comprising the CDR sequences of SEQ ID NOs: 4, 5, and 6, and an anti-CD3 antigen binding domain comprising a first binding subunit comprising the CDR sequences of SEQ ID NO: 9, SEQ ID NO: 10 or 11, and SEQ ID NO: 12, 13, 14, 15, or 16, and a second binding subunit comprising the CDR sequences of SEQ ID NOs: 17, 18, and 19. In some variations, the anti-CD19 antigen binding domain comprises a first binding subunit comprising the CDR sequences of SEQ ID NOs: 1, 2, and 3. and a second binding subunit comprising the CDR sequences of SEQ ID NOs: 4. 5, and 6, and the anti-CD3 antigen binding domain comprises a first anti-CD3 binding subunit comprising the CDR sequences of SEQ ID NOs: 9, 10, and 12 and a second anti-CD3 binding subunit comprising the CDR sequences of SEQ ID NOs: 17. 18. and 19. In some variations, the anti-CD19 antigen binding domain comprises a first binding subunit comprising the CDR sequences of SEQ ID NOs: 1, 2, and 3, and a second binding subunit comprising the CDR sequences of SEQ ID NOs: 4, 5, and 6, and the anti-CD3 antigen binding domain comprises a first anti-CD3 binding subunit comprising the CDR sequences of SEQ ID NOs: 9, 10, and 13 and a second anti-CD3 binding subunit comprising the CDR sequences of SEQ ID NOs: 17, 18, and 19. In some variations, the anti-CD19 antigen binding domain comprises a first binding subunit comprising the CDR sequences of SEQ ID NOs: 1, 2, and 3, and a second binding subunit comprising the CDR sequences of SEQ ID NOs: 4, 5, and 6, and the anti-CD3 antigen binding domain comprises a first anti-CD3 binding subunit comprising the CDR sequences of SEQ ID NOs: 9, 11, and 12 and a second anti-CD3 binding subunit comprising the CDR sequences of SEQ ID NOs: 17, 18, and 19. In some variations, the anti-CD19 antigen binding domain comprises a first binding subunit comprising the CDR sequences of SEQ ID NOs: 1, 2, and 3, and a second binding subunit comprising the CDR sequences of SEQ ID NOs: 4, 5, and 6, and the anti-CD3 antigen binding domain comprises a first anti-CD3 binding subunit comprising the CDR sequences of SEQ IDP190500. W0.01 NOs: 9. 11. and 13 and a second anti-CD3 binding subunit comprising the CDR sequences of SEQ ID NOs: 17, 18, and 19. In some variations, the anti-CD19 antigen binding domain comprises a first binding subunit comprising the CDR sequences of SEQ ID NOs: 1, 2, and 3, and a second binding subunit comprising the CDR sequences of SEQ ID NOs: 4, 5, and 6, and the anti-CD3 antigen binding domain comprises a first anti-CD3 binding subunit comprising the CDR sequences of SEQ ID NOs: 9, 10, and 14 and a second anti-CD3 binding subunit comprising the CDR sequences of SEQ ID NOs: 17, 18, and 19. In some variations, the antiCD 19 antigen binding domain comprises a first binding subunit comprising the CDR sequences of SEQ ID NOs: 1, 2, and 3, and a second binding subunit comprising the CDR sequences of SEQ ID NOs: 4, 5, and 6, and the anti-CD3 antigen binding domain comprises a first anti-CD3 binding subunit comprising the CDR sequences of SEQ ID NOs: 9, 10, and 15 and a second anti-CD3 binding subunit comprising the CDR sequences of SEQ ID NOs: 17, 18, and 19. In some variations, the anti-CD19 antigen binding domain comprises a first binding subunit comprising the CDR sequences of SEQ ID NOs: 1, 2, and 3, and a second binding subunit comprising the CDR sequences of SEQ ID NOs: 4, 5, and 6, and the anti-CD3 antigen binding domain comprises a first anti-CD3 binding subunit comprising the CDR sequences of SEQ ID NOs: 9, 11, and 14 and a second anti-CD3 binding subunit comprising the CDR sequences of SEQ ID NOs: 17, 18, and 19. In some variations, the anti-CD19 antigen binding domain comprises a first binding subunit comprising the CDR sequences of SEQ ID NOs: 1, 2, and 3, and a second binding subunit comprising the CDR sequences of SEQ ID NOs: 4, 5, and 6, and the anti-CD3 antigen binding domain comprises a first anti-CD3 binding subunit comprising the CDR sequences of SEQ ID NOs: 9, 11, and 15 and a second anti-CD3 binding subunit comprising the CDR sequences of SEQ ID NOs: 17. 18. and 19. In some variations, the anti-CD19 antigen binding domain comprises a first binding subunit comprising the CDR sequences of SEQ ID NOs: 1, 2, and 3, and a second binding subunit comprising the CDR sequences of SEQ ID NOs: 4, 5, and 6, and the anti-CD3 antigen binding domain comprises a first anti-CD3 binding subunit comprising the CDR sequences of SEQ ID NOs: 9, 10, and 16 and a second anti-CD3 binding subunit comprising the CDR sequences of SEQ ID NOs: 17, 18, and 19.Anti-CD19 x CD3 Antigen Binding Domain Framework Regions

[0152] In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules comprise an anti-CD19 antigen binding domain comprising a framework region 1 (FR1) comprising at least 95% identity to an amino acid sequence of SEQ ID NO: 20, a framework region 2 (FR2) comprising at least 95% identity7to the amino acid sequence of SEQ ID NO: 21. a framework region 3 (FR3) comprising at least 95% identity to theP190500. W0.01 amino acid sequence of SEQ ID NO: 22, a framework region 4 (FR4) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 23, a framework region 5 (FR5) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 24, a framework region 6 (FR6) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 25, a framework region 7 (FR7) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 26, and a framework region 8 (FR8) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 27. At least 95% sequence identity includes, for example, 96%, 97%, 98%, and 99% sequence identity.

[0153] In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules comprise an anti-CD19 antigen binding domain comprising an amino acid sequence comprising at least 95% identity to SEQ ID NO: 20 N-terminal to a first binding subunit CDR1, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 21 between the first binding subunit CDR1 and a first binding subunit CDR2, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 22 between the first binding subunit CDR2 and a first binding subunit CDR3, an amino acid sequence comprising at least 95% identity' to SEQ ID NO: 23 C-terminal to the first binding subunit CDR3, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 24 N-terminal to a second binding subunit CDR1, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 25 between the second binding subunit CDR1 and a second binding subunit CDR2, an amino acid sequence comprising at least 95% identity' to SEQ ID NO: 26 between the second binding subunit CDR2 and a second binding subunit CDR3, and an amino acid sequence comprising at least 95% identity’ to SEQ ID NO: 27 C-terminal to the second binding subunit CDR3. At least 95% sequence identity includes, for example, 96%, 97%, 98%, and 99% sequence identity.

[0154] In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules comprise an anti-CD19 antigen binding domain comprising a framework region 1 (FR1) comprising an amino acid sequence of SEQ ID NO: 20, a framework region 2 (FR2) comprising the amino acid sequence of SEQ ID NO: 21, a framework region 3 (FR3) comprising the amino acid sequence of SEQ ID NO: 22, a framework region 4 (FR4) comprising the amino acid sequence of SEQ ID NO: 23, a framework region 5 (FR5) comprising the amino acid sequence of SEQ ID NO: 24, a framework region 6 (FR6) comprising the amino acid sequence of SEQ ID NO: 25, a framework region 7 (FR7) comprising the amino acid sequence of SEQ ID NO: 26, and a framework region 8 (FR8) comprising the amino acid sequence of SEQ ID NO: 27.

[0155] In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules comprise an anti-CD19 antigen binding domain comprising an aminoP190500. W0.01 acid sequence comprising SEQ ID NO: 20 N-terminal to a first binding subunit CDR1. an amino acid sequence comprising SEQ ID NO: 21 between the first binding subunit CDR1 and a first binding subunit CDR2, an amino acid sequence comprising SEQ ID NO: 22 between the first binding subunit CDR2 and a first binding subunit CDR3, an amino acid sequence comprising SEQ ID NO: 23 C-terminal to the first binding subunit CDR3, an amino acid sequence comprising SEQ ID NO: 24 N-terminal to a second binding subunit CDR1, an amino acid sequence comprising SEQ ID NO: 25 between the second binding subunit CDR1 and a second binding subunit CDR2, an amino acid sequence comprising SEQ ID NO: 26 between the second binding subunit CDR2 and a second binding subunit CDR3. and an amino acid sequence comprising SEQ ID NO: 27 C-terminal to the second binding subunit CDR3.Anti-CD19 Antigen Binding Domain Variable Regions

[0156] In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules comprise a first anti-CD19 binding subunit and a first anti-CD3 binding subunit that are heavy chain variable regions, and a second anti-CDl 9 binding subunit and a second anti-CD3 binding subunit that are light chain variable regions. In some variations, the bi-specific anti-CDl 9 and anti-CD3 antigen binding molecules comprise an anti-CDl 9 antigen binding domain comprising a heavy chain variable region comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 7, and a light chain variable region comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 8. At least 95% sequence identity includes, for example, 96%, 97%, 98%, and 99% sequence identity. In some variations, the bi-specific anti-CDl 9 and anti-CD3 antigen binding molecules comprise an anti-CDl 9 antigen binding domain comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 7, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8.Anti-CD3 Antigen binding domain Framework Regions

[0157] In some variations, the herein disclosed bi-specific anti-CDl 9 and anti-CD3 antigen binding molecules comprise an anti-CD3 antigen binding domain comprising a framework region 1 (FR1) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 28, a framework region 2 (FR2) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 29, a framework region 3 (FR3) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 30, a framework region 4 (FR4) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 31, a framework region 5 (FR5) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 32, a framework region 6 (FR6)P190500. W0.01 comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 33, a framework region 7 (FR7) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 34, and a framework region 8 (FR8) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 35. At least 95% sequence identity includes, for example, 96%, 97%, 98%, and 99% sequence identity.

[0158] In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules comprise an anti-CD3 antigen binding domain comprising an amino acid sequence comprising at least 95% identity to SEQ ID NO: 28 N-terminal to a first binding subunit CDR1, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 29 between the first binding subunit CDR1 and a first binding subunit CDR2, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 30 between the first binding subunit CDR2 and a first binding subunit CDR3, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 31 C-terminal to the first binding subunit CDR3, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 32 N-terminal to a second binding subunit CDR1, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 33 between the second binding subunit CDR1 and a second binding subunit CDR2, an amino acid sequence comprising at least 95% identity to SEQ ID NO: 34 between the second binding subunit CDR2 and a second binding subunit CDR3, and an amino acid sequence comprising at least 95% identity to SEQ ID NO: 35 C-terminal to the second binding subunit CDR3. At least 95% sequence identity includes, for example, 96%, 97%, 98%, and 99% sequence identity.

[0159] In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules comprise an anti-CD3 antigen binding domain comprising a framework region 1 (FR1) comprising the amino acid sequence of SEQ ID NO: 28. a framework region 2 (FR2) comprising the amino acid sequence of SEQ ID NO: 29, a framework region 3 (FR3) comprising the amino acid sequence of SEQ ID NO: 30, a framework region 4 (FR4) comprising the amino acid sequence of SEQ ID NO: 31, a framework region 5 (FR5) comprising the amino acid sequence of SEQ ID NO: 32. a framework region 6 (FR6) comprising the amino acid sequence of SEQ ID NO: 33, a framework region 7 (FR7) comprising the amino acid sequence of SEQ ID NO: 34, and a framework region 8 (FR8) comprising the amino acid sequence of SEQ ID NO: 35.

[0160] In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules comprise an anti-CD3 antigen binding domain comprising an amino acid sequence comprising SEQ ID NO: 28 N-terminal to a first binding subunit CDR1, an amino acid sequence comprising SEQ ID NO: 29 between the first binding subunit CDR1 and a first binding subunit CDR2. an amino acid sequence comprising SEQ ID NO: 30 between the firstP190500. W0.01 domain CDR2 and a first binding subunit CDR3, an amino acid sequence comprising SEQ ID NO: 31 C-terminal to the first binding subunit CDR3, an amino acid sequence comprising SEQ ID NO: 32 N-terminal to a second binding subunit CDR1, an amino acid sequence comprising SEQ ID NO: 33 between the second binding subunit CDR1 and a second domain CDR2, an amino acid sequence comprising SEQ ID NO: 34 between the second binding subunit CDR2 and a second binding subunit CDR3, and an amino acid sequence comprising SEQ ID NO: 35 C-terminal to the second binding subunit CDR3.Anti-CD3 Antigen binding domain Variable Regions

[0161] In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules comprise a first anti-CD19 binding subunit and a first anti-CD3 binding subunit that are heavy chain variable regions, and a second anti-CD19 binding subunit and a second anti-CD3 binding subunit that are light chain variable regions. In some variations, the bi-specific anti-CD19 and anti-CD3 antigen binding molecules comprise an anti-CD3 antigen binding domain comprising a light chain variable region comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 180, and a heavy chain variable region comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 179. 181, 182, 183, 184, 185. 186, 187, 188, or 189. At least 95% sequence identity includes, for example, 96%, 97%, 98%, and 99% sequence identity. In some variations, the bi-specific anti-CD19 and anti-CD3 antigen binding molecules comprise an anti-CD3 antigen binding domain comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO: 180, and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 179. 181, 182, 183, 184, 185. 186, 187. 188, or 189.Anti-CD3 Antigen binding domain Single-Chain Variable Fragments

[0162] In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules comprise an anti-CD3 antigen binding domain comprising a singlechain variable fragment comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, or 209. At least 95% sequence identity includes, for example, 96%, 97%, 98%, and 99% sequence identity. In some variations, the bi-specific anti-CD19 and anti-CD3 antigen binding molecules comprise an anti-CD3 antigen binding domain comprising a single-chain variable fragment comprising the amino acid sequence of SEQ ID NO: 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, or 209.P190500. W0.01 Bispecific Formats

[0163] The herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules can be a 1+1 Fab-scFv-Fc, 2+1 Fab2--scFv-Fc, 1+1 Common Light Chain, 2+1 Common Light Chain, 2+1 mAb-scFv, 2+1 stackFab2-scFv-Fc, Dual scFv, One-arm scFv-mAb, caveman. Bispecific mAb, One-arm central-scFv, mAb-Fv, central-Fv, or Trident. In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules are a 1+1 Fab-scFv-Fc. In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules are a 2+1 Fab2--scFv-Fc. In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules are a 1+1 Common Light Chain. In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules are a 2+1 Common Light Chain. In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules are a 2+1 mAb-scFv. In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules are a 2+1 stackFab2-scFv-Fc. In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules are a Dual scFv. In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules are a One-arm scFv-mAb. In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules are a scFv-mAb. In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules are a Bispecific mAb. In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules are a One-arm central-scFv. In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules are a mAb-Fv. In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules are a central-Fv. In some variations, the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules are a Trident.

[0164] The 1+1 Fab-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecules can comprise the amino acid sequences of SEQ ID NOs: 210, 211, and 212, SEQ ID NOs: 213, 214. and 215, SEQ ID NOs: 213, 216, and 215, SEQ ID NOs: 213, 217, and 215, or SEQ ID NOs: 213, 218, and 215. In some variations, the 1+1 Fab-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 210, 211, and 212. In some variations, the 1+1 Fab-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 213, 214, and 215. In some variations, the 1+1 Fab-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 213, 216, and 215. In some variations, the 1+1 Fab-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of 213, 217, and 215. In some variations, the 1+1P190500. W0.01 Fab-scFv-Fc bi -specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 213, 218, and 215.

[0165] The 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecules can comprise the amino acid sequences of SEQ ID NOs: 210, 219, and 212, SEQ ID NOs: 210, 220. and 212, SEQ ID NOs: 210, 221, and 212, SEQ ID NOs: 210, 222, and 212, SEQ ID NOs: 210, 223, and 212, SEQ ID NOs: 210, 224, and 212, SEQ ID NOs: 210, 225, and 212, SEQ ID NOs: 210, 226, and 212, SEQ ID NOs: 210, 227, and 212, SEQ ID NOs: 210, 228, and 212, SEQ ID NOs: 210, 229, and 212, SEQ ID NOs: 210, 230, and 212, SEQ ID NOs: 210, 231, and 212, SEQ ID NOs: 232, 233, and 215, SEQ ID NOs: 232. 234, and 215, SEQ ID NOs: 232, 235, and 215, SEQ ID NOs: 232, 236, and 215, SEQ ID NOs: 232, 237, and 215, SEQ ID NOs: 232, 238, and 215, SEQ ID NOs: 232, 239, and 215, SEQ ID NOs: 232, 240, and 215, SEQ ID NOs: 232, 241, and 215, SEQ ID NOs: 232, 242, and 215, SEQ ID NOs: 232, 243, and 215, SEQ ID NOs: 232. 244, and 215, SEQ ID NOs: 232, 245. and 215, SEQ ID NOs: 213, 246, and 215, SEQ ID NOs: 213. 247, and 215, SEQ ID NOs: 213, 248. and 215, or SEQ ID NOs: 213. 249, and 215. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 210, 219, and 212. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 210, 220, and 212. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 210, 221, and 212. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 210, 222, and 212. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 210, 223, and 212. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 210, 224, and 212. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific antiCD 19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 210, 225, and 212. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 210, 226, and 212. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 210, 227, and 212. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 210, 228, and 212. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 210, 229, and 212. In someP190500. W0.01 variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 210, 230, and 212. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 210, 231, and 212. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 232, 233, and 215. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 232, 234, and 215. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific antiCD 19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 232, 235, and 215. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 232, 236, and 215. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 232, 237, and 215. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 232, 238, and 215. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 232, 239, and 215. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 232, 240, and 215. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 232, 241, and 215. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 232, 242, and 215. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 232, 243, and 215. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 232, 244. and 215. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 232, 245, and 215. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 213, 246, and 215. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 213, 247, and 215. In some variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 213, 248, and 215. In someP190500. W0.01 variations, the 2 + 1 Fab2-scFv-Fc bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprises the amino acid sequences of SEQ ID NOs: 213, 249, and 215.Stability

[0166] The anti-CD3 antigen binding domain of the bispecific antigen binding molecule can include, for example, a non-canonical deamidation motif GNF at positions 96-98 (Kabat numbering) in the first binding subunit CDR3. Consequentially, variants are made to remove this deamidation liability as well as to engineer stabilizing disulfide bridges. N97 is important for binding, but variants at N97 abrogated binding affinity (with the exception of N97Q which retains some binding). Variants at G96 is also important to binding, and G96E or G96T was found to improve binding affinity7(albeit with decrease in stability by ~2-4°C).

[0167] The disulfide bridges can increase the thermodynamic stability7of the anti-CD3 antigen binding domain of the bispecific antigen binding molecule. In some variations, the disulfide bridges can increase the denaturation temperature by at least 2°C). In some variations, the disulfide bridges can increase the denaturation temperature by' at least 3°C). In some variations, the disulfide bridges can increase the denaturation temperature by at least 4°C).

[0168] The anti-CD3 antigen binding domain of the bispecific antigen binding molecule can include, for example a D100G substitution to improve affinity7. Further variants are engineered to fix the deamidation and incorporate disulfide bridges while maintaining an affinity' ladder and improved stability7of the anti-CD3 antigen binding domain. Sequences for these variants are depicted in, for example, FIG. 17A-FIG. 17J, and affinity (in the context of 2 + 1 Fab2-scFv-Fc molecules of FIG. 20A-FIG. 20AD) are depicted in FIG. 30. The re-engineered anti-CD3 antigen binding domain can have binding affinity similar to a non-engineered anti-CD3 antigen binding molecule, albeit with removal of the deamidation motif and introduction of stabilizing disulfide bridges.

[0169] The anti-CD19 antigen binding domain of the bispecific antigen binding molecules are, for example, re-humanized and demonstrate enhanced stability7. Germline identities and 9mers were increased across both the VH and VL, while maintaining binding affinity' (FIG. 29). Further, the re-humanized anti-CD19 antigen binding domain has +7°C improved Tm (FIG. 29). For example, FIG. 29 shows that the anti-CD19 antigen binding domain has a Tm of 78.5 °C compared to a reference anti-CD19 molecule with a Tmof 71.5 °C.

[0170] An A100Q substitution in the VL FR4 alone of the anti-CD19 antigen binding domain of the bispecific antigen binding molecules improves stability7by 1°C and removed a predicted MHC -binding peptide (data not shown). Additionally, a VL CDR2 methionine that may be liable to oxidation is substituted with alanine (FIG. 28B). The M51A substitution doesP190500. W0.01 not damage binding affinity. The re-engineered anti-CD19 antigen binding domain can contain, for example, 33 amino acid substitutions relative to the parental sequence.

[0171] The herein disclosed anti-CD3 and anti-CD19 bispecific antigen binding molecules exhibit comparable CD19+ Ramos cell killing and T-cell activation compared to the reference bispecific molecules that lack the sequence changes that mediate the improved stabilityNucleic Acids, Vectors, and Host Cells

[0172] In one embodiment of the present invention, nucleic acids are created that encode the antigen binding molecules, and particularly antibodies described herein, and that may then be cloned into vectors host cells. A nucleic acid composition may refer to one or multiple polynucleotides.

[0173] Thus, nucleic acids, and particularly DNA, may be made that encode each protein sequence. These practices are carried out using well-known procedures. For example, a variety of methods that may find use in the present invention are described in Molecular Cloning - A Laboratory Manual, 3rd Ed. (Maniatis, Cold Spring Harbor Laboratory Press, New York, 2012), incorporated herein by reference in its entirety7.

[0174] The antibodies of the present invention may be produced by culturing a host cell transformed with nucleic acid, preferably an expression vector, containing nucleic acid encoding the antibodies, under the appropriate conditions to induce or cause expression of the protein. The conditions appropriate for expression will vary7with the choice of the expression vector and the host cell, and will be easily ascertained by one skilled in the art through routine experimentation. A wide variety of appropriate host cells may be used, including but not limited to mammalian cells, bacteria, insect cells, and yeast. For example, a variety of cell lines that may find use in the present invention are described in the ATCC® cell line catalog, available from the American Type Culture Collection.

[0175] The nucleic acids that encode the antibodies may be incorporated into an expression vector in order to express components of the antibody. A variety of expression vectors may be utilized for protein expression. Expression vectors may comprise self-replicating extra-chromosomal vectors or vectors which integrate into a host genome. Expression vectors are constructed to be compatible w ith the host cell ty pe. Thus expression vectors which find use in the present invention include but are not limited to those which enable protein expression in mammalian cells, bacteria, insect cells, yeast, and in in vitro systems. As is known in the art, a variety of expression vectors are available, commercially or otherwise, that may find use in the present invention for expressing antibodies.P190500. W0.01

[0176] As is known in the art. the nucleic acids encoding the components of the binding domains and antibodies disclosed herein can be incorporated into expression vectors as is known in the art, and depending on the host cells used to produce antibodies, including heterodimeric antibodies. Expression vectors typically comprise a protein operably linked with control or regulatory sequences, selectable markers, any fusion partners, and / or additional elements. Nucleic acids are operably linked to any number of regulatory elements (promoters, origin of replication, selectable markers, ribosomal binding sites, inducers, etc.). The expression vectors can be extra-chromosomal or integrating vectors. Generally, vectors include transcriptional and translational regulatory nucleic acid operably linked to the nucleic acid encoding the antibody, and are t pically appropriate to the host cell used to express the protein.

[0177] As will be appreciated by those in the art, the nucleic acid compositions will depend on the format and scaffold of the heterodimeric protein. Thus, for example, when the format requires three amino acid sequences, such as for the 1 + 1 Fab-scFv-Fc or 2 + 1 Fab2-scFv-Fc formats, three polynucleotides can be incorporated into one or more expression vectors for expression. In some embodiments, each polynucleotide is incorporated into a different expression vector. In other embodiments, polynucleotides can be incorporated into the same expression vector.

[0178] The polynucleotides and / or expression vectors of the invention are then transformed into any number of different types of host cells as is well known in the art, including mammalian, bacterial, yeast, insect and / or fungal cells, with mammalian cells (e.g., CHO cells), finding use in many embodiments.

[0179] In some embodiments, polynucleotides encoding each monomer are each contained within a single expression vector, generally under different or the same promoter controls. In embodiments of particular use in the present invention, each of these polynucleotides are contained on different expression vectors.

[0180] The antibodies provided herein are made by culturing host cells comprising the expression vector(s) as is well known in the art. Once produced, traditional antibody purification steps are done, including an ion exchange chromatography step. As discussed herein, having the pls of the two monomers differ by at least 0.5 can allow separation by ion exchange chromatography or isoelectric focusing, or other methods sensitive to isoelectric point. That is, the inclusion of pl substitutions that alter the isoelectric point (pl) of each monomer so that such that each monomer has a different pl and the heterodimer also has a distinct pl, thus facilitating isoelectric purification of the "1 + 1 Fab-scFv-Fc" heterodimer (e.g., anionic exchange columns, cationic exchange columns). These substitutions also aid in the determination and monitoring ofP190500. W0.01 any contaminating dual scFv-Fc and mAb homodimers post-purification (e.g., 1EF gels, clEF, and analytical IEX columns).

[0181] Proteins may be isolated or purified in a variety of ways known to those skilled in the art. Standard purification methods include chromatographic techniques, including ion exchange, hydrophobic interaction, affinity, sizing or gel filtration, and reversed-phase, carried out at atmospheric pressure or at high pressure using systems such as FPLC and HPLC.Purification methods also include electrophoretic, immunological, precipitation, dialysis, and chromatofocusing techniques. Ultrafiltration and diafiltration techniques, in conjunction with protein concentration, are also useful. As is well known in the art. a variety of natural proteins bind Fc and antibodies, and these proteins can find use in the present invention for purification of antibodies. For example, the bacterial proteins A and G bind to the Fc region. Likewise, the bacterial protein L binds to the Fab region of some antibodies, as of course does the antibody’s target antigen. Purification can often be enabled by a particular fusion partner. For example, antibodies may be purified using glutathione resin if a GST fusion is employed, Ni+2 affinity chromatography if a His-tag is employed, or immobilized anti-flag antibody if a flag-tag is used. For general guidance in suitable purification techniques, see, e.g. incorporated entirely by reference Protein Purification: Principles and Practice, 3rd Ed., Scopes, Springer-Verlag, NY, 1994. incorporated entirety by reference. The degree of purification necessary’ will vary’ depending on the screen or use of the antibodies. In some instances no purification is necessary'. If the antibodies are secreted, screening may take place directly from the media. As is well known in the art, some methods of selection do not involve purification of proteins. Thus, for example, if a library of antibodies is made into a phage display library’, protein purification may not be performed.Uses of the Bispecific Antigen Binding Molecules

[0182] Disclosed herein are methods of depleting B-cells in a subject, the method comprising administering any of the herein disclosed bi-specific anti-CD19 and anti-CD3 antigen binding molecules to the subject. Disclosed herein are compositions comprising any of the herein disclosed bispecific antibodies for use in a method of depleting B-cells in a subject, the method comprising administering the composition to the subject. Disclosed herein are any of the herein disclosed bispecific antibodies for use in the preparation of a medicament useful for depleting B-cells in a subject.

[0183] The methods for depleting B-cells can be useful in treating B cell-mediated disorders, including but not limited to allergic responses, autoimmune diseases, inflammatory’ diseases, and hematological malignancies.P190500. W0.01

[0184] The herein disclosed methods and uses can deplete peripheral B-cells, bone marrow B-cells, and lymph node B-cells. In some embodiments, peripheral B-cells are depleted. In some embodiments, bone marrow B-cells are depleted. In some embodiments, lymph node B-cells are depleted.

[0185] The bi-specific binding molecules can be administered to the subject by any suitable means of administration including, for example, subcutaneous administration or intravenous administration. In some embodiments, the bi-specific binding molecules are administered subcutaneously. In some embodiments, the bi-specific binding molecules are administered intravenously.

[0186] Disclosed herein are methods of treating an autoimmune, inflammatory disorder, or cancer in a subject, the method comprising administering any of the herein disclosed the bi-specific anti-CD19 and anti-CD3 antigen binding molecules to the subject. In some embodiments, an autoimmune disorder is treated. In some embodiments, an inflammatory disease is treated. In some variations, cancer is treated. Specifically, the treatment methods disclosed herein comprise administration to a patient in need of such administration a therapeutic amount of the antigen binding molecule as described herein.

[0187] Disclosed herein are compositions comprising any of the herein disclosed bispecific antibodies for use in a method of treating an autoimmune disorder, inflammatory disease, or cancer in a subject, the method comprising administering the composition to the subject. Disclosed herein are any of the herein disclosed bi-specific antibodies for use in the preparation of a medicament useful for treating an autoimmune disorder in a subject. In some embodiments, an autoimmune disorder is treated. In some embodiments, an inflammatory disease is treated. In some embodiments, cancer is treated.

[0188] In some variations, the antibodies disclosed herein can be for use in the treatment of an autoimmune disorder (or any specific autoimmune disorder described herein). In some variations, the antibodies disclosed herein can be for use in the treatment of an inflammatory disease (or any specific inflammatory disease described herein). In some variations, the antibodies disclosed herein can be for use in the treatment of cancer (or any specific cancer described herein).

[0189] In one variation, the autoimmune disease is allogenic islet graft rejection. In another variation, the autoimmune disease is alopecia areata. In another variation, the autoimmune disease is ankylosing spondylitis. In another variation, the autoimmune disease is antiphospholipid syndrome. In another variation, the autoimmune disease is autoimmune Addison's disease. In another variation, the autoimmune disease is antineutrophil cytoplasmic autoantibodies (ANCA). In another variation, the autoimmune disease is autoimmune diseases ofP190500. W0.01 the adrenal gland. In some variations, the autoimmune disease is anti-MDA5 syndrome. In another variation, the autoimmune disease is antisynthetase syndrome. In another variation, the autoimmune disease is autoimmune autonomic ganglionopathy (AAG). In another variation, the autoimmune disease is autoimmune encephalitis (anti-NMDAR, anti-LGIl). In another variation, the autoimmune disease is autoimmune hemolytic anemia. In another variation, the autoimmune disease is autoimmune hepatitis. In another variation, the autoimmune disease is autoimmune myocarditis. In another variation, the autoimmune disease is autoimmune neutropenia. In another variation, the autoimmune disease is autoimmune oophoritis and orchitis. In another variation, the autoimmune disease is autoimmune thrombocytopenia. In another variation, the autoimmune disease is autoimmune urticaria. In another variation, the autoimmune disease is Behcet's disease. In another variation, the autoimmune disease is bullous pemphigoid. In another variation, the autoimmune disease is cardiomyopathy. In another variation, the autoimmune disease is Castleman's syndrome. In another variation, the autoimmune disease is celiac sprucedermatitis. In another variation, the autoimmune disease is chronic fatigue immune disfunction syndrome. In another variation, the autoimmune disease is chronic inflammatory demyelinating polyneuropathy. In another variation, the autoimmune disease is Churg-Strauss syndrome. In another variation, the autoimmune disease is cicatrical pemphigoid. In another variation, the autoimmune disease is cold agglutinin disease. In another variation, the autoimmune disease is CREST syndrome. In another variation, the autoimmune disease is cold agglutinin disease. In another variation, the autoimmune disease is Crohn's disease. In another variation, the autoimmune disease is dermatomyositis. In another variation, the autoimmune disease is discoid lupus. In another variation, the autoimmune disease is essential mixed cryoglobulinemia. In another variation, the autoimmune disease is Evans syndrome. In another variation, the autoimmune disease is factor VIII deficiency. In another variation, the autoimmune disease is fibromyalgia-fibromyositis. In another variation, the autoimmune disease is glomerulonephritis. In another variation, the autoimmune disease is Grave's disease. In another variation, the autoimmune disease is Guillain-Barre. In another variation, the autoimmune disease is Goodpasture's syndrome. In another variation, the autoimmune disease is graft-versus-host disease (GVHD). In another variation, the autoimmune disease is Hashimoto's thyroiditis. In another variation, the autoimmune disease is hemophilia A. In another variation, the autoimmune disease is idiopathic pulmonary’ fibrosis. In another variation, the autoimmune disease is immune mediated thrombocytopenia. In another variation, the autoimmune disease is immune thrombocytopenic purpura (ITP). In another variation, the autoimmune disease is IgA neuropathy. In another variation, the autoimmune disease is IgM polyneuropathies. In another variation, the autoimmune disease is IgA vasculitis (Henoch-Schonlein purpura). In anotherP190500. W0.01 variation, the autoimmune disease is lgG4-related disease. In another variation, the autoimmune disease is immune mediated thrombocytopenia. In another variation, the autoimmune disease is juvenile arthritis. In another variation, the autoimmune disease is Kawasaki's disease. In another variation, the autoimmune disease is lichen planus. In another variation, the autoimmune disease is lupus erthematosis. In another variation, the autoimmune disease is Meniere's disease. In another variation, the autoimmune disease is mixed connective tissue disease. In another variation, the autoimmune disease is multiple sclerosis. In another variation, the autoimmune disease is type 1 diabetes mellitus. In another variation, the autoimmune disease is myasthenia gravis. In another variation, the autoimmune disease is Neuromyelitis optica spectrum disorder (NMOSD). In another variation, the autoimmune disease is Overlap myositis. In another variation, the autoimmune disease is pemphigus vulgaris. In another variation, the autoimmune disease is pernicious anemia. In another variation, the autoimmune disease is polyarteritis nodosa. In another variation, the autoimmune disease is polychrondritis. In another variation, the autoimmune disease is polyglandular syndromes. In another variation, the autoimmune disease is polymyalgia rheumatica. In another variation, the autoimmune disease is polymyositis and dermatomyositis. In another variation, the autoimmune disease is primary' agammaglobinulinemia. In another variation, the autoimmune disease is primary biliary' cirrhosis. In another variation, the autoimmune disease is primary biliary cholangitis (CD19+plasma-cell dominant). In another variation, the autoimmune disease is psoriasis. In another variation, the autoimmune disease is psoriatic arthritis. In another variation, the autoimmune disease is Reynauld's phenomenon. In another variation, the autoimmune disease is Reiter's syndrome. In another variation, the autoimmune disease is rheumatoid arthritis. In another variation, the autoimmune disease is sarcoidosis. In another variation, the autoimmune disease is scleroderma. In another variation, the autoimmune disease is Sjogren's syndrome. In another variation, the autoimmune disease is solid organ transplant rejection. In another variation, the autoimmune disease is stiff-man syndrome. In another variation, the autoimmune disease is systemic lupus erythematosus (SLE). in another variation, the autoimmune disease is type 1 diabetes mellitus (early or new-onset). In another variation, the autoimmune disease is systemic sclerosis (diffuse cutaneous subtype). In another variation, the autoimmune disease is takayasu arteritis. In another variation, the autoimmune disease is temporal arteristis / giant cell arteritis. In another variation, the autoimmune disease is thrombotic thrombocytopenia purpura. In another variation, the autoimmune disease is ulcerative colitis. In another variation, the autoimmune disease is uveitis. In another variation, the autoimmune disease is vasculitides such as dermatitis herpetiformis vasculitis. In another variation, the autoimmune disease is vitiligo. In another variation, the autoimmune disease is Wegner's granulomatosis.P190500. W0.01

[0190] In one variation, the inflammatory disorder is acute respiratory distress syndrome (ARDS). In another variation, the inflammatory disorder is acute septic arthritis. In another variation, the inflammatory disorder is adjuvant arthritis (Prakken et al., Springer Semin Immunopathol., 2003 August; 25(l):47-63, incorporated entirely by reference), juvenile idiopathic arthritis (de Kleer et al.. Arthritis Rheum. 2003 July; 47(7):2001-10, incorporated entirely by reference). In another variation, the inflammatory disorder is allergic encephalomyelitis. In another variation, the inflammatory disorder is allergic rhinitis. In another variation, the inflammatory disorder is allergic vasculitis. In another variation, the inflammatory disorder is allergy. In another variation, the inflammatory disorder is asthma. In another variation, the inflammatory disorder is atherosclerosis. In another variation, the inflammatory disorder is chronic inflammation due to chronic bacterial or viral infections. In another variation, the inflammatory disorder is chronic obstructive pulmonary' disease (COPD). In another variation, the inflammatory disorder is coronary artery disease. In another variation, the inflammatory disorder is encephalitis. In another variation, the inflammatory disorder is inflammatory' bowel disease. In another variation, the inflammatory' disorder is inflammatory osteolysis. In another variation, the inflammatory' disorder is inflammation associated with acute and delayed hypersensitivity reactions. In another variation, the inflammatory disorder is inflammation associated with tumors. In another variation, the inflammatory disorder is peripheral nerve injury or demyelinating diseases. In another variation, the inflammatory disorder is inflammation associated with tissue trauma such as bums and ischemia. In another variation, the inflammatory' disorder is inflammation due to meningitis. In another variation, the inflammatory disorder is multiple organ injury syndrome. In another variation, the inflammatory disorder is pulmonary fibrosis. In another variation, the inflammatory disorder is sepsis and septic shock. In another variation, the inflammatory disorder is Stevens-Johnson syndrome. In another variation, the inflammatory' disorder is undifferentiated arthropy. In another variation, the inflammatory disorder is undifferentiated spondyloarthropathy.

[0191] In some variations, a cancer is treated. The cancer can be a non-small cell lung cancer, small cell lung cancer, renal cell cancer, colorectal cancer, ovarian cancer, breast cancer, pancreatic cancer, gastric carcinoma, bladder cancer, esophageal cancer, mesothelioma, melanoma, head and neck cancer, thyroid cancer, sarcoma, prostate cancer, glioblastoma, cervical cancer, thymic carcinoma, leukemia, lymphomas, myelomas, mycoses fungoids. merkel cell cancer, and other hematologic malignancies, such as classical Hodgkin lymphoma (CHL), primary' mediastinal large B-cell lymphoma, T-cell / histiocyte-rich B-cell lymphoma, EBV-positive and -negative PTLD, and EBV-associated diffuse large B-cell lymphoma (DLBCL), plasmablastic lymphoma, extranodal NK / T-cell lymphoma, nasopharyngeal carcinoma, andP190500. W0.01 HHV8-associated primary effusion lymphoma, Hodgkin's lymphoma, neoplasm of the central nervous system (CNS), such as primary CNS lymphoma, spinal axis tumor, or brain stem glioma.

[0192] Cancers can include hematologic malignancies, such as non-Hodgkin's lymphomas (NHL), including refractory / resistant NHL. NHL cancers include but are not limited to Burkitt's lymphoma (BL), small lymphocytic lymphoma (SLL), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), follicular lymphoma (FL), marginal zone B-cell lymphoma (MZL), hairy cell leukemia (HCL) and lymphoplasmacytic leukemia (LPL), mucosa-associated lymphoid tissue (MALT), extranodal marginal zone B-cell lymphoma of MALT, nodal marginal zone B cell lymphoma, mediastinal large cell lymphoma, intravascular large cell lymphoma, primary effusion lymphoma, precursor B-lymphoblastic leukemia / lymphoma, precursor T- and NK-cells lymphoma (precursor T lymphoblastic lymphoma, blastic NK cell lymphoma), tumors of the mature T and NK cells, including peripheral T-cell lymphoma and leukemia (PTL), adult T-cell leukemia / T-cell lymphomas and large granular lymphocytic leukemia, T-cell chronic lymphocytic leukemia / prolymphocytic leukemia, T-cell large granular lymphocytic leukemia, aggressive NK-cell leukemia, extranodal T- / NK cell lymphoma, enteropathy-type T-cell lymphoma, hepatosplenic T-cell lymphoma, anaplastic large cell lymphoma (ALCL), angiocetric and angioimmunoblastic T-cell lymphoma, mycosis fungoides / Sezary syndrome, and cutaneous T-cell lymphoma (CTCL).

[0193] In some variations, the cancer is a CD19-related cancer, such as B cell ly mphoma, optionally Hodgkin lymphoma or non-Hodgkin lymphoma. Non-Hodgkin lymphoma can include large B-cell lymphoma (DLBCL), FL, MLL, MZL, MALT, small lymphocytic lymphoma (such as CLL), MCL. Acute Lymphoblastic Leukemia (ALL), or Waldenstrom's Macroglobulinemia (WM). In some variations, the CD19-related cancer is CD 19 macrophagerich hepatocellular carcinoma (experimental T AM-targeting). In some variations, the Correlated cancer is CD19+multiple myeloma (subsets with retained B-cell lineage antigens). In some variations, the CD19-related cancer is CD19+testicular lymphoma (relapsed cases). In some variations, the CD19-related cancer is CD19-expressing transformed marginal zone lymphoma.

[0194] In some variations, the cancer is a non-small cell lung cancer. In some variations, the cancer is a small cell lung cancer. In some variations, the cancer is a renal cell cancer. In some variations, the cancer is a colorectal cancer. In some variations, the cancer is a ovarian cancer. In some variations, the cancer is a breast cancer. In some variations, the cancer is a pancreatic cancer. In some variations, the cancer is a gastric carcinoma. In some variations, the cancer is a bladder cancer. In some variations, the cancer is a esophageal cancer. In someP190500. W0.01 variations, the cancer is a mesothelioma. In some variations, the cancer is a melanoma. In some variations, the cancer is a head and neck cancer. In some variations, the cancer is a thyroid cancer. In some variations, the cancer is a sarcoma. In some variations, the cancer is a prostate cancer. In some variations, the cancer is a glioblastoma. In some variations, the cancer is a cervical cancer. In some variations, the cancer is a thymic carcinoma. In some variations, the cancer is a leukemia. In some variations, the cancer is a lymphomas. In some variations, the cancer is a myeloma. In some variations, the cancer is a mycoses fungoids. In some variations, the cancer is a myeloma. In some variations, the cancer is a myeloid neoplasias. In some variations, the cancer is a merkel cell cancer. In some variations, the cancer is a primary mediastinal large B-cell lymphoma. In some variations, the cancer is a T-cell / histiocyte-rich B-cell lymphoma. In some variations, the cancer is gray zone lymphoma. In some variations, the cancer is diffuse large B-cell lymphoma (DLCL). In some variations, the cancer is diffuse large B-cell lymphoma (DLBCL). In some variations, the cancer is an EBV-positive PTLD. In some variations, the cancer is an EBV-negative PDLL. In some variations, the cancer is an EBV-associated DLBCL. In some variations, the cancer is a plasmablastic lymphoma. In some variations, the cancer is an extranodal NK / T-cell lymphoma. In some variations, the cancer is a nasophary ngeal carcinoma. In some variations, the cancer is an HHV8-associated primary effusion lymphoma. In some variations, the cancer is a Hodgkin's lymphoma. In some variations, the cancer is a Hodgkin's lymphoma including AML with maturation. In some variations, the cancer is a neoplasm of the central nervous system (CNS). In some variations, the cancer is a spinal axis tumor. In some variations, the cancer is a brain stem glioma. In some variations, the cancer is a post-transplant lymphoproliferative disorder (PTLD). In some variations, the cancer is thymoma. In some variations, the cancer is transformed follicular lymphoma. In some variations, the cancer is a tumor of lymphocyte precursor cells.

[0195] In some variations, the cancer is a non-Hodgkin's lymphomas (NHL). In some variations, the NHL is a refractory / resistant NHL. In some variations, the NHL is Burkitt's lymphoma (BL). In some variations, the NHL is small lymphocytic lymphoma (SLL). In some variations, the NHL is chronic lymphocytic leukemia (CLL). In some variations, the NHL is a mantle cell lymphoma (MCL). In some variations, the NHL is follicular lymphoma (FL). In some variations, the NHL is marginal zone B-cell lymphoma (MZL). In some variations, the NHL is hairy cell leukemia (HCL) and lymphoplasmacytic leukemia (LPL). In some variations, the NHL is mucosa-associated lymphoid tissue (MALT). In some variations, the NHL is extranodal marginal zone B-cell lymphoma of MALT. In some variations, the NHL is nodal marginal zone B cell lymphoma. In some variations, the NHL is mediastinal large cell lymphoma. In some variations, the NHL is intravascular large cell lymphoma. In some variations, the NHL isP190500. W0.01 primary effusion lymphoma. In some variations, the NHL is precursor B-lymphoblastic leukemia / lymphoma. In some variations, the NHL is precursor T- and NK-cells lymphoma (such as precursor T lymphoblastic lymphoma. In some variations, the NHL is blastic NK cell lymphoma). In some variations, the NHL is peripheral T-cell lymphoma and leukemia (PTL). In some variations, the NHL is large granular lymphocytic leukemia. In some variations, the NHL is T-cell chronic lymphocytic leukemia / prolymphocytic leukemia. In some variations, the NHL is T-cell large granular lymphocytic leukemia. In some variations, the NHL is aggressive NK-cell leukemia. In some variations, the NHL is extranodal T- / NK cell lymphoma. In some variations, the NHL is enteropathy-type T-cell lymphoma. In some variations, the NHL is hepatosplenic T-cell lymphoma. In some variations, the NHL is an aplastic large cell lymphoma (ALCL). In some variations, the NHL is an anangiocetric and angioimmunoblastic T-cell lymphoma. In some variations, the NHL is a mycosis fungoides / Sezary syndrome. In some variations, the NHL is a cutaneous T-cell lymphoma (CTCL).

[0196] The bi-specific binding molecules can be administered to the subject by any suitable means of administration including, for example, subcutaneous administration or intravenous administration. In some embodiments, the bi-specific binding molecules are administered subcutaneously. In some embodiments, the bi-specific binding molecules are administered intravenously.EXAMPLES

[0197] The following examples are provided to further describe some of the variations disclosed herein. The examples are intended to illustrate, not to limit, the disclosed variations.Example 1: CD19 Binding Domains

[0198] Sequences for human, mouse, and cynomolgus CD19 are depicted in FIG. 2 and are useful for the development of cross-reactive CD19 antigen binding domains for ease of clinical development. Sequences for CD 19 binding domains which may find use in the CD 19 x CD3 bsAbs of the disclosure are known in the art. illustrative sequences which are depicted in FIG. 14. CD19 binding domains were also engineered (as will be described in Example 6) for use in the CD19 x CD3 bsAbs of the disclosure, sequences for which are depicted in FIG. 15.Example 2: CD3 Binding DomainsP190500. W0.01

[0199] Sequences for human, mouse, and cynomolgus CD3 are depicted in FIG. 1 and are useful for the development of cross-reactive CD3 antigen binding domains for ease of clinical development. Sequences for CD3 binding domains which may find use in the CD 19 x CD3 bsAbs of the disclosure are known in the art, illustrative sequences which are depicted in FIG. 16A-FIG.16F. Novel CD3 binding domains were also engineered (as will be described in Example 7) for use in the CD 19 x CD3 bsAbs of the disclosure, sequences for which are depicted in FIG. 17A-FIG. 17J.Example 3: Engineering CD19 x CD3 bsAbs3A: 1 + 1 Fab-scFv-Fc Format

[0200] One example format utilizing Fab domains and scFv is the 1 + 1 Fab-scFv-Fc format (depicted schematically in FIG. 18A) which comprises a first monomer comprising a single-chain Fv (“scFv’’) with a first antigen binding specificity covalently attached to a first heterodimeric Fc domain i.e., scFv-domain linker-CH2-CH3, a second monomer comprising a heavy chain i.e., VH-CHl-hinge-CH2-CH3, wherein the CH2-CH3 is a second heterodimeric Fc domain complementary to the first heterodimeric Fc domain, and a light chain (LC) transfected separately so that a Fab domain having a second antigen binding specificity is formed with the heavy chain variable region.3B: 2 + 1 Fabi-scFv-Fc Format

[0201] Another format utilizing two Fab domains and scFv is the 2 + 1 Fab2-scFv-Fc format (depicted schematically in FIG. 18B) which comprises a first monomer comprising a VH-CH1 domain covalently attached to an scFv (having a first antigen binding specificity) covalently attached to a first heterodimeric Fc domain i.e., VH-CHl-first domain linked-scFv-second domain linker-CH2-CH3, a second monomer comprising a heavy chain i.e.. VH-CH1-hinge-CH2-CH3. wherein the CH2-CH3 is a second heterodimeric Fc domain complementary to the first heterodimeric Fc domain, and a light chain (LC) transfected separately so that a Fab domain having a second antigen binding specificity is formed with the heavy chain variable region.

[0202] Any number of heterodimerization approaches as is known in the art could find use in this (and other) bispecific formats, in combination with any number of approaches for purifying heterodimers from contaminating homodimers, including those depicted in FIG. A-FIG.3F. Any of the number of linkers as is known in the art may find use in linking the VH and VL domains of the scFv. Finally, it may be useful to maximize serum half-life of the bsAbs, andP190500. W0.01 any of the number of half-life extending variants as is known in the art may find use in these bsAbs.3

[0203] In particular, the bsAbs may utilize Backbones 1 or 11 in FIG. 9. The backbones utilize the L368D / K370S (on the HC): S364K / E357Q (on the VH-scFv-Fc or scFv-Fc) heterodimeric Fc variants. The HC side further includes pl variants N208D / Q295E / N384D / Q418E / N421D to increase negative charge of the heavy chain. The scFv utilizes a positively charged (GKPGS)4 linker between the VH and VL domains to increase positive charge of the scFv-Fc chain. Collectively, these two approaches enable easy purification of heterodimers from contaminating homodimers. The FcyR ablation variants utilized in this platform are the E233P / L234V / L235A / G236_ / S267K substitutions on both the HC and the VH-scFv-Fc or scFv-Fc monomers. In some cases, bsAbs includes the M428L / N434S half-life extension variants. Sequences for illustrative CD19 x CD3 bsAbs in the 1 + 1 Fab-scFv-Fc and in the 2 + 1 Fab2-scFv-Fc formats are depicted respectively in FIG. 19A-FIG. 19C and FIG. 20A-FIG. 20AD.

[0204] Although the above provide particularly useful backbones, any number of heterodimerization approaches as is known in the art could find use in the bispecific formats, in combination with any number of approaches for purifying heterodimers from contaminating homodimers, including those depicted in FIG. 3A-FIG.3F and FIG. 4. Any of the number of linkers as is known in the art may find use in linking the VH and VL domains of the scFv.Finally, it may be useful to maximize serum half-life of the bsAbs, and any of the number of half-life extending variants as is known in the art may find use in these bsAbs. Regardless of bsAb format, the CD28 bispecific antibodies are monovalent for CD28 and incorporate Fc variants engineered to ablate FcyR binding to avoid potential superagonism. Such Fc variants include those depicted in FIG. 5A and FIG. 5B.Example 4: CD19 x CD3 bsAbs deplete CD19+cells in vitro

[0205] In a first experiment, first generation CD 19 x CD3 bsAbs (utilizing the CD 19-OG binding domain and the H1.3O_L1.47 or H1.32_L1.47 CD3 binding domains; sequences for which are depicted in FIG. 19A-FIG. 19C and FIG. 20A-FIG. 20 AD) were incubated with CD19+Ramos lymphoma cell line and CD19 depleted human PBMCs at a 10:1 effector: target ratio for 48 hours. The data as depicted in FIG. 21 show that the CD 19 x CD3 bsAbs exhibit highly efficacious cytotoxicity for CD19+human lymphoma line.Example 5: CD19 x CD3 bsAbs deplete peripheral B cell in vivoP190500. W0.01

[0206] Next, activity of the first generation CD 19 x CD3 bsAbs were investigated in vivo. Cynomolgus monkeys were dosed with various concentrations of the bsAbs, and peripheral, bone marrow, and lymph node B cell depletion was assessed (along with peripheral T cell activation, IL-6 release, and pharmacokinetics). The data as depicted in FIG. 22, FIG. 23, FIG. 25 A, and FIG. 25B. show that B cell depletion was achieved with all of the CD19 x CD3 bsAbs, although most robust with the 2 + 1 Fab2-scFv-Fc format with CD3 High (XENP26982) which almost completely depletes B cells out to 14 days after dosing. T cell activation was similar among the bsAbs, although IL-6 release was consistently higher for bsAbs utilizing CD3 High (FIG. 24A and FIG. 24B). In data not shown, both CD4+and CD8+T cells exhibit similar trends in activation (CD69 and CD25). As depicted in FIG. 26, bsAbs could be measured in serum up to 4 weeks after dosing, although pharmacokinetics showed signs of ADA- or TMDD-mediated rapid clearance at -7-14 days. Notably as depicted in FIG. 27A and FIG. 27B, rapid clearance of CD 19 x CD3 bsAb coincide with early B cell recovery.Example 6: Re-engineering CD19 Binding Domain

[0207] As described in Example 5, PK showed potential signs of ADA (albeit in cynomolgus monkeys), and so it may be important to modulate immunogenicity of the bsAbs. Accordingly, the CD19-OG binding domain was re-humanized (FIG. 28 A and FIG. 28B). As depicted in FIG. 29, germline identities and 9mers were increased across both the VH and VL. while maintaining binding affinity. Another important property is the stability of the binding domains, especially if the CD 19 x CD3 bsAbs are to be used in a high concentration subcutaneous formulation. As depicted in FIG. 29, the re-humanized CD 19 binding domain had +7°C improved Tm. Notably, the A100Q substitution in the VL FR4 alone improved stability by 1°C and removed a predicted MHC-binding peptide (data not shown). Additionally, a VL CDR2 methionine that may be liable to oxidation was substituted with alanine. Notably, the M51A substitution did not damage binding affinity. Collectively, the re-engineered CD19 binding domain contains 33 amino acid substitutions relative to the parental sequence.Example 7: Re-engineering CD3 Binding DomainAs described above, improving stability of the binding domains is important if the CD 19 x CD3 bsAbs are to be used in high concentrations subcutaneous formulations. Accordingly, the CD3 binding domains were engineered with disulfide bridges to improve stability in the context of an scFv. Data as depicted in Figure 51 show that the engineered disulfide bridges increaseP190500. W0.01 Tml by ~2-4°C. While the engineered disulfide bridges did result in increased stability, there was also some loss of affinity as depicted in Figure 52.Additionally, the CD3 binding domains utilized in Examples 4 and 5 include a non-canonical deamidation motif GNF at positions 96-98 (Kabat numbering) in the VH CDR3. In a first round of engineering, variants were made to remove this deamidation liability as well as to engineer stabilizing disulfide bridges. In data not shown from initial engineering, N97 was determined to be important for binding, but it was found that variants at N97 abrogated binding affinity (with the exception of N97Q which retains some binding). Variants at G96 was also important to binding, and G96E or G96T was found to improve binding affinity’ (albeit with decrease in stability by ~2-4°C, as depicted in Figure 51). And finally it was found that D100G improves affinity. Based on these (and the baseline that Y52cA and SlOOaE were affinity' modulating variants already included in Hl.32 and Hl.89 respectively relative to Hl.30), further variants were engineered to fix the deamidation and incorporate disulfide bridges while maintaining an affinity ladder similar to the original 3x CD3 binding domains and improved stability'. Sequences for these variants are depicted in FIG. 17A-FIG. 17J, and affinity (in the context of 2 + 1 Fab2-scFv-Fc molecules of FIG. 20A-FIG. 20AD) are depicted in FIG. 30. The data show that some of the re-engineered CD3 binding domains had binding affinity similar to the original CD3 binding domains, albeit with removal of the deamidation motif and introduction of stabilizing disulfide bridges.Example 8: Optimizing CD19 x CD3 bsAbs

[0208] CD19 x CD3 bsAbs were enhanced using the novel CD19 binding domains described in Example 6, novel CD3 binding domains in Example 7. and half-life extension Fc variants, sequences for which are depicted in FIG. 19A-FIG. 19C and FIG. 20A-FIG. 20AD.Example 8A: In vitro assessmentActivity of the first generation and enhanced CD 19 x CD3 bsAbs were assessed to determine whether the re-engineered CD 19 and CD3 binding domains impacted activity. As before, the bsAbs were incubated with CD19+Ramos cells as well as human or cynomolgus PBMCs. Data as depicted in FIG. 31, FIG. 32A-FIG. 32D, FIG. 33, and FIG. 34 show that bsAbs with the re-engineered binding domains have equivalent activity to the first generation bsAbs (with respect to target cell kill and T cell activation).Example 8B: In vivo assessmentP190500. W0.01

[0209] The activity of the enhanced CD 19 x CD3 bsAbs were investigated in vivo. In a first study, cynomolgus monkeys were dosed intravenously with various concentrations (low IX dose and high 3X dose) of the bsAbs, and peripheral, bone marrow, and lymph node B cell plasma depletion and pharmacokinetics was assessed, data for which are depicted in FIG. 35 -FIG. 43, 46, and 49 (cyno 3501 and 3506 are from separate studies). As before, the study was impacted by ADA across the bispecific antibodies, so animals impacted by ADA are denoted in FIG. 35. Half-life estimated were generally consistent between compartmental and noncompartmental analysis. Noting only the animals without ADA, XENP49657 has better half-life than XENP49654 (13.2 and 16.6 days vs. 13.0 days). Notably in cynomolgus monkey 1001 and 2002 which were not impacted by ADA, respectively dosed with bsAbs having old and new CD3 binding domain, depletion of B cells and plasma cells were similar. For cynomolgus monkeys 2002 and 3501 which were not impacted by ADA, both dosed with XENP49657 respectively at IX and 3X dose, B cell depletion in peripheral blood was sustained for at least 28 days.Although XENP49665 also demonstrated effective depletion of B cells, it was dosed at much higher relative concentration, while XENP49659 demonstrated less B cell depletion despite 10X dose. Further as depicted in FIG. 38 and FIG. 39, the CD19 x CD3 bsAbs, particularly XENP49657, promoted deep B cell depletion in bone marrow and lymph nodes. As depicted in FIG. 42, the bsAbs also deplete CD 19+ plasma cells in bone marrow. As depicted in FIG. 43, XENP49657 significantly reduces serum immunoglobulin. As depicted in FIG. 46, XENP49657 exhibited half-life up to ~15 days. Notably as depicted in FIG. 49, single intravenous dose XENP49657 results in deep B-cell depletion for at least 42 days.

[0210] In a second study, cynomolgus monkeys were dosed subcutaneously with high 3X dose of XENP49657. and peripheral B cell depletion and pharmacokinetics were assessed, data for which are depicted in FIG. 47 and FIG. 48. Consistent with intravenous dosing, single subcutaneous dose XENP49657 results in deep B-cell depletion sustained for at least 42 days, and half-life up to ~ 15 days.

[0211] As depicted in FIG. 50, XENP49657 promotes depletion of CD19+ plasma cells in bone marrow.Example 9: Further Biophysical Characterization of Enhanced CD19 x CD3 bsAbs

[0212] Affinity for human and cyno CD19 and CD3 were determined as follows. For CD19, human and cyno CD19-His were captured onto a CD5 chip pre-conjugated with anti-His capture mAb. Each antigen was captured at different density in order to achieve the desired response ranges suitable for kinetics. Dilution series of antibody were followed over each surface at 25°C. For CD3, amine couple human and cyno cd3 / e were used, and bispecific antibodiesP190500. W0.01 were flowed at various concentrations. Melting temperature for the bispecific antibodies were also determined using DSC methods. Temperature was ramped from 25°C to 95°C at a rate of 1°C / min, and date were fit to a non-2 state model, with 3 transitions when appropriate. Data are collectively shown in FIG. 44.

[0213] Finally (data not shown), CD 19 x CD3 bsAb XENP49654 having the old CD3 binding domain was found to aggregate at higher concentrations, while XENP49657 and XENP49665 having stability-enhanced CD3 binding domains did not show such behavior.Example 10: CD19 x CD3 bsAbs Deplete B Cells from Patients with RA

[0214] 0.3 x 106human PBMCs from healthy donors or donors with rheumatoid arthritis were incubated with varying concentrations of XENP49657 for 24 hours at 37°C. Cells were stained for 45 minutes at 4°C followed by fixation and analysis by flow cytometry.Supernatant was used for cytokine analysis. Data, as depicted in FIG. 53, show that XENP49657 effectively depleted B cells from both healthy donors and donors with rheumatoid arthritis.Example 11: CD19 x CD3 bsAbs Deplete B Cells from Patients with SLE

[0215] 0.3 x 106human PBMCs from healthy donors or donors with systemic lupus erythematosus were incubated with varying concentrations of XENP49657 for 24 hours at 37°C. Cells were stained for 45 minutes at 4°C followed by fixation and analysis by flow cytometry. Supernatant was used for cytokine analysis. Data, as depicted in Figure 54, show that XENP49657 effectively depleted B cells from both healthy donors and donors with systemic lupus erythematosus.

[0216] Those skilled in the art will appreciate that numerous changes and modifications can be made to the preferred variations or embodiments disclosed herein and that such changes and modifications can be made without departing from the spirit of the disclosure. It is, therefore, intended that the appended claims cover all such equivalent variations as fall within the true spirit and scope of the disclosure.Sequence TableAnti-CD19 SEQ ID SYVMHFirst Subunit NO: 1CDR1P190500. W0.01 Anti-CD19 SEQ ID YINPYNDGTKYNEKFQGFirst Subunit NO: 2CDR2Anti-CD19 SEQ ID GTYYYGTRVFDYFirst Subunit NO: 3CDR3Anti-CD19 SEQ ID RS SKSLQNVNGNTYLYSecond NO: 4Subunit CDR1Anti-CD19 SEQ ID RASNLNSSecond NO: 5Subunit CDR2Anti-CD19 SEQ ID MQHLEYPITSecond NO: 6Subunit CDR3Anti-CD19 SEQ ID QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYVMHWVR First Subunit NO: 7 QAPGQGLEWMGYINPYNDGTKYNEKFQGRVTITSDKSTS TAYMELSSLRSEDTAVYYCARGTYYYGTRVFDYWGQGT LVTVSSAnti-CD19 SEQ ID DIVMTQSPLSLPVTPGEPASISCRSSKSLQNVNGNTYLYWF Second NO: 8 LQKPGQSPQLLIYRASNLNSGVPDRFSGSGSGTDFTLKISR Subunit VEAEDVGVYYCMQHLEYPITFGQGTKLEIKAnti-CD3 First SEQ ID TYAMNSubunit CDR1 NO: 9Anti-CD3 SEQ ID RIRSKYNNYATYYADSVKGNO: 10First SubunitCDR2aAnti-CD3 SEQ ID RIRSKANNYATYYADSVKGNO: 11First SubunitCDR2bAnti-CD3 First SEQ ID HTNFGDSYVSWFAYSubunit NO: 12CDR3aAnti-CD3 First SEQ ID HTNFGGSYVSWFAYSubunit NO: 13CDR3bP190500. W0.01 Anti-CD3 First SEQ ID HENFGDSYVSWFAYSubunit NO: 14CDR3cAnti-CD3 First SEQ ID HENFGGSYVSWFAYSubunit NO: 15CDR3dAnti-CD3 First SEQ ID HTNFGGEYVSWFAYSubunit NO: 16CDR3eAnti-CD3 SEQ ID GSSTGAVTTSNYANSecond NO: 17Subunit CDR1Anti-CD3 SEQ ID GTNKRAPSecond NO: 18Subunit CDR2Anti-CD3 SEQ ID ALWYSNHWVSecond NO: 19Subunit CDR3Anti-CD19 SEQ ID QVQLVQSGAEVKKPGASVKVSCKASGYTFTFR1 NO: 20Anti-CD19 SEQ ID WVRQAPGQGLEWMGFR2 NO: 21Anti-CD19 SEQ ID RVTITSDKSTSTAYMELSSLRSEDTAVYYCARFR3 NO: 22Anti-CD19 SEQ ID WGQGTLVTVSSFR4 NO: 23Anti-CD19 SEQ ID DIVMTQSPLSLPVTPGEPASISCFR5 NO: 24Anti-CD19 SEQ ID WFLQKPGQSPQLLIYFR6 NO: 25Anti-CD19 SEQ ID GVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFR7 NO: 26Anti-CD19 SEQ ID FGQGTKLEIKFR8 NO: 27Anti-CD3 FR1 SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFTFSNO: 28Anti-CD3 FR2 SEQ ID WVRQAPGKCLEWVGNO: 29P190500. W0.01 Anti-CD3 FR3 SEQ ID RFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRNO: 30Anti-CD3 FR4 SEQ ID WGQGTLVTVSSNO: 31Anti-CD3 FR5 SEQ ID QAVVTQEPSLTVSPGGTVTLTCNO: 32Anti-CD3 FR6 SEQ ID WVQQKPGKSPRGLIGNO: 33Anti-CD3 FR7 SEQ ID GVPARFSGSLLGGKAALTISGAQPEDEADYYCNO: 34Anti-CD3 FR8 SEQ ID FGCGTKLTVLNO: 35Anti-CD3 SEQ ID GKPGSGKPGSGKPGSGKPGSLinker NO: 36Human CD3 SEQ ID MQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKV sequence NO: 37 SISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDE DHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVC ENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAK AKPVTRGAGAGGRQRGQNKERPPPVPNPDYEPIRKGQRD LYSGLNQRRIHuman CD3, SEQ ID DGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHND extracellular NO: 38 KNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGS domain KPEDANFYLYLRARVCENCMEMDMouse CD3 SEQ ID MRWNTFWGILCLSLLAVGTCQDDAENIEYKVSISGTSVEL sequence NO: 39 TCPLDSDENLKWEKNGQELPQKHDKHLVLQDFSEVEDSG YYVCYTPASNKNTYLYLKARVCEYCVEVDLTAVAIIIIVDI CITLGLLMVIYYWSKNRKAKAKPVTRGTGAGSRPRGQNK ERPPP VPNPDYEPIRKGQRDLYS GLNQRAVMouse CD3, SEQ ID DAENIEYKVSISGTSVELTCPLDSDENLKWEKNGQELPQK extracellular NO: 40 HDKHLVLQDFSEVEDSGYYVCYTPASNKNTYLYLKARVC domain EYCVEVDCynomolgus SEQ ID MQSGTRWRVLGLCLLSIGVWGQDGNEEMGSITQTPYQVS CD3 sequence NO: 41 ISGTTVILTCSQHLGSEAQWQHNGKNKEDSGDRLFLPEFSE MEQS GY Y VC YPRGSNPED ASHHL YLKARVCEN CMEMD V MAVATIVIVDICITLGLLLLVYYWSKNRKAKAKPVTRGAG AGGRQRGQNKERPPPVPNPDYEPIRKGQQDLYSGLNQRRICynomolgus SEQ ID QDGNEEMGSITQTPYQVSISGTTVILTCSQHLGSEAQWQH CD3, NO: 42 NGKNKEDSGDRLFLPEFSEMEQSGYYVCYPRGSNPEDASH extracellular HLYLKARVCENCMEMDdomainP190500. W0.01 Human CD 19 SEQ ID MPPPRLLFFLLFLTPMEVRPEEPLVVKVEEGDNAVLQCLK sequence NO: 43 GTSDGPTQQLTWSRESPLKPFLKLSLGLPGLGIHMRPLAIW LFIFNVSQQMGGFYLCQPGPPSEKAWQPGWTVNVEGSGE LFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLMSPKLYVW AKDRPEIWEGEPPCLPPRDSLNQSLSQDLTMAPGSTLWLS CGVPPDSVSRGPLSWTHVHPKGPKSLLSLELKDDRPARDM WVMETGLLLPRATAQDAGKYYCHRGNLTMSFHLEITARP VLWHWLLRTGGWKVSAVTLAYLIFCLCSLVGILHLQRAL VLRRKRKRMTDPTRRFFKVTPPPGSGPQNQYGNVLSLPTP TSGLGRAQRWAAGLGGTAPSYGNPSSDVQADGALGSRSP PGVGPEEEEGEGYEEPDSEEDSEFYENDSNLGQDQLSQDG SGYENPEDEPLGPEDEDSFSNAESYENEDEELTQPVARTM DFLSPHGSAWDPSREATSLGSQSYEDMRGILYAAPQLRSIR GQPGPNHEEDADSYENMDNPDGPDPAWGGGGRMGTWST RHuman CD 19 SEQ ID PEEPLVVKVEEGDNAVLQCLKGTSDGPTQQLTWSRESPLK sequence, NO: 44 PFLKLSLGLPGLGIHMRPLAIWLFIFNVSQQMGGFYLCQPG extracellular PPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKN domain RSSEGPSSPSGKLMSPKLYVWAKDRPEIWEGEPPCLPPRDS LNQSLSQDLTMAPGSTLWLSCGVPPDSVSRGPLSWTHVHP KGPKSLLSLELKDDRPARDMWVMETGLLLPRATAQDAG KYYCHRGNLTMSFHLEITARPVLWHWLLRTGGWKMouse CD 19 SEQ ID MPSPLPVSFLLFLTLVGGRPQKSLLVEVEEGGNVVLPCLPD sequence NO: 45 SSPVSSEKLAWYRGNQSTPFLELSPGSPGLGLHVGSLGILL VIVNVSDHMGGFYLCQKRPPFKDIWQPAWTVNVEDSGEM FRWNASDVRDLDCDLRNRSSGSHRSTSGSQLYVWAKDHP KVWGTKPVCAPRGSSLNQSLINQDLTVAPGSTLWLSCGVP PVPVAKGSISWTHVHPRRPNVSLLSLSLGGEHPVREMWV WGSLLLLPQATALDEGTYYCLRGNLTIERHVKVIARSAV WLWLLRTGGWIVPVVTLVYVIFCMVSLVAFLYCQRAFILR RKRKRMTDPARRFFKVTPPSGNGTQNQYGNVLSLPTSTSG QAHAQRWAAGLGSVPGSYGNPRIQVQDTGAQSHETGLEE EGEAYEEPDSEEGSEFYENDSNLGQDQVSQDGSGYENPED EPMGPEEEDSFSNAESYENADEELAQPVGRMMDFLSPHGS AWDPSREASSLGSQSYEDMRGILYAAPQLHSIQSGPSHEE DADSYENMDKSDDLEPAWEGEGHMGTWGTTMouse CD 19 SEQ ID RPQKSLLVEVEEGGNVVLPCLPDSSPVSSEKLAWYRGNQS sequence, NO: 46 TPFLELSPGSPGLGLHVGSLGILLVIVNVSDHMGGFYLCQK extracellular RPPFKDIWQPAWTVNVEDSGEMFRWNASDVRDLDCDLR domain NRSSGSHRSTSGSQLYVWAKDHPKVWGTKPVCAPRGSSL NQSLINQDLTVAPGSTLWLSCGVPPVPVAKGSISWTHVHP RRPNVSLLSLSLGGEHPVREMWVWGSLLLLPQATALDEG TYYCLRGNLTIERHVKVIARSAVWLWLLRTGGMacaca SEQ ID MPPPCLLFFLLFLTPMEVRPQEPLVVKVEEGDNAVLQCLE fascicularis NO: 47 GTSDGPTQQLVWCRDSPFEPFLNLSLGLPGMGIRMGPLGIWLLIFNVSNQTGGFYLCQPGLPSEKAWQPGWTVSVEGSGP190500. W0.01 CD19 ELFRWNVSDLGGLGCGLKNRSSEGPSSPSGKLNSSQLYVW sequence AKDRPEMWEGEPVCGPPRDSLNQSLSQDLTMAPGSTLWL SCGVPPDSVSRGPLSWTHVRPKGPKSSLLSLELKDDRPDR DMWVVDTGLLLTRATAQDAGKYYCHRGNWTKSFYLEIT ARPALWHWLLRIGGWKVPAVTLTYLIFCLCSLVGILQLQR ALVLRRKRKRMTDPTRRFFKVTPPPGSGPQNQYGNVLSLP TPTSGLGRAQRWAAGLGGTAPSYGNPSSDVQVDGAVGSR SPPGAGPEEEEGEGYEEPDSEEGSEFYENDSNFGQDQLSQD GSGYENPEDEPLGPEDEDSFSNAESYENEDEELTQPVART MDFLSPHGSAWDPSREATSLGSQSYEDMRGLLYAAPQLR TIRGQPGPNHEEDADSYENMDNPDGPDPAWGGGGRMGT WSARMacaca SEQ ID PQEPLVVKVEEGDNAVLQCLEGTSDGPTQQLVWCRDSPF fascicularis NO: 48 EPFLNLSLGLPGMGIRMGPLGIWLLIFNVSNQTGGFYLCQP CD19 GLPSEKAWQPGWTVSVEGSGELFRWNVSDLGGLGCGLK sequence, NRSSEGPSSPSGKLNSSQLYVWAKDRPEMWEGEPVCGPPR extracellular DSLNQSLSQDLTMAPGSTLWLSCGVPPDSVSRGPLSWTHV domain RPKGPKSSLLSLELKDDRPDRDMWVVDTGLLLTRATAQD (predicted) AGKYYCHRGNWTKSFYLEITARPALWHWLLRIGGWK Gly-Ser 15 SEQ ID GGGGSGGGGSGGGGSlinker NO: 49Whitlow linker SEQ ID GSTSGSGKPGSGEGSTKGNO: 506paxA_l (+A) SEQ ID IRPRAIGGSKPRVAlinker NO: 51+B linker SEQ ID GKGGSGKGGSGKGGSNO: 52+C linker SEQ ID GGKGSGGKGSGGKGSNO: 53+D linker SEQ ID GGGKSGGGKSGGGKSNO: 54+E linker SEQ ID GKGKSGKGKSGKGKSNO: 55+F linker SEQ ID GGGKSGGKGSGKGGSNO: 56+G linker SEQ ID GKPGSGKPGSGKPGSNO: 57+H linker SEQ ID GKPGSGKPGSGKPGSGKPGSNO: 58+1 linker SEQ ID GKGKSGKGKSGKGKSGKGKSNO: 59P190500. W0.01 Gly-Ser 20 SEQ ID GGGGSGGGGSGGGGSGGGGSlinker NO: 603hsc_2 (-A) SEQ ID STAGDTHLGGEDFDlinker NO: 61-B linker SEQ ID GEGGSGEGGSGEGGSNO: 62-C linker SEQ ID GEGGSGEGGSGEGGSNO: 63-D linker SEQ ID GGGES GGGES GGGESNO: 64-E linker SEQ ID GEGESGEGESGEGESNO: 65-F linker SEQ ID GGGES GGEGS GEGGSNO: 66-G linker SEQ ID GEGESGEGESGEGESGEGESNO: 67Addl linker SEQ ID GGGGSGGGGSGGGGSNO: 68Add2 linker SEQ ID GGGGSGGGGSGGGGSGGGGSNO: 69Add3 linker SEQ ID GSTSGSGKPGSGEGSTKGNO: 70Add4 linker SEQ ID PRGASKSGSASQTGSAPGSNO: 71Add5 linker SEQ ID GTAAAGAGAAGGAAAGAAGNO: 72Add6 linker SEQ ID GTSGSSGSGSGGSGSGGGGNO: 73Add7 linker SEQ ID GKPGSGKPGSGKPGSGKPGSNO: 74EB linker SEQ ID GGSEGKSSGSGSESKSTGGSNO: 75Staple linker SEQ ID GGGSGGSGGCPPCGGSGGNO: 76(GGGGS)i SEQ ID GGGGSP190500. W0.01 (GGGGS)2 SEQ ID GGGGSGGGGSNO: 78(GGGGS)3 SEQ ID GGGGSGGGGSGGGGSNO: 79(GGGGS)4 SEQ ID GGGGSGGGGSGGGGSGGGGSNO: 80(GGGGS)5 SEQ ID GGGGSGGGGSGGGGSGGGGSGGGGSNO: 81(GGGGS)6 SEQ ID GGGGSGGGGSGGGGSGGGGSGGGGSGGGGSNO: 82(GGGGS)7 SEQ ID GGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGS NO: 83(GGGGA)I SEQ ID GGGGANO: 84(GGGGA)2SEQ ID GGGGAGGGGANO: 85(GGGGA)3 SEQ ID GGGGAGGGGAGGGGANO: 86(GGGGA)4 SEQ ID GGGGAGGGGAGGGGAGGGGANO: 87(GGGGA)5SEQ ID GGGAGGGGAGGGGAGGGGAGGGGANO: 88(GGGGA)6 SEQ ID GGGGAGGGGAGGGGAGGGGAGGGGAGGGGANO: 89(GGGGA)7 SEQ ID GGGGAGGGGAGGGGAGGGGAGGGGAGGGGAGGGGA NO: 9030AA-linker SEQ ID DPALVHQRPAPPGGGGSGGGGSGGGGSGGGNO: 91(GKPGS)i SEQ ID GKPGSNO: 92(GKPGS)s SEQ ID GKPGSGKPGSGKPGSGKPGSGKPGSNO: 93(GKPGS)S SEQ ID GKPGSGKPGSGKPGSGKPGSGKPGSGKPGSNO: 94(GGGES)I SEQ ID GGGESNO: 95P190500. W0.01 ‘'Full hinge” SEQ ID EPKSCDKTHTCPPCPNO: 96‘'Lower half SEQ ID KTHTCPPCPhinge” NO: 97‘'Full hinge SEQ ID EPKSSDKTHTCPPCPC220S variant” NO: 98“Flex lower SEQ ID GGGGSGGGGSKTHTCPPCPhalf hinge” NO: 99“Charged SEQ ID GKPGSGKPGSKTHTCPPCPlower half NO: 100hinge 1”“Charged SEQ ID GKPGSKTHTCPPCPlower half NO: 101hinge2”“Upper half SEQ ID EPKSChinge” NO: 102“Flex upper SEQ ID EPKSCGGGGSGGGGShalf hinge” NO: 103“Charged SEQ ID EPKSCGKPGSGKPGSupper half NO: 104hinge 1”“Charged SEQ ID EPKSCGKPGSupper half NO: 105hinge2”Heterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone 1 NO: 106 VKFNWYVDGVEVHNAKTKPREEEYNSTYRVVSVLTVLH monomer 1 (-) QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREEMTKNQVSLTCDVSGFYPSDIAVEWESDGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWEQGDVFSCSVMHE ALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone 1 NO: 107 VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH monomer 2 (+) QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREQMTKNQVKLTCLVKGFYPSD1AVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone 2 NO: 108 VKFNWYVDGVEVHNAKTKPREEEYNSTYRVVSVLTVLH monomer 1 (-) QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCDVSGFYPSDIAVEWESDGQPENNP190500. W0.01YKTTPPVLDSDGSFFLYSKLTVDKSRWEQGDVFSCSVMHE ALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone NO: 109 VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH monomer 2 (+) QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREEMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone 3 NO: 110 VKFNWYVDGVEVHNAKTKPREEEYNSTYRVVSVLTVLH monomer 1 (-) QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREEMTKNQVSLTCEVSGFYPSDIAVEWESDGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWEQGDVFSCSVMHE ALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone 3 NO: 111 VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH monomer 2 (+) QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREEMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone 4 NO: 112 VKFNWYVDGVEVHNAKTKPREEEYNSTYRVVSVLTVLH monomer 1 (-) QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREEMTENEVSLTCLVKGFYPSDIAVEWESDGQPENN YKTTPPVLDSDGSFFLYSKLEVDKSRWEQGDVFSCSVMHE ALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone 4 NO: 113 VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH monomer 2 (+) QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSKGSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone 5 NO: 114 VKFNWYVDGVEVHNAKTKPREEEYNSTYRVVSVLTVLH monomer 1 (-) QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCDVSGFYPSDIAVEWESDGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWEQGDVFSCSVMHE ALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone 5 NO: 115 VKFNWYVDGVEVHN AKTKPREEQYN STYRV V S VLTVLH monomer 2 (+) QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDQLTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPGKP190500. W0.01 Heterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone 6 NO: 116 VKFNWYVDGVEVHNAKTKPREEEYASTYRVVSVLTVLH monomer 1 (-) QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREEMTKNQVSLTCDVSGFYPSDIAVEWESDGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWEQGDVFSCSVMHE ALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone 6 NO: 117 VKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLH monomer 2 (+) QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone 7 NO: 118 VKFNWYVDGVEVHNAKTKPREEEYSSTYRVVSVLTVLHQ monomer 1 (-) DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREEMTKNQVSLTCDVSGFYPSDIAVEWESDGQPENNY KTTPPVLDSDGSFFLYSKLTVDKSRWEQGDVFSCSVMHEA LHNHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone 7 NO: 119 VKFNWYVDGVEVHNAKTKPREEQYSSTYRVVSVLTVLH monomer 2 (+) QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDP Fc Backbone 8 NO: 120 EVQFNWYVDGVEVHNAKTKPREEEFNSTYRVVSVLTVLH monomer 1 (-) QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPSQEEMTKNQVSLTCDVSGFYPSDIAVEWESDGQPENN YKTTPPVLDSDGSFFLYSRLTVDKSRWEEGDVFSCSVMHE ALHNHYTQKSLSLSLGKHeterodimeric SEQ ID APEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDP Fc Backbone 8 NO: 121 EVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLH monomer 2 (+) QDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYT LPPSQEQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPEN NYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVM HEALHNHYTQKSLSLSLGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE Fc Backbone 9 NO: 122 VQFNWYVDGVEVHNAKTKPREEEFNSTFRVVSVLTVVHQ monomer 1 (-) DWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTL PPSREEMTKNQVSLTCDVSGFYPSDIAVEWESDGQPENNY KTTPPMLDSDGSFFLYSKLTVDKSRWEQGDVFSCSVMHE ALHNHYTQKSLSLSPGKP190500. W0.01 Heterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE Fc Backbone 9 NO: 123 VQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH monomer 2 (+) QDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYT LPPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone NO: 124 VQFNWYVDGVEVHNAKTKPREEEFNSTFRVVSVLTVVHQ 10 monomer 1 DWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTL (-) PPSREEMTKNQVSLTCDVSGFYPSDIAVEWESDGQPENNY KTTPPMLDSDGSFFLYSKLTVDKSRWEQGDVFSCSVMHE ALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone NO: 125 VQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH 10 monomer 2 QDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYT (+) LPPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone NO: 126 VKFNWYVDGVEVHNAKTKPREEEYNSTYRVVSVLTVLH 11 monomer 1 QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT (-) LPPSREEMTKNQVSLTCDVSGFYPSDIAVEWESDGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWEQGDVFSCSVLHE ALHSHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone NO: 127 VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH 11 monomer 2 QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT (+) LPPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHSHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone NO: 128 VKFKWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH 12 monomer 1 QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT (-) LPPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone NO: 129 VKFNWYVDGVEVHNAKTKPREEEYNSTYRVVSVLTVLH 12 monomer 2 QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT (+) LPPSREEMTKNQVSLTCDVSGFYPSDIAVEWESDGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWEQGDVFSCSVMHE ALHNHYTQKSLSLSPGKP190500. W0.01 Heterodimeric SEQ ID APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDP Fc Backbone NO: 130 EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL 13 monomer 1 HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQV YTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPE NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDP Fc Backbone NO: 131 EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL 13 monomer 2 HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQV YTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPE NNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSV MHEALHNRFTQKSLSLSPGKHeterodimeric SEQ ID APEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVSVSHEDP Fc Backbone NO: 132 EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL 14 monomer 1 HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQV YTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPE NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGKHeterodimeric SEQ ID APEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVSVSHEDP Fc Backbone NO: 133 EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL 14 monomer 2 HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQV YTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPE NNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSV MHEALHNRFTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone NO: 134 VKFNWYVDGVEVHNAKTKPREEEYNSTYRVVSVLTVLH 15 monomer 1 QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCDVSGFYPSDIAVEWESDGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWEQGDVFSCSVLHE ALHSHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone NO: 135 VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH 15 monomer 2 QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDQLTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHSHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone NO: 136 VKFNWYVDGVEVHNAKTKPREEEYNSTYRVVSVLTVLH 16 monomer 1 QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREEMTKNQVSLTCDVSGFYPSDIAVEWESDGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWEQGDVFSCSVLHE ALHAHYTQKSLSLSPGKP190500. W0.01 Heterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone NO: 137 VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH 16 monomer 2 QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHAHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone NO: 138 VKFNWYVDGVEVHNAKTKPREEEYNSTYRVVSVLTVLH 17 monomer 1 QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCDVSGFYPSDIAVEWESDGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWEQGDVFSCSVLHE ALHAHYTQKSLSLSPGKHeterodimeric SEQ ID APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPE Fc Backbone NO: 139 VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH 17 monomer 2 QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSRDQLTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHAHYTQKSLSLSPGKIgGl CH1(+) SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS NO: 140 WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDKKVIgGl CHl(-) SEQ ID ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS NO: 141 WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT YICNVNHKPSDTKVDKKVIgG2 CHl(+) SEQ ID ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSW NO: 142 NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYT CNVDHKPSNTKVDKTVIgG2 CHl(-) SEQ ID ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSW NO: 143 NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYT CNVDHKPSDTKVDKTVIgG4 CH1(+) SEQ ID ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSW NO: 144 NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYT CNVDHKPSNTKVDKRVIgG4 CHl(-) SEQ ID ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSW NO: 145 NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYT CNVDHKPSDTKVDKRVIgGl hinge SEQ ID EPKSCDKTHTCPPCPNO: 146IgG2 hinge SEQ ID ERKCCVECPPCPNO: 147P190500. W0.01 IgG4 hinge SEQ ID ESKYGPPCPSCPNO: 148Light Chain SEQ ID RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQ Constant NO: 149 WKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE Domain - KHKVY ACEVTHQGL S S P VTKSFNRGECKappaLight Chain SEQ ID GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVA Constant NO: 150 WKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWK Domain - SHRSYSCQVTHEGSTVEKTVAPTECSLambdaAnti-CD19- SEQ ID EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ OG First NO: 151 APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY Subunit MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT vssAnti-CD19- SEQ ID DIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNTYLYW OG Second NO: 152 FQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISS Subunit LEPEDFAVYYCMQHLEYPITFGAGTKLEIKAnti-CD19- SEQ ID RMSNLNSOG Second NO: 153Subunit CDR2CD3 High - SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ |anti- NO: 154 APGKGLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK CD3]_H1.30_ NTLYLQMNSLRAEDTAVYYCVRHGNFGDSYVSWFAYWG L1.47_scFv QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVP ARF SGSLLGGKAALTIS GAQPEDEADYYC A LWYSNHWVFGGGTKLTVLAnti-CD3 VHa SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ NO: 155 APGKGLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK NTLYLQMNSLRAEDTAVYYCVRHGNFGDSYVSWFAYWG QGTLVTVSSAnti-CD3 scFv SEQ ID HGNFGDSYVSWFAYvhCDR3a NO: 156Anti-CD3 VLa SEQ ID QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ NO: 157 QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG AQPEDEADYYCALWYSNHWVFGGGTKLTVL CD3 High-Int SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ #1 - [anti- NO: 158 APGKGLEWVGRIRSKANNYATYYADSVKGRFTISRDDSK CD3]_H1.32_ NTLYLQMNSLRAEDTAVYYCVRHGNFGDSYVSWFAYWG L1.47_scFv QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIGP190500. W0.01GTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCA LWYSNHWVFGGGTKLTVLAnti-CD3 VHb SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ NO: 159 APGKGLEWVGRIRSKANNYATYYADSVKGRFTISRDDSK NTLYLQMNSLRAEDTAVYYCVRHGNFGDSYVSWFAYWG QGTLVTVSSAnti-CD3 scFv SEQ ID HGNFGDEYVSWFAYvhCDR3b NO: 160Anti-CD3 scFv SEQ ID HGNFGDPYVSWFAYvhCDR3c NO: 161Anti-CD3 scFv SEQ ID HGNFGDSYVSWFDYvhCDR3d NO: 162Anti-CD3 scFv SEQ ID TYAMSvhCDRl NO: 163Anti-CD3 scFv SEQ ID HENFGGEYVSWFAYvhCDR3e NO: 164CD3 High-Int SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ #2 - [anti- NO: 165 APGKGLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK CD3]_H1.89_ NTLYLQMNSLRAEDTAVYYCVRHGNFGDEYVSWFAYWG L1.47 scFv QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCA LWYSNHWVFGGGTKLTVLAnti-CD3 VHc SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ NO: 166 APGKGLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK NTLYLQMNSLRAEDTAVYYCVRHGNFGDEYVSWFAYWG QGTLVTVSS CD3 High-Int SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ - [anti- NO: 167 APGKGLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK CD3]_H1.90_ NTLYLQMNSLRAEDTAVYYCVRHGNFGDPYVSWFAYWG L1.47_scFv QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCA LWYSNHWVFGGGTKLTVLAnti-CD3 VHd SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ NO: 168 APGKGLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK NTLYLQMNSLRAEDTAVYYCVRHGNFGDPYVSWFAYWG QGTLVTVSSAnti-CD3 VHe SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ NO: 169 APGKGLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKP190500. W0.01NTLYLQMNSLRAEDTAVYYCVRHGNFGDSYVSWFDYWG QGTLVTVSSAnti-CD3- SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ Intermediate - NO: 170 APGKGLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK [anti- NTLYLQMNSLRAEDTAVYYCVRHGNFGDSYVSWFDYWG CD3]_H1.33_ QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT L1.47_scFv VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCA LWYSNHWVFGGGTKLTVLAnti-CD3 VHf SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMSWVRQ NO: 171 APGKGLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK NTLYLQMNSLRAEDTAVYYCVRHGNFGDSYVSWFAYWG QGTLVTVSS CD3 Low - EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMSWVRQ [anti- APGKGLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK CD3] Hl.31 NTLYLQMNSLRAEDTAVYYCVRHGNFGDSYVSWFAYWG SEQ IDL1.47_scFv QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT NO: 172VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVP ARF SGSLLGGKAALTIS GAQPEDEADYYC A LWYSNHWVFGGGTKLTVL CD3 High[VL- QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ VH] - [anti- QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG CD3]_L1.47_ AQPEDEADYYCALWYSNHWVFGGGTKLTVLGKPGSGKP SEQ IDH1.30_scFv GSGKPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFT NO: 173FSTYAMNWVRQAPGKGLEWVGRIRSKYNNYATYYADSV KGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFG DSYVSWFAYWGQGTLVTVSS CD3 High-lnt QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ #1[VL-VH] - QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG [anti- AQPEDEADYYCALWYSNHWVFGGGTKLTVLGKPGSGKP SEQ ID CD3]_L1.47_ GSGKPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFT NO: 174H1.32_scFv FSTYAMNWVRQAPGKGLEWVGRIRSKANNYATYYADSV KGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFG DSYVSWFAYWGQGTLVTVSS CD3 High-lnt QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ #2[VL-VH] - QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG [anti- AQPEDEADYYCALWYSNHWVFGGGTKLTVLGKPGSGKP SEQ ID CD3] L1.47 GSGKPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFT NO: 175H1.89_scFv FSTYAMNWVRQAPGKGLEWVGRIRSKYNNYATYYADSV KGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFG DEYVSWFAYWGQGTLVTVSS CD3 High- SEQ ID QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ Int[VL-VH] - NO: 176 QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG[anti- AQPEDEADYYCALWYSNHWVFGGGTKLTVLGKPGSGKPP190500. W0.01 CD3]_L1.47_ GSGKPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFT H1.90_scFv FSTYAMNWVRQAPGKGLEWVGRIRSKYNNYATYYADSV KGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFG DPYVSWFAYWGQGTLVTVSSAnti-CD3- QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ Intermediate[V QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG L-VH] - [anti- AQPEDEADYYCALWYSNHWVFGGGTKLTVLGKPGSGKP SEQ ID CD3]_L1.47_ GSGKPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFT NO: 177H1.33_scFv FSTYAMNWVRQAPGKGLEWVGRIRSKYNNYATYYADSV KGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFG DSYV S WFDYWGQGTLVTVS S CD3 High- QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ Int[VL-VH] - QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG [anli- AQPEDEADYYCALWYSNHWVFGGGTKLTVLGKPGSGKP SEQ ID CD3]_L1.47_ GSGKPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFT NO: 178Hl.31 scFv FSTYAMSWVRQAPGKGLEWVGRIRSKYNNYATYYADSV KGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFG DSYVSWFAYWGQGTLVTVSSAnti-CD3 VHg EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ SEQ ID APGKCLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK NO: 179 NTLYLQMNSLRAEDTAVYYCVRHTNFGDSYVSWFAYWG QGTLVTVSSAnti-CD3 VLb QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ SEQ ID QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG NO: 180AQPEDEADYYCALWYSNHWVFGCGTKLTVLAnti-CD3 VHh EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ SEQ ID APGKCLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK NO: 181 NTLYLQMNSLRAEDTAVYYCVRHTNFGGSYVSWFAYWG QGTLVTVSSAnti-CD3 VHi EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ SEQ ID APGKCLEWVGRIRSKANNYATYYADSVKGRFTISRDDSK NO: 182 NTLYLQMNSLRAEDTAVYYCVRHTNFGDSYVSWFAYWG QGTLVTVSSAnti-CD3 VHj EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ SEQ ID APGKCLEWVGRIRSKANNYATYYADSVKGRFTISRDDSK NO: 183 NTLYLQMNSLRAEDTAVYYCVRHTNFGGSYVSWFAYWG QGTLVTVSSAnti-CD3 VHk EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ SEQ ID APGKCLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK NO: 184 NTLYLQMNSLRAEDTAVYYCVRHENFGGSYVSWFAYWGQGTLVTVSSP190500. W0.01 Anti-CD3 VH1 EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ SEQ ID APGKCLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK NO: 185 NTLYLQMNSLRAEDTAVYYCVRHENFGDSYVSWFAYWG QGTLVTVSSAnti-CD3 EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ VHm SEQ ID APGKCLEWVGRIRSKANNYATYYADSVKGRFTISRDDSK NO: 186 NTLYLQMNSLRAEDTAVYYCVRHENFGDSYVSWFAYWG QGTLVTVSSAnti-CD3 VHn EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ SEQ ID APGKCLEWVGRIRSKANNYATYYADSVKGRFTISRDDSK NO: 187 NTLYLQMNSLRAEDTAVYYCVRHENFGGSYVSWFAYWG QGTLVTVSSAnti-CD3 VHo EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ SEQ ID APGKCLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK NO: 188 NTLYLQMNSLRAEDTAVYYCVRHTNFGGEYVSWFAYWG QGTLVTVSSAnti-CD3 VHp EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ SEQ ID APGKCLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK NO: 189 NTLYLQMNSLRAEDTAVYYCVRHENFGGEYVSWFAYWG QGTLVTVSSAnti-CD3 - EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ H1.214_L1.47 APGKCLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK VH G44C V NTLYLQMNSLRAEDTAVYYCVRHTNFGDSYVSWFAYWG SEQ ID L GlOOC scF QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT NO: 190V VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCA LWYSNHWVFGC GTKLTVLAnti-CD3 - EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ H1.270_L1.47 APGKCLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK VH G44C V NTLYLQMNSLRAEDTAVYYCVRHTNFGGSYVSWFAYWG SEQ ID L G100C scF QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT NO: 191V VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCA LWYSNHWVFGC GTKLTVLAnti-CD3 - EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ H1.271_L1.47 APGKCLEWVGRIRSKANWATYYADSVKGRFTISRDDSK VH G44C V NTLYLQMNSLRAEDTAVYYCVRHTNFGDSYVSWFAYWG SEQ IDL_G100C_scF QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT NO: 192V VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCA LWYSNHWVFGCGTKLTVLAnti-CD3 - SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ H1.272_L1.47 NO: 193 APGKCLEWVGRIRSKANNYATYYADSVKGRFTISRDDSKP190500. W0.01 VH G44C V NTLYLQMNSLRAEDTAVYYCVRHTNFGGSYVSWFAYWG L GlOOC scF QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT V VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVP ARF SGSLLGGKAALTIS GAQPEDEADYYC A LWYSNHWVFGC GTKLTVLAnti-CD3 - EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ H1.273_L1.47 APGKCLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK VH G44C V NTLYLQMNSLRAEDTAVYYCVRHENFGDSYVSWFAYWG SEQ ID L GlOOC scF QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT NO: 194V VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCA LWYSNHWVFGC GTKLTVLAnti-CD3 - EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ H1.274_L1.47 APGKCLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK VH G44C V NTLYLQMNSLRAEDTAVYYCVRHENFGGSYVSWFAYWG SEQ ID L GlOOC scF QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT NO: 195V VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCA LWYSNHWVFGC GTKLTVLAnti-CD3 - EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ H1.275_L1.47 APGKCLEWVGRIRSKANNYATYYADSVKGRFTISRDDSK VH G44C V NTLYLQMNSLRAEDTAVYYCVRHENFGDSYVSWFAYWG SEQ ID L GlOOC scF QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT NO: 196V VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVP ARFSGSLLGGKAALTISGAQPEDEADYYCA LWYSNHWVFGCGTKLTVLAnti-CD3 - EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ H1.276_L1.47 APGKCLEWVGRIRSKANNYATYYADSVKGRFTISRDDSK VH G44C V NTLYLQMNSLRAEDTAVYYCVRHENFGGSYVSWFAYWG SEQ ID L GlOOC scF QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT NO: 197V VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVP ARFSGSLLGGKAALTISGAQPEDEADYYCA LWYSNHWVFGCGTKLTVLAnti-CD3 - EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ H1.279_L1.47 APGKCLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK VH G44C V NTLYLQMNSLRAEDTAVYYCVRHTNFGGEYVSWFAYWG SEQ IDL_G100C_scF QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT NO: 198V VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVP ARF SGSLLGGKAALTIS GAQPEDEADYYC A LWYSNHWVFGC GTKLTVLAnti-CD3 - EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ H1.280_L1.47 SEQ ID APGKCLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK VH G44C V NO: 199 NTLYLQMNSLRAEDTAVYYCVRHENFGGEYVSWFAYWGQGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIGP190500. W0.01 L GlOOC scF GTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCA V LWYSNHWVFGCGTKLTVLAnti-CD3 - QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ L1.47_H1.214 QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG VH G44C V AQPEDEADYYCALWYSNHWVFGCGTKLTVLGKPGSGKP SEQ ID L GlOOC scF GSGKPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFT NO: 200V FSTYAMNWVRQAPGKCLEWVGRIRSKYNNYATYYADSV KGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHTNFG DSYVSWFAYWGQGTLVTVSSAnti-CD3 - QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ L1.47_H1.27O QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG VH G44C V AQPEDEADYYCALWYSNHWVFGCGTKLTVLGKPGSGKP SEQ ID L GlOOC scF GSGKPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFT NO: 201V FSTYAMNWVRQAPGKCLEWVGRIRSKYNNYATYYADSV KGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHTNFG GSYV S WF AYWGQGTLVTVS SAnti-CD3 - QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ L1.47_H1.271 QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG VH G44C V AQPEDEADYYCALWYSNHWVFGCGTKLTVLGKPGSGKP SEQ ID L GlOOC scF GSGKPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFT NO: 202V FSTYAMNWVRQAPGKCLEWVGRIRSKANNYATYYADSV KGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHTNFG DSYVSWFAYWGQGTLVTVSSAnti-CD3 - QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ L1.47_H1.272 QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG VH G44C V AQPEDEADYYCALWYSNHWVFGCGTKLTVLGKPGSGKP SEQ IDL_G100C_scF GSGKPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFT NO: 203V FSTYAMNWVRQAPGKCLEWVGRIRSKANNYATYYADSV KGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHTNFG GSYVSWFAYWGQGTLVTVSSAnti-CD3 - QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ L1.47_H1.273 QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG VH G44C V AQPEDEADYYCALWYSNHWVFGCGTKLTVLGKPGSGKP SEQ ID L GlOOC scF GSGKPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFT NO: 204V FSTYAMNWVRQAPGKCLEWVGRIRSKYNNYATYYADSV KGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHENFG DSYVSWFAYWGQGTLVTVSSAnti-CD3 - QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ L1.47_H1.274 QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG VH G44C V AQPEDEADYYCALWYSNHWVFGCGTKLTVLGKPGSGKP SEQ ID L GlOOC scF GSGKPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFT NO: 205V FSTYAMNWVRQAPGKCLEWVGRIRSKYNNYATYYADSV KGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHENFG GSYVSWFAYWGQGTLVTVSSP190500. W0.01 Anti-CD3 - QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ L1.47_H1.275 QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG VH G44C V AQPEDEADYYCALWYSNHWVFGCGTKLTVLGKPGSGKP SEQ ID L GlOOC scF GSGKPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFT NO: 206V FSTYAMNWVRQAPGKCLEWVGRIRSKANNYATYYADSV KGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHENFG DSYVSWFAYWGQGTLVTVSSAnti-CD3 - QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ L1.47_H1.276 QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG VH G44C V AQPEDEADYYCALWYSNHWVFGCGTKLTVLGKPGSGKP SEQ IDL_G100C_scF GSGKPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFT NO: 207V FSTYAMNWVRQAPGKCLEWVGRIRSKANNYATYYADSV KGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHENFG GSYVSWFAYWGQGTLVTVSSAnti-CD3 - QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ L1.47_H1.279 QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG VH G44C V AQPEDEADYYCALWYSNHWVFGCGTKLTVLGKPGSGKP SEQ IDL_G100C_scF GSGKPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFT NO: 208c FSTYAMNWVRQAPGKCLEWVGRIRSKYNNYATYYADSV KGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHTNFG GEYVSWFAYWGQGTLVTVSSAnti-CD3 - QAVVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQ LI.47_HI.280 QKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISG VH G44C V AQPEDEADYYCALWYSNHWVFGCGTKLTVLGKPGSGKP SEQ IDL_G100C_scF GSGKPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFT NO: 209c FSTYAMNWVRQAPGKCLEWVGRIRSKYNNYATYYADSV KGRFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHENFG GEYVSWFAYWGQGTLVTVSSChain 1 - QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYVMHWVR CD19- QAPGQGLEWMGYINPYNDGTKYNEKFQGRVTITSDKSTS NEW VH Ig TAYMELSSLRSEDTAVYYCARGTYYYGTRVFDYWGQGT Gl_pl(- LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE )_Isosteric_A_ PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL PVA_ / S267K / SEQ ID GTQTYICNVNHKPSDTKVDKKVEPKSCDKTHTCPPCPAPP L368D / K370S / NO: 210 VAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVK M428L / N434S FNWYVDGVEVHNAKTKPREEEYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLP PSREEMTKNQVSLTCDVSGFYPSDIAVEWESDGQPENNYK TTPPVLDSDGSFFLYSKLTVDKSRWEQGDVFSCSVLHEAL HSHYTQKSLSLSPGKChain 2 - EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ [CD3]_H1.30_ SEQ ID APGKGLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK L1.47_scFv(G NO: 211 NTLYLQMNSLRAEDTAVYYCVRHGNFGDSYVSWFAYWG KPGS)4_Fc(2 QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT16)_IgGl_C22 VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIGP190500. W0.01 0S / PVAJS267 GTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCA K / S364K / E357 LWYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPPVAG Q / M428L / N43 PSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNW 4S YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE QMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTTP PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSH YTQKSLSLSPGKChain 3 - DIVMTQSPLSLPVTPGEPASISCRSSKSLQNVNGNTYLYWF 4G7_L4.201 LQKPGQSPQLLIYRASNLNSGVPDRFSGSGSGTDFTLKISR Light Chain SEQ ID VEAEDVGVYYCMQHLEYPITFGQGTKLEIKRTVAAPSVFI NO: 212 FPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQS GNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLS SPVTKSFNRGECChain 1 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG VHa lgG MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT 1 VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT SEQ ID QTYICNVNHKPSDTKVDKKVEPKSCDKTHTCPPCPAPPVA NO: 213 GPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFN WYVDGVEVHNAKTKPREEEYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR EEMTKNQVSLTCDVSGFYPSDIAVEWESDGQPENNYKTTP PVLDSDGSFFLYSKLTVDKSRWEQGDVFSCSVMHEALHN HYTQKSLSLSPGKChain 2 - EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ [CD3]_H1.89_ APGKGLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK L1.47_scFv(G NTLYLQMNSLRAEDTAVYYCVRHGNFGDEYVSWFAYWG KPGS)4_Fc(2 QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT 16)_IgGl VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCA SEQ ID LWYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPPVAG NO: 214PSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNW YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE QMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTTP PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHN HYTQKSLSLSPGKChain 3 - DIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNTYLYW 4G7 L1.155 FQQKPGQSPQLLIYRMSNLNSGVPDRFSGSGSGTEFTLTISS Light Chain SEQ ID LEPEDFAVYYCMQHLEYPITFGAGTKLEIKRTVAAPSVFIF NO: 215 PPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSG NSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVT HQGLSSPVTKSFNRGECP190500. W0.01 Chain 2 - EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ [CD3]_H1.33_ APGKGLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK L1.47_scFv(G NTLYLQMNSLRAEDTAVYYCVRHGNFGDSYVSWFDYWG KPGS)4_Fc(2 QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT 16)_IgGl VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCA SEQ ID LWYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPPVAG NO: 216PSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNW YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE QMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTTP PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHN HYTQKSLSLSPGKChain 2 - EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ [CD3]_H1.32_ APGKGLEWVGRIRSKANNYATYYADSVKGRFTISRDDSK L1.47_scFv(G NTLYLQMNSLRAEDTAVYYCVRHGNFGDSYVSWFAYWG KPGS)4_Fc(2 QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT 16)_IgGl VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVP ARF SGSLLGGKAALTIS GAQPEDEADYYC A SEQ ID LWYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPPVAG NO: 217PSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNW YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE QMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTTP PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHN HYTQKSLSLSPGKChain 2 - EVQLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQ [CD3]_H1.30_ APGKGLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSK L1.47_scFv(G NTLYLQMNSLRAEDTAVYYCVRHGNFGDSYVSWFAYWG KPGS)4_Fc(2 QGTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLT 16)_IgGl VSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIG GTNKRAPGVP ARFSGSLLGGKAALTISGAQPEDEADYYCA SEQ ID LWYSNHWVFGGGTKLTVLEPKSSDKTHTCPPCPAPPVAG NO: 218PSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNW YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE QMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTTP PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHN HYTQKSLSLSPGKChain 2 - QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYVMHWVR CD19- QAPGQGLEWMGYINPYNDGTKYNEKFQGRVTITSDKSTS NEW VH (G SEQ ID TAYMELSSLRSEDTAVYYCARGTYYYGTRVFDYWGQGT 4S)2_[CD3]_L NO: 219 LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE 1.47_H1.280_ PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL VH G44C VL GTQTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQAG100C scFv( VVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKP190500. W0.01 GKPGS)4_(G4 PGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQP S)2_Fc(222)_I EDEAD YYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSG gGl KPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFST YAMNWVRQAPGKCLEWVGRIRSKYNNYATYYADSVKG RFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHENFGGE YVSWFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAP PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHSHYTQKSLSLSPGKChain 2 - QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYVMHWVR CD19- QAPGQGLEWMGYINPYNDGTKYNEKFQGRVTITSDKSTS NEW VH (G TAYMELSSLRSEDTAVYYCARGTYYYGTRVFDYWGQGT 4S)2_[CD3]_L LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE 1.47_H1.279_ PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL VH G44C VL GTQTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQA _G100C_scFv( VVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQK GKPGS)4_(G4 PGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQP S)2_Fc(222)_I EDEAD YYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSG SEQ IDgGl KPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFST NO: 220YAMNWVRQAPGKCLEWVGRIRSKYNNYATYYADSVKG RFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHTNFGGE YVSWFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAP PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHSHYTQKSLSLSPGKChain 2 - QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYVMHWVR CD19- QAPGQGLEWMGYINPYNDGTKYNEKFQGRVTITSDKSTS NEW VH (G TAYMELSSLRSEDTAVYYCARGTYYYGTRVFDYWGQGT 4S)2_[CD3]_L LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE 1.47_H1.276_ PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL VH G44C VL GTQTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQA _G100C_scFv( VVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQK GKPGS)4_(G4 SEQ ID PGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQP S)2_Fc(222)_I NO: 221 EDEAD YYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSG gGl KPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFST YAMNWVRQAPGKCLEWVGRIRSKANNYATYYADSVKG RFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHENFGGS YVSWFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAP PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLP190500. W0.01PPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHSHYTQKSLSLSPGKChain 2 - QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYVMHWVR CD19- QAPGQGLEWMGYINPYNDGTKYNEKFQGRVTITSDKSTS NEW_VH_(G TAYMELSSLRSEDTAVYYCARGTYYYGTRVFDYWGQGT 4S)2_[CD3]_L LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE 1.47_H1.275_ PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL VH G44C VL GTQTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQA _G100C_scFv( VVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQK GKPGS)4_(G4 PGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQP S)2_Fc(222)_I EDEAD YYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSG SEQ IDgGl KPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFST NO: 222YAMNWVRQAPGKCLEWVGRIRSKANNYATYYADSVKG RFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHENFGDS YVSWFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAP PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHSHYTQKSLSLSPGKChain 2 - QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYVMHWVR CD19- QAPGQGLEWMGYINPYNDGTKYNEKFQGRVTITSDKSTS NEW VH (G TAYMELSSLRSEDTAVYYCARGTYYYGTRVFDYWGQGT 4S)2_[CD3]_L LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE 1.47_H1.274_ PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL VH G44C VL GTQTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQA _G100C_scFv( VVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQK GKPGS)4_(G4 PGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQP S)2_Fc(222)_I EDEAD YYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSG SEQ IDgGl KPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFST NO: 223YAMNWVRQAPGKCLEWVGRIRSKYNNYATYYADSVKG RFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHENFGGS YVSWFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAP PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHSHYTQKSLSLSPGKChain 2 - QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYVMHWVR CD19- SEQ ID QAPGQGLEWMGYINPYNDGTKYNEKFQGRVTITSDKSTS NEW VH (G NO: 224 TAYMELSSLRSEDTAVYYCARGTYYYGTRVFDYWGQGT 4S)2_[CD3]_L LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE1.47_H1.273_ PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLP190500. W0.01 VH_G44C_VL GTQTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQA _G100C_scFv( VVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQK GKPGS)4_(G4 PGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQP S)2_Fc(222)_I EDEAD YYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSG gGl KPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFST YAMNWVRQAPGKCLEWVGRIRSKYNNYATYYADSVKG RFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHENFGDS YVSWFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAP PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHSHYTQKSLSLSPGKChain 2 - QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYVMHWVR CD19- QAPGQGLEWMGYINPYNDGTKYNEKFQGRVTITSDKSTS NEW VH (G TAYMELSSLRSEDTAVYYCARGTYYYGTRVFDYWGQGT 4S)2_[CD3]_L LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE 1.47_H1.272_ PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL VH G44C VL GTQTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQA _G100C_scFv( VVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQK GKPGS)4_(G4 PGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQP S)2_Fc(222)_I EDEAD YYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSG SEQ IDgGl KPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFST NO: 225YAMNWVRQAPGKCLEWVGRIRSKANNYATYYADSVKG RFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHTNFGGS YVSWFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAP PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHSHYTQKSLSLSPGKChain 2 - QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYVMHWVR CD19- QAPGQGLEWMGYINPYNDGTKYNEKFQGRVTITSDKSTS NEW VH (G TAYMELSSLRSEDTAVYYCARGTYYYGTRVFDYWGQGT 4S)2_[CD3]_L LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE 1.47_H1.271_ PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL VH G44C VL GTQTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQA _G100C_scFv( SEQ ID VVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQK GKPGS)4_(G4 NO: 226 PGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQP S)2_Fc(222)_I EDEAD YYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSG gGl KPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFST YAMNWVRQAPGKCLEWVGRIRSKANNYATYYADSVKG RFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHTNFGDS YVSWFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVP190500. W0.01KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHSHYTQKSLSLSPGKChain 2 - QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYVMHWVR CD19- QAPGQGLEWMGYINPYNDGTKYNEKFQGRVTITSDKSTS NEW VH _(G TAYMELSSLRSEDTAVYYCARGTYYYGTRVFDYWGQGT 4S)2_[CD3]_L LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE 1.47_H1.270_ PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL VH_G44C_VL GTQTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQA _G100C_scFv( VVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQK GKPGS)4_(G4 PGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQP S)2_Fc(222)_I EDEAD YYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSG SEQ IDgGl KPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFST NO: 227YAMNWVRQAPGKCLEWVGRIRSKYNNYATYYADSVKG RFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHTNFGGS YVSWFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAP PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHSHYTQKSLSLSPGKChain 2 - QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYVMHWVR CD19- QAPGQGLEWMGYINPYNDGTKYNEKFQGRVTITSDKSTS NEW VH (G TAYMELSSLRSEDTAVYYCARGTYYYGTRVFDYWGQGT 4S)2_[CD3]_L LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE 1.47_H1.214_ PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL VH G44C VL GTQTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQA _G100C_scFv( VVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQK GKPGS)4_(G4 PGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQP S)2_Fc(222)_I EDEADYYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSG SEQ IDgGl KPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFST NO: 228YAMNWVRQAPGKCLEWVGRIRSKYNNYATYYADSVKG RFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHTNFGDS YVSWFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAP PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHSHYTQKSLSLSPGKChain 2 - SEQ ID QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYVMHWVR CD19- NO: 229 QAPGQGLEWMGYINPYNDGTKYNEKFQGRVTITSDKSTSNEW VHJG TAYMELSSLRSEDTAVYYCARGTYYYGTRVFDYWGQGTP190500. W0.01 4S)2_[CD3]_L LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE 1.47_H1.89_sc PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL Fv(GKPGS)4_ GTQTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQA Fc(216)_IgGl VVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQK PGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQP EDEAD YYCALWYSNHWVFGGGTKLTVLGKPGSGKPGSG KPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFST YAMNWVRQAPGKGLEWVGRIRSKYNNYATYYADSVKG RFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFGDE YVSWFAYWGQGTLVTVSSEPKSSDKTHTCPPCPAPPVAGP SVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWY VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREQ MTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSH YTQKSLSLSPGKChain 2 - QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYVMHWVR CD19- QAPGQGLEWMGYINPYNDGTKYNEKFQGRVTITSDKSTS NEW VH (G TAYMELSSLRSEDTAVYYCARGTYYYGTRVFDYWGQGT 4S)2_[CD3]_L LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE 1.47_H1.32_sc PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL Fv(GKPGS)4_ GTQTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQA (G4S)2_Fc(22 VVTQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQK 2)_IgGl PGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQP EDEAD YYCALWYSNHWVFGGGTKLTVLGKPGSGKPGSG SEQ ID KPGSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFST NO: 230YAMNWVRQAPGKGLEWVGRIRSKANNYATYYADSVKG RFTISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFGDS YVSWFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAP PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHE ALHSHYTQKSLSLSPGKChain 2 - QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYVMHWVR CD19- QAPGQGLEWMGYINPYNDGTKYNEKFQGRVTITSDKSTS NEW VH (G TAYMELSSLRSEDTAVYYCARGTYYYGTRVFDYWGQGT 4S)2_[CD3]_H LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE 1.30_L1.47_sc PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL Fv(GKPGS)4_ SEQ ID GTQTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSEV (G4S)2_Fc(22 NO: 231 QLVESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAP 2)_IgGl GKGLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNT LYLQMNSLRAEDTAVYYCVRHGNFGDSYVSWFAYWGQ GTLVTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLTV SPGGTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCALP190500. W0.01WYSNHWVFGGGTKLTVLGGGGSGGGGSKTHTCPPCPAPP VAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVK FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLP PSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNY KTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEA LHSHYTQKSLSLSPGKChain 1 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG VHb lgG MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT 1 VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT SEQ ID QTYICNVNHKPSDTKVDKKVEPKSCDKTHTCPPCPAPPVA NO: 232 GPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFN WYVDGVEVHNAKTKPREEEYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR EEMTKNQVSLTCDVSGFYPSDIAVEWESDGQPENNYKTTP PVLDSDGSFFLYSKLTVDKSRWEQGDVFSCSVLHEALHSH YTQKSLSLSPGKChain 2 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG_VH_(G4S MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT )2_[CD3]_L1. VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT 47 H1.280 V VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT H_G44C_VL_ QTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQAVV G100C scFv( TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPG GKPGS)4_(G4 KSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPED S)2_Fc(222)_I EADYYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSGKP SEQ IDgGl GSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSTYA NO: 233MNWVRQAPGKCLEWVGRIRSKYNNYATYYADSVKGRFT ISRDDSKNTLYLQMNSLRAEDTAVYYCVRHENFGGEYVS WFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAPPVA GPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR EQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHS HYTQKSLSLSPGKChain 2 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG VH (G4S MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT )2_[CD3]_L1. SEQ ID VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT 47 H1.279 V NO: 234 VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT H_G44C_VL_ QTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQAVV G100C_scFv( TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPGGKPGS)4 (G4 KSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPEDP190500. W0.01 S)2_Fc(222)_I EADYYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSGKP gGl GSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSTYA MNWVRQAPGKCLEWVGRIRSKYNNYATYYADSVKGRFT ISRDDSKNTLYLQMNSLRAEDTAVYYCVRHTNFGGEYVS WFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAPPVA GPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR EQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHS HYTQKSLSLSPGKChain 2 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG_VH (G4S MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT )2_[CD3]_L1. VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT 47 H1.276 V VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT H_G44C_VL_ QTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQAVV G100C_scFv( TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPG GKPGS)4_(G4 KSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPED S)2_Fc(222)_I EADYYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSGKP SEQ IDgGl GSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSTYA NO: 235MNWVRQAPGKCLEWVGRIRSKANNYATYYADSVKGRFT ISRDDSKNTLYLQMNSLRAEDTAVYYCVRHENFGGSYVS WFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAPPVA GPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR EQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHS HYTQKSLSLSPGKChain 2 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG_VH_(G4S MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT )2_[CD3]_L1. VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT 47 HL275 V VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT H_G44C_VL_ QTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQAVV G100C_scFv( TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPG GKPGS)4_(G4 SEQ ID KSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPED S)2_Fc(222)_I NO: 236 EADYYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSGKP gGl GSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSTYA MNWVRQAPGKCLEWVGRIRSKANNYATYYADSVKGRFT ISRDDSKNTLYLQMNSLRAEDTAVYYCVRHENFGDSYVS WFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAPPVA GPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTTP190500. W0.01PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHS HYTQKSLSLSPGKChain 2 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG_VH_(G4S MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT )2_[CD3]_L1. VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT 47 H1.274 V VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT H_G44C_VL_ QTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQAVV G100C_scFv( TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPG GKPGS)4_(G4 KSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPED S)2_Fc(222)_I EADYYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSGKP SEQ IDgGl GSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSTYA NO: 237MNWVRQAPGKCLEWVGRIRSKYNNYATYYADSVKGRFT ISRDDSKNTLYLQMNSLRAEDTAVYYCVRHENFGGSYVS WFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAPPVA GPSVFLFPPKPKDTLM1SRTPEVTCVVVDVKHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR EQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHS HYTQKSLSLSPGKChain 2 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG_VH_(G4S MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT )2_[CD3]_L1. VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT 47 Hl.273 V VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT H_G44C_VL_ QTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQAVV G100C_scFv( TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPG GKPGS)4_(G4 KSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPED S)2_Fc(222)_I EADYYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSGKP SEQ IDgGl GSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSTYA NO: 238MNWVRQAPGKCLEWVGRIRSKYNNYATYYADSVKGRFT ISRDDSKNTLYLQMNSLRAEDTAVYYCVRHENFGDSYVS WFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAPPVA GPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR EQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHS HYTQKSLSLSPGKChain 2 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG_VH_(G4S SEQ ID MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT )2_[CD3]_L1. NO: 239 VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT 47 HL272 V VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTH G44C VL QTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQAVVP190500. W0.01 G100C_scFv( TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPG GKPGS)4_(G4 KSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPED S)2_Fc(222)_I EADYYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSGKP gGl GSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSTYA MNWVRQAPGKCLEWVGRIRSKANNYATYYADSVKGRFT ISRDDSKNTLYLQMNSLRAEDTAVYYCVRHTNFGGSYVS WFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAPPVA GPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR EQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHS HYTQKSLSLSPGKChain 2 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG_VH_(G4S MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT )2_[CD3]_L1. VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT 47 H1.271 V VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT H_G44C_VL_ QTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQAVV G100C_scFv( TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPG GKPGS)4_(G4 KSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPED S)2_Fc(222)_I EADYYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSGKP SEQ IDgGl GSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSTYA NO: 240MNWVRQAPGKCLEWVGRIRSKANNYATYYADSVKGRFT ISRDDSKNTLYLQMNSLRAEDTAVYYCVRHTNFGDSYVS WFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAPPVA GPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR EQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHS HYTQKSLSLSPGKChain 2 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG_VH_(G4S MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT )2_[CD3]_L1. VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT 47 H1.270 V VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT H_G44C_VL_ QTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQAVV G100C_scFv( SEQ ID TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPG GKPGS)4_(G4 NO: 241 KSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPED S)2_Fc(222)_I EADYYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSGKP gGl GSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSTYA MNWVRQAPGKCLEWVGRIRSKYNNYATYYADSVKGRFT ISRDDSKNTLYLQMNSLRAEDTAVYYCVRHTNFGGSYVS WFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAPPVA GPSVFLFPPKPKDTLM1SRTPEVTCVVVDVKHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLP190500. W0.01NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR EQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHS HYTQKSLSLSPGKChain 2 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG_VH_(G4S MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT )2_[CD3]_L1. VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT 47 H1.214 V VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT H_G44C_VL_ QTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQAVV G100C_scFv( TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPG GKPGS)4_(G4 KSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPED S)2_Fc(222)_I EADYYCALWYSNHWVFGCGTKLTVLGKPGSGKPGSGKP SEQ IDgGl GSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSTYA NO: 242MNWVRQAPGKCLEWVGRIRSKYNNYATYYADSVKGRFT ISRDDSKNTLYLQMNSLRAEDTAVYYCVRHTNFGDSYVS WFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAPPVA GPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR EQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHS HYTQKSLSLSPGKChain 2 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG VH (G4S MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT )2_[CD3]_L1. VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT 47_H1.89_scF VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT v(GKPGS)4_F QTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQAVV c(216)_IgGl TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPG KSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPED EADYYCALWYSNHWVFGGGTKLTVLGKPGSGKPGSGKP SEQ ID GSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSTYA NO: 243MNWVRQAPGKGLEWVGRIRSKYNNYATYYADSVKGRFT ISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFGDEYVS WFAYWGQGTLVTVSSEPKSSDKTHTCPPCPAPPVAGPSVF LFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFNWYVD GVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREQMT KNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYT QKSLSLSPGKChain 2 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- SEQ ID APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG_VH_(G4S NO: 244 MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT)2 [CD3] LI. VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTP190500. W0.01 47_H1.32_scF VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT v(GKPGS)4_( QTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSQAVV G4S)2_Fc(222 TQEPSLTVSPGGTVTLTCGSSTGAVTTSNYANWVQQKPG )_IgGl KSPRGLIGGTNKRAPGVPARFSGSLLGGKAALTISGAQPED EADYYCALWYSNHWVFGGGTKLTVLGKPGSGKPGSGKP GSGKPGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSTYA MNWVRQAPGKGLEWVGRIRSKANNYATYYADSVKGRFT ISRDDSKNTLYLQMNSLRAEDTAVYYCVRHGNFGDSYVS WFAYWGQGTLVTVSSGGGGSGGGGSKTHTCPPCPAPPVA GPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL NGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSR EQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHS HYTQKSLSLSPGKChain 2 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG_VH_(G4S MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT )2_[CD3]_H1. VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT 30_L1.47_scF VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT v(GKPGS)4_( QTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSEVQL G4S)2_Fc(222 VESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK )_IgGl GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLY LQMNSLRAEDTAVYYCVRHGNFGDSYVSWFAYWGQGTL SEQ ID VTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLTVSPG NO: 245GTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIGGTN KRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCALW YSNHWVFGGGTKLTVLGGGGSGGGGSKTHTCPPCPAPPV AGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKF NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS REQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALH SHYTQKSLSLSPGKChain 2 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG_VH_(G4S MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT )2_[CD3]_H1. VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT 89_L1.47_scF VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT v(GKPGS)4_F SEQ ID QTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSEVQL c(222)_IgGl NO: 246 VESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLY LQMNSLRAEDTAVYYCVRHGNFGDEYVSWFAYWGQGTL VTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLTVSPG GTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIGGTN KRAPGVPARFSGSLLGGKAAETISGAQPEDEADYYCALWYSNHWVFGGGTKLTVLGGGGSGGGGSKTHTCPPCPAPPVP190500. W0.01AGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKF NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS REQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPGKChain 2 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG_VH (G4S MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT )2_[CD3]_H1. VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT 33_L1.47_scF VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT v(GKPGS)4_F QTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSEVQL c(222)_IgGl VESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLY LQMNSLRAEDTAVYYCVRHGNFGDSYVSWFDYWGQGTL SEQ ID VTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLTVSPG NO: 247GTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIGGTN KRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCALW YSNHWVFGGGTKLTVLGGGGSGGGGSKTHTCPPCPAPPV AGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKF NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS REQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPGKChain 2 - EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD19- APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAY OG_VH_(G4S MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT )2_[CD3]_H1. VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT 32_L1.47_scF VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT v(GKPGS)4_F QTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSEVQL c(222)_IgGl VESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK GLEWVGRIRSKANNYATYYADSVKGRFTISRDDSKNTLY LQMNSLRAEDTAVYYCVRHGNFGDSYVSWFAYWGQGTL SEQ ID VTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLTVSPG NO: 248GTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIGGTN KRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCALW YSNHWVFGGGTKLTVLGGGGSGGGGSKTHTCPPCPAPPV AGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKF NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS REQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPGKChain 2 - SEQ ID EVQLVESGGGLVKPGGSLKLSCAASGYTFTSYVMHWVRQ CD 19- NO: 249 APGKGLEWIGYINPYNDGTKYNEKFQGRVTISSDKSISTAYP190500. W0.01 OG_VH_(G4S MELSSLRSEDTAMYYCARGTYYYGTRVFDYWGQGTLVT )2_[CD3]_H1. VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT 30_L1.47_scF VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT v(GKPGS)4_F QTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSEVQL c(222)_IgGl VESGGGLVQPGGSLRLSCAASGFTFSTYAMNWVRQAPGK GLEWVGRIRSKYNNYATYYADSVKGRFTISRDDSKNTLY LQMNSLRAEDTAVYYCVRHGNFGDSYVSWFAYWGQGTL VTVSSGKPGSGKPGSGKPGSGKPGSQAVVTQEPSLTVSPG GTVTLTCGSSTGAVTTSNYANWVQQKPGKSPRGLIGGTN KRAPGVPARFSGSLLGGKAALTISGAQPEDEADYYCALW YSNHWVFGGGTKLTVLGGGGSGGGGSKTHTCPPCPAPPV AGPSVFLFPPKPKDTLMISRTPEVTCVVVDVKHEDPEVKF NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS REQMTKNQVKLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPGK

Claims

P190500. W0.01 What is claimed:

1. A bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprising:an anti-CD19 antigen binding domain comprisinga first anti-CD19 binding subunit comprising CDR amino acid sequences of SEQ ID NOs: 1, 2, and 3, anda second anti-CD19 binding subunit comprising CDR amino acid sequences of SEQ ID NOs: 4, 5, and 6; andan anti-CD3 antigen binding domain comprisinga first anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NO: 9, SEQ ID NO: 10 or 11, and SEQ ID NO: 12, 13, 14, 15, or 16, and an anti-CD3 second binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19.

2. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of claim 1, wherein:a) the first anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 9, 10, and 12 and the second anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19;b) the first anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 9, 10, and 13 and the second anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NOs:

17.

18. and 19:c) the first anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 9, 11, and 12 and the second anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19;d) the first anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 9, 11, and 13 and the second anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19;P190500. W0.01 e) the first anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 9, 10, and 14 and the second anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19;f) the first anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 9, 10, and 15 and the second anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NOs:

17.

18. and 19:g) the first anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 9, 11, and 14 and the second binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19;h) the first anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 9, 11, and 15 and the second anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19; andi) the first anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 9, 10, and 16 and the second anti-CD3 binding subunit binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19.

3. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of any one of claims 1 - 2, wherein:the first anti-CD19 binding subunit and the first anti-CD3 binding subunit are heavy chain variable regions; andthe second anti-CD19 binding subunit and the second anti-CD3 binding subunit are light chain variable regions.

4. A bi-specific anti-CD19 and anti-CD3 antigen binding molecule, wherein the anti-CD19 antigen binding domain comprises:a first anti-CD19 binding subunit comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 7; anda second anti-CD19 binding subunit comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 8.P190500. W0.01 5. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of claim 4, wherein the anti-CD19 antigen binding domain comprises:a first anti-CD19 binding subunit comprising the amino acid sequence of SEQ ID NO: 7; anda second anti-CD19 binding subunit comprising the amino acid sequence of SEQ ID NO: 8.

6. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 1-5, wherein the anti-CD3 antigen binding domain comprises:a light chain variable region comprising at least 95% sequence identity to an amino acid sequence of SEQ ID NO: 180; anda heavy chain variable region comprising at least 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 179, 181, 182, 183. 184, 185. 186, 187, 188, and 189.

7. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of claim 6, wherein the anti-CD19 antigen binding domain comprises:the light chain variable region comprising an amino acid sequence of SEQ ID NO: 180; andthe heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 179, 181, 182, 183, 184, 185, 186, 187, 188, and 189.

8. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of claim 7, wherein the anti-CD3 antigen binding domain comprises a single-chain variable fragment comprising at least 95% sequence identity to an amino acid sequence of SEQ ID NO: 190, 191, 192, 193. 194, 195, 196. 197, 198. 199, 200, 201. 202, 203. 204, 205, 206. 207, 208. or 209.

9. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of claim 8, wherein the anti-CD3 antigen binding domain comprises a single-chain variable fragment comprising anP190500. W0.01 amino acid sequence of SEQ ID NO:

190. 191, 192. 193, 194.

195. 196, 197. 198, 199. 200, 201, 202, 203, 204, 205, 206, 207, 208, or 209.

10. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of any one of claims 1-9, wherein the bi-specific antibody is a 1+1 Fab-scFv-Fc, 2+1 Fab2--scFv-Fc, 1+1 Common Light Chain, 2+1 Common Light Chain. 2+1 mAb-scFv, 2+1 stackFab2-scFv-Fc. Dual scFv, One-arm scFv-mAb, scFv-mAb, Bispecific mAb, One-arm central-scFv, mAb-Fv, central-Fv, or Trident.

11. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of any one of claims 1-10, comprising:a first monomer comprising, from N- to C-terminus, a scFv-domain linker-CH2-CH3, whereinthe scFv comprises an antigen binding domain that binds CD3; and the CH2-CH3 is first variant Fc domain;a second monomer comprising a VH-CHl-hinge-CH2-CH3.wherein the CH2-CH3 is a second variant Fc domain; anda light chain that together with the second monomer forms a Fab domain that comprises an antigen binding domain that binds CD 19.

12. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of any one of claims 1-10, comprising:a first monomer comprising, from N- to C-terminus, a VH-CH1 - first domain linker -scFv- second domain linker-CH2-CH3, whereinthe scFv comprises an antigen binding domain that binds CD3; and the CH2-CH3 is first variant Fc domain;a second monomer comprising a VH-CHl-hinge-CH2-CH3, wherein the CH2-CH3 is a second variant Fc domain;a first light chain that together with the first monomer forms a Fab domain that comprises an antigen binding domain that binds CD 19; anda second light chain that together with the second monomer forms a Fab domain that comprises an antigen binding domain that binds CD 19.- I ll -P190500. W0.01 13. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of claim 11 or 12. wherein the scFv comprises an antigen binding domain comprising:a light chain variable region comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 180; anda heavy chain variable region comprising at least 95% sequence identity’ to a sequence selected from the group consisting of SEQ ID NO:

179. 181, 182. 183, 184. 185, 186, 187, 188, and 189.

14. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of claim 11 or 12, wherein the scFv comprises an amino acid sequence having at least 95% sequence identity to a sequence selected from the group consisting of SEQ ID NO: 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, and 209.

15. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of any one of claims 11-14, wherein the Fab domain comprises:the VH comprising an amino acid sequence having at least 95% sequence identity' to SEQ ID NO: 7; andthe light chain comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 8.

16. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of claim any one of claims 11-15, wherein the scFv comprises a sequence selected from the group consisting of SEQ ID NO: 190-209.

17. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of claim any one of claims 11-16, wherein the VH comprises SEQ ID NO: 7 and the Fab domain comprises a VL domain comprising SEQ ID NO: 8.

18. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of any one of claims 11-17, whereinthe first variant Fc domain is selected from the group consisting of SEQ ID NOs: 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, and 138; andP190500. W0.01 the second variant Fc domain is selected from the group consisting of SEQ ID NOs: 107.109, 111, 113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, and 139.

19. Ihe bi-specific anti-CD19 and anti-CD3 antigen binding molecule of any one of claims 11-18, wherein the first variant Fc domain and the second Fc domain comprise a pair selected from the group consisting of SEQ ID NOs: 106 and 107, SEQ ID NOs: 108 and 109, SEQ ID NOs: 110 and 111, SEQ ID NOs: 112 and 113, SEQ ID NOs: 114 and 115, SEQ ID NOs: 116 and 117, SEQ ID NOs: 118 and 119, SEQ ID NOs: 120 and 121, SEQ ID NOs: 122 and 123, SEQ ID NOs: 124 and 125, SEQ ID NOs: 126 and 127, SEQ ID NOs: 128 and 129, SEQ ID NOs: 130 and 131, SEQ ID NOs: 132 and 133, SEQ IDNOs: 134 and 135, SEQ ID NOs: 136 and 137. and SEQ ID NOs: and 138 and 139.

20. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 11-17,wherein the first variant Fc domain and second variant Fc domain are based on humanlgGl,wherein the first variant Fc domain comprises amino acid substitutions L368D / K370S and a Q295E / N384D / Q418E / N421D,the second variant Fc domain comprises amino acid substitutions S364K / E357Q; andboth the first variant Fc domain and second variant Fc domain comprise the amino acid substitutions E233P / L234V / L235A / G236del / S267K.

21. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 11-17,wherein the first variant Fc domain and second variant Fc domain are based on human IgGl.wherein the first variant Fc domain comprises amino acid substitutions L368D / K370S and Q295E / N384D / Q418E / N421D,wherein the second variant Fc domain comprises the amino acid substitution S364K; andP190500. W0.01 wherein both the first variant Fc domain and second variant Fc domain comprise the amino acid substitutions E233P / L234V / L235A / G236del / S267K.

22. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 11-17,wherein the first variant Fc domain and second variant Fc domain are based on human IgGl,wherein the first variant Fc domain comprises amino acid substitutions L368E / K370S and Q295E / N384D / Q418E / N421D,wherein the second variant Fc domain comprises the amino acid substitution S364K; andwherein both the first variant Fc domain and second variant Fc domain comprise the amino acid substitutions E233P / L234V / L235A / G236del / S267K.

23. The bi -specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 11-17,wherein the first variant Fc domain and second variant Fc domain are based on human IgGl, wherein the first variant Fc domain comprises amino acid substitutions L368E / K370S and Q295E / N384D / Q418E / N421D,wherein the second variant Fc domain comprises the amino acid substitution S364K; andwherein both the first variant Fc domain and second variant Fc domain comprise the amino acid substitutions E233P / L234V / L235A / G236del / S267K.24 T he bi-specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 11-17,wherein the first variant Fc domain and second variant Fc domain are based on human IgGl,wherein the first variant Fc domain comprises amino acid substitutions K360E / Q362E / T411E and Q295E / N384D / Q418E / N421D,wherein the second variant Fc domain comprises the amino acid substitution D401K; andP190500. W0.01 wherein both the first variant Fc domain and second variant Fc domain comprise the amino acid substitutions E233P / L234V / L235A / G236del / S267K.

25. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 11-17,wherein the first variant Fc domain and second variant Fc domain are based on human IgGl,wherein the first variant Fc domain comprises amino acid substitutions L368D / K370S and Q295E / N384D / Q418E / N421D,wherein the second variant Fc domain comprises amino acid substitutions S364K / E357Q; andwherein both the first variant Fc domain and second variant Fc domain comprise the amino acid substitutions E233P / L234V / L235A / G236del / S267K.

26. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 11-17,wherein the first variant Fc domain and second variant Fc domain are based on human IgGl,wherein the first variant Fc domain comprises amino acid substitutions L368D / K370S and Q295E / N384D / Q418E / N421D,wherein the second variant Fc domain comprises amino acid substitutions S364K / E357Q; andwherein both the first variant Fc domain and second variant Fc domain comprise the amino acid substitutions E233P / L234V / L235A / G236del / S267K.

27. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 11-17,wherein the first variant Fc domain and second variant Fc domain are based on human IgG4,wherein the first variant Fc domain comprises amino acid substitutions L368D / K370S and Q295E / N384D / Q418E / N421D,wherein the second variant Fc domain comprises amino acid substitutions S364K / E357Q; andP190500. W0.01 wherein both the first variant Fc domain and second variant Fc domain comprise the amino acid substitution S228P (according to EU numbering, S241P in Kabat).

28. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 11-17,wherein the first variant Fc domain and second variant Fc domain are based on human IgG2,wherein the first variant Fc domain comprises amino acid substitutions L368D / K370S and Q295E / N384D / Q418E / N421D, andwherein the second variant Fc domain comprises amino acid substitutions S364K / E357Q.

29. The bi -specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 11-17,wherein the first variant Fc domain and second variant Fc domain are based on based on human IgG2,wherein the first variant Fc domain comprises amino acid substitutions L368D / K370S and Q295E / N384D / Q418E / N421D,wherein the second variant Fc domain comprises amino acid substitutions S364K / E357Q; andwherein both the first variant Fc domain and second variant Fc domain comprise the amino acid substitution S267K.

30. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 11-17,wherein the first variant Fc domain and second variant Fc domain are based on human IgGl.wherein the first variant Fc domain comprises amino acid substitutions L368D / K370S and Q295E / N384D / Q418E / N421D,wherein the second variant Fc domain comprises amino acid substitutions S364K / E357Q, andwherein both the first variant Fc domain and second variant Fc domain comprise the amino acid substitutions E233P / L234V / L235A / G236del / S267K and M428L / N434S.P190500. W0.0131. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 11-17,wherein the first variant Fc domain and second variant Fc domain are based on human IgGl,wherein the first variant Fc domain comprises amino acid substitutions L368D / K370S, wherein the second variant Fc domain comprises amino acid substitutions S364K / E357Q and P217R / P229R / N276K;andwherein both the first variant Fc domain and second variant Fc domain comprise the amino acid substitutions E233P / L234V / L235A / G236del / S267K.

32. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 11-17,wherein the first variant Fc domain and second variant Fc domain are based on human IgGl.wherein the first variant Fc domain comprises amino acid substitution T366W, wherein the second variant Fc domain comprises amino acid substitutions T366S / L368A / Y407V and H435R / Y436F;andwherein both the first variant Fc domain and second variant Fc domain comprise the amino acid substitutions L234A / L235A / D265S.

33. T he bi-specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 11-17,wherein the first variant Fc domain and second variant Fc domain are based on human IgGl.wherein the first variant Fc domain comprises amino acid substitution 356E / 358M, wherein the second variant Fc domain comprises amino acid substitutions T366S / L368A / Y407V and H435R / Y436F;andP190500. W0.01 wherein both the first variant Fc domain and second variant Fc domain comprise the amino acid substitutions L234A / L235A / D265S and M252Y / S254T / T256E.

34. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 11-17,wherein the first variant Fc domain and second variant Fc domain are based on human IgGl,wherein the first variant Fc domain comprises amino acid substitutions L368D / K370S and Q295E / N384D / Q418E / N421D,wherein the second variant Fc domain comprises amino acid substitutions S364K / E357Q; andwherein both the first variant Fc domain and second variant Fc domain comprise the amino acid substitutions E233P / L234V / L235A / G236del / S267K and M428L / N434S.

35. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 11-17,wherein the first variant Fc domain and second variant Fc domain are based on human IgGl,wherein the first variant Fc domain comprises amino acid substitutions L368D / K370S and Q295E / N384D / Q418E / N421D,wherein the second variant Fc domain comprises amino acid substitutions S364K / E357Q; andwherein both the first variant Fc domain and second variant Fc domain comprise the amino acid substitutions E233P / L234V / L235A / G236del / S267K and M428L / N434A.

36. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of one of claims 11-17,wherein the first variant Fc domain and second variant Fc domain are based on human IgGl,wherein the first variant Fc domain comprises amino acid substitutions L368D / K370S and Q295E / N384D / Q418E / N421D,P190500. W0.01 wherein the second variant Fc domain comprises amino acid substitutions S364K / E357Q; andwherein both the first variant Fc domain and second variant Fc domain comprise the amino acid substitutions E233P / L234V / L235A / G236del / S267K and M428L / N434A.

37. A bi-specific anti-CD19 and anti-CD3 antigen binding molecule, wherein the binding molecule is a 1+1 Fab-scFv-Fc and comprises an amino acid sequence selected from the group consisting of:SEQ ID NOs: 210, 211, and 212;SEQ ID NOs: 213, 214, and 215;SEQ ID NOs: 213, 216, and 215;SEQ ID NOs: 213, 217, and 215; andSEQ ID NOs: 213, 218, and 215.

38. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of claim 12, wherein the binding molecule is a 2 + 1 Fab2-scFv-Fc and comprises an amino acid sequence selected from the group consisting of:SEQ ID NOs: 210, 219, and 212;SEQ ID NOs: 210, 220, and 212;SEQ ID NOs: 210, 221, and 212;SEQ ID NOs: 210, 222, and 212;SEQ ID NOs: 210, 223, and 212;SEQ ID NOs: 210, 224, and 212;SEQ ID NOs: 210, 225, and 212;SEQ ID NOs: 210, 226, and 212;SEQ ID NOs: 210, 227, and 212;SEQ ID NOs: 210, 228, and 212;SEQ ID NOs: 210, 229, and 212;P190500. W0.01 SEQ ID NOs: 210, 230, and 212;SEQ ID NOs: 210, 231, and 212;SEQ ID NOs: 232, 233, and 215;SEQ ID NOs: 232, 234, and 215;SEQ ID NOs: 232, 235, and 215;SEQ ID NOs: 232, 236, and 215;SEQ ID NOs: 232, 237, and 215;SEQ ID NOs: 232, 238, and 215;SEQ ID NOs: 232, 239, and 215;SEQ ID NOs: 232, 240, and 215;SEQ ID NOs: 232, 241, and 215;SEQ ID NOs: 232, 242, and 215;SEQ ID NOs: 232, 243, and 215;SEQ ID NOs: 232, 244, and 215;SEQ ID NOs: 232, 245, and 215;SEQ ID NOs: 213, 246, and 215;SEQ ID NOs: 213, 247, and 215;SEQ ID NOs: 213, 248, and 215; andSEQ ID NOs: 213, 249, and 215.

39. A bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprising:a means for binding CD 19; anda first anti-CD3 binding subunit comprising the CDR amino acid sequences of SEQ ID NO: 9, SEQ ID NO: 10 or 11, and SEQ ID NO: 12, 13, 14, 15, or 16, and an anti-CD3 second binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19.P190500. W0.01 40. A bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprising:a means for binding CD3; andan anti-CD19 antigen binding domain comprising:a first binding subunit comprising CDR amino acid sequences of SEQ ID NOs: 1, 2, and 3; anda second binding subunit comprising CDR amino acid sequences of SEQ ID NOs: 4, 5, and 6.

41. A method of depleting B-cells in a subject, the method comprising administering the bi-specific anti-CD19 and anti-CD3 antigen binding molecule of any one of claims 1-40 to the subject.

42. The method of claim 41, wherein the B-cells are peripheral B-cells, bone marrow B-cells, or lymph node B-cells.

43. The method of claim 41 or 42, wherein the administering is intravenous or subcutaneous.

44. A method of treating an autoimmune disorder in a subject, the method comprising administering the bi-specific anti-CD19 and anti-CD3 antigen binding molecule of any one of claims 1-40 to the subject.

45. The method of claim 41 or 42. wherein the administering is intravenous or subcutaneous.

46. A method of treating an inflammatory disease in a subject, the method comprising administering the bi-specific anti-CD19 and anti-CD3 antigen binding molecule of any one of claims 1-40 to the subject.

47. A method of treating cancer in a subject, the method comprising administering a therapeutically effective amount of the bi-specific anti-CD19 and anti-CD3 antigen binding molecule of any one of claims 1-40 to the subject.

48. The method of claim 46 or claim 47, wherein the administering is intravenous or subcutaneous.

49. An anti-CD3 antigen binding molecule, comprising an antigen binding domain comprising:P190500. W0.01 a first binding subunit comprising complementarity determining region (CDR) amino acid sequences of SEQ ID NO: 9, SEQ ID NO: 10 or 11, and SEQ ID NO: 12, 13, 14, 15, or 16; anda second binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19.

50. The anti-CD3 antigen binding molecule of claim 49, wherein the first binding subunit and the second binding subunits are selected from the group consisting of:a) the first binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 9, 10, and 12 and the second binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19;b) the first binding subunit comprising the CDR amino acid sequences of SEQ ID NOs:

9.

10. and 13 and the second binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19;c) the first binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 9, 11, and 12 and the second binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19;d) the first binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 9, 11, and 13 and the second binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19;e) the first binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 9, 10, and 14 and the second binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19;1) the first binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 9, 10, and 15 and the second binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19;g) the first binding subunit comprising the CDR amino acid sequences of SEQ ID NOs:

9. 11, and 14 and the second binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19;P190500. W0.01 h) the first binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 9, 11, and 15 and the second binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19; andi) the first binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 9, 10, and 16 and the second binding subunit comprising the CDR amino acid sequences of SEQ ID NOs: 17, 18, and 19.

51. The anti-CD3 antigen binding molecule of claim 49 or claim 50, further comprising a linker comprising at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 36 between the first binding subunit and the second binding subunit.

52. The anti-CD3 antigen binding molecule of claim 51, further comprising a linker comprising the amino acid sequence of SEQ ID NO: 36 between the first binding subunit and the second binding subunit.

53. The anti-CD3 antigen binding molecule of any one of claims 49 - 52, wherein the first binding subunit is a heavy chain variable region, and the second binding subunit is a light chain variable region.

54. The anti-CD3 antigen binding molecule of claim 53, whereinthe second binding subunit comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 180; andthe first binding subunit comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 179, 181, 182, 183, 184. 185, 186, 187, 188, or 189.

55. The anti-CD3 antigen binding molecule of claim 54, whereinthe light chain variable region comprises the amino acid sequence of SEQ ID NO: 180; andP190500. W0.01 the heavy chain variable region comprises the amino acid sequence selected from the group consisting of SEQ ID NO: 179, 181, 182, 183, 184, 185, 186, 187, 188, and 189.

56. The anti-CD3 antigen binding molecule of claim 55, wherein the first light chain variable region and the second light chain variable region together comprise a single-chain variable fragment (scFv), the scFv comprising at least 95% sequence identity to the amino acid sequence selected from the group consisting of SEQ ID NO: 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, and 209.

57. The anti-CD3 antigen binding molecule of claim 56, wherein the scFv comprises the amino acid sequence selected from the group consisting of SEQ ID NO: 190, 191, 192. 193, 194, 195, 196. 197, 198. 199, 200, 201. 202, 203. 204, 205, 206, 207, 208. and 209.

58. An anti-CD3 antigen binding molecule, comprising:a first binding subunit and a second binding subunit;wherein the first binding subunit comprises a framework region having at least 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 28, 29, 30, and 31, and / orwherein the second binding subunit comprises a framework region having at least 95% identity to an amino acid sequence selected from the group consisting of 32, 33, 34, and 35.

59. The anti-CD3 antigen binding molecule of claim 58, comprising:a first binding subunit and a second binding subunit; andwherein the first binding subunit comprises a framework region selected from the group consisting of SEQ ID NO: 28, 29, 30, and 31, and / orwherein the second binding subunit comprises a framework region selected from the group consisting of 32.

33.

34. and 35.

60. The anti-CD3 antigen binding molecule of claim 59,P190500. W0.01 wherein the first binding subunit compnses:a framework region 1 (FR1) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 28;a framework region 2 (FR2) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 29;a framework region 3 (FR3) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 30; anda framework region 4 (FR4) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 31; andwherein the second binding subunit comprises:a framework region 5 (FR5) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 32;a framework region 6 (FR6) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 33;a framework region 7 (FR7) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 34; anda framework region 8 (FR8) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 35.

61. The anti-CD3 antigen binding molecule of claim 60,wherein the first binding subunit comprises:a framework region 1 (FR1) comprising the amino acid sequence of SEQ ID NO: 28;a framework region 2 (FR2) comprising the amino acid sequence of SEQ ID NO: 29;a framework region 3 (FR3) comprising the amino acid sequence of SEQ ID NO: 30; anda framework region 4 (FR4) comprising the amino acid sequence of SEQ ID NO: 31; andP190500. W0.01 wherein the second binding subunit comprises: a framework region 5 (FR5) comprising the amino acid sequence of SEQ ID NO: 32;a framework region 6 (FR6) comprising the amino acid sequence of SEQ ID NO: 33;a framework region 7 (FR7) comprising the amino acid sequence of SEQ ID NO: 34; anda framework region 8 (FR8) comprising the amino acid sequence of SEQ ID NO: 35.

62. An anti-CD3 antigen binding molecule comprising:a first binding subunit comprising:an amino acid sequence comprising at least 95% identity to SEQ ID NO: 28 N- terminal to a first binding subunit CDR1;an amino acid sequence comprising at least 95% identity to SEQ ID NO: 29 between the first binding subunit CDR1 and a first binding subunit CDR2; an amino acid sequence comprising at least 95% identity to SEQ ID NO: 30 between the first binding subunit CDR2 and a first binding subunit CDR3; and an amino acid sequence comprising at least 95% identity to SEQ ID NO: 31 C- terminal to the first binding subunit CDR3; anda second binding subunit comprising:an amino acid sequence comprising at least 95% identity to SEQ ID NO: 32 N- terminal to a second binding subunit CDR1;an amino acid sequence comprising at least 95% identity’ to SEQ ID NO: 33 between the second binding subunit CDR1 and a second binding subunit CDR2; an amino acid sequence comprising at least 95% identity to SEQ ID NO: 34 between the second binding subunit CDR2 and a second binding subunit CDR3; andan amino acid sequence comprising at least 95% identity to SEQ ID NO: 35 C- terminal to the second binding subunit CDR3.P190500. W0.0163. The anti-CD3 antigen binding molecule of claim 62, comprising:a first binding subunit comprising:an amino acid sequence comprising SEQ ID NO: 28 N-terminal to a first binding subunit CDR1;an amino acid sequence comprising SEQ ID NO: 29 between the first binding subunit CDR1 and a first binding subunit CDR2;an amino acid sequence comprising SEQ ID NO: 30 between the first binding subunit CDR2 and a first binding subunit CDR3; andan amino acid sequence comprising SEQ ID NO: 31 C-terminal to the first binding subunit CDR3;a second binding subunit comprising:an amino acid sequence comprising SEQ ID NO: 32 N-terminal to a second binding subunit CDR1;an amino acid sequence comprising SEQ ID NO: 33 between the second binding subunit CDR1 and a second binding subunit CDR2;an amino acid sequence comprising SEQ ID NO: 34 between the second binding subunit CDR2 and a second binding subunit CDR3; andan amino acid sequence comprising SEQ ID NO: 35 C-terminal to the second binding subunit CDR3.

64. The anti-CD3 antigen binding molecule of one of claims 62 or 63, wherein:the first binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 9; the first binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 10 or;the first binding subunit CDR3 comprises the amino acid sequence of SEQ ID NO: 12,, 14, 15, or 16;the second binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 17;P190500. W0.01 the second binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 18; andthe second binding subunit CDR3 comprises the amino acid sequence of SEQ ID NO: 19.

65. The anti-CD3 antigen binding molecule any one of claims 62-64, wherein a) the first binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 9; the first binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 10; the first binding subunit CDR3 comprise the amino acid sequence of SEQ ID NO: 12; andthe second binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 17; the second binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 18; andthe second binding subunit CDR3 comprises the amino acid sequence of SEQ ID NO: 19; b) the first binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 9; the first binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 10; the first binding subunit CDR3 comprise the amino acid sequence of SEQ ID NO: 13; andthe second binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 17; the second binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 18; andthe second binding subunit CDR3 comprises the amino acid sequence of SEQ ID NO: 19; c) the first binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 9; the first binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 11; the first binding subunit CDR3 comprise the amino acid sequence of SEQ ID NO: 13; andthe second binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 17;P190500. W0.01 the second binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 18; andthe second binding subunit CDR3 comprises the amino acid sequence of SEQ ID NO: 19; d) the first binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 9; the first binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 10; the first binding subunit CDR3 comprise the amino acid sequence of SEQ ID NO: 14; andthe second binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 17; the second binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 18; andthe second binding subunit CDR3 comprises the amino acid sequence of SEQ ID NO: 19; e) the first binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 9; the first binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 10; the first binding subunit CDR3 comprise the amino acid sequence of SEQ ID NO: 15; andthe second binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 17; the second binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 18; andthe second binding subunit CDR3 comprises the amino acid sequence of SEQ ID NO: 19; 1) the first binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 9; the first binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 11; the first binding subunit CDR3 comprise the amino acid sequence of SEQ ID NO: 14; andthe second binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 17; the second binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 18; andthe second binding subunit CDR3 comprises the amino acid sequence of SEQ ID NO: 19;P190500. W0.01g) the first binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 9; the first binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 1 1; the first binding subunit CDR3 comprise the amino acid sequence of SEQ ID NO: 15; andthe second binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 17; the second binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 18; andthe second binding subunit CDR3 comprises the amino acid sequence of SEQ ID NO: 19; h) the first binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 9; the first binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 10; the first binding subunit CDR3 comprise the amino acid sequence of SEQ ID NO: 16; andthe second binding subunit CDR1 comprises the amino acid sequence of SEQ ID NO: 17; the second binding subunit CDR2 comprises the amino acid sequence of SEQ ID NO: 18; andthe second binding subunit CDR3 comprises the amino acid sequence of SEQ ID NO: 19.

66. An anti-CD19 antigen binding molecule, comprising an antigen binding domain comprising:a first binding subunit comprising CDR amino acid sequences of SEQ ID NOs: 1, 2, and 3; anda second binding subunit comprising CDR amino acid sequences of SEQ ID NOs: 4, 5, and 6.

67. The anti-CD19 antigen binding molecule of claim 66. comprising a linker between the first binding subunit and the second binding subunit.

68. The anti-CD19 antigen binding molecule of claim 66 or 67, wherein the first binding subunit is a heavy chain variable region, and the second binding subunit is a light chain variable region.P190500. W0.01 69. The anti-CD19 antigen binding molecule of claim 68. whereinthe heavy chain variable region comprises a heavy chain HCDR1 comprising the amino acid sequence of SEQ ID NO: 1, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 3; andthe light chain variable region comprises a light chain LCDR1 comprising the amino acid sequence of SEQ ID NO: 4, a LCDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 6.

70. The anti-CD19 antigen binding molecule of claim 69, whereinthe heavy chain variable region comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 7; andthe light chain variable region comprises at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 8.

71. The anti-CD19 antigen binding molecule of claim 70. whereinthe heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 7; andthe light chain variable region comprises the amino acid sequence of SEQ ID NO: 8.

72. An anti-CD19 antigen binding molecule, comprising:a first binding subunit and a second binding subunit; andwherein the first binding subunit comprises a framework region having at least 95% identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 20, 21, 22, and 23, and / orwherein the second binding subunit comprises a framew ork region having at least 95% identity to an amino acid sequence selected from the group consisting of 24, 25, 26, and 27.

73. The anti-CD19 antigen binding molecule of claim 72, comprising:a first binding subunit and a second binding subunit; andP190500. W0.01 wherein the first binding subunit compnses a framework region selected from the group consisting of SEQ ID NO: 20, 21, 22, and 23, and / orwherein the second binding subunit comprises a framework region selected from the group consisting of 24, 25, 26, and 27.

74. The anti-CD19 antigen binding molecule of claim 73, comprising:wherein the first binding subunit comprises:a framework region 1 (FR1) comprising at least 95% identity to an amino acid sequence of SEQ ID NO: 20;a framework region 2 (FR2) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 21;a framework region 3 (FR3) comprising at least 95% identity7to the amino acid sequence of SEQ ID NO: 22; anda framework region 4 (FR4) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 23; andwherein the second binding subunit comprises: a framework region 5 (FR5) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 24;a framework region 6 (FR6) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 25;a framework region 7 (FR7) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 26; anda framework region 8 (FR8) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 27.

75. The anti-CD19 antigen binding molecule of claim 74, comprising:wherein the first binding subunit comprises:a framework region 1 (FR1) comprising an amino acid sequence of SEQ ID NO: 20;P190500. W0.01 a framework region 2 (FR2) comprising the amino acid sequence of SEQ ID NO: 21;a framework region 3 (FR3) comprising the amino acid sequence of SEQ ID NO: 22; anda framework region 4 (FR4) comprising the amino acid sequence of SEQ ID NO: 23; andwherein the second binding subunit comprises:a framework region 5 (FR5) comprising the ammo acid sequence of SEQ ID NO: 24;a framework region 6 (FR6) comprising the amino acid sequence of SEQ ID NO: 25;a framework region 7 (FR7) comprising the amino acid sequence of SEQ ID NO: 26; anda framework region 8 (FR8) comprising the amino acid sequence of SEQ ID NO: 27.

76. An anti-CD19 antigen binding molecule, comprising:a first binding subunit comprising:an amino acid sequence comprising at least 95% identity to SEQ ID NO: 20 N- terminal to a first binding subunit CDREan amino acid sequence comprising at least 95% identity to SEQ ID NO: 21 between the first binding subunit CDR1 and a first binding subunit CDR2; an amino acid sequence comprising at least 95% identity to SEQ ID NO: 22 between the first binding subunit CDR2 and a first binding subunit CDR3; and an amino acid sequence comprising at least 95% identity' to SEQ ID NO: 23 C- terminal to the first binding subunit CDR3; anda second binding subunit comprising:an amino acid sequence comprising at least 95% identity to SEQ ID NO: 24 N- terminal to a second binding subunit CDR1;P190500. W0.01 an amino acid sequence comprising at least 95% identity to SEQ ID NO: 25 between the second binding subunit CDR1 and a second binding subunit CDR2; an amino acid sequence comprising at least 95% identity to SEQ ID NO: 26 between the second binding subunit CDR2 and a second binding subunit CDR3; andan amino acid sequence comprising at least 95% identity to SEQ ID NO: 27 C- terminal to the second binding subunit CDR3.

77. An anti-CD19 antigen binding molecule comprising:a first binding subunit comprising:an amino acid sequence comprising SEQ ID NO: 20 N-terminal to a first binding subunit CDR1;an amino acid sequence comprising SEQ ID NO: 21 between the first binding subunit CDR1 and a first binding subunit CDR2;an amino acid sequence comprising SEQ ID NO: 22 between the first binding subunit CDR2 and a first binding subunit CDR3; andan amino acid sequence comprising SEQ ID NO: 23 C-terminal to the first binding subunit CDR3; anda second binding subunit comprising:an amino acid sequence comprising SEQ ID NO: 24 N-terminal to a second binding subunit CDR1;an amino acid sequence comprising SEQ ID NO: 25 between the second binding subunit CDR1 and a second binding subunit CDR2;an amino acid sequence comprising SEQ ID NO: 26 between the second binding subunit CDR2 and a second binding subunit CDR3; andan amino acid sequence comprising SEQ ID NO: 27 C-terminal to the second binding subunit CDR3.

78. The anti-CD19 antigen binding molecule of claim 76 or 77, wherein:P190500. W0.01 the first binding subunit comprises the CDR amino acid sequences of SEQ ID NOs: 1, 2. and 3; andthe second binding subunit comprises the CDR amino acid sequences of SEQ ID NOs: 4, 5, and 6.

79. A bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprising:a first anti-CDl 9 binding subunit comprising:a framework region 1 (FR1) comprising at least 95% identity to an amino acid sequence of SEQ ID NO: 20;a framework region 2 (FR2) comprising at least 95% identity' to the amino acid sequence of SEQ ID NO: 21;a framework region 3 (FR3) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 22; anda framework region 4 (FR4) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 23; anda second anti-CDl 9 binding subunit comprising:a framework region 5 (FR5) comprising at least 95% identity' to the amino acid sequence of SEQ ID NO: 24;a framework region 6 (FR6) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 25;a framework region 7 (FR7) comprising at least 95% identity to the amino acid sequence of SEQ ID NO: 26; anda framework region 8 (FR8) comprising at least 95% identity' to the amino acid sequence of SEQ ID NO: 27.

80. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of claim 79. wherein:the first anti-CDl 9 binding subunit comprises:P190500. W0.01 an amino acid sequence comprising at least 95% identity to SEQ ID NO: 20 N- terminal to a first binding subunit CDR1;an amino acid sequence comprising at least 95% identity to SEQ ID NO: 21 between the first binding subunit CDR1 and a first binding subunit CDR2; an amino acid sequence comprising at least 95% identity to SEQ ID NO: 22 between the first binding subunit CDR2 and a first binding subunit CDR3; and an amino acid sequence comprising at least 95% identity to SEQ ID NO: 23 C- terminal to the first binding subunit CDR3; andthe second anti-CD19 binding subunit comprises:an amino acid sequence comprising at least 95% identity to SEQ ID NO: 24 N- terminal to a second binding subunit CDR1;an amino acid sequence comprising at least 95% identity to SEQ ID NO: 25 between the second binding subunit CDR1 and a second binding subunit CDR2; an amino acid sequence comprising at least 95% identity to SEQ ID NO: 26 between the second binding subunit CDR2 and a second binding subunit CDR3; andan amino acid sequence comprising at least 95% identity' to SEQ ID NO: 27 C- terminal to the second binding subunit CDR3.

81. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of claim 79. wherein:the first anti-CD19 binding subunit comprises:a framework region 1 (FR1) comprising an amino acid sequence of SEQ ID NO: 20;a framework region 2 (FR2) comprising the amino acid sequence of SEQ ID NO: 21;a framework region 3 (FR3) comprising the amino acid sequence of SEQ ID NO: 22; andP190500. W0.01 a framework region 4 (FR4) comprising the amino acid sequence of SEQ ID NO: 23; andthe second anti-CD19 binding subunit comprises:a framework region 5 (FR5) comprising the amino acid sequence of SEQ ID NO: 24;a framework region 6 (FR6) comprising the amino acid sequence of SEQ ID NO: 25;a framework region 7 (FR7) comprising the ammo acid sequence of SEQ ID NO: 26; anda framework region 8 (FR8) comprising the amino acid sequence of SEQ ID NO: 27.

82. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of claim 80. wherein the anti-CD19 antigen binding domain comprises:an amino acid sequence comprising SEQ ID NO: 20 N-terminal to a first anti-CD19 binding subunit CDR1;an amino acid sequence comprising SEQ ID NO: 21 between the first anti-CD19 binding subunit CDR1 and a first binding subunit CDR2;an amino acid sequence comprising SEQ ID NO: 22 between the first anti-CD19 binding subunit CDR2 and a first binding subunit CDR3;an amino acid sequence comprising SEQ ID NO: 23 C-terminal to the first anti-CD19 binding subunit CDR3;an amino acid sequence comprising SEQ ID NO: 24 N-terminal to a second anti-CD19 binding subunit CDR1;an amino acid sequence comprising SEQ ID NO: 25 between the second anti-CD19 binding subunit CDR1 and a second binding subunit CDR2:an amino acid sequence comprising SEQ ID NO: 26 between the second anti-CD19 binding subunit CDR2 and a second binding subunit CDR3; andP190500. W0.01 an amino acid sequence comprising SEQ ID NO: 27 C-terminal to the second anti-CD19 binding subunit CDR3.

83. A bi-specific anti-CD19 and anti-CD3 antigen binding molecule comprising:a first anti-CD3 binding subunit comprising:a framework region 1 (FR1) comprising at least 95% identity' to an amino acid sequence of SEQ ID NO: 28;a framework region 2 (FR2) comprising at least 95% identity to an amino acid sequence of SEQ ID NO: 29;a framework region 3 (FR3) comprising at least 95% identity to an amino acid sequence of SEQ ID NO: 30; anda frameyvork region 4 (FR4) comprising at least 95% identity' to an amino acid sequence of SEQ ID NO: 31; anda second anti-CD3 binding subunit comprising:a framework region 5 (FR5) comprising at least 95% identity to an amino acid sequence of SEQ ID NO: 32;a framework region 6 (FR6) comprising at least 95% identity to an amino acid sequence of SEQ ID NO: 33;a frameyvork region 7 (FR7) comprising at least 95% identity' to an amino acid sequence of SEQ ID NO: 34; anda framework region 8 (FR8) comprising at least 95% identity to an amino acid sequence of SEQ ID NO: 35.

84. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of claim 83 comprising:a first anti-CD3 binding subunit comprising:an amino acid sequence comprising at least 95% identity to SEQ ID NO: 28 N- terminal to a first binding subunit CDR1;P190500. W0.01 an amino acid sequence comprising at least 95% identity to SEQ ID NO: 29 between the first binding subunit CDR1 and a first antigen binding domain CDR2;an amino acid sequence comprising at least 95% identity to SEQ ID NO: 30 between the first binding subunit CDR2 and a first binding subunit CDR3; and an amino acid sequence comprising at least 95% identity to SEQ ID NO: 31 C- terminal to the first binding subunit CDR3; anda second anti-CD3 binding subunit comprising:an amino acid sequence comprising at least 95% identity to SEQ ID NO: 32 N- terminal to a second binding subunit CDR1;an amino acid sequence comprising at least 95% identity to SEQ ID NO: 33 between the second binding subunit CDR1 and a second binding subunit CDR2; an amino acid sequence comprising at least 95% identity to SEQ ID NO: 34 between the second binding subunit CDR2 and a second binding subunit CDR3; andan amino acid sequence comprising at least 95% identity to SEQ ID NO: 35 C- terminal to the second binding subunit CDR3.

85. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of claim 84. wherein the anti-CD3 antigen binding domain comprises:the first anti-CD3 binding subunit comprises:a framework region 1 (FR1) comprising an amino acid sequence of SEQ ID NO: 28;a framework region 2 (FR2) comprising an amino acid sequence of SEQ ID NO: 29;a framework region 3 (FR3) comprising an amino acid sequence of SEQ ID NO: 30; anda framework region 4 (FR4) comprising an amino acid sequence of SEQ ID NO:P190500. W0.01 the second anti-CD3 binding subunit comprises:a framework region 5 (FR5) comprising an amino acid sequence of SEQ ID NO: 32;a framework region 6 (FR6) comprising an amino acid sequence of SEQ ID NO: 33;a framework region 7 (FR7) comprising an amino acid sequence of SEQ ID NO: 34; anda framework region 8 (FR8) comprising an amino acid sequence of SEQ ID NO: 35.

86. The bi-specific anti-CD19 and anti-CD3 antigen binding molecule of claim 85 comprising:a first anti-CD3 binding subunit comprising:an amino acid sequence comprising SEQ ID NO: 28 N-terminal to a first binding subunit CDR1;an amino acid sequence comprising SEQ ID NO: 29 between the first binding subunit CDR1 and a first antigen binding domain CDR2;an amino acid sequence comprising SEQ ID NO: 30 between the first binding subunit CDR2 and a first binding subunit CDR3: andan amino acid sequence comprising SEQ ID NO: 31 C-terminal to the first binding subunit CDR3; anda second anti-CD3 binding subunit comprising:an amino acid sequence comprising SEQ ID NO: 32 N-terminal to a second binding subunit CDR1;an amino acid sequence comprising SEQ ID NO: 33 between the second binding subunit CDR1 and a second domain CDR2;an amino acid sequence comprising SEQ ID NO: 34 between the second binding subunit CDR2 and a second binding subunit CDR3; andan amino acid sequence comprising SEQ ID NO: 35 C-terminal to the second binding subunit CDR3.P190500. W0.01 87. A nucleic acid encoding a first anti-CD19 binding subunit of one of claims 66 - 71.

88. The nucleic acid according to claim 87, encoding a second anti-CD19 subunit of one of claims 66 - 71.

89. A nucleic acid encoding a second anti-CD19 binding subunit of one of claims 66 -71.

90. A nucleic acid encoding a first anti-CD3 binding subunit of one of claims 49 - 57.

91. The nucleic acid according to claim 89, encoding a second anti-CD3 subunit of one of claims 49 - 57.

92. A nucleic acid encoding a second anti-CD3 binding subunit of one of claims 49 - 57.

93. A nucleic acid encoding an amino acid sequence selected from the group consisting of SEQ ID NO: 210 - 218.

94. A nucleic acid encoding an amino acid sequence selected from the group consisting ofSEQ ID NOs: 210, 211, and 212;SEQ ID NOs: 213, 214, and 215;SEQ ID NOs: 213, 216, and 215;SEQ ID NOs: 213, 217, and 215; andSEQ ID NOs: 213, 218, and 215.

95. A nucleic acid encoding an amino acid sequence selected from the group consisting of SEQ ID NO: 219 - 249.

96. A nucleic acid encoding an amino acid sequence selected from the group consisting ofSEQ ID NOs: 210, 219, and 212;SEQ ID NOs: 210, 220, and 212;SEQ ID NOs: 210, 221, and 212;P190500. W0.01 SEQ ID NOs: 210, 222. and 212;SEQ ID NOs: 210, 223, and 212;SEQ ID NOs: 210, 224, and 212;SEQ ID NOs. 210, 225, and 212;SEQ ID NOs: 210, 226, and 212;SEQ ID NOs: 210, 227, and 212;SEQ ID NOs: 210, 228, and 212;SEQ ID NOs: 210, 229. and 212;SEQ ID NOs: 210, 230, and 212;SEQ ID NOs: 210, 231, and 212;SEQ ID NOs: 232, 233, and 215;SEQ ID NOs: 232, 234. and 215;SEQ ID NOs: 232, 235, and 215;SEQ ID NOs: 232, 236, and 215;SEQ ID NOs: 232, 237, and 215;SEQ ID NOs: 232, 238, and 215;SEQ ID NOs: 232, 239, and 215;SEQ ID NOs. 232, 240, and 215;SEQ ID NOs: 232, 241, and 215;SEQ ID NOs: 232, 242, and 215;SEQ ID NOs: 232, 243, and 215;SEQ ID NOs: 232, 244, and 215;SEQ ID NOs: 232, 245, and 215;SEQ ID NOs: 213, 246. and 215;SEQ ID NOs: 213, 247, and 215;P190500. W0.01 SEQ ID NOs: 213, 248. and 215; andSEQ ID NOs: 213, 249, and 215.

97. An expression vector encoding any one of claims 87-96.

98. A host cell comprising the expression vector of claim 97.

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