Antigen binding constructs directed to integrin αvβ6

Abstract

Provided herein are antigen binding constructs, minibodies, and scFv-Fc fusions. Specifically, provided herein are αVβ6 specific antigen binding constructs, minibodies, and scFv-Fc fusions.

Description

IGNAB.064WO PATENT ANTIGEN BINDING CONSTRUCTS DIRECTED TO INTEGRIN ĮVȕ6 REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Patent Application No. 63 / 583697, filed September 19, 2023, the disclosure of which is hereby incorporated by reference in their entirety. REFERENCE TO SEQUENCE LISTING

[0002] The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled IGNAB_064WO_SEQLIST.xml, which was created and last modified on September 4, 2024, which is 140,930 bytes in size. The information in the electronic Sequence Listing is hereby incorporated by reference in its entirety. BACKGROUND

[0003] The present disclosure generally relates to antigen binding constructs, minibodies, and scFv-Fc fusions. Specifically, integrin ĮVȕ6 specific antigen binding constructs, minibodies, and scFv-Fc fusions. SUMMARY

[0004] Some embodiments provided herein are described by way of the following embodiments and also provided as possible combinations or overlapping embodiments:

[0005] Some embodiments herein are directed to an antigen binding construct. In some embodiments, the antigen binding construct disclosed herein comprises: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO. 35 or 36; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is any one of SEQ ID NOs. 38-40; and a variable heavy (VH) domain comprising: a HCDR1 that is any one of SEQ ID NO.43-46; a HCDR2 that is any one of SEQ ID NO.47-48, 119, or 136; and a HCDR3 that is SEQ ID NO.49 or 50.

[0006] Some embodiments herein are directed to an antigen binding construct. In some embodiments, the antigen binding construct disclosed herein comprises: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO.35; a LCDR2 that is SEQ ID NO.37; a LCDR3 that is SEQ ID NO. 38; and a variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO.43; a HCDR2 that is SEQ ID NO.48; and a HCDR3 that is SEQ ID NO.

[0007] Some embodiments herein are directed to an antigen binding construct. In some embodiments, the antigen binding construct disclosed herein comprises: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO.35; a LCDR2 that is SEQ ID NO.37; a LCDR3 that is SEQ ID NO. 39; and a variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO.43; a HCDR2 that is SEQ ID NO.48; and a HCDR3 that is SEQ ID NO. 50.

[0008] Some embodiments herein are directed to an antigen binding construct. In some embodiments, the antigen binding construct disclosed herein comprises: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO.35; a LCDR2 that is SEQ ID NO.37; a LCDR3 that is SEQ ID NO. 39; and a variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO. 43; a HCDR2 that is SEQ ID NO. 136; and a HCDR3 that is SEQ ID NO.50.

[0009] Some embodiments herein are directed to an antigen binding construct. In some embodiments, the antigen binding construct disclosed herein comprises: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO.36; a LCDR2 that is SEQ ID NO.37; a LCDR3 that is SEQ ID NO. 40; and a variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO. 43; a HCDR2 that is SEQ ID NO. 119; and a HCDR3 that is SEQ ID NO.50.

[0010] Some embodiments herein are directed to a single-chain variable fragment- Fc fusion (scFv-Fc) that binds to ĮVȕ6. In some embodiments, the scFv-Fc comprises a variable light (VL) domain linked to a variable heavy (VH) domain, the VL domain comprising: a LCDR1 that is SEQ ID NO.35 or 36; a LCDR2 that is any one of SEQ ID NO.37; a LCDR3 that is any one of SEQ ID NO. 38-40; and the variable heavy (VH) domain comprising: a HCDR1 that is any one of SEQ ID NO.43-46; a HCDR2 that is any one of SEQ ID NO. 47- 48, 119, or 136; and a HCDR3 that is SEQ ID NO. 49 or 50.

[0011] Some embodiments herein are directed to a single-chain variable fragment- Fc fusion (scFv-Fc) that binds to ĮVȕ6. In some embodiments, the scFv-Fc comprises a variable light (VL) domain linked to a variable heavy (VH) domain, the VL domain comprising: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO. 35; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 38; and a variable heavy (VH) domaincomprising: a HCDR1 that is SEQ ID NO. 43; a HCDR2 that is SEQ ID NO. 48; and a HCDR3 that is SEQ ID NO.50.

[0012] Some embodiments herein are directed to a single-chain variable fragment- Fc fusion (scFv-Fc) that binds to ĮVȕ6. In some embodiments, the scFv-Fc comprises a variable light (VL) domain linked to a variable heavy (VH) domain, the VL domain comprising: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO. 35; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 39; and a variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO. 43; a HCDR2 that is SEQ ID NO. 48; and a HCDR3 that is SEQ ID NO.50.

[0013] Some embodiments herein are directed to a minibody that binds to integrin ĮVȕ6. In some embodiments, the minibody comprises: a single-chain variable fragment (scFv) that binds to ĮVȕ6, the scFv comprising a variable light (VL) domain linked to a variable heavy (VH) domain, the VL domain comprising: a LCDR1 that is SEQ ID NO. 35 or 36; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is any one of SEQ ID NO. 38-40; and the variable heavy (VH) domain comprising: a HCDR1 that is any one of SEQ ID NO. 43-46; a HCDR2 that is any one of SEQ ID NO. 47-48, 119, or 136; and a HCDR3 that is SEQ ID NO. 49 or 50; a hinge-extension domain comprising a hinge region; and a IgG CH3 sequence.

[0014] Some embodiments herein are directed to a minibody that binds to integrin ĮVȕ6. In some embodiments, the minibody comprises: a single-chain variable fragment (scFv) that binds to ĮVȕ6, the scFv comprising a variable light (VL) domain linked to a variable heavy (VH) domain, the VL domain comprising: a LCDR1 that is SEQ ID NO. 35; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 38; and the variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO. 43; a HCDR2 that is SEQ ID NO. 48; and a HCDR3 that is SEQ ID NO. 50; a hinge-extension domain comprising a hinge region; and a IgG CH3 sequence.

[0015] Some embodiments herein are directed to a minibody that binds to integrin ĮVȕ6. In some embodiments, the minibody comprises: a single-chain variable fragment (scFv) that binds to ĮVȕ6, the scFv comprising a variable light (VL) domain linked to a variable heavy (VH) domain, the VL domain comprising: a LCDR1 that is SEQ ID NO. 35; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 39; and the variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO. 43; a HCDR2 that is SEQ ID NO. 48; and aHCDR3 that is SEQ ID NO. 50; a hinge-extension domain comprising a hinge region; and a IgG CH3 sequence.

[0016] Some embodiments herein are directed to a minibody that binds to integrin ĮVȕ6. In some embodiments, the minibody comprises: a single-chain variable fragment (scFv) that binds to ĮVȕ6, the scFv comprising a variable light (VL) domain linked to a variable heavy (VH) domain, the VLdomain comprising: a LCDR1 that is SEQ ID NO. 36; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 40; and the variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO. 46; a HCDR2 that is SEQ ID NO. 119; and a HCDR3 that is SEQ ID NO. 50; a hinge-extension domain comprising a hinge region; and a IgG CH3 sequence.

[0017] Some embodiments herein are directed to a minibody, scFv or scFv-Fc fusion comprising: a variable light (VL) domain of SEQ ID NO. 1 linked to a variable heavy (VH) domain of SEQ ID NO.18.

[0018] Provided herein is an antigen binding construct comprising: a variable light (VL) domain comprising: an LCDR1 that is an LCDR1 in SEQ ID NO:2 or 3: an LCDR2 that is an LCDR2 in SEQ ID NO:2 or 3; and an LCDR3 that is an LCDR3 in SEQ ID NO:2 or 3, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:2 or 3; a variable heavy (VH) domain comprising: an HCDR1 that is an HCDR1 in SEQ ID NO:19 or 20: an HCDR2 that is an HCDR2 in SEQ ID NO:19 or 20; and an HCDR3 that is an HCDR3 in SEQ ID NO:19 or 20, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:19 or 20.

[0019] Also provided is an antigen binding construct comprising: a variable light (VL) domain comprising: an LCDR1 that is an LCDR1 in SEQ ID NO:8: an LCDR2 that is an LCDR2 in SEQ ID NO:8; and an LCDR3 that is an LCDR3 in SEQ ID NO:8, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:8; a variable heavy (VH) domain comprising: an HCDR1 that is an HCDR1 in SEQ ID NO:25: an HCDR2 that is an HCDR2 in SEQ ID NO:25; and an HCDR3 that is an HCDR3 in SEQ ID NO:25, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:25.

[0020] Further provided is an antigen binding construct comprising: a variable light (VL) domain comprising: an LCDR1 that is an LCDR1 in SEQ ID NO:9: an LCDR2 that is anLCDR2 in SEQ ID NO:9; and an LCDR3 that is an LCDR3 in SEQ ID NO:9, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:9; a variable heavy (VH) domain comprising: an HCDR1 that is an HCDR1 in SEQ ID NO:26: an HCDR2 that is an HCDR2 in SEQ ID NO:26; and an HCDR3 that is an HCDR3 in SEQ ID NO:26, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:26.

[0021] Provided herein is an antigen binding construct comprising: a variable light (VL) domain comprising: an LCDR1 that is an LCDR1 in SEQ ID NO:8: an LCDR2 that is an LCDR2 in SEQ ID NO:8; and an LCDR3 that is an LCDR3 in SEQ ID NO:8, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:8; a variable heavy (VH) domain comprising: an HCDR1 that is an HCDR1 in SEQ ID NO:139: an HCDR2 that is an HCDR2 in SEQ ID NO:139; and an HCDR3 that is an HCDR3 in SEQ ID NO:139, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:139.

[0022] Also provided is antigen binding construct comprising: a variable light (VL) domain comprising: an LCDR1 that is an LCDR1 in SEQ ID NO:11: an LCDR2 that is an LCDR2 in SEQ ID NO:11; and an LCDR3 that is an LCDR3 in SEQ ID NO:11, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:11; a variable heavy (VH) domain comprising: an HCDR1 that is an HCDR1 in SEQ ID NO:28: an HCDR2 that is an HCDR2 in SEQ ID NO:28; and an HCDR3 that is an HCDR3 in SEQ ID NO:28, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:28.

[0023] Also provided herein is a pharmaceutical composition comprising any one of the antigen binding construct, minibody, scFv, or scFv-Fc fusion of the present disclosure; and a therapeutic agent, toxic payload, and / or a detectable marker.

[0024] Provided herein is a method of detecting the presence or absence of a target molecule, comprising: applying any one of the antigen binding construct, minibody, scFv, or scFv-Fc fusion or the pharmaceutical composition of the present disclosure to a sample; and detecting binding (or lack thereof) of the antigen binding construct, minibody, scFv, or scFv- Fc fusion to the target molecule in the sample.

[0025] Further provided is a method of treating cancer, comprising administering any one of the antigen binding construct, minibody, scFv, or scFv-Fc fusion or the pharmaceutical composition of the present disclosure to a subject in need thereof, optionally wherein the method comprises administering any one of the minibody or the pharmaceutical composition of the present disclosure to the subject. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] FIG. 1 is an illustrative representation of some non-limiting embodiments of an antibody and antigen binding fragments thereof.

[0027] FIG. 2 is an illustrative representation of some non-limiting embodiments of a single-chain variable fragment (scFv).

[0028] FIG. 3 is an illustrative representation of some non-limiting embodiments of a minibody.

[0029] FIG. 4 is an illustrative representation of some non-limiting embodiments of a single-chain variable fragment (scFv) Fc fusion (scFv-Fc).

[0030] FIG. 5 is flow chart illustrating some non-limiting embodiments of a method for reducing tumor size.

[0031] FIG. 6 is flow chart illustrating some non-limiting embodiments of a method of treating cancer.

[0032] FIG. 7 depicts an alignment of some non-limiting embodiments of ĮVȕ6 VL domain sequences disclosed herein.

[0033] FIG. 8 depicts an alignment of some non-limiting embodiments of ĮVȕ6 VH domain sequences disclosed herein.

[0034] FIG.9 depicts some non-limiting embodiments of an alignment of IgG CH2 domain sequences disclosed herein.

[0035] FIG. 10 depicts some non-limiting embodiments of an alignment of IgG CH3 domain sequences disclosed herein.

[0036] FIG. 11 depicts an alignment of some non-limiting embodiments of Fc domain sequences disclosed herein.

[0037] FIG. 12 shows some non-limiting embodiments of a polypeptide sequence of LCDR and HCDR sequences disclosed herein.

[0038] FIG. 13 shows some non-limiting embodiments of a polypeptide sequence of ĮVȕ6 binding antibody VH and VL domains. (SEQ ID NO.1 and 18).

[0039] FIG. 14 shows some non-limiting embodiments of a polypeptide sequence of antigen binding minibodies. (SEQ ID NO. 103-108).

[0040] FIG. 15 shows some non-limiting embodiments of a polypeptide sequence of a VH and VL with embodiments of HCDR and LCDR sequences highlighted.

[0041] FIG. 16 shows some non-limiting embodiments of an IAB56M2-14 variable region numbered according to the Aho numbering scheme.

[0042] FIG.17 shows some non-limiting embodiments of minibody scaffolds and mutations in the scaffold that reduce uptake of the minibody by healthy organs.

[0043] FIG. 18A shows some non-limiting embodiments of charge distribution of the scFv domain of IAB56M2-14 minibodies of the present disclosure.

[0044] FIG. 18A shows some non-limiting embodiments of charge distribution of the scFv domain of IAB56M2-14 minibodies of the present disclosure.

[0045] FIG. 18B shows some non-limiting embodiments of charge distribution of the scFv domain of IAB56M2-19 minibodies of the present disclosure.

[0046] FIG. 18C shows some non-limiting embodiments of charge distribution of the scFv domain of IAB56M2-27 minibodies of the present disclosure.

[0047] FIG. 19A shows some non-limiting embodiments of differences in charge distribution of the CH3 domain of minibodies of the present disclosure.

[0048] FIG. 19B shows some non-limiting embodiments of a 3D model of scFv IAB56M2-14 indicating the location of exemplary positive patches and negative patches over the exposed surface. Black: positively charged potential; White: negatively charged potential.

[0049] FIG. 19C shows some non-limiting embodiments of a 3D model of scFv IAB56M2-14 indicating the location of exemplary positive patches and negative patches over the exposed surface. Black: positively charged potential; White: negatively charged potential.

[0050] FIG. 19D shows some non-limiting embodiments of a 3D model of scFv IAB56M2-14 indicating the location of exemplary positive patches and negative patches over the exposed surface. Black: positively charged potential; White: negatively charged potential.

[0051] FIG. 20 shows some non-limiting embodiments of a polypeptide sequence of antigen binding minibodies. (SEQ ID NO. 109-118).

[0052] FIG. 21 shows some non-limiting embodiments of a polypeptide sequence of antigen binding scFv-Fc fusions. (SEQ ID NO. 72-81).

[0053] FIG. 22A shows some embodiments of EC50 determination of parental murine antibody 12B3 by ELISA.

[0054] FIG. 22B shows some embodiments of parental murine antibody 12B3 binding to A431 cells.

[0055] FIG. 22C shows some embodiments of parental murine antibody 12B3 binding to HT29 cells.

[0056] FIG. 22D shows some embodiments of parental murine antibody 12B3 binding to BxPC3 cells.

[0057] FIG. 22E shows some embodiments of parental murine antibody 12B3 binding to OVMANA cells.

[0058] FIG. 22F shows some embodiments of parental murine antibody 12B3 binding kinetics determined using biolayer interferometry.

[0059] FIG. 23 shows some embodiments of internalization of parental murine antibody 12B3 in BT20, A431, BxPC3, and HT-29 cells.

[0060] FIG. 24 shows some embodiments of parental murine antibody 12B3 non- blocking of ĮVȕ6 binding to latency associated protein (LAP).

[0061] FIG. 25 shows some embodiments of parental murine antibody 12B3 non- blocking ĮVȕ6 binding to fibronectin.

[0062] FIG.26 shows some embodiments of parental murine antibody 12B3 cross- reactivity with non- ĮVȕ6 integrins.

[0063] FIG.27 is a graph and corresponding table showing some embodiments of EC50determinations of antigen binding minibodies by ELISA.

[0064] FIG.28 is a graph and corresponding table showing some embodiments of EC50 determinations of antigen binding minibodies by ELISA.

[0065] FIG.29 is a graph and corresponding tables showing some embodiments of equilibrium dissociation constant determinations of antigen binding minibodies.

[0066] FIG.30A is a graph showing some embodiments of EC50 determinations of antigen binding minibodies in a BxPC3 pancreatic adenocarcinoma cell line.

[0067] FIG.30B is a graph showing some embodiments of EC50determinations of antigen binding minibodies in an A431 epidermoid carcinoma cell line.

[0068] FIG.30C is a graph showing some embodiments of EC50 determinations of antigen binding minibodies in a BT20 breast carcinoma cell line.

[0069] FIG.30D is a graph showing some embodiments of EC50 determinations of antigen binding minibodies in a HT29 colorectal adenocarcinoma cell line.

[0070] FIG. 31 is a graph showing some embodiments of EC50determinations of antigen binding minibodies by ELISA.

[0071] FIG.32 is a graph and corresponding table showing some embodiments of EC50 determinations of antigen binding minibodies by ELISA.

[0072] FIG.33 is a graph and corresponding tables showing some embodiments of equilibrium dissociation constant determinations of antigen binding minibodies.

[0073] FIG.34 is a graph and corresponding tables showing some embodiments of equilibrium dissociation constant determinations of antigen binding minibodies.

[0074] FIG.35 is a graph and corresponding table showing some embodiments of EC50determinations of antigen binding minibodies by flow cytometry.

[0075] FIG. 36A is a graph showing some embodiments of quantification of minibody in-vivo uptake in kidneys.

[0076] FIG. 36B is a graph showing some embodiments of quantification of minibody in-vivo uptake in blood, lungs, muscle, bone, heart, stomach, small intestine, large intestine, and pancreas.

[0077] FIG. 36C is a graph showing some embodiments of quantification of minibody in-vivo uptake.

[0078] FIG. 36D is a graph showing some embodiments of quantification of minibody in-vivo uptake in tumor xenografts in mice.

[0079] FIG. 36E is a graph showing some embodiments of quantification of the ratio between minibody uptake in tumor and kidney organs.

[0080] FIG. 37A is a graph showing some embodiments of differences in tumor size following treatment with (squares) or without (circles) antigen binding minibodies.

[0081] FIG. 37B is a graph showing some embodiments of quantification of differences in tumor size following treatment with or without antigen binding minibodies.

[0082] FIG.37C is a Kaplan-Meier plot showing differences in survival in animals treated with (squares) or without (circle) antigen binding minibodies.

[0083] FIG.38 shows some embodiments of the sequence of human IgG1 Fc with EU numbering.

[0084] FIG. 39A shows some non-limiting embodiments of modeling of surface electrostatic potential of a IAB56FC2-54 Fc domain.

[0085] FIG. 39B shows some non-limiting embodiments of modeling of surface electrostatic potential of a IAB56FC2-58 Fc domain.

[0086] FIG.40 is a graph showing quantification of scFv-Fc fusion in-vivo uptake in blood, liver, kidney, spleen, and tumor with or without electrostatic mutations.

[0087] FIG. 41A is a graph and corresponding table showing some non-limiting embodiments of EC50 determinations of antigen binding minibodies by ELISA.

[0088] FIG. 41B is a graph and corresponding table showing some non-limiting embodiments of EC50determinations of antigen binding minibodies by flow cytometry.

[0089] FIG. 42 shows some non-limiting embodiments of a polypeptide sequence of antigen binding minibodies. (SEQ ID NO. 133-135).

[0090] FIG. 43A shows some non-limiting embodiments of the polypeptide sequence of the ĮV integrin subunit.

[0091] FIG. 43B shows some non-limiting embodiments of the polypeptide sequence of the ȕ6 integrin subunit. DETAILED DESCRIPTION

[0092] Integrins are transmembrane receptors that are central to the biology of many human pathologies. Classically mediating cell-extracellular matrix and cell-cell interaction, and with a role as local activators of TGF, they influence cancer, fibrosis, thrombosis and inflammation. In some embodiments, their ligand binding and some regulatory sites are extracellular and sensitive to pharmacological intervention, consistent with the at least seven clinically approved drugs that target integrins as a primary mechanism of action.

[0093] Integrins are heterodimeric cell-surface adhesion molecules found on all nucleated cells. They can integrate processes in the intracellular compartment with the extracellular environment. The 18-alpha (Į) and 8-beta (ȕ) subunits form 24 differentheterodimers, and each can have functional and tissue specificity. Most have multiple activation states, heterogeneous glycosylation and many have multiple splice variants. See Raab-Westphal et al. (2017) Cancers 2017, 9, 110; doi:10.3390 / cancers9090110.

[0094] Integrin ĮVȕ6 is one of the integrin heterodimers. Originally identified on airway epithelial cells, and as a fibronectin receptor on colon carcinoma cells, it was shown in a landmark study to be a local activator of latent TGF. Several companies are developing antibody and small molecule inhibitors of ĮVȕ6. The humanized monoclonal antibody STX- 100 (BG00011) from Biogen-Idec has been in Phase 2 trials for idiopathic pulmonary fibrosis and for nephropathy. Exemplary antibody amino acid sequences are found in Biogen patent applications WO2003100033 and WO2007008712. GSK3008348 (GSK (Brentford, UK)) is an ĮVȕ6 PET-imaging agent in clinical trial for therapy of idiopathic pulmonary fibrosis (NCT03069989). An antibody-drug conjugate targeting ĮVȕ6 is reportedly in development (15H3: Seattle Genetics (Bothell, WA, USA)) but clinical trial results have not been reported. In some embodiments, the antigen binding construct binds ĮVȕ6 integrin. In some embodiments, ĮVȕ6 integrin comprises the polypeptide sequences depicted in FIG.43A-43B. In some embodiments, ĮVȕ6 integrin comprises an ĮV subunit comprising a polypeptide sequence comprising SEQ ID NO: 141. In some embodiments, ĮVȕ6 integrin comprises a ȕ6 subunit comprising a polypeptide sequence comprising SEQ ID NO: 142. In some embodiments, the antigen binding construct of the present disclosure binds to the ȕ6 subunit of ĮVȕ6 integrin. In some embodiments, the antigen binding construct of the present disclosure binds to a polypeptide having, having about, or having at least 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity to SEQ ID NO:142.

[0095] An attractive modality for therapeutic agents towards cell surface expressed targets is to use antibodies, antibody fragments, or antibody constructs (comprising selected domains of antibodies which may be re-ordered or re-organized and mutated at specific sequence locations). The commonality of these biologic agents is that they can be designed to be extremely selective for specific targets, historically termed antigens. A highly specific antigen binding domain can be found in each agent. The agents are sometimes known as antigen binding fragments. ImaginAb, Inc. (Inglewood CA) has been active in development of antigen binding fragments for diagnostics and / or therapeutics as may be seen in WIPO patentpublications WO 2011 / 069019, WO 2014 / 164553, WO 2013 / 188693, WO 2017 / 027325, WO 2018 / 147960, WO 2020 / 069433, WO 2019 / 236684, and WO 2023 / 283643.

[0096] Some embodiments herein are directed to an antigen binding construct (e.g., antibody or antigen binding fragment thereof). In some embodiments, the antigen binding construct (e.g., antibody or antigen binding fragment thereof) binds ĮVȕ6. In some embodiments, the antigen binding construct (e.g., antibody or antigen binding fragment thereof) is a scFv, minibody, nanobody, or scFv-Fc fusion. In some embodiments, the antigen binding construct comprises: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO.35 or 36; a LCDR2 that is SEQ ID NO.37; a LCDR3 that is any one of SEQ ID NO. 38-40; and a variable heavy (VH) domain comprising: a HCDR1 that is any one of SEQ ID NO.43-46; a HCDR2 that is any one of SEQ ID NO. 47-48, 119, or 136; and a HCDR3 that is SEQ ID NO. 49 or 50. In some embodiments, the antigen binding construct further comprises an IgG CH2 sequence. In some embodiments, the antigen binding construct comprises an IgG CH3 sequence. In some embodiments, the antigen binding construct comprises a Fc domain. In some embodiments, the antigen binding construct comprises a hinge-extension domain comprising a hinge region. In some embodiments, the antigen binding construct comprises a linker. In some embodiments, the scFv-Fc comprises a signal peptide.

[0097] Some embodiments herein are directed to a minibody that binds to integrin ĮVȕ6. In some embodiments, the minibody comprises a single-chain variable fragment (scFv) that binds to ĮVȕ6, the scFv comprising a variable light (VL) domain linked to a variable heavy (VH) domain, the VL domain comprising: a LCDR1 that is SEQ ID NO.36; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 40; and the variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO. 46; a HCDR2 that is SEQ ID NO. 119; and a HCDR3 that is SEQ ID NO. 50; a hinge-extension domain comprising a hinge region; and a IgG CH3 sequence. In some embodiments, the antigen binding construct comprises a linker. In some embodiments, the scFv-Fc comprises a signal peptide.

[0098] Some embodiments herein are directed to a minibody, scFv or scFv-Fc fusion. In some embodiments, the minibody, scFv or scFv-Fc fusion comprises: a variable light (VL) domain of SEQ ID NO. 1 linked to a variable heavy (VH) domain of SEQ ID NO.18.Definitions

[0099] All terms have their ordinary and customary meaning as understood by one of ordinary skill in the art, in view of the present disclosure.

[0100] The term “antigen binding construct” includes all varieties of antibodies, including binding fragments thereof. Further included are constructs that include 1, 2, 3, 4, 5, and / or 6 CDRs. In some embodiments, tandem scFvs can be provided, which can provide two arms with bivalent binding. In some embodiments, these CDRs can be distributed between their appropriate framework regions in a traditional antibody. In some embodiments, the CDRs can be contained within a heavy and / or light chain variable region. In some embodiments, the CDRs can be within a heavy chain and / or a light chain. In some embodiments, the CDRs can be within a single peptide chain. Unless otherwise denoted herein, the antigen binding constructs described herein bind to the noted target molecule. The term “target” or “target molecule” denotes the protein to which the antigen binding construct binds. Examples of target proteins are known in the art.

[0101] The term “antibody” includes, but is not limited to, genetically engineered or otherwise modified forms of immunoglobulins, such as intrabodies, chimeric antibodies, fully human antibodies, humanized antibodies, antibody fragments, single-chain variable fragment (scFv), and heteroconjugate antibodies (for example, bispecific antibodies, diabodies, triabodies, tetrabodies, and nanobodies, etc.). The term “antibody” includes minibodies and diabodies. The term “antibody” includes a polypeptide of the immunoglobulin family or a polypeptide comprising fragments of an immunoglobulin that is capable of noncovalently, reversibly, and in a specific manner binding a corresponding antigen. An exemplary antibody structural unit comprises a tetramer. In some embodiments, a full-length antibody can be composed of two identical pairs of polypeptide chains, each pair having one “light” and one “heavy” chain (connected through a disulfide bond). The recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon, hinge, and mu constant region genes, as well as the myriad immunoglobulin variable region genes. For full length chains, the light chains are classified as either kappa or lambda. For full length chains, the heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which in turn define the immunoglobulin classes, IgG, IgM, IgA, IgD, and IgE, respectively. The N-terminus of each chain defines a variable region of up to about 149 or more amino acids primarilyresponsible for antigen recognition. The terms variable light chain (VL) and variable heavy chain (VH) refer to these regions of light and heavy chains respectively. As used in this application, an “antibody” encompasses all variations of antibody and fragments thereof. Also, the term “antibody” includes camelid derived immunoglobulins like single heavy-chain antibodies and nanobodies®. Thus, within the scope of this concept are full length antibodies, chimeric antibodies, humanized antibodies, single chain antibodies (scFv), Fab, Fab', and multimeric versions of these fragments (for example, F(ab')2) with the same binding specificity, scFv-Fc, single domain fragments (e.g., nanobodies®), peptibodies, nanobodies®, nanobody®-Fc, minibodies, and diabodies. In some embodiments, the antibody binds specifically to a desired target.

[0102] The term "complementarity-determining domains" or "complementarity- determining regions ("CDRs") interchangeably refer to the hypervariable regions of VL and VH. The CDRs are the target molecule-binding site of the antibody chains that harbors specificity for such target molecule. In some embodiments, there are three CDRs (CDR1-3, numbered sequentially from the N-terminus) in each VLand / or VH, constituting about 15-20% of the variable domains. The CDRs are structurally complementary to the epitope of the target molecule and are thus directly responsible for the binding specificity. The remaining stretches of the VL or VH, the so-called FRs, exhibit less variation in amino acid sequence (Kuby, Immunology, 4th ed., Chapter 4. W.H. Freeman & Co., New York, 2000).

[0103] The positions of the CDRs and framework regions can be determined using various well known definitions in the art, for example, Kabat (Wu, T. T. et al., “An analysis of the sequences of the variable regions of Bence Jones proteins and myeloma light chains and their implications for antibody complementarity,” J. Exp. Med., Vol.132, No.2, pp.211-250, 1970; Kabat, E. A. et al., “Sequences of Proteins of Immunological Interest,” 5th Ed., NIH Publication No. 91–3242, Bethesda, MD, 1991); Chotia (Chothia C. et al., “Canonical structures for the hypervariable regions of immunoglobulins,” J. Mol. Biol., Vol. 196, No. 4, pp. 901-917, 1987; Chothia C. et al., “Conformations of immunoglobulin hypervariable regions,” Nature, Vol. 342, No. 6252, pp. 877-883, 1989; Chothia C. et al., "Structural repertoire of the human VH segments," J. Mol. Biol., Vol. 227, No.3, pp.799-817, 1992; Al- Lazikani B. et al., “Standard conformations for the canonical structures of immunoglobulins,” J. Mol. Biol., Vol.273, No.4, pp.927-748, 1997), ImMunoGeneTics database (IMGT) (on theworldwide web at imgt.org / ) (Giudicelli, V. et al., “IMGT / LIGM-DB, the IMGT® comprehensive database of immunoglobulin and T cell receptor nucleotide sequences,” Nucleic Acids Res., Vol. 34 (Database Issue), pp. D781-D784, 2006; Lefranc, M. P. et al., “IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains,” Dev. Comp. Immunol., Vol.27, No.1, pp.55-77, 2003; Brochet, X. et al., “IMGT / V-QUEST: the highly customized and integrated system for IG and TR standardized V-J and V-D-J sequence analysis,” Nucleic Acids Res., Vol. 36 (Web Server Issue), pp. W503-508, 2008); AbM (Martin, A. C. et al., “Modeling antibody hypervariable loops: a combined algorithm.” Proc. Natl. Acad. Sci. U.S.A., Vol.86, No.23, pp. 9268-9272, 1989); Aho (Honegger A, Plückthun A. Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool. J Mol Biol. (2001) 309:657–70); Gelfand (Gelfand IM, Kister a E. Analysis of the relation between the sequence and secondary and three-dimensional structures of immunoglobulin molecules. Proc Natl Acad Sci USA. (1995) 92:10884–8); North (North B, Lehmann A, Dunbrack RLJ. A new clustering of antibody CDR loop conformations. J Mol Biol. (2011) 406:228–56); the contact definition (MacCallum, R. M. et al., “Antibody-antigen interactions: contact analysis and binding site topography,” J. Mol. Biol., Vol.262, No.5, pp.732-745, 1996), and / or the automatic modeling and analysis tool (Honegger, A. et al., Accessible on the world wide web at bioc.uzh.ch / plueckthun / antibody / Numbering / ). In some embodiments, the polypeptide is numbered from the beginning of the polypeptide signal sequence. In some embodiments, the polypeptide is numbered according from the beginning of the polypeptide and not including the signal sequence. It is understood that K47, Q50 and K53 in Aho numbering correspond to K45, Q48 and K51 in IMGT numbering, respectively.

[0104] An "antibody variable light chain" or an "antibody variable heavy chain" as used herein refers to a polypeptide comprising the VLor VH, respectively. The endogenous VLis encoded by the gene segments V (variable) and J (junctional), and the endogenous VH by V, D (diversity), and J. Each of VL or VH includes the CDRs as well as the framework regions. In this application, antibody variable light chains and / or antibody variable heavy chains may, from time to time, be collectively referred to as "antibody chains." These terms encompass antibody chains containing mutations that do not disrupt the basic structure of VLor VH, as one skilled in the art will readily recognize. In some embodiments, full length heavy and / or lightchains are contemplated. In some embodiments, only the variable region of the heavy and / or light chains are contemplated as being present.

[0105] The term “hinge” denotes at least a part of a hinge region for an antigen binding construct, such as an antibody, a minibody, a scFv-Fc, or a nanobody®-Fc. A hinge region can include a combination of the upper hinge, core (or middle) hinge and lower hinge regions. In some embodiments, the hinge is defined according to any of the antibody hinge definitions. Native IgG1, IgG2, and IgG4 antibodies have hinge regions having of 12-15 amino acids. IgG3 has a hinge region, having 62 amino acids, including 21 prolines and 11 cysteines. The functional hinge region of naturally occurring antibodies, deduced from crystallographic studies, extends from amino acid residues 216-237 of the IgG1 H chain (EU numbering; ref. 12) and includes a small segment of the N terminus of the CH2 domain in the lower hinge, with the lower hinge being the N terminus of CH2 domain. The hinge can be divided into three regions; the "upper hinge," the "core," and the "lower hinge".

[0106] The term “artificial” or “non-natural” when modifying a hinge (or a subpart thereof) denotes that the sequence in question is not present, in the noted state, in nature. In the present context the hinges have been altered from their native state, so that their sequences are no longer those found in wild-type antibodies. As will be appreciated by those of skill in the art, minibodies do not naturally occur in nature, and thus, any construct which is a minibody construct is also not found in nature. In some embodiments, any of the hinge subparts or full hinge sequences can be artificial hinge sequences, as long as the sequence (or resulting combination for the hinge) does not occur in nature.

[0107] The term “full hinge region” or “entire hinge region” denotes the presence of the entire upper, core, and lower hinge regions as a single construct. The upper, core, and lower regions can be positioned immediately adjacent to one another, or additional residues can be added between, or N- or C-terminal to the regions. In some embodiments, the native lower hinge can be replaced with an extension sequence. In some embodiments, one can combine a native lower hinge with the extension sequence. In some embodiments, an extension or other set of sequences can be added after the upper and / or core sequences.

[0108] The phrase “effective hinge region” denotes that an adequate amount of part of at least one of the upper, core and lower hinge regions is present to allow the hinge region to be effective for its intended purpose. Thus, the phrase encompasses variants of hinge regionsand fragments of the various hinge regions. In some embodiments, the function of the hinge region is one or more of the following: to link the scFv with the CH3 domain, provide flexibility and spacing for the two scFvs to bind to the target properly, to link two half molecules together, to provide overall stability to the molecule, and / or to provide a site for site-specific conjugation due to its solvent exposure. In some embodiments, the hinge should be close to natural as to reduce potential immunogenicity. In some embodiments, the upper hinge provides flexibility to scFv (starts at residue 216 in native IgGs), the middle hinge provides stability, and the lower hinge mediates flexibility to CH3 (starts at residue 231 in native IgGs).

[0109] The term “upper hinge” denotes the first part of the hinge that starts at the end of the scFv. The upper hinge includes the amino acids from the end of the scFv up to, but not including the first cysteine residue in the core hinge. As above, the term “effective upper hinge” denotes that enough of the sequence is present to allow the section to function as an upper hinge; the term encompasses functional variants and fragments of the designated hinge section.

[0110] The term “core hinge” denotes the second part of the hinge region that is C- terminal to the upper hinge. The core hinge contains the inter-chain disulfide bridges and a high content of prolines. As above, the term “effective core hinge” denotes that enough of the sequence is present to allow the section to function as a core hinge; the term encompasses functional variants and fragments of the designated hinge section.

[0111] The term “lower hinge” denotes the third part of the hinge region that is C terminal to the core hinge. In the context of a minibody or antibody fragment, the lower hinge connects to the CH3 domain. As above, the term “effective lower hinge” denotes that enough of the sequence is present to allow the section to function as a lower hinge; the term encompasses functional variants and fragments of the designated hinge section. The term “lower hinge” as used herein can encompass various amino acid sequences including naturally occurring IgG lower hinge sequences and artificial extension sequences in place of one another or a combination thereof provided herein. In some embodiments, the various extensions can be considered to be a lower hinge region in its entirety or a replacement.

[0112] Antibodies can exist as intact immunoglobulins or as a number of fragments produced by digestion with various peptidases. Thus, for example, pepsin digests an antibody below the disulfide linkages in the hinge region to produce F(ab)'2, a dimer of Fab' which itselfis a light chain (VL-CL) joined to VH-CH1 by a disulfide bond. The F(ab)'2may be reduced under mild conditions to break the disulfide linkage in the hinge region, thereby converting the F(ab)'2 dimer into an Fab' monomer. The Fab' monomer is a Fab with part of the hinge region. (Paul, W. E., “Fundamental Immunology,” 3d Ed., New York: Raven Press, 1993). While various antibody fragments are defined in terms of the digestion of an intact antibody, one of skill will appreciate that such fragments may be synthesized de novo either chemically or by using recombinant DNA methodology. Thus, the term "antibody," as used herein, also includes antibody fragments either produced by the modification of whole antibodies, or those synthesized de novo using recombinant DNA methodologies (for example, single chain Fv) or those identified using phage display libraries (see, for example, McCafferty, J. et al., “Phage antibodies: filamentous phage displaying antibody variable domains,” Nature, Vol. 348, No. 66301, pp. 552-554, 1990).For preparation of monoclonal or polyclonal antibodies, any technique known in the art can be used (see, for example, Kohler, G. et al., “Continuous cultures of fused cells secreting antibody of predefined specificity,” Nature, Vol. 256, No. 5517, pp. 495-497, 1975; Kozbor, D. et al., “The production of monoclonal antibodies from human lymphocytes,” Immunology Today, Vol. 4, No. 3, pp. 72-79, 1983; Cole, et al., “Monoclonal Antibodies and Cancer Therapy,” Alan R. Liss, Inc., pp.77-96, 1985; Wang, S., “Advances in the production of human monoclonal antibodies,” Antibody Technology Journal, Vol.1, pp.1-4, 2011; Sharon, J. et al., “Recombinant polyclonal antibodies for cancer therapy,” J. Cell Biochem., Vol.96, No.2, pp.305-313, 2005;; Haurum, J. S., “Recombinant polyclonal antibodies: the next generation of antibody therapeutics?,” Drug Discov. Today, Vol. 11, No. 13-14, pp.655-660, 2006). Techniques for the production of single chain antibodies (U.S. Pat. No.4,946,778) can be adapted to produce antibodies to polypeptides of the present disclosure. Also, transgenic mice, or other organisms such as other mammals, may be used to express fully human monoclonal antibodies. Furthermore, E. Coli or yeast may be used to express and manufacture recombinant antibodies and antibody fragments (Simmons L. C., Reilly D., Klimowski L., Shantha Raju T., Meng G., Sims P., Hong K., Shields R. L., Damico L. A., Rancatore P., Yansura D. G.; Expression of full-length immunoglobulins in Escherichia coli: rapid and efficient production of aglycosylated antibodies, J Immunol Methods. 2002 May 1;263(1-2):133-47; Kulagina N, Besseau S, Godon C, Goldman G. H., Papon N., Courdavault V., Yeasts as biopharmaceutical production platforms; Front. Fungal Biol., 22 September2021). Alternatively, phage display and yeast display technologies can be used to identify high affinity binders to selected antigens (see, for example, McCafferty et al., supra; Marks, J. D. et al., “By-passing immunization: building high affinity human antibodies by chain shuffling,” Biotechnology (N. Y.), Vol.10, No.7, pp.779-783, 1992; Feldhaus M. J., Siegel R. W., Yeast display of antibody fragments: a discovery and characterization platform, J Immunol Methods, 2004 Jul;290(1-2):69-80). Alternatively, antibodies can be produced through B-cell screening technologies from human hosts (Pedrioli A., Oxenius A., Single B cell technologies for monoclonal antibody discovery, Trends in Immunology, (2021) volume 42, issue 12, p1143- 1158). Furthermore, antibodies can be derived from immunization of camelid animals or screening of camelid phage libraries (Harmsen M. M., De Haard H. J., Properties, production, and applications of camelid single-domain antibody fragments, Appl Microbiol Biotechnol. 2007; 77(1): 13–22). Alternatively, antibodies can be derived from in-silico screening simulations through deep learning and artificial intelligence algorithms (Graves J., Byerly J., Priego E., Makkapati N., Vince Parish S., Brenda Medellin and Monica Berrondo, A Review of Deep Learning Methods for Antibodies, Antibodies 2020, 9, 12).

[0113] Methods for humanizing or primatizing non-human antibodies are well known in the art. Generally, a humanized antibody has one or more amino acid residues introduced into it from a source which is non-human. These non-human amino acid residues are often referred to as import residues, which are typically taken from an import variable domain. In some embodiments, the terms “donor” and “acceptor” sequences can be employed. Humanization can be essentially performed following the method of Winter and co-workers (see, for example, Jones, P. T. et al., “Replacing the complementarity-determining regions in a human antibody with those from a mouse,” Nature, Vol. 321, No. 6069, pp. 522-525, 1986; Riechmann, L. et al., “Reshaping human antibodies for therapy,” Nature, Vol. 332, No.6162, pp. 323-327, 1988; Verhoeyen, M. et al., “Reshaping human antibodies: grafting an antilysozyme activity,” Science, Vol. 239, No. 4847, pp. 1534-1536, 1988; Presta, L. G., “Antibody engineering,”, Curr. Op. Struct. Biol., Vol. 2, No. 4, pp. 593-596, 1992), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. Accordingly, such humanized antibodies are chimeric antibodies (U.S. Pat. No. 4,816,567), wherein substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species. In practice, humanizedantibodies are typically human antibodies in which some complementarity determining region ("CDR") residues and possibly some framework ("FR") residues are substituted by residues from analogous sites in rodent antibodies.

[0114] The term "Fc region" or “Fc domain” or “Fc” denotes a C-terminal region of an immunoglobulin heavy chain. The "Fc region" may be a native sequence Fc region or a variant Fc region (e.g., a variant having one or more mutations that reduces an effector function, e.g., FcJR binding, and / or binding to the Fc neonatal receptor (FcRn), etc.). The Fc region of an immunoglobulin (e.g., IgG) generally comprises two constant domains, CH2 and CH3. The Fc region can include a hinge region or sequence, as described herein. An Fc region can be present in dimer or monomeric form. In some embodiments, the Fc region is a human Fc region, or a variant thereof.

[0115] A "chimeric antibody" is an antibody molecule in which (a) the constant region, or a portion thereof, is altered, replaced or exchanged so that the antigen binding site (variable region) is linked to a constant region of a different or altered class, effector function and / or species, or an entirely different molecule which confers new properties to the chimeric antibody, for example, an enzyme, toxin, hormone, growth factor, and drug; or (b) the variable region, or a portion thereof, is altered, replaced or exchanged with a variable region having a different or altered antigen specificity.

[0116] The term “antibody fragment” includes but is not limited to one or more antigen binding fragments of antibodies alone or in combination with other molecules, including, but not limited to Fab', F(ab')2, Fab, Fv, rIgG (reduced IgG), scFv fragments, scFv- Fc, single domain fragments (e.g., nanobodies® or single domain fragments), peptibodies, nanobodies®, nanobody®-Fc, minibodies, and diabodies. The term “scFv” refers to a single chain Fv (“fragment variable”) antibody in which the variable domains of the heavy chain and of the light chain of a traditional two chain antibody have been joined to form one chain.

[0117] The term “artificial” or “non-natural” when modifying a CDR or FR (or a subpart thereof) denotes that the sequence in question is not present, in the noted state, in nature. In the present context the CDRs or FRs have been altered from their native state, so that their sequences are no longer those found in wild-type antibodies. As will be appreciated by those of skill in the art, minibodies and cys-diabodies do not naturally occur in nature, and thus, any construct which is a minibody or a cys-diabody construct is also not found in nature.This also applies to at least some of the constructs found in and / or incorporating the sequences of any of the CDR or FR sequence tables provided herein. In some embodiments, any of the CDR or FR sequences in the figures or tables can be artificial CDR or FR sequences, as long as the sequence (or resulting combination for the CDR or FR) does not occur in nature.

[0118] A “minibody” is an antibody format that has a smaller molecular weight than the full-length antibody while maintaining the bivalent binding property against an antigen. Because of its smaller size, absence of CH2 domain that binds Fc-gamma and FcRn receptors, absence of glycosylation, the minibody has a faster clearance from the system and potentially enhanced penetration when targeting tumor tissue. With the ability for strong targeting combined with rapid clearance, the minibody is advantageous for diagnostic imaging and delivery of radioactive payloads for which prolonged circulation times may result in adverse patient dosing or dosimetry. In some embodiments, it can also be advantageous for delivery of a cytotoxic payload due to the above-mentioned features such as tumor penetration and faster clearance. A “minibody” as described herein, encompasses a homodimer, wherein each monomer is a single-chain variable fragment (scFv) linked to a human IgG CH3 domain by a hinge sequence. In some embodiments, a minibody is a bivalent or bispecific, covalently bound homodimer of ~80 kDa. In some embodiments, each monomer (half-molecule) is comprised of a variable heavy (VH) domain linked to the corresponding variable light (VL) domain by an approximate 15-18 amino acid Gly-Ser-rich linker sequence. As used herein, “minibody” and “antigen binding minibody” are used interchangeably.

[0119] The term “extension sequence” (e.g., in a diabody context) denotes a region that connects a first VHdomain to a second VHdomain, or a first VLto a second VLdomain, in for example, a diabody. Extension sequences can connect the domains through the C- terminus of each domain. In some embodiments, extension sequences connect the domains through covalent bonds. In some embodiments, the extension sequence will include one or more cysteine, allowing for one or more disulfide bonds to be formed between two such extension sequences. A non-limiting example of an extension sequence includes -(Gly)2-(Cys). In some embodiments, the extension sequence includes 1, 2, 3, or more cysteines per monomer chain. While the extension sequence will be towards the C-terminus of the constructs, it need not be the absolute last amino acid in the variable domain. That is, the extension sequence can be positioned slightly N-terminal to the C-terminus. For example, the extension sequence canbe placed within the 10 amino acids of the C-terminus of the monomer. Similarly, additional sequence can be placed between the native C-terminus and where the extension sequence starts. The extension sequence can connect VH to VH or VL to VL through a disulfide bond.

[0120] As used herein, “pharmaceutically acceptable” has its plain and ordinary meaning as understood in light of the specification and refers to carriers, excipients, and / or stabilizers that are nontoxic to the cell or mammal being exposed thereto at the dosages and concentrations employed or that have an acceptable level of toxicity. A “pharmaceutically acceptable” “diluent,” “excipient,” and / or “carrier” as used herein have their plain and ordinary meaning as understood in light of the specification and are intended to include any and all solvents, dispersion media, coatings, antibacterial or antifungal agents, isotonic or absorption delaying agents, compatible with administration to humans, primates, cats, dogs, or other vertebrate hosts. Typically, a pharmaceutically acceptable diluent, excipient, and / or carrier is a diluent, excipient, and / or carrier approved by a regulatory agency of a Federal, a state government, or other regulatory agency, or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, including humans as well as non-human mammals, such as cats and dogs. The term diluent, excipient, and / or “carrier” can refer to a diluent, adjuvant, excipient, or vehicle with which the pharmaceutical composition is administered. Such pharmaceutical diluent, excipient, and / or carriers, which can be incorporated in any one or more of the compositions described herein, include sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin. Water, saline solutions or aqueous dextrose and glycerol solutions can be employed as liquid diluents, excipients, and / or carriers. Suitable pharmaceutical diluents and / or excipients, which can be incorporated in any one or more of the compositions described herein, also include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, or ethanol. The physiologically acceptable carrier may also comprise one or more of the following: antioxidants, such as ascorbic acid, low molecular weight (less than about 10 residues) polypeptides, proteins, such as serum albumin, gelatin, immunoglobulins, hydrophilic polymers such as polyvinylpyrrolidone, amino acids, carbohydrates such as glucose, mannose, or dextrins, chelating agents such as EDTA, sugar alcohols such as mannitol or sorbitol, salt-forming counterions such as sodium, and nonionic surfactants such asTWEEN®, polyethylene glycol (PEG), PLURONICS® or preservatives such as an essential oil, methyl paraben, propyl paraben, or sodium salt of parabens. In some embodiments, the preservative is bronidiol. The composition, if desired, can also contain minor amounts of wetting, bulking, emulsifying agents, or pH buffering agents. These compositions can take the form of solutions, suspensions, emulsion, sustained release formulations and the like. The formulation should suit the mode of administration.

[0121] Additional excipients with desirable properties include but are not limited to preservatives, adjuvants, stabilizers, solvents, buffers, diluents, solubilizing agents, detergents, surfactants, chelating agents, antioxidants, alcohols, ketones, aldehydes, ethylenediaminetetraacetic acid (EDTA), citric acid, salts, sodium chloride, sodium bicarbonate, sodium phosphate, sodium borate, sodium citrate, potassium chloride, potassium phosphate, magnesium sulfate sugars, dextrose, fructose, mannose, lactose, galactose, sucrose, sorbitol, cellulose, serum, amino acids, polysorbate 20, polysorbate 80, sodium deoxycholate, sodium taurodeoxycholate, magnesium stearate, octylphenol ethoxylate, benzethonium chloride, thimerosal, gelatin, esters, ethers, 2-phenoxyethanol, urea, or vitamins, or any combination thereof. In some embodiments, the formulation includes an at least one agent that acts to reduce radiolysis (also known as “radioprotectors”) or is a kidney protecting agent. Non-limiting examples of radiolysis reducing agents include Gentisic acid, Acetylcholine, AET, ACE inhibitors, acteoside, alpha-tocopherol acetate, amifostine, ascorbic acid, aspirin, atorvastatin, beta-carotene, Bowman-Birk proteinase inhibitor, Caffeic acid, Captopril, carbaminoylcholine, Carvacrol, Celecoxib, coenzyme Q10, COX2 inhibitors / NSAIDs, curcumin, cysteine, cysteamine, cystamine, dendrodine analog, Dithiolthione, Dopamine, enalapril, epigallocatechin-3-gallate, Epinephrine, 17-ȕ-estradiol, GANRA-5, Genistein, green tea abstract, growth factors, guanine nucleotides, Halofuginone, Hmg-CoA reductase inhibitors (statins), heroin, histamine, ibuprofen, inapoyl-E-glucoside, Isoflavone, isofraxidin, kukoamine A, lactoferrin amifostine, lipoic acid, lovastatin, luteolin-7-O-(2-apiosyl)- glucoside, 2-mercaptoethylguanidine, melatonin, methacholine, morphine, N-acetyl cysteine, Oltipraz, palifermin, phenethyl ester, polyphenols, pravastatin, protease inhibitors, quercetin- 3-O-rhamnoside-7-O-glucoside, quercetin-3-O-rhamnoside, ramipril, Resveratrol, rutin, serotonin, Simvastatin, Sodium ascorbate, superoxide dismutase, TGF- signaling inhibitors, tocopherols, vitamin C, vitamin E, watermelon juice, black grape juice, and thiols such asglutathione. Kidney protecting agents include free lysine, arginine, probenecid, gelofusin, and other compositions. Some excipients may be in residual amounts or contaminants from the process of manufacturing, including but not limited to serum, albumin, ovalbumin, antibiotics, inactivating agents, formaldehyde, glutaraldehyde, ȕ-propiolactone, gelatin, cell debris, nucleic acids, peptides, amino acids, or growth medium components or any combination thereof. The amount of the excipient may be found in the composition at a percentage that is, is about, is at least, is at least about, is not more than, or is not more than about, 0%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 100% w / w or any percentage by weight in a range defined by any two of the aforementioned numbers.

[0122] As used herein, a “carrier” has its plain and ordinary meaning as understood in light of the specification and refers to a compound, particle, solid, semi-solid, liquid, or diluent that facilitates the passage, delivery and / or incorporation of a compound to cells, tissues and / or bodily organs.

[0123] As used herein, a “diluent” has its plain and ordinary meaning as understood in light of the specification and refers to an ingredient in a pharmaceutical composition that lacks pharmacological activity but may be pharmaceutically necessary or desirable. For example, a diluent may be used to increase the bulk of a potent drug whose mass is too small for manufacture and / or administration. It may also be a liquid for the dissolution of a drug to be administered by injection, ingestion or inhalation. A common form of diluent in the art is a buffered aqueous solution such as, without limitation, phosphate buffered saline that mimics the composition of human blood.

[0124] The term “target molecule dependent disorder” “or “target molecule associated disorder” includes any disorder in which the target molecule plays a role in the disorder itself. In some embodiments, this denotes over-expression of the target molecule. In some embodiments, the disorders can include any of the disorders discussed herein. In some embodiments, the disorder can be any for which there is a target molecule that can be targeted by binding, whose binding will result in the detection and / or treatment of the disorder.

[0125] The term “treating” or “treatment” of a condition can refer to preventing the condition, slowing the onset and / or rate of development of the condition, reducing the risk of developing the condition, preventing and / or delaying the development of symptoms associatedwith the condition, reducing or ending symptoms associated with the condition, generating a complete or partial regression of the condition, or some combination thereof. The term “prevent” does not require the absolute prohibition of the disorder or disease. Examples of disorders or diseases include fibrosis, cancer, tumor and neoplasms, autoimmune disease, cardiovascular, neurodegenerative, metabolic and endocrine disorders, inflammatory, immunity, genetic disorders, infectious disorders, hematological, congenital disorders, musculoskeletal, oral and gastrointestinal, renal and urogenital disorders, reproductive disorders, respiratory disorders, skin and / or epithelial, as well as disorders with disputed or unknown etiology.

[0126] “Tumor,” as used herein, refers to all neoplastic cell growth and proliferation, whether malignant or benign, and all pre-cancerous and cancerous cells and tissues. The terms “cancer,” “cancerous,” “cell proliferative disorder,” “proliferative disorder” and “tumor” are not mutually exclusive as referred to herein. The term “neoplasia” encompasses the term tumor.

[0127] The terms “cancer” and “cancerous” refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth. Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies. More particular examples of such cancers include lung cancer including small-cell lung cancer, non-small cell lung cancer and lung adenocarcinomas with neuroendocrine features; neuroendocrine prostate cancer, melanoma, gliomas, low-grade gliomas and glioblastoma, medullary thyroid cancer, carcinoid tumors, neuroendocrine tumors in the pancreas, bladder cancer, testicular cancer squamous cell cancer (e.g. epithelial squamous cell cancer), neuroendocrine neoplasms, such as neuroendocrine tumors of unknown primary, neuroendocrine neoplasms of the small bowel, carotid body, adrenal gland, colorectal gynecological organ, abdomen, esophagus, GI tract, bile duct, nervous system, appendix, liver, anal, thymus, ileocecal junction, head and neck, breast, peritoneum and retroperitoneum, kidney, thyroid, stomach, bone,; adenocarcinomas, such as adenocarcinoma of the lung and squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastric or stomach cancer including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, cancer of the urinary tract, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial or uterine carcinoma,salivary gland carcinoma, kidney or renal cancer, prostate cancer, vulval cancer, thyroid cancer, bone cancer, hepatic carcinoma, anal carcinoma, penile carcinoma, melanoma, multiple myeloma and B-cell lymphoma, brain, as well as head and neck cancer, and associated metastases. The term cancer includes adult and pediatric solid cancers. In some embodiments, the cancer can be a solid tumor. In some embodiments, the cancer is a highly fibrotic tumor or cancer. In some embodiments, the cancer is a desmoplasia.

[0128] A “therapeutically effective amount” or a “therapeutically effective dose” is an amount that produces a desired therapeutic effect in a subject, such as preventing, treating a target condition, delaying the onset of the disorder and / or symptoms, and / or alleviating symptoms associated with the condition. This amount will vary depending upon a variety of factors, including but not limited to the characteristics of the therapeutic compound (including activity, pharmacokinetics, pharmacodynamics, biodistribution and bioavailability), the physiological condition of the subject (including age, sex, disease type and stage, general physical condition, responsiveness to a given dosage, and type of medication), the nature of the pharmaceutically acceptable carrier or carriers in the formulation, and / or the route of administration. One skilled in the clinical and pharmacological arts will be able to determine a therapeutically effective amount through routine experimentation, for example by monitoring a subject's response to administration of a compound and adjusting the dosage accordingly, given the present disclosure. For additional guidance, see Remington: The Science and Practice of Pharmacy 21stEdition, Univ. of Sciences in Philadelphia (USIP), Lippincott Williams & Wilkins, Philadelphia, PA, 2005.

[0129] “Label”, “detectable label” or “detectable marker” are used interchangeably herein and refer to a detectable compound or composition which is conjugated directly or indirectly associated with the antibody so as to generate a “labeled” antibody. The label may be detectable by itself (e.g., radioisotope labels or fluorescent labels) or, in the case of an enzymatic label, may catalyze chemical alteration of a substrate compound or composition which is detectable.

[0130] The term “payload” denotes an atom or molecule or other entity that is associated (covalently or otherwise) to an antigen binding construct. It includes labels or markers for aspects for diagnostics for example, as well as toxins, cytotoxic agents, chemotherapeutic agents for various therapies. In some embodiments, the payload involves aschelator so as to attach the antigen binding construct to the molecule or atom to be delivered via or colocalized via the antigen binding construct.

[0131] The term “cytotoxic agent” as used herein refers to a substance that inhibits or prevents a cellular function and / or causes cell death or destruction. The term is intended to include non-radioactive isotopes (ADC), radioactive isotopes (e.g.,177Lu,227Th,211At,131I,125I,90Y,186Re,188Re,153Sm,212Bi,213Bi,32P,149Tb,161Tb,212Pb, and radioactive isotopes of Lu), chemotherapeutic agents (as defined elsewhere herein). Other cytotoxic agents are described below. A tumoricidal agent causes destruction of tumor cells.

[0132] A “toxin” is any substance capable of having a detrimental effect on the growth or proliferation of a cell. Non-radioactive payloads include those commonly used for antibody drug conjugates (ADC) and fragment drug conjugates (FDC), such as toxins belonging to the families of auristatins, maytansines, maytansinoids, calicheamicins, duocarymycins, pyrrolobenzodiazepines dimers and amatoxins.

[0133] A “therapeutic ion” refers to an electrically charged particle that that is useful in the treatment of a disorder related to a target molecule. Examples of therapeutic ions include18F,18F-FDG,32P,33P,45Ti,47Sc,52Fe,59Fe,62Cu,64Cu,67Cu,67Ga,68Ga,75Sc,77As,86Y,90Y,89Sr,89Zr,94Tc,94Tc,99mTc,99Mo,105Pd,105Rh,111Ag,111In,123I,124I,125I,131I,142Pr,143Pr,149Pm,149Tb,153Sm,154-158Gd,161Tb,166Dy,166Ho,169Er,175Lu,177Lu,186Re,188Re,189Re,194Ir,198Au,199Au,211At,211Pb,212Bi,212Pb,213Bi,223Ra,227Th, and225Ac. These are also options of therapeutic agents.

[0134] A “chemotherapeutic agent” is a chemical compound useful in the treatment of cancer. Examples of chemotherapeutic agents include alkylating agents such as thiotepa and CYTOXANTM cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylolomelamine; acetogenins (especially bullatacin and bullatacinone); delta-9-tetrahydrocannabinol (dronabinol, MARINOLTM); beta-lapachone; lapachol; colchicines; betulinic acid; a camptothecin (including the synthetic analogue topotecan (HYCAMTINTM), CPT-11 (irinotecan, CAMPTOSARTM), acetylcamptothecin, scopolectin, and 9-aminocamptothecin); bryostatin; callystatin; CC-1065 (including its adozelesin, carzelesin and bizelesin syntheticanalogues); podophyllotoxin; podophyllinic acid; teniposide; cryptophycins (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogues, KW-2189 and CB1-TM1); eleutherobin; pancratistatin; a sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; antibiotics such as the enediyne antibiotics (e. g., calicheamicin, especially calicheamicin gammall and calicheamicin omegall (see, e.g., Agnew, Chem Intl. Ed. Engl., 33: 183-186 (1994)); dynemicin, including dynemicin A; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, carminomycin, carzinophilin, chromomycins, dactinomycin, daunorubicin, detorubicin, 6- diazo-5-oxo-L-norleucine, ADRIAMYCINTM doxorubicin (including morpholino- doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, porfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; metabolic inhibitor such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfornithine; elliptinium acetate; an epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; 2-ethylhydrazide; procarbazine; PSK.RTM. polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; sizofiran;spirogermanium; tenuazonic acid; triaziquone; 2,2’,2’’-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A and anguidine); urethan; vindesine (ELDISINETM, FILDESINTM); dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside (“Ara-C”); thiotepa; taxoids, e.g., TAXOL.RTM. paclitaxel (Bristol-Myers Squibb Oncology, Princeton, N.J.), ABRAXANETM Cremophor- free, albumin-engineered nanoparticle formulation of paclitaxel (American Pharmaceutical Partners, Schaumberg, Ill.), and TAXOTERETM docetaxel (Rhone-Poulenc Rorer, Antony, France); chloranbucil; gemcitabine (GEMZARTM); 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine (VELBANTM); platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine (ONCOVINTM); oxaliplatin; leucovovin; vinorelbine (NAVELBINETM); novantrone; edatrexate; daunomycin; aminopterin; ibandronate; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; capecitabine (XELODATM); pharmaceutically acceptable salts, acids or derivatives of any of the above; as well as combinations of two or more of the above such as CHOP, an abbreviation for a combined therapy of cyclophosphamide, doxorubicin, vincristine, and prednisolone, and FOLFOX, an abbreviation for a treatment regimen with oxaliplatin (ELOXATINTM) combined with 5-FU and leucovovin. These are also options of therapeutic agents.

[0135] “Radiotherapy” also sometimes called “radiopharmaceutical therapy” or “RPT”, or “radioimmunotherapy” (RIT), means treatment using a radioisotope with a therapeutic purpose. Embodiments of the invention are radiopharmaceutical agents and useful in RPT. It includes radiation therapy intended to have abscopal effect as described in Yang Liu, Yinping Dong, Li Kong, Fang Shi, Hui Zhu & Jinming Yu; “Abscopal effect of radiotherapy combined with immune checkpoint inhibitors”; Journal of Hematology & Oncology volume 11, Article number: 104 (2018); and in Melek Tugce Yilmaz, Aysenur Elmali, and Gozde Yazici; “Abscopal Effect, From Myth to Reality: From Radiation Oncologists' Perspective”; Cureus. 2019 Jan; 11(1).

[0136] As used herein, the term “pretargeting” has its plain and ordinary meaning and may refer to target site localization of a targeting moiety conjugated with one member of a ligand / anti-ligand pair; after a time period sufficient for optimal target-to-non-target accumulation of this targeting moiety conjugate, an active agent conjugated to the oppositemember of the ligand / anti-ligand pair is administered and is bound (directly or indirectly) to the targeting moiety conjugate at the target site.

[0137] As used herein, the term “chelation” has it’s plain and ordinary meaning and also may refer to the formation of multiple coordination bonds between organic molecules and a transition metal ion leading to sequestration of the metal. In some embodiments, the chelating ligand comprises ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), dodecane tetraacetic acid (DOTA), 1,4,7,10- tetraazacyclododececane,1-(glutaric acid)-4,7,10-triacetic acid (DOTAGA), 1,4,7- triazacyclononane-1,4,7-triacetic acid (NOTA), 1,4,7-triazacyclononane, 1-glutaric acid-5,7 acetic acid (NODAGA), 4-[2-(bis-carboxymethyl-amino)-ethyl]-7-carboxymethyl- [1,4,7]triazonan-1-yl-acetic acid (NETA), deferoxamine (Df, which may also be referred to as DFO), porphyrins, polyamines, crown ethers, bis-thiosemicarbazones, polyoximes, and like groups.

[0138] The terms "subject," "patient," and "individual" interchangeably refer to an entity that is being examined and / or treated. The term “mammal” is used in its usual biological sense. Thus, it specifically includes, but is not limited to, primates, including simians (chimpanzees, apes, monkeys), humans, cattle, horses, sheep, goats, swine, rabbits, dogs, cats, rodents, rats, mice, or guinea pigs.

[0139] The term "co-administer" refers to the administration of two active agents in the blood of an individual or in a sample to be tested. Active agents that are co-administered in combination or sequentially delivered. “In combination” means that two (or more) different compositions are delivered to the subject during the course of the subject's affliction with the disorder, e.g., the two or more compositions are delivered after the subject has been diagnosed or selected as one having the disorder and before the disorder has been cured or eliminated. In some embodiments the subject is selected to receive any one or more of the compositions described herein by diagnostic analysis or clinical evaluation or both. In some embodiments, the delivery of one therapy is still occurring when the delivery of the second begins, so that there is overlap. This is sometimes referred to herein as "simultaneous" or "concomitant" or "concurrent delivery". In other embodiments, the delivery of one therapy ends before the delivery of the other therapy begins. This is sometimes referred to herein as "successive" or "sequential delivery." In embodiments of either case, the therapy is more effective because ofcombined administration. For example, the second therapy is a more effective, e.g., an equivalent effect is seen with less of the second therapy, or the second therapy reduces symptoms to a greater extent, than would be seen if the second therapy were administered in the absence of the first therapy, or the analogous situation is seen with the first therapy. In some embodiments, delivery is such that the reduction in a symptom, or other parameter related to the disorder is greater than what would be observed with one therapy delivered in the absence of the other. The effect of the two therapies can be partially additive, wholly additive, or greater than additive (e.g., synergistic). The delivery can be such that an effect of the first therapy delivered is still detectable when the second is delivered.

[0140] As used herein, the term “palliative care” has its plain and ordinary meaning and may refer to specialized medical care focused on providing relief from pain and other symptoms of a serious illness and / or side effects from medical treatments. The availability of palliative care does not depend on whether the serious illness can be cured. Palliative care may be offered alongside one or more other treatments.

[0141] The phrase "specifically (or selectively) bind," when used in the context of describing the interaction between an antigen, for example, a protein, to an antibody or antibody-derived binding agent, refers to a binding reaction that is determinative of the presence of the antigen in a heterogeneous population of proteins and other biologics, for example, in a biological sample, for example, a blood, serum, plasma or tissue sample. Thus, under designated immunoassay conditions, in some embodiments, the antibodies or binding agents with a particular binding specificity bind to a particular antigen at least two times the background and do not substantially bind in a significant amount to other antigens present in the sample. Specific binding to an antibody or binding agent under such conditions may require the antibody or agent to have been selected for its specificity for a particular protein. A variety of immunoassay formats may be used to select antibodies specifically immunoreactive with a particular protein. For example, solid-phase ELISA immunoassays are routinely used to select antibodies specifically immunoreactive with a protein (see, for example, Harlow, E. & Lane D., “Using Antibodies, A Laboratory Manual,” Cold Spring Harbor Laboratory Press, 1998, for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity). Typically, a specific or selective binding reaction will produce a signal atleast twice over the background signal and more typically at least than 10 to 100 times over the background.

[0142] The term "equilibrium dissociation constant (KD, M)" refers to the dissociation rate constant (kd, time-1) divided by the association rate constant (ka, time-1M-1). Equilibrium dissociation constants can be measured using any known method in the art. The antibodies of the present disclosure generally will have an equilibrium dissociation constant of less (that is superior binding) than about 10-7or 10-8M, for example, less than about 10-9M or 10-10M, in some embodiments, less than about 10-11M, 10-12M, or 10-13M.

[0143] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non- naturally occurring amino acid polymer.

[0144] The term "nucleic acid" or "polynucleotide" refers to deoxyribonucleic acids (DNA) or ribonucleic acids (RNA) and polymers thereof in either single- or double- stranded form. Unless specifically limited, the term encompasses nucleic acids containing known analogues of natural nucleotides that have similar binding properties as the reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides. Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (for example, degenerate codon substitutions), alleles, orthologs, SNPs, and complementary sequences as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and / or deoxyinosine residues (Batzer, M. A. et al., “Enhanced evolutionary PCR using oligonucleotides with inosine at the 3'-terminus,” Nucleic Acid Res., Vol.19, No.18, pp.5081, 1991; Ohtsuka, E. et al., “An alternative approach to deoxyoligonucleotides as hybridization probes by insertion of deoxyinosine at ambiguous codon positions,” J. Biol. Chem., Vol. 260, No. 5, pp. 2605-2608, 1985; Rossolini, G. M. et al., “Use of deoxyinosine-containing primers vs degenerate primers for polymerase chain reaction based on ambiguous sequence information,” Mol. Cell. Probes, Vol.8, No. 2, pp.91-98, 1994).

[0145] The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, for example, hydroxyproline, gamma-carboxyglutamate, and O-phosphoserine. Amino acid analogs refer to compounds that have the same basic chemical structure as a naturally occurring amino acid, for example, an alpha-carbon that is bound to a hydrogen, a carboxyl group, an amino group, and an R group, for example, homoserine, norleucine, methionine sulfoxide, methionine methyl sulfonium. Such analogs have modified R groups (for example, norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Amino acid mimetics refers to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that functions in a manner similar to a naturally occurring amino acid.

[0146] The term "conservatively modified variants" applies to both amino acid and nucleic acid sequences. With respect to particular nucleic acid sequences, conservatively modified variants refers to those nucleic acids which encode identical or essentially identical amino acid sequences, or where the nucleic acid does not encode an amino acid sequence, to essentially identical sequences. Because of the degeneracy of the genetic code, a large number of functionally identical nucleic acids encode any given protein. For instance, the codons GCA, GCC, GCG and GCU all encode the amino acid alanine. Thus, at every position where an alanine is specified by a codon, the codon can be altered to any of the corresponding codons described without altering the encoded polypeptide. Such nucleic acid variations are "silent variations," which are one species of conservatively modified variations. Every nucleic acid sequence herein which encodes a polypeptide also describes every possible silent variation of the nucleic acid. One of skill will recognize that each codon in a nucleic acid (except AUG, which is ordinarily the only codon for methionine, and TGG, which is ordinarily the only codon for tryptophan) can be modified to yield a functionally identical molecule. Accordingly, each silent variation of a nucleic acid that encodes a polypeptide is implicit in each described sequence.

[0147] As to amino acid sequences, one of skill will recognize that individual substitutions, deletions or additions to a nucleic acid, peptide, polypeptide, or protein sequencewhich alters, adds or deletes a single amino acid or a small percentage of amino acids in the encoded sequence is a "conservatively modified variant" where the alteration results in the substitution of an amino acid with a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are well known in the art. Such conservatively modified variants are in addition to and do not exclude polymorphic variants, interspecies homologs, and alleles of the present disclosure.

[0148] The following eight groups each contain amino acids that are conservative substitutions for one another: 1) Alanine (A), Glycine (G); 2) Aspartic acid (D), Glutamic acid (E); 3) Asparagine (N), Glutamine (Q); 4) Arginine (R), Lysine (K); 5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V); 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W); 7) Serine (S), Threonine (T); and 8) Cysteine (C), Methionine (M) (see, for example, Creighton, T. E., “Proteins - Structures and Molecular Properties,” W. H. Freeman & Co. Ltd., 1984).

[0149] The term "percentage of sequence identity" can be determined by comparing two optimally aligned sequences over a comparison window, wherein the portion of the polynucleotide sequence in the comparison window may comprise additions or deletions (i.e., gaps) as compared to the reference sequence (for example, a polypeptide of the present disclosure), which does not comprise additions or deletions, for optimal alignment of the two sequences. The percentage is calculated by determining the number of positions at which the identical nucleic acid base or amino acid residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison and multiplying the result by 100 to yield the percentage of sequence identity.

[0150] The terms "identical" or percent "identity," in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences that are the same sequences. Two sequences are "substantially identical" if two sequences have a specified percentage of amino acid residues or nucleotides that are the same (for example, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity over a specified region, or, when not specified, over the entire sequence of a reference sequence), when compared and aligned for maximum correspondence over a comparison window, or designated region as measured using one of the following sequence comparison algorithms or by manual alignment and visual inspection. Some embodiments provided herein provide polypeptides orpolynucleotides that are substantially identical to the polypeptides or polynucleotides, respectively, exemplified herein. Optionally, the identity exists over a region that is at least about 15, 25 or 50 nucleotides in length, or more preferably over a region that is 100 to 500 or 1000 or more nucleotides in length, or over the full length of the reference sequence. With respect to amino acid sequences, identity or substantial identity can exist over a region that is at least 5, 10, 15 or 20 amino acids in length, or optionally at least about 25, 30, 35, 40, 50, 75 or 100 amino acids in length, or optionally at least about 150, 200 or 250 amino acids in length, or over the full length of the reference sequence. With respect to shorter amino acid sequences, for example, amino acid sequences of 20 or fewer amino acids, in some embodiments, substantial identity exists when one or two amino acid residues are conservatively substituted, according to the conservative substitutions defined herein.

[0151] In some embodiments, the percent identity is over the CDR and / or FR regions noted herein. In such situations, the percent identity of the CDR or FR can be identified separately from the rest of the protein or nucleic acid sequence. Thus, two CDRs or FRs can have a specified percentage of amino acid residues or nucleotides that are the same (for example, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity over a specified region, or, when not specified, over the entire sequence of a reference sequence), while allowing for the remainder of the protein to either stay 100% identical to the comparison protein, our while also allowing the remainder of the protein to also have variation by a specified percent identity.

[0152] For sequence comparison, typically one sequence acts as a reference sequence, to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. Default program parameters can be used, or alternative parameters can be designated. The sequence comparison algorithm then calculates the percent sequence identities for the test sequences relative to the reference sequence, based on the program parameters.

[0153] A "comparison window", as used herein, includes reference to a segment of any one of the number of contiguous positions selected from the group consisting of from 20 to 600, usually about 50 to about 200, more usually about 100 to about 150 in which a sequence may be compared to a reference sequence of the same number of contiguous positions after thetwo sequences are optimally aligned. Methods of alignment of sequences for comparison are well known in the art. Optimal alignment of sequences for comparison can be conducted, for example, by the local homology algorithm of Smith and Waterman (1970) Adv. Appl. Math. 2:482c, by the homology alignment algorithm of Needleman, S. B. et al., “A general method applicable to the search for similarities in the amino acid sequence of two proteins,” J. Mol. Biol., Vol. 48, No. 3, pp. 443-453, 1970, by the search for similarity method of Pearson, W. R. et al., “Improved tools for biological sequence comparison,” Proc. Natl. Acad. Sci. U.S.A., Vol. 85, No. 8, pp. 2444-2448, 1988, by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by manual alignment and visual inspection (see, for example, Ausubel, F. M. et al., Current Protocols in Molecular Biology, Supplement, 1995).

[0154] Two examples of algorithms that are suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul, S. F. et al., “Gapped BLAST and PSI-BLAST: a new generation of protein database search programs,” Nucleic Acids Res., Vol.25, No.17, pp.3389-3402, 1977, and Altschul, S. F. et al., “Basic local alignment search tool,” J. Mol. Biol., Vol. 215, No. 3, pp. 403-410, 1990, respectively. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information. This algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul, S. F. et al., supra). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for mismatching residues; always <0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a word length (W) of 11, an expectation (E) or 10, M=5, N=-4 and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a word length of 3, and expectation (E) of 10, and the BLOSUM62 scoring matrix (see, Henikoff, S. et al., “Amino acid substitution matrices from protein blocks,” Proc. Natl. Acad. Sci. U.S.A., Vol. 89, No. 22, pp. 10915-10919, 1992) alignments (B) of 50, expectation (E) of 10, M=5, N=-4, and a comparison of both strands.

[0155] The BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, for example, Karlin, S. et al., “Applications and statistics for multiple high-scoring segments in molecular sequences,” Proc. Natl. Acad. Sci. U.S.A., Vol. 90, No. 12, pp. 5873-5787, 1993). One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by which a match between two nucleotide or amino acid sequences would occur by chance. For example, a nucleic acid is considered similar to a reference sequence if the smallest sum probability in a comparison of the test nucleic acid to the reference nucleic acid is less than about 0.2, more preferably less than about 0.01, and most preferably less than about 0.001.

[0156] An indication that two nucleic acid sequences or polypeptides are substantially identical is that the polypeptide encoded by the first nucleic acid is immunologically cross reactive with the antibodies raised against the polypeptide encoded by the second nucleic acid, as described below. Thus, in some embodiments, a polypeptide is typically substantially identical to a second polypeptide, for example, where the two peptides differ only by conservative substitutions. Another indication that two nucleic acid sequences are substantially identical is that the two molecules or their complements hybridize to each other under stringent conditions, as described below. Yet another indication that two nucleic acid sequences are substantially identical is that the same primers can be used to amplify the sequence. Antigen Binding Constructs and Methods of Use Thereof:

[0157] Provided herein is an antigen binding construct that comprises: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO. 35 or 36; a LCDR2 that is SEQID NO.37; a LCDR3 that is any one of SEQ ID NOs.38-40; and a variable heavy (VH) domain comprising: a HCDR1 that is any one of SEQ ID NO.43-46; a HCDR2 that is any one of SEQ ID NO. 47-48, 119, or 136; and a HCDR3 that is SEQ ID NO. 49 or 50.

[0158] Provided herein is an antigen binding construct that comprises: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO. 35; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 38; and a variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO. 43; a HCDR2 that is SEQ ID NO. 48; and a HCDR3 that is SEQ ID NO. 50.

[0159] Provided herein is an antigen binding construct that comprises: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO. 35; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 39; and a variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO. 43; a HCDR2 that is SEQ ID NO. 48; and a HCDR3 that is SEQ ID NO. 50.

[0160] Provided herein is an antigen binding construct that comprises: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO. 35; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 39; and a variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO.43; a HCDR2 that is SEQ ID NO.136; and a HCDR3 that is SEQ ID NO. 50.

[0161] Provided herein is an antigen binding construct that comprises: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO. 36; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 40; and a variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO.43; a HCDR2 that is SEQ ID NO.119; and a HCDR3 that is SEQ ID NO. 50.

[0162] In some embodiments, the antigen binding construct is or comprises a single-chain variable fragment-Fc fusion (scFv-Fc) that binds to ĮVȕ6, the scFv-Fc comprising the variable light (VL) domain linked to the variable heavy (VH) domain. In some embodiments, the antigen binding construct is or comprises a minibody that binds to integrin ĮVȕ6, the minibody comprising: a single-chain variable fragment (scFv) that binds to ĮVȕ6, the scFv comprising the variable light (VL) domain linked to the variable heavy (VH) domain.

[0163] FIG. 1 is an illustrative representation of some non-limiting embodiments of an antibody and antigen binding fragments thereof. Some embodiments herein are directedto an antibody or antigen binding fragment thereof 100. In some embodiments, the antibody or antigen binding fragment thereof is a ĮVȕ6 antibody or antigen binding fragment thereof. In some embodiments, the antibody of antigen binding fragment thereof 101 comprises: a variable light (VL) domain 102 and a variable heavy (VH) domain 103. In some embodiments, the VL domain comprises a sequence that is any one of SEQ ID NO. 2-17. In some embodiments, the VH domain comprises a sequence that is any one of SEQ ID NO. 19-34, 139 or 140. In some embodiments the VL and VH domains comprise one or more CDRs 104. In some embodiments, the VL domain comprises an LCDR1105, LCDR2106, and LCDR3 107. In some embodiments, the LCDR1 has a sequence that is SEQ ID NO.35 or 36. In some embodiments, the LCDR2 has a sequence that is SEQ ID NO. 37. In some embodiments, the LCDR3 has a sequence that is any one of SEQ ID NO. 38-40. In some embodiments, the VH domain comprises an HCDR1108, HCDR2109, and HCDR3110. In some embodiments, the HCDR1 has a sequence that is any one of SEQ ID NO. 43-46. In some embodiments, the HCDR2 has a sequence that is any one of SEQ ID NO.47-48, 119, 136. In some embodiments, the HCDR3 has a sequence that is SEQ ID NO.49 or 50. In some embodiments, the antibody or antigen binding fragment thereof comprises a hinge domain 111 comprising a hinge region. In some embodiments, the hinge region comprises an upper hinge, a core hinge, and a lower hinge region. In some embodiments, the upper hinge comprises a sequence that is SEQ ID NO.51 or 52. In some embodiments, the core hinge comprises a sequence that is SEQ ID NO. 53. In some embodiments, the lower hinge region comprises a sequence that is SEQ ID NO. 54. In some embodiments, the antibody or antigen binding fragment thereof comprises a linker. In some embodiments, the linker comprises a sequence that is SEQ ID NO. 55. In some embodiments, the antibody or antigen binding fragment thereof comprises a signal peptide. In some embodiments, the signal peptide comprises a sequence that is SEQ ID NO.56. In some embodiments, the antibody or antigen binding fragment thereof comprises an IgG CH2 domain 112. In some embodiments, the CH2 domain comprises a sequence that is any one of SEQ ID NO.47-76. In some embodiments, the antibody or antigen binding fragment thereof comprises an IgG CH3 domain 113. In some embodiments, the CH3 domain comprises any one of SEQ ID NO. 77-80. In some embodiments, the CH2 and / or CH3 domain comprises one or more substitutions. In some embodiments, the one or more substitutions in the CH2 and / or CH3 domain alters biodistribution of the antibody or antigen binding fragment thereof. In someembodiments, the one or more substitutions in the CH2 and / or CH3 domain alter distribution of the antibody or antigen binding fragment thereof to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the antibody or antigen binding fragment thereof comprises a Fc domain 114. In some embodiments, the antibody or antigen binding fragment thereof comprises a Fc domain comprises a sequence that is any one of SEQ ID NO. 83-102. In some embodiments, the Fc domain comprises one or more substitutions. In some embodiments, the one or more substitutions in the Fc domain alters biodistribution of the antibody or antigen binding fragment thereof. In some embodiments, the one or more substitutions in the Fc domain alters distribution of the antibody or antigen binding fragment thereof to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof.

[0164] In some embodiments, the antibody or antigen binding fragment thereof further comprise a payload. In some embodiments, the payload comprises a therapeutic agent, toxin, and / or a detectable marker. In some embodiments, the detectable marker is a radio label. In some embodiments, the antibody or antigen binding fragment thereof is administered to a subject having one or more diseases. In some embodiments, the disease is a disease of the blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, administration of the antibody or antigen binding fragment thereof reduces or maintains the severity of one or more symptoms of the disease. In some embodiments, administration of the antibody or antigen binding fragment thereof eliminates on or more symptoms of the disease. In some embodiments, the antibody or antigen binding fragment thereof is administered as palliative care.

[0165] Some embodiments herein are directed to an antibody or antigen binding fragment thereof having altered biodistribution. In some embodiments, the antibody or antigen binding fragment thereof having altered biodistribution is a ĮVȕ6 antibody or antigen binding fragment thereof. In some embodiments, the antibody or antigen binding fragment thereof comprises a hinge-extension domain comprising a hinge region. In some embodiments, the hinge region comprises an upper hinge, a core hinge, and a lower hinge region. In someembodiments, the upper hinge comprises a sequence that is SEQ ID NO. 51 or 52. In some embodiments, the core hinge comprises a sequence that is SEQ ID NO. 53. In some embodiments, the lower hinge region comprises a sequence that is SEQ ID NO. 54. In some embodiments, the antibody or antigen binding fragment thereof comprises a linker. In some embodiments, the linker comprises a sequence that is SEQ ID NO.55. In some embodiments, the antibody or antigen binding fragment thereof comprises a signal peptide. In some embodiments, the signal peptide comprises a sequence that is SEQ ID NO. 56. In some embodiments, the antibody or antigen binding fragment thereof comprises an IgG CH2 domain. In some embodiments, the CH2 domain comprises a sequence that is any one of SEQ ID NO. 47-76. In some embodiments, the antibody or antigen binding fragment thereof comprises an IgG CH3 domain. In some embodiments, the CH3 domain comprises any one of SEQ ID NO. 77-80. In some embodiments, the CH2 and / or CH3 domain comprises one or more substitutions. In some embodiments, the one or more substitutions in the CH2 and / or CH3 domain alters biodistribution of the antibody or antigen binding fragment thereof. In some embodiments, the one or more substitutions in the CH2 and / or CH3 domain alter distribution of the antibody or antigen binding fragment thereof to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the antibody or antigen binding fragment thereof comprises a Fc domain. In some embodiments, the antibody or antigen binding fragment thereof comprises a Fc domain having a sequence that is any one of SEQ ID NO. 83-102. In some embodiments, the Fc domain comprises one or more substitutions. In some embodiments, the one or more substitutions in the Fc domain alters biodistribution of the antibody or antigen binding fragment thereof. In some embodiments, the one or more substitutions in the Fc domain alters distribution of the antibody or antigen binding fragment thereof to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof.

[0166] In some embodiments, the antibody or antigen binding fragment thereof further comprise a payload. In some embodiments, the payload comprises a therapeutic agent, toxin, and / or a detectable marker. In some embodiments, the detectable marker is a radio label. In some embodiments, the antibody or antigen binding fragment thereof is administered to asubject having one or more diseases. In some embodiments, the disease is a disease of the blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, administration of the antibody or antigen binding fragment thereof reduces or maintains the severity of one or more symptoms of the disease. In some embodiments, administration of the antibody or antigen binding fragment thereof eliminates on or more symptoms of the disease. In some embodiments, the antibody or antigen binding fragment thereof is administered as palliative care.

[0167] FIG. 2 is an illustrative representation of some embodiments of a single- chain variable fragment (scFv). Some embodiments herein are directed to a single-chain variable fragment (scFv) 200. In some embodiments, the scFv is a ĮVȕ6 binding scFv. In some embodiments, the scFv comprises: a variable light (VL) domain 201 and a variable heavy (VH) domain 205. In some embodiments, the VL domain is N-terminal to the VH domain in the scFv. In some embodiments, the VH domain is N-terminal to the VL domain in the scFv. In some embodiments, the VL domain comprises a sequence that is any one of SEQ ID NO. 1- 17. In some embodiments, the VH domain comprises a sequence that is any one of SEQ ID NO.18-34, 139 or 140. In some embodiments the VL and VH domains comprise one or more CDRs 202-208. In some embodiments, the VL domain comprises an LCDR1 202, LCDR2 203, and LCDR3204. In some embodiments, the LCDR1 has a sequence that is SEQ ID NO. 35 or 36. In some embodiments, the LCDR2 has a sequence that is SEQ ID NO.37. In some embodiments, the LCDR3 has a sequence that is any one of SEQ ID NO. 38-40. In some embodiments, the VH domain comprises an HCDR1206, HCDR2207, and HCDR3208. In some embodiments, the HCDR1 has a sequence that is any one of SEQ ID NO.43-46. In some embodiments, the HCDR2 has a sequence that is any one of SEQ ID NO.47-48, 119, or 136. In some embodiments, the HCDR3 has a sequence that is SEQ ID NO. 49 or 50. In some embodiments, the scFv comprises a hinge-extension domain comprising a hinge region. In some embodiments, the hinge region comprises an upper hinge, a core hinge, and a lower hinge region. In some embodiments, the upper hinge comprises a sequence that is SEQ ID NO. 51 or 52. In some embodiments, the core hinge comprises a sequence that is SEQ ID NO.53. In some embodiments, the lower hinge region comprises a sequence that is SEQ ID NO. 54. In some embodiments, the scFv comprises a linker 209. In some embodiments, the linkercomprises a sequence that is SEQ ID NO. 55. In some embodiments, the scFv comprises a signal peptide. In some embodiments, the signal peptide comprises a sequence that is SEQ ID NO. 56. In some embodiments, the scFv further comprise a payload. In some embodiments, the payload comprises a therapeutic agent, toxin, and / or a detectable marker. In some embodiments, the detectable marker is a radio label. In some embodiments, the scFv is administered to a subject having one or more diseases. In some embodiments, the disease is a disease of the blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, administration of the scFv reduces or maintains the severity of one or more symptoms of the disease. In some embodiments, administration of the scFv eliminates on or more symptoms of the disease. In some embodiments, the scFv is administered as palliative care.

[0168] Some embodiments herein are directed to a single-chain variable fragment (scFv) having altered biodistribution. In some embodiments, the scFv having altered biodistribution is a ĮVȕ6 binding scFv. In some embodiments, the scFv comprises a variable light (VL) domain comprising a framework region (LFR). In some embodiments, the VL domain comprises a LFR 1, 2, 3, and 4. In some embodiments, the LFR2 domain comprises a sequence that is SEQ ID NO.41 or 42. In some embodiments, the LFR2 domain comprises a sequence that is X1X2X3X4X5X6X7(SEQ ID NO. 81), wherein X2, X3, X5, and X6 are each independently a negatively charged or a non-charged amino acid, and X1, X4, and X7 are each a lysine (K). In some embodiments, X1, X4, and / or X7, are substituted (e.g., is not K). In some embodiments, substitution of X1, X4, and / or X7, in the LFR2 domain alter distribution of the scFv to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the scFv comprises a linker. In some embodiments, the linker comprises a sequence that is SEQ ID NO.55. In some embodiments, the scFv comprises a signal peptide. In some embodiments, the signal peptide comprises a sequence that is SEQ ID NO. 56. In some embodiments, the scFv is administered to a subject having one or more diseases. In some embodiments, the disease is a disease of the blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the scFv further comprise a payload. In someembodiments, the payload comprises a therapeutic agent, toxin, and / or a detectable marker. In some embodiments, the detectable marker is a radio label. In some embodiments, administration of the scFv reduces or maintains the severity of one or more symptoms of the disease. In some embodiments, administration of the scFv eliminates on or more symptoms of the disease. In some embodiments, the scFv is administered as palliative care.

[0169] FIG.3 is an illustrative representation of some embodiments of a minibody. Some embodiments herein are directed to a minibody 300. In some embodiments, the minibody is a ĮVȕ6 binding minibody. In some embodiments, the minibody 300 comprises: a variable light (VL) domain 301 and a variable heavy (VH) domain 302. In some embodiments, the minibody 300 comprises a scFv comprising the VL and VH domains. In some embodiments, the VL domain is N-terminal to the VH domain in the minibody or the scFv portion thereof. In some embodiments, the VH domain is N-terminal to the VL domain in the minibody or the scFv portion thereof. In some embodiments, the VL domain comprises a sequence that is any one of SEQ ID NO.2-17. In some embodiments, the VH domain comprises a sequence that is any one of SEQ ID NO. 19-34, 139 or 140. In some embodiments the VL and VH domains comprise one or more CDRs 303. In some embodiments, the VL domain comprises an LCDR1 304, LCDR2305, and LCDR3306. In some embodiments, the LCDR1 has a sequence that is SEQ ID NO.35 or 36. In some embodiments, the LCDR2 has a sequence that is SEQ ID NO. 37. In some embodiments, the LCDR3 has a sequence that is any one of SEQ ID NO. 38-40. In some embodiments, the VH domain comprises an HCDR1307, HCDR2308, and HCDR3 309. In some embodiments, the HCDR1 has a sequence that is any one of SEQ ID NO.43-46. In some embodiments, the HCDR2 has a sequence that is any one of SEQ ID NO.47-48, 119, or 136. In some embodiments, the HCDR3 has a sequence that is SEQ ID NO. 49 or 50. In some embodiments, the antibody or antigen binding fragment thereof comprises a hinge- extension domain 310 comprising a hinge region. In some embodiments, the hinge region comprises an upper hinge, a core hinge, and a lower hinge region. In some embodiments, the upper hinge comprises a sequence that is SEQ ID NO. 51 or 52. In some embodiments, the core hinge comprises a sequence that is SEQ ID NO. 53. In some embodiments, the lower hinge region comprises a sequence that is SEQ ID NO. 54. In some embodiments, the minibody comprises a linker. In some embodiments, the linker comprises a sequence that is SEQ ID NO. 55. In some embodiments, the minibody comprises a signal peptide. In someembodiments, the signal peptide comprises a sequence that is SEQ ID NO. 56. In some embodiments, the minibody comprises an IgG CH3 domain 311. In some embodiments, the CH3 domain comprises any one of SEQ ID NO. 77-80. In some embodiments, the CH3 domain comprises one or more substitutions. In some embodiments, the one or more substitutions in the CH3 domain alters biodistribution of the minibody. In some embodiments, the one or more substitutions in the CH3 domain alters distribution of the minibody to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the minibody further comprise a payload. In some embodiments, the payload comprises a therapeutic agent, toxin, and / or a detectable marker. In some embodiments, the detectable marker is a radio label. In some embodiments, the minibody is administered to a subject having one or more diseases. In some embodiments, the disease is a disease of the blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, administration of the minibody reduces or maintains the severity of one or more symptoms of the disease. In some embodiments, administration of the minibody eliminates on or more symptoms of the disease. In some embodiments, the minibody is administered as palliative care.

[0170] Some embodiments herein are directed to a minibody having altered biodistribution. In some embodiments, the minibody having altered biodistribution is a ĮVȕ6 binding minibody. In some embodiments, the minibody comprises a variable light (VL) domain comprising a framework region (LFR). In some embodiments, the VL domain comprises a LFR 1, 2, 3, and 4. In some embodiments, the LFR2 domain comprises a sequence that is SEQ ID NO.41 or 42. In some embodiments, the LFR2 domain comprises a sequence that is X1X2X3X4X5X6X7(SEQ ID NO.81), wherein X2, X3, X5, and X6are each independently a negatively charged or a non-charged amino acid, and X1, X4, and X7 are each a lysine (K). In some embodiments, X1, X4, and / or X7, are substituted (e.g., is not K). In some embodiments, substitution of X1, X4, and / or X7, in the LFR2 domain alter distribution of the minibody to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the minibody comprises a hinge-extension domain comprising a hingeregion. In some embodiments, the hinge region comprises an upper hinge, a core hinge, and a lower hinge region. In some embodiments, the upper hinge comprises a sequence that is SEQ ID NO. 51 or 52. In some embodiments, the core hinge comprises a sequence that is SEQ ID NO. 53. In some embodiments, the lower hinge region comprises a sequence that is SEQ ID NO. 54. In some embodiments, the minibody comprises a linker. In some embodiments, the linker comprises a sequence that is SEQ ID NO. 55. In some embodiments, the minibody comprises a signal peptide. In some embodiments, the signal peptide comprises a sequence that is SEQ ID NO.56. In some embodiments, the minibody comprises an IgG CH3 domain. In some embodiments, the CH3 domain comprises any one of SEQ ID NO. 77-80. In some embodiments, the CH3 domain comprises one or more substitutions. In some embodiments, the one or more substitutions in the CH3 domain alters biodistribution of the minibody. In some embodiments, the one or more substitutions in the CH3 domain alter distribution of the minibody to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the minibody further comprise a payload. In some embodiments, the payload comprises a therapeutic agent, toxin, and / or a detectable marker. In some embodiments, the detectable marker is a radio label. In some embodiments, the minibody is administered to a subject having one or more diseases. In some embodiments, the disease is a disease of the blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, administration of the minibody reduces or maintains the severity of one or more symptoms of the disease. In some embodiments, administration of the minibody eliminates on or more symptoms of the disease.

[0171] FIG. 4 is an illustrative representation of some embodiments of a single- chain variable fragment (scFv) Fc fusion (scFv-Fc). Some embodiments herein are directed to a single-chain variable fragment Fc fusion (scFv-Fc) 400. In some embodiments, the scFv-Fc fusion is a ĮVȕ6 binding scFv-Fc fusion. In some embodiments, the scFv-Fc fusion comprises a scFv 401 that comprises a variable light (VL) domain 402 and a variable heavy (VH) domain 403. In some embodiments, the VL domain is N-terminal to the VH domain in the scFv. In some embodiments, the VH domain is N-terminal to the VL domain in the scFv. In some embodiments, the VL domain comprises a sequence that is any one of SEQ ID NO. 1-17. Insome embodiments, the VH domain comprises a sequence that is any one of SEQ ID NO.18- 34, 139 or 140. In some embodiments the VL and VH domains comprise one or more CDRs 404. In some embodiments, the VL domain comprises an LCDR1 405, LCDR2 406, and LCDR3 407. In some embodiments, the LCDR1 has a sequence that is SEQ ID NO.35 or 36. In some embodiments, the LCDR2 has a sequence that is SEQ ID NO. 37. In some embodiments, the LCDR3 has a sequence that is any one of SEQ ID NO. 38-40. In some embodiments, the VH domain comprises an HCDR1408, HCDR2409, and HCDR3410. In some embodiments, the HCDR1 has a sequence that is any one of SEQ ID NO.43-46. In some embodiments, the HCDR2 has a sequence that is any one of SEQ ID NO.47-48, 119, or 136. In some embodiments, the HCDR3 has a sequence that is SEQ ID NO. 49 or 50. In some embodiments, the scFv-Fc fusion comprises a hinge-extension domain 411 comprising a hinge region. In some embodiments, the hinge region comprises an upper hinge, a core hinge, and a lower hinge region. In some embodiments, the upper hinge comprises a sequence that is SEQ ID NO.51 or 52. In some embodiments, the core hinge comprises a sequence that is SEQ ID NO. 53. In some embodiments, the lower hinge region comprises a sequence that is SEQ ID NO. 54. In some embodiments, the scFv-Fc fusion comprises a linker 412. In some embodiments, the linker comprises a sequence that is SEQ ID NO.55. In some embodiments, the scFv-Fc fusion comprises a signal peptide. In some embodiments, the signal peptide comprises a sequence that is SEQ ID NO. 56. In some embodiments, the scFv-Fc fusion comprises a Fc domain 413. In some embodiments, the scFv-Fc fusion comprises a Fc domain comprising a sequence that is any one of SEQ ID NO. 83-102. In some embodiments, the Fc domain comprises one or more substitutions, e.g., relative to a reference sequence, such as SEQ ID NO:83. In some embodiments, the one or more substitutions in the Fc domain alters biodistribution of the scFv-Fc fusion, e.g., relative to the reference sequence. In some embodiments, the one or more substitutions in the Fc domain alters distribution of the scFv-Fc fusion to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof, e.g., relative to the reference sequence. In some embodiments, the scFv-Fc fusion comprises an IgG CH2 domain 414. In some embodiments, the CH2 domain comprises a sequence that is any one of SEQ ID NO.47-76. In some embodiments, the scFv-Fc fusion comprises an IgG CH3 domain 415. In some embodiments, the CH3 domain comprises any one of SEQ ID NO.77-80. In some embodiments, the CH2 and / or CH3 domain comprises one or more substitutions. In some embodiments, the one or more substitutions in the CH2 and / or CH3 domain alters biodistribution of the antibody or antigen binding fragment thereof. In some embodiments, the one or more substitutions in the CH2 and / or CH3 domain alter distribution of the scFv-Fc fusion to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. Some embodiments herein are directed to a single-chain variable fragment Fc fusion (scFv-Fc fusion) having altered biodistribution. In some embodiments, the scFv-Fc fusion having altered biodistribution is a ĮVȕ6 binding scFv-Fc fusion. In some embodiments, the scFv-Fc fusion comprises a variable light (VL) domain comprising a framework region (LFR). In some embodiments, the VL domain comprises a LFR 1, 2, 3, and 4. In some embodiments, the LFR2 domain comprises a sequence that is SEQ ID NO. 41 or 42. In some embodiments, the LFR2 domain comprises a sequence that is X1X2X3X4X5X6X7(SEQ ID NO.81), wherein X2, X3, X5, and X6are each independently a negatively charged or a non-charged amino acid, and X1, X4, and X7are each a lysine (K). In some embodiments, X1, X4, and / or X7, are substituted (e.g., is not K). In some embodiments, substitution of X1, X4, and / or X7, in the LFR2 domain alter distribution of the scFv-Fc fusion to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the scFv-Fc fusion comprises a hinge-extension domain comprising a hinge region. In some embodiments, the hinge region comprises an upper hinge, a core hinge, and a lower hinge region. In some embodiments, the upper hinge comprises a sequence that is SEQ ID NO.51 or 52. In some embodiments, the core hinge comprises a sequence that is SEQ ID NO. 53. In some embodiments, the lower hinge region comprises a sequence that is SEQ ID NO.54. In some embodiments, the scFv-Fc fusion comprises a linker. In some embodiments, the linker comprises a sequence that is SEQ ID NO. 55. In some embodiments, the scFv-Fc fusion comprises a signal peptide. In some embodiments, the signal peptide comprises a sequence that is SEQ ID NO.56. In some embodiments, the scFv-Fc fusion comprises an IgG CH2 domain. In some embodiments, the CH2 domain comprises a sequence that is any one of SEQ ID NO.47-76. In some embodiments, the scFv-Fc fusion comprises an IgG CH3 domain. In some embodiments, the CH3 domain comprises any one of SEQ ID NO. 77-80. In someembodiments, the CH2 and / or CH3 domain comprises one or more substitutions. In some embodiments, the one or more substitutions in the CH2 and / or CH3 domain alters biodistribution of the antibody or antigen binding fragment thereof. In some embodiments, the one or more substitutions in the CH2 and / or CH3 domain alter distribution of the scFv-Fc fusion to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the scFv-Fc fusion comprises a Fc domain. In some embodiments, the scFv-Fc fusion comprises a Fc domain having a sequence that is any one of SEQ ID NO. 83- 102. In some embodiments, the Fc domain comprises one or more substitutions. In some embodiments, the one or more substitutions in the Fc domain alters biodistribution of the scFv- Fc fusion. In some embodiments, the one or more substitutions in the Fc domain alters distribution of the scFv-Fc fusion to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the scFv-Fc fusion further comprise a payload. In some embodiments, the payload comprises a therapeutic agent, toxin, and / or a detectable marker. In some embodiments, the detectable marker is a radio label. In some embodiments, the scFv-Fc fusion is administered to a subject having one or more diseases. In some embodiments, the disease is a disease of the blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, administration of the scFv-Fc fusion reduces or maintains the severity of one or more symptoms of the disease. In some embodiments, administration of the scFv-Fc fusion eliminates on or more symptoms of the disease. In some embodiments, the scFv-Fc fusion is administered as palliative care.

[0172] FIG. 5 is flow chart illustrating some embodiments of a method for reducing tumor size. Some embodiments herein are directed to a method of reducing tumor size in a subject 500. In some embodiments, the method comprises administering an antibody or antigen binding fragment thereof to a subject having a tumor 501. In some embodiments, the antibody or antigen binding fragment thereof is an ĮVȕ6 antibody or ĮVȕ6binding fragment thereof. In some embodiments, the antibody or antigen binding fragment thereof comprises: a variable light (VL) domain and a variable heavy (VH) domain. In some embodiments, the VL domain comprises a sequence that is any one of SEQ ID NO. 1-17. Insome embodiments, the VH domain comprises a sequence that is any one of SEQ ID NO.18- 34, 139 or 140. In some embodiments the VL and VH domains comprise one or more CDRs. In some embodiments, the VL domain comprises an LCDR1, LCDR2, and LCDR3. In some embodiments, the LCDR1 has a sequence that is SEQ ID NO.35 or 36. In some embodiments, the LCDR2 has a sequence that is SEQ ID NO. 37. In some embodiments, the LCDR3 has a sequence that is any one of SEQ ID NO. 38-40. In some embodiments, the VL domain comprises a framework region (LFR). In some embodiments, the VL domain comprises a LFR 1, 2, 3, and 4. In some embodiments, the LFR2 domain comprises a sequence that is SEQ ID NO. 41 or 42. In some embodiments, the LFR2 domain comprises a sequence that is X1X2X3X4X5X6X7(SEQ ID NO. 81), wherein X2, X3, X5, and X6 are each independently a negatively charged or a non-charged amino acid, and X1, X4, and X7 are each a lysine (K). In some embodiments, X1, X4, and / or X7, are substituted (e.g., is not K). In some embodiments, substitution of X1, X4, and / or X7, in the LFR2 domain alter distribution of the antibody or antigen binding fragment thereof to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the VH domain comprises an HCDR1, HCDR2, and HCDR3. In some embodiments, the HCDR1 has a sequence that is any one of SEQ ID NO.43-46. In some embodiments, the HCDR2 has a sequence that is any one of SEQ ID NO.47-48, 119, or 136. In some embodiments, the HCDR3 has a sequence that is SEQ ID NO. 49 or 50. In some embodiments, the antibody or antigen binding fragment thereof comprises a hinge-extension domain comprising a hinge region. In some embodiments, the hinge region comprises an upper hinge, a core hinge, and a lower hinge region. In some embodiments, the upper hinge comprises a sequence that is SEQ ID NO. 51 or 52. In some embodiments, the core hinge comprises a sequence that is SEQ ID NO. 53. In some embodiments, the lower hinge region comprises a sequence that is SEQ ID NO. 54. In some embodiments, the antibody or antigen binding fragment thereof comprises a linker. In some embodiments, the linker comprises a sequence that is SEQ ID NO.55. In some embodiments, the antibody or antigen binding fragment thereof comprises a signal peptide. In some embodiments, the signal peptide comprises a sequence that is SEQ ID NO. 56. In some embodiments, the antibody or antigen binding fragment thereof comprises an IgG CH2 domain. In some embodiments, the CH2 domain comprises a sequence that is any one of SEQID NO. 47-76. In some embodiments, the antibody or antigen binding fragment thereof comprises an IgG CH3 domain. In some embodiments, the CH3 domain comprises any one of SEQ ID NO. 77-80. In some embodiments, the CH2 and / or CH3 domain comprises one or more substitutions. In some embodiments, the one or more substitutions in the CH2 and / or CH3 domain alters biodistribution of the antibody or antigen binding fragment thereof. In some embodiments, the one or more substitutions in the CH2 and / or CH3 domain alter distribution of the antibody or antigen binding fragment thereof to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the antibody or antigen binding fragment thereof comprises a Fc domain. In some embodiments, the antibody or antigen binding fragment thereof comprises a Fc domain comprises a sequence that is any one of SEQ ID NO. 83-102. In some embodiments, the Fc domain comprises one or more substitutions. In some embodiments, the one or more substitutions in the Fc domain alters biodistribution of the antibody or antigen binding fragment thereof. In some embodiments, the one or more substitutions in the Fc domain alters distribution of the antibody or antigen binding fragment thereof to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the one or more substitutions in the Fc domain alters the ratio of biodistribution of the antibody or antigen binding fragment thereof in a subject’s tumor as compared to biodistribution of the antibody or antigen binding fragment thereof to the subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, administration of the antibody or antigen binding fragment thereof reduces tumor size as compared to tumor size in the subject prior to administration of the antibody or antigen binding fragment thereof. In some embodiments, reduction in tumor size reduces or maintains the severity of one or more symptoms of the disease. In some embodiments, reduction of tumor size eliminates on or more symptoms of the disease.

[0173] FIG.6 is flow chart illustrating some embodiments of a method of treating cancer. Some embodiments herein are directed to a method of treating cancer 600. In some embodiments, the method comprises administering an antibody or antigen binding fragmentthereof to a subject having a tumor 601. In some embodiments, the antibody or antigen binding fragment thereof is an ĮVȕ6 antibody or ĮVȕ6 binding fragment thereof. In some embodiments, the antibody or antigen binding fragment thereof comprises: a variable light (VL) domain and a variable heavy (VH) domain. In some embodiments, the VL domain comprises a sequence that is any one of SEQ ID NO. 1-17. In some embodiments, the VH domain comprises a sequence that is any one of SEQ ID NO. 18-34, 139 or 140. In some embodiments the VL and VH domains comprise one or more CDRs. In some embodiments, the VL domain comprises an LCDR1, LCDR2, and LCDR3. In some embodiments, the LCDR1 has a sequence that is SEQ ID NO.35 or 36. In some embodiments, the LCDR2 has a sequence that is SEQ ID NO. 37. In some embodiments, the LCDR3 has a sequence that is any one of SEQ ID NO.38-40. In some embodiments, the VL domain comprises a framework region (LFR). In some embodiments, the VL domain comprises a LFR 1, 2, 3, and 4. In some embodiments, the LFR2 domain comprises a sequence that is SEQ ID NO. 41 or 42. In some embodiments, the LFR2 domain comprises a sequence that is X1X2X3X4X5X6X7(SEQ ID NO. 81), wherein X2, X3, X5, and X6are each independently a negatively charged or a non-charged amino acid, and X1, X4, and X7are each a lysine (K). In some embodiments, X1, X4, and / or X7, are substituted (e.g., is not K). In some embodiments, substitution of X1, X4, and / or X7, in the LFR2 domain alter distribution of the antibody or antigen binding fragment thereof to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the VH domain comprises an HCDR1, HCDR2, and HCDR3. In some embodiments, the HCDR1 has a sequence that is any one of SEQ ID NO. 43-46. In some embodiments, the HCDR2 has a sequence that is any one of SEQ ID NO.47-48, 119, or 136. In some embodiments, the HCDR3 has a sequence that is SEQ ID NO. 49 or 50. In some embodiments, the antibody or antigen binding fragment thereof comprises a hinge-extension domain comprising a hinge region. In some embodiments, the hinge region comprises an upper hinge, a core hinge, and a lower hinge region. In some embodiments, the upper hinge comprises a sequence that is SEQ ID NO. 51 or 52. In some embodiments, the core hinge comprises a sequence that is SEQ ID NO. 53. In some embodiments, the lower hinge region comprises a sequence that is SEQ ID NO. 54. In some embodiments, the antibody or antigen binding fragment thereof comprises a linker. In some embodiments, the linker comprises a sequencethat is SEQ ID NO. 55. In some embodiments, the antibody or antigen binding fragment thereof comprises a signal peptide. In some embodiments, the signal peptide comprises a sequence that is SEQ ID NO. 56. In some embodiments, the antibody or antigen binding fragment thereof comprises an IgG CH2 domain. In some embodiments, the CH2 domain comprises a sequence that is any one of SEQ ID NO. 47-76. In some embodiments, the antibody or antigen binding fragment thereof comprises an IgG CH3 domain. In some embodiments, the CH3 domain comprises any one of SEQ ID NO. 77-80. In some embodiments, the CH2 and / or CH3 domain comprises one or more substitutions. In some embodiments, the one or more substitutions in the CH2 and / or CH3 domain alters biodistribution of the antibody or antigen binding fragment thereof. In some embodiments, the one or more substitutions in the CH2 and / or CH3 domain alter distribution of the antibody or antigen binding fragment thereof to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the antibody or antigen binding fragment thereof comprises a Fc domain. In some embodiments, the antibody or antigen binding fragment thereof comprises a Fc domain comprises a sequence that is any one of SEQ ID NO.83-102. In some embodiments, the Fc domain comprises one or more substitutions. In some embodiments, the one or more substitutions in the Fc domain alters biodistribution of the antibody or antigen binding fragment thereof. In some embodiments, the one or more substitutions in the Fc domain alters distribution of the antibody or antigen binding fragment thereof to a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the one or more substitutions in the Fc domain alters the ratio of biodistribution of the antibody or antigen binding fragment thereof in a subject’s tumor as compared to biodistribution of the antibody or antigen binding fragment thereof to the subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, administration of the antibody or antigen binding fragment thereof treats the subject’s cancer. In some embodiments, treating the subject’s cancer comprises reducing and / or eliminating one or more symptoms of the subject’s cancer, increasing the subject’s probability of survival, and / or reducing tumor size, as compared to the severity or presence ofone or more symptoms, tumor size, and / or the subject’s probability of survival prior to administration of the antibody or antigen binding fragment thereof. In some embodiments, administration of the antibody or antigen binding fragment thereof reduces tumor size as compared to tumor size in the subject prior to administration of the antibody or antigen binding fragment thereof. In some embodiments, administration of the antibody or antigen binding fragment thereof reduces or maintains the severity of one or more symptoms of the disease. In some embodiments, administration of the antibody or antigen binding fragment thereof eliminates on or more symptoms of the disease. In some embodiments, the administration of the antibody or antigen binding fragment thereof is administered as palliative care.

[0174] In some embodiments, the ĮVȕ6 antibody or antigen binding fragment thereof comprises a VL domain having a sequence that is any one of SEQ ID NO. 1-17. In some embodiments, the VL domain comprises a sequence having, having about, or having at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identity to any one of SEQ ID NO.1-17, or a percentage identity that is in a range defined by any two of the preceding values. For example, in some embodiments, the VL domain comprises a sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%- 90%, 70%-80%, 70%-75%, 57%-100%, 75%-95%, 75%-90%, 75%-80%, 80%-100%, 80%- 95%, 80%-90%, 90%-100%, 90%-95%, or 95%-100% identity to any one of SEQ ID NO. 1- 17. In some embodiments, the ĮVȕ6 antibody or antigen binding fragment thereof comprises a LFR2 having SEQ ID NO. 41 or 42. In some embodiments, the ĮVȕ6 antibody or antigen binding fragment thereof comprises a VL domain comprising the polypeptide sequence of X1X2X3X4X5X6X7(SEQ ID NO. 81), wherein X1is a positively charged amino acid, wherein X2, X3, X5, and X6are each independently a negatively charged or a non-charged amino acid, and wherein X4and X7are each independently any amino acid with the proviso that at least one is a positively charged amino acid, wherein the sequence is outside of any CDR of the antibody or antigen binding fragment thereof. In some embodiments, the sequence is within a VL framework region (FR) of the antibody. In some embodiments, the sequence is within a LFR2 of the antibody. In some embodiments, the ĮVȕ6 antibody or antigen binding fragment thereof comprises a LFR2 having SEQ ID NO. 41 or 42. In some embodiments, the ĮVȕ6 antibody or antigen binding fragment thereof comprises a LFR2 in Table 2. In someembodiments, the ĮVȕ6 antibody or antigen binding fragment thereof comprises a LFR in Table 2. In some embodiments, one or more mutations in the LFR2 alter biodistribution of the antibody or antigen binding fragment thereof in a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the VL is based on a human germline sequence of IGKV1-39*01. In some embodiments, the VL is based on a human germline sequence of IGKJ4*01.

[0175] Table 1 depicts some embodiments of VL domain sequences disclosed herein. In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys- diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable light (VL) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:1, or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:1 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable light (VL) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:2, or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:2 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%- 95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable light (VL) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:3, or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:3 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variablelight (VL) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:4, or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:4 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%- 100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable light (VL) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:5, or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:5 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%- 95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable light (VL) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:6, or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:6 in a range defined by any two of the preceding values (e.g., 70%- 100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv- Fc fusion, nanobody®-Fc) includes a variable light (VL) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:7, or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:7 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%- 100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable light (VL) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:8, or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:8 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%- 95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable light (VL) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:9, or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:9 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable light (VL) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:10, or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:10 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable light (VL) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:11, or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:11 in a range defined by any two of the preceding values (e.g., 70%- 100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv- Fc fusion, nanobody®-Fc) includes a variable light (VL) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:12, or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:12 in a range defined by any two of thepreceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%- 100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable light (VL) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:13, oroptionally the amino acid sequence has percent sequence identity to SEQ ID NO:13 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable light (VL) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:14, or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:14 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable light (VL) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:15, or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:15 in a range defined by any two of the preceding values (e.g., 70%- 100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv- Fc fusion, nanobody®-Fc) includes a variable light (VL) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:16, or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:16 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%- 100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody,cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable light (VL) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:16, or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:17 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%- 95%, 90%-100%, 95%-100%, etc.). Table 1

[0176] Table 2 depicts some embodiments of LFR2 sequences disclosed herein. Table 2.

[0177] In some embodiments, the ĮVȕ6 antibody or antigen binding fragment thereof comprises a VL domain having a sequence in Table 1. In some embodiments, the VL domain comprises a sequence having, having about, or having at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identity to a VL domain sequence in Table 1, or a percentage identity that is in a range defined by any two of the preceding values. For example, in some embodiments, the VL domain comprises a sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 70%- 75%, 57%-100%, 75%-95%, 75%-90%, 75%-80%, 80%-100%, 80%-95%, 80%-90%, 90%- 100%, 90%-95%, or 95%-100% identity to a VL domain sequence in Table 1. In some embodiments, the ĮVȕ6 antibody or antigen binding fragment thereof comprises a LFR2 having SEQ ID NO. 41 or 42. In some embodiments, the VL is based on a human germline sequence of IGKV1-39*01. In some embodiments, the VL is based on a human germline sequence of IGKJ4*01.

[0178] FIG. 7 depicts an alignment of some embodiments of ĮVȕ6 VL domain sequences disclosed herein. In some embodiments, the ĮVȕ6 antibody or antigen binding fragment thereof comprises a VL domain having a VL domain sequence in FIG. 7. In some embodiments, the ĮVȕ6 antibody or antigen binding fragment thereof comprises a VL domain having, having about, or having at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identity to a VL domain sequence in FIG. 7, or has a percentage identity that is in a range defined by any two of the preceding values. For example, in some embodiments, the ĮVȕ6 antibody or antigen binding fragment thereof comprises a VL domain having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%,70%-75%, 57%-100%, 75%-95%, 75%-90%, 75%-80%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-95%, or 95%-100% identity to a VL domain sequence in FIG. 7. In some embodiments, the antibody or antigen binding fragment thereof comprises a LFR2 having SEQ ID NO. 41 or 42. In some embodiments, the VL is based on a human germline sequence of IGKV1-39*01. In some embodiments, the VL is based on a human germline sequence of IGKJ4*01.

[0179] In some embodiments, the VL domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VL domain in SEQ ID NO. 9, or a percentage identity that is in a range defined by any two of the preceding values. For example, in some embodiments, the VL domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VL domain in SEQ ID NO.9. In some embodiments, the antibody or antigen binding fragment thereof comprises a LFR2 having SEQ ID NO. 41 or 42. In some embodiments, the VL is based on a human germline sequence of IGKV1-39*01. In some embodiments, the VL is based on a human germline sequence of IGKJ4*01.

[0180] In some embodiments, the antibody or antigen binding fragment thereof comprises a VL domain comprising the polypeptide sequence X1X2X3X4X5X6X7(SEQ ID NO. 81), wherein X1is a positively charged amino acid, wherein X2, X3, X5, and X6are each independently a negatively charged or a non-charged amino acid, and wherein X4and X7are each independently any amino acid with the proviso that at least one is a positively charged amino acid. In some embodiments, X1is lysine (K), arginine (R), or histidine (H). In some embodiments, X2, X3, X5, and X6 are each independently a glycine (G), alanine (A), valine (V), cysteine (C), proline (P), leucine (L), isoleucine (I), methionine (M), tryptophan (W), phenylalanine (F), serine (S), threonine (T), tyrosine (Y), asparagine (N), glutamine (Q), aspartic acid (D), or glutamic acid (E). In some embodiments, the sequence is within a VL framework region (FR) of the antibody. In some embodiments, X4and X7are each independently a lysine (K), arginine (R), histidine (H), glycine (G), alanine (A), valine (V),cysteine (C), proline (P), leucine (L), isoleucine (I), methionine (M), tryptophan (W), phenylalanine (F), serine (S), threonine (T), tyrosine (Y), asparagine (N), glutamine (Q), aspartic acid (D), or glutamic acid (E) with the proviso that at least one is lysine (K), arginine (R), or histidine (H). In some embodiments, X1, X4, and X7, are lysine (K). In some embodiments, the antibody or antigen binding fragment thereof comprises a VL domain comprising the polypeptide sequence X1X2X3X4X5X6X7(SEQ ID NO. 81), wherein X1, X4, and X7are each a lysine (K) (SEQ ID NO.82). In some embodiments, the sequence is outside of any CDR of the antibody or antigen binding fragment thereof. In some embodiments, the sequence is within a LFR2 of the antibody or antigen binding fragment thereof. In some embodiments, the sequence is within a LFR2 of a ĮVȕ6 antibody or antigen binding fragment thereof. In some embodiments, one or more mutations in the LFR2 alter biodistribution of the antibody or antigen binding fragment thereof in a subject’s blood, lungs, muscle, bone (bone marrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination thereof. In some embodiments, the sequence is in a LFR2 having SEQ ID NO.41 or 42. In some embodiments, the sequence is in an antibody or antigen binding fragment thereof comprising a LFR2 in Table 2. In some embodiments, the sequence is in a VL is based on a human germline sequence of IGKV1-39*01. In some embodiments, the sequence is in a VL is based on a human germline sequence of IGKJ4*01.

[0181] In some embodiments, the VL domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VL domain in SEQ ID NO. 1, wherein the VL domain comprises a K47X and / or K53X mutation (Aho), wherein X is any non-lysine (and non- arginine) residue. For example, in some embodiments, the VL domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%- 80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%- 100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%- 97%, or 97%-100%, identity to a VL domain in SEQ ID NO. 1, wherein the VL domain comprises a K47X and / or K53X mutation (Aho), wherein X is any non-lysine (and non- arginine) residue. In some embodiments, X is a histidine (H), glycine (G), alanine (A), valine (V), cysteine (C), proline (P), leucine (L), isoleucine (I), methionine (M), tryptophan (W),phenylalanine (F), serine (S), threonine (T), tyrosine (Y), asparagine (N), glutamine (Q), aspartic acid (D), or glutamic acid (E). In some embodiments, the VL domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VL domain in SEQ ID NO. 1, wherein the VL domain comprises a non-lysine and non-arginine residue at position 47 and / or 53 (Aho). In some embodiments, the VL domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VL domain in SEQ ID NO. 1, wherein the VL domain comprises a glutamine (Q) at position 47 and / or 53 (Aho). In any antigen-binding construct herein, in some embodiments, the numbering of an amino acid residue within the VL domain can be represented using an alternative numbering convention than Aho. For example, in some embodiments, positions 47, 50, and 53 of the VL domain according to Aho numbering corresponds to positions 45, 48, and 51, respectively, according to IMGT numbering. In some embodiments, the sequence is outside of any CDR of the antibody or antigen binding construct thereof.

[0182] In some embodiments, the VL domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VL domain in SEQ ID NO. 1, wherein the VL domain comprises a K53Q mutation (Aho). For example, in some embodiments, the VL domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%- 95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%- 95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%- 95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VL domain in SEQ ID NO. 1, wherein the VL domain comprises a K53Q mutation (Aho).

[0183] In some embodiments, the VL domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VL domain in SEQ ID NO. 9, wherein the VL domaincomprises a Q to K substitution at a position corresponding to residue number 46 and / or residue number 49 of SEQ ID NO.9. For example, in some embodiments, the VL domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%- 90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%- 90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%- 100%, 95%-97%, or 97%-100%, identity to a VL domain in SEQ ID NO. 9, wherein the VL domain comprises a Q to K substitution at a position corresponding to residue number 46 and / or residue number 49 of SEQ ID NO. 9.

[0184] In some embodiments, the VL domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VL domain in SEQ ID NO. 1 wherein the VL domain comprises a E29Q mutation (Aho). For example, in some embodiments, the VL domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%- 95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%- 95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%- 95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VL domain in SEQ ID NO. 1, wherein the VL domain comprises a E29Q mutation (Aho).

[0185] In some embodiments, the VL domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VL domain in SEQ ID NO. 9, wherein the VL domain comprises an E to Q substitution at a position corresponding to residue number 27 of SEQ ID NO.9. For example, in some embodiments, the VL domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%- 100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%- 97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VL domain in SEQ ID NO.9, wherein the VL domain comprises an E to Q substitution at a position corresponding to residue number 46 and / or residue number 27 of SEQ ID NO.9.

[0186] In some embodiments, the VL domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VL domain in SEQ ID NO. 9, wherein the VL domain comprises a S to A substitution at a position corresponding to residue number 47 of SEQ ID NO.9. For example, in some embodiments, the VL domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%- 100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%- 97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VL domain in SEQ ID NO. 9, wherein the VL domain comprises a S to A substitution at a position corresponding to residue number 47 of SEQ ID NO.9.

[0187] In some embodiments, the VL domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VL domain in SEQ ID NO. 9, wherein the VL domain comprises a G to K substitution at a position corresponding to residue number 96 of SEQ ID NO.9. For example, in some embodiments, the VL domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%- 100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%- 97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VL domain in SEQ ID NO. 9, wherein the VL domain comprises a G to K substitution at a position corresponding to residue number 96 of SEQ ID NO. 9.

[0188] In some embodiments, the VL domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VL domain in SEQ ID NO. 1, wherein the VL domain comprises a E111S mutation (Aho). For example, in some embodiments, the VL domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%- 95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%- 95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VL domain in SEQ ID NO. 1, wherein the VL domain comprises a E111S mutation (Aho).

[0189] In some embodiments, the VL domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VL domain in SEQ ID NO. 9, wherein the VL domain comprises an E to S substitution at a position corresponding to residue number 97 of SEQ ID NO.9. For example, in some embodiments, the VL domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%- 100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%- 97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VL domain in SEQ ID NO.9, wherein the VL domain comprises an E to S substitution at a position corresponding to residue number 46 and / or residue number 97 of SEQ ID NO.9.

[0190] In some embodiments, the VL domain comprises any one or more of the VL domain mutations (e.g., any 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, or all of the non-overlapping VL domain mutations) provided herein.

[0191] In some embodiments, the antibody or antigen binding fragment thereof comprises a VL domain comprising an LCDR1, LCDR2, and LCDR3. In some embodiments, the LCDR1 is an LCDR1 having the sequence of SEQ ID NO.35 or 36. In some embodiments, the LCDR1 is an LCDR1 having the sequence of SEQ ID NO. 35 wherein the sequence comprises an E to Q substitution at a position corresponding to residue number 4 of SEQ ID NO.35. In some embodiments, the LCDR2 is an LCDR2 having the sequence of SEQ ID NO. 37. In some embodiments, the LCDR3 is an LCDR3 having the sequence of any one of SEQ ID NO. 38-40. In some embodiments, the LCDR3 is an LCDR3 having the sequence of SEQ ID NO.38, wherein the sequence comprises a K to G substitution at a position corresponding to residue 4 of SEQ ID NO. 38. In some embodiments, the LCDR3 is an LCDR3 having the sequence of SEQ ID NO. 38, wherein the sequence comprises a K to G substitution at a position corresponding to residue 4 and an E to S substitution at a position corresponding to residue 5 of SEQ ID NO. 38. In some embodiments, the LCDR3 is an LCDR3 having the sequence of SEQ ID NO. 39, wherein the sequence comprises an E to S substitution at aposition corresponding to residue 5 of SEQ ID NO. 39. Table 3 depicts some embodiments of LCDR sequences disclosed herein. In some embodiments, the antibody or antigen binding fragment thereof comprises a VL domain with an LCDR1 sequence that is an LCDR1 sequence in Table 3. In some embodiments, the antibody or antigen binding fragment thereof comprises a VL domain with an LCDR2 sequence that is an LCDR2 sequence in Table 3. In some embodiments, the antibody or antigen binding fragment thereof comprises a VL domain with an LCDR3 sequence that is an LCDR3 sequence in Table 3. Table 3

[0192] In some embodiments, the antibody or antigen binding fragment thereof comprises a variable light (VL) domain comprising a LCDR1 that is SEQ ID NO. 35; a LCDR2 that is SEQ ID NO.37; and a LCDR3 that is SEQ ID NO.39. In some embodiments, the antibody or antigen binding fragment thereof comprises a variable light (VL) domain that has a sequence with, with at least, or with about 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VL domain comprising: a LCDR1 that is SEQ ID NO.35; a LCDR2 that is SEQ ID NO.37; and a LCDR3 that is SEQ ID NO.39. For example, in some embodiments, the antibody or antigen binding fragment thereof comprises a VL domain that has a sequence with between about, or in a range of, 70%-100%, 70%-95%, 70%- 90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%- 90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%- 100%, 95%-97%, or 97%-100%, identity to a VL domain comprising: a LCDR1 that is SEQ ID NO.35; a LCDR2 that is SEQ ID NO.37; and a LCDR3 that is SEQ ID NO. 39.

[0193] Provided herein is an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) that includes a variable light (VL) domain that includes a LCDR1 that is an LCDR1 in any one of SEQ ID NOs:1-17, a LCDR2 that is an LCDR2 in any one of SEQ ID NOs:1-17, and a LCDR3 that is an LCDR3 in any one of SEQ ID NOs:1-17. In some embodiments, the LCDR1, LCDR2 and LCDR3 are combined as they are arranged in each one of SEQ ID NOs:1-17. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:1, a LCDR2 that is an LCDR2 in SEQ ID NO:1, and a LCDR3 that is an LCDR3 in SEQ ID NO:1. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:2 or 3, a LCDR2 that is an LCDR2 in SEQ ID NO:2 or 3, and a LCDR3 that is an LCDR3 in SEQ ID NO:2 or 3. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:4 or 5, a LCDR2 that is an LCDR2 in SEQ ID NO:4 or 5, and a LCDR3 that is an LCDR3 in SEQ ID NO:4 or 5. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:6 or 7, a LCDR2 that is an LCDR2 in SEQ ID NO:6 or 7, and a LCDR3 that is an LCDR3 in SEQ ID NO:6 or 7. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:8, a LCDR2 that is an LCDR2 in SEQ ID NO:8, and a LCDR3 that is an LCDR3 in SEQ ID NO:8. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:9, a LCDR2 that is an LCDR2 in SEQ ID NO:9, and a LCDR3 that is an LCDR3 in SEQ ID NO:9. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:10, a LCDR2 that is an LCDR2 in SEQ ID NO:10, and a LCDR3 that is an LCDR3 in SEQ ID NO:10. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:11, a LCDR2 that is an LCDR2 in SEQ ID NO:11, and a LCDR3 that is an LCDR3 in SEQ ID NO:11. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:12, a LCDR2 that is an LCDR2 in SEQ ID NO:12, and a LCDR3 that is an LCDR3 in SEQ ID NO:12. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:13, a LCDR2 that is an LCDR2 in SEQ ID NO:13, and a LCDR3 that is anLCDR3 in SEQ ID NO:13. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:14, a LCDR2 that is an LCDR2 in SEQ ID NO:14, and a LCDR3 that is an LCDR3 in SEQ ID NO:14. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:15, a LCDR2 that is an LCDR2 in SEQ ID NO:15, and a LCDR3 that is an LCDR3 in SEQ ID NO:15. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:16, a LCDR2 that is an LCDR2 in SEQ ID NO:16, and a LCDR3 that is an LCDR3 in SEQ ID NO:16. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:17, a LCDR2 that is an LCDR2 in SEQ ID NO:17, and a LCDR3 that is an LCDR3 in SEQ ID NO:17.

[0194] In some embodiments, the ĮVȕ6 antibody or antigen binding fragment thereof comprises a VH domain having a sequence that is any one of SEQ ID NO.18-34, 139 or 140. In some embodiments, the VH domain comprises a sequence having, having about, or having at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identity to any one of SEQ ID NO. 18-34, 139 or 140, or a percentage identity that is in a range defined by any two of the preceding values. For example, in some embodiments, the VH domain comprises a sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 70%-75%, 57%-100%, 75%-95%, 75%-90%, 75%-80%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-95%, or 95%-100% identity to any one of SEQ ID NO. 18-34, 139, or 140. In some embodiments, the VH is based on a human germline sequence of IGHV1-69*01. In some embodiments, the VH is based on a human germline sequence of IGHJ4*01.

[0195] FIG. 8 depicts an alignment of some embodiments of ĮVȕ6 VH domain sequences disclosed herein. In some embodiments, the ĮVȕ6 antibody or antigen binding fragment thereof comprises a VH domain having a sequence in FIG.8. In some embodiments, the VH domain comprises a sequence having, having about, or having at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identity to a VH domain having a sequence in FIG. 8, or a percentage identity that is in a range defined by any two ofthe preceding values. For example, in some embodiments, the VH domain comprises a sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 70%-75%, 57%-100%, 75%-95%, 75%-90%, 75%-80%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-95%, or 95%-100% identity to a VH domain having a sequence in FIG. 8. In some embodiments, the VH is based on a human germline sequence of IGHV1-69*01. In some embodiments, the VH is based on a human germline sequence of IGHJ4*01.

[0196] Table 4 depicts some embodiments of VH domain sequences disclosed herein. In some embodiments, the ĮVȕ6 antibody or antigen binding fragment thereof comprises a VH domain having a sequence in Table 4. In some embodiments, the VH domain comprises a sequence having, having about, or having at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identity to a VH domain sequence in Table 4, or a percentage identity that is in a range defined by any two of the preceding values. For example, in some embodiments, the VH domain comprises a sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 70%-75%, 57%- 100%, 75%-95%, 75%-90%, 75%-80%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%- 95%, or 95%-100% identity to a VH domain sequence in Table 4. In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:18,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:18 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:19,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:19 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:20,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:20 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%- 95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:21,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:21 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:22,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:22 in a range defined by any two of the preceding values (e.g., 70%- 100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv- Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:23,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:23 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%- 100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain thatincludes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:24,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:24 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%- 95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:25,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:25 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:26,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:26 in a range defined by any two of the preceding values (e.g., 70%- 100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv- Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:27,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:27 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%- 100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:28,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:28 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%- 95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:29,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:29 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:30,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:30 in a range defined by any two of the preceding values (e.g., 70%- 100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv- Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:31,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:31 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%- 100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:32,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:32 in a rangedefined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%- 95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:33,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:33 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:34,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:34 in a range defined by any two of the preceding values (e.g., 70%- 100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv- Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:139,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:139 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%- 100%, etc.). In some embodiments, an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a variable heavy (VH) domain that includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:140,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:140 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%- 95%, 90%-100%, 95%-100%, etc.).Table 4

[0197] Also provided herein is an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) that includes a variable light (VL) domain that includes an amino acid sequence of a VL in Table 1; and a variable heavy (VH) domain that includes an amino acid sequence of a VH in Table 4. In some embodiments, the VH sequence from Table 4 and the VL sequence from Table 1 are paired as they are combined in any one of the antigen binding constructs set forth in FIGs.13, 14, 15, 20, or 42.

[0198] In some embodiments, the antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a VL that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to any one of SEQ ID NOs:1-17,or optionally the amino acid sequence has percent sequence identity to the any one of SEQ ID NOs:1-17 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%- 100%, etc.); and a VH that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to any one of SEQ ID NOs:18-34, 139, or 140,or optionally the amino acid sequence has percent sequence identity to the any one of SEQ ID NOs: 18-34, 139, or 140 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, the VH sequence from Table 4 and the VL sequence from Table 1 are paired asthey are combined in any one of the antigen binding constructs set forth in FIGs. 13, 14, 15, 20, or 42.

[0199] In some embodiments, the antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a VL that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:1,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:1 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%- 95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.); and a VH that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:18,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO: 18 in a range defined by any two of the preceding values (e.g., 70%- 100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, the antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv- Fc fusion, nanobody®-Fc) includes a VL that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:2 or 3,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:2 or 3 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.); and a VH that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:19 or 20,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO: 19 or 20 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%- 95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, the antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a VL that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:4 or 5,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:4 or 5 in a range defined by any two ofthe preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%- 100%, etc.); and a VH that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:21 or 22,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO: 21 or 22 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%- 95%, 90%-100%, 95%-100%, etc.). In some embodiments, the antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a VL that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:6 or 7,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:6 or 7 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.); and a VH that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:23 or 24,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO: 23 or 24 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%- 100%, etc.). In some embodiments, the antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a VL that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:8,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:8 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%- 100%, 80%-95%, 90%-100%, 95%-100%, etc.); and a VH that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:25,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO: 25 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, the antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc)includes a VL that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:9,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:9 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%- 100%, etc.); and a VH that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:26,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO: 26 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%- 100%, 95%-100%, etc.). In some embodiments, the antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a VL that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:10,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:10 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.); and a VH that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:27,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO: 27 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, the antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a VL that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:11,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:11 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.); and a VH that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQID NO:28,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO: 28 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%- 100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, the antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a VL that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:12,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:12 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.); and a VH that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:29,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO: 29 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, the antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a VL that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:11,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:11 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.); and a VH that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:140,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO: 140 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, the antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a VL that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:13,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:13 in a range defined by any two of thepreceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%- 100%, etc.); and a VH that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:30,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO: 30 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%- 100%, 95%-100%, etc.). In some embodiments, the antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a VL that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:14,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:14 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.); and a VH that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:31,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO: 31 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, the antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a VL that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:15,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:15 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.); and a VH that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:32,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO: 32 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%- 100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, the antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includesa VL that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:16,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:16 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.); and a VH that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:33,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO: 33 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, the antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a VL that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:17,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:17 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.); and a VH that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:34,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO: 34 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%- 100%, 80%-95%, 90%-100%, 95%-100%, etc.). In some embodiments, the antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a VL that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:8,or optionally the amino acid sequence has percent sequence identity to SEQ ID NO:8 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%-100%, etc.); and a VH that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:139,or optionally the amino acidsequence has percent sequence identity to SEQ ID NO: 139 in a range defined by any two of the preceding values (e.g., 70%-100%, 75%-95%, 80%-100%, 80%-95%, 90%-100%, 95%- 100%, etc.).

[0200] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K73D mutation (Aho). For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%- 95%, 70%-90%, 70%-80%, 70%-75%, 57%-100%, 75%-95%, 75%-90%, 75%-80%, 80%- 100%, 80%-95%, 80%-90%, 90%-100%, 90%-95%, or 95%-100% identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a K73D mutation (Aho).

[0201] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K73S mutation (Aho). For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%- 95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%- 95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%- 95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K73S mutation (Aho).

[0202] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K13V mutation (Aho). For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%- 95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%- 95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K13V mutation (Aho).

[0203] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K14Q mutation (Aho). For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%- 95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%- 95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%- 95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K14Q mutation (Aho).

[0204] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K13V and K14Q mutation (Aho). For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a K13V and K14Q mutation (Aho).

[0205] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a D83S mutation (Aho). For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%- 95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%- 95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a D83S mutation (Aho).

[0206] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a E84N mutation (Aho). For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%- 95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%- 95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%- 95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a E84N mutation (Aho).

[0207] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a D83S and E84N mutation (Aho). For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a D83S and E84N mutation (Aho).

[0208] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a E92S mutation (Aho). For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%- 95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%- 95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a E92S mutation (Aho).

[0209] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K24A mutation (Aho). For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%- 95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%- 95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%- 95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K24A mutation (Aho).

[0210] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a R97T A mutation (Aho). For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%- 95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%- 95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%- 95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VH domain in SEQ ID NO. 18 wherein the VH domain comprises a R97T mutation (Aho).

[0211] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises an E to Q mutation at a position corresponding to residue 1 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a E to Q mutation at a position corresponding to residue 1 of SEQ ID NO.18.

[0212] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a Q to V mutation at a position corresponding to residue 5 of SEQ ID NO. 18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a Q to V mutation at a position corresponding to residue 5 of SEQ ID NO.18.

[0213] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a P to A mutation at a position corresponding to residue 9 of SEQ ID NO. 18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a P to A mutation at a position corresponding to residue 9 of SEQ ID NO.18.

[0214] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a L to V mutation at a position corresponding to residue 11 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%,75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a L to V mutation at a position corresponding to residue 11 of SEQ ID NO. 18.

[0215] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a V to K mutation at a position corresponding to residue 12 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises ato K mutation at a position corresponding to residue 12 of SEQ ID NO. 18.

[0216] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K to Q mutation at a position corresponding to residue 13 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a K to Q mutation at a position corresponding to residue 13 of SEQ ID NO. 18.

[0217] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises an A to S mutation at a position corresponding to residue 16 of SEQ ID NO. 18.For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises an A to S mutation at a position corresponding to residue 16 of SEQ ID NO.18.

[0218] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises an I to V mutation at a position corresponding to residue 20 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises an I to V mutation at a position corresponding to residue 20 of SEQ ID NO. 18.

[0219] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K to A mutation at a position corresponding to residue 23 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a K to A mutation at a position corresponding to residue 23 of SEQ ID NO. 18.

[0220] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two ofthe preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises an A to G mutation at a position corresponding to residue 26 of SEQ ID NO. 18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises an A to G mutation at a position corresponding to residue 26 of SEQ ID NO.18.

[0221] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises an A to S mutation at a position corresponding to residue 31 of SEQ ID NO. 18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises an A to S mutation at a position corresponding to residue 31 of SEQ ID NO.18.

[0222] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K to R mutation at a position corresponding to residue 38 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a K to R mutation at a position corresponding to residue 38 of SEQ ID NO.18.

[0223] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a S to A mutation at a position corresponding to residue 40 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a S to A mutation at a position corresponding to residue 40 of SEQ ID NO. 18.

[0224] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a H to P mutation at a position corresponding to residue 41 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a H to P mutation at a position corresponding to residue 41 of SEQ ID NO. 18.

[0225] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K to Q mutation at a position corresponding to residue 43 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a K to Q mutation at a position corresponding to residue 43 of SEQ ID NO. 18.

[0226] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a S to G mutation at a position corresponding to residue 44 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a S to G mutation at a position corresponding to residue 44 of SEQ ID NO. 18.

[0227] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises an I to M mutation at a position corresponding to residue 48 of SEQ ID NO. 18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises an I to M mutation at a position corresponding to residue 48 of SEQ ID NO.18.

[0228] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a N to A mutation at a position corresponding to residue 61 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%,75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a N to A mutation at a position corresponding to residue 61 of SEQ ID NO. 18.

[0229] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K to D mutation at a position corresponding to residue 63 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a K to D mutation at a position corresponding to residue 63 of SEQ ID NO. 18.

[0230] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K to S mutation at a position corresponding to residue 63 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a K to S mutation at a position corresponding to residue 63 of SEQ ID NO. 18.

[0231] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K to Q mutation at a position corresponding to residue 65 of SEQ ID NO.18. Forexample, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a K to Q mutation at a position corresponding to residue 65 of SEQ ID NO. 18.

[0232] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K to R mutation at a position corresponding to residue 67 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a K to R mutation at a position corresponding to residue 67 of SEQ ID NO.18.

[0233] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises an A to V mutation at a position corresponding to residue 68 of SEQ ID NO. 18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises an A to V mutation at a position corresponding to residue 68 of SEQ ID NO.18.

[0234] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two ofthe preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a L to I mutation at a position corresponding to residue 70 of SEQ ID NO. 18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a L to I mutation at a position corresponding to residue 70 of SEQ ID NO. 18.

[0235] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a V to A mutation at a position corresponding to residue 72 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a V to A mutation at a position corresponding to residue 72 of SEQ ID NO. 18.

[0236] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K to E mutation at a position corresponding to residue 74 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a K to E mutation at a position corresponding to residue 74 of SEQ ID NO.18.

[0237] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a K to N mutation at a position corresponding to residue 74 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a K to N mutation at a position corresponding to residue 74 of SEQ ID NO. 18.

[0238] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a S to T mutation at a position corresponding to residue 76 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a S to T mutation at a position corresponding to residue 76 of SEQ ID NO.18.

[0239] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a H to Y mutation at a position corresponding to residue 80 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a H to Y mutation at a position corresponding to residue 80 of SEQ ID NO. 18.

[0240] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a D to E mutation at a position corresponding to residue 82 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a D to E mutation at a position corresponding to residue 82 of SEQ ID NO.18.

[0241] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a D to S mutation at a position corresponding to residue 82 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a D to S mutation at a position corresponding to residue 82 of SEQ ID NO. 18.

[0242] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a L to S mutation at a position corresponding to residue 84 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%,75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a L to S mutation at a position corresponding to residue 84 of SEQ ID NO. 18.

[0243] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a T to R mutation at a position corresponding to residue 87 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a T to R mutation at a position corresponding to residue 87 of SEQ ID NO.18.

[0244] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a S to T mutation at a position corresponding to residue 91 of SEQ ID NO.18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a S to T mutation at a position corresponding to residue 91 of SEQ ID NO.18.

[0245] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a G to A mutation at a position corresponding to residue 97 of SEQ ID NO.18. Forexample, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO.18, wherein the VH domain comprises a G to A mutation at a position corresponding to residue 97 of SEQ ID NO. 18.

[0246] In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a S to L mutation at a position corresponding to residue 117 of SEQ ID NO. 18. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%- 100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises a S to L mutation at a position corresponding to residue 117 of SEQ ID NO. 18.

[0247] In some embodiments, the VH domain comprises any one or more of the VH domain mutations (e.g., any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 45 or all of the non-overlapping VH domain mutations) provided herein.

[0248] In some embodiments, the antibody or antigen binding fragment thereof comprises a VH domain comprising an HCDR1, HCDR2, and HCDR3. In some embodiments, the HCDR1 is an HCDR1 having the sequence of any one of SEQ ID NO. 43-46. In some embodiments, the HCDR1 is an HCDR1 having the sequence of SEQ ID NO.43 wherein the sequence comprises an A to G substitution at a position corresponding to residue number 4 of SEQ ID NO. 43. In some embodiments, the HCDR1 is an HCDR1 having the sequence of SEQ ID NO. 43 wherein the sequence comprises an A to S substitution at a position corresponding to residue number 9 of SEQ ID NO. 43. In some embodiments, the HCDR2 is an HCDR2 having the sequence of any one of SEQ ID NO. 47-48, 119, or 136. In some embodiments, the HCDR2 is an HCDR2 having the sequence of SEQ ID NO.47 wherein the sequence comprises a N to A substitution at a position corresponding to residue number 12 ofSEQ ID NO. 47. In some embodiments, the HCDR3 is an HCDR3 having the sequence of SEQ ID NO. 49 or 50. In some embodiments, the HCDR3 is a HCDR3 having the sequence of SEQ ID NO. 49, wherein the sequence comprises a G to A substitution at a position corresponding to residue 1 of SEQ ID NO. 49. Table 5 depicts some embodiments of HCDR sequences disclosed herein. In some embodiments, the antibody or antigen binding fragment thereof comprises a VH domain with a HCDR1 sequence that is a HCDR1 sequence in Table 5. In some embodiments, the antibody or antigen binding fragment thereof comprises a VH domain with a HCDR2 sequence that is a HCDR2 sequence in Table 5. In some embodiments, the antibody or antigen binding fragment thereof comprises a VH domain with an HCDR3 sequence that is a HCDR3 sequence in Table 5. In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain comprising a HCDR1, HCDR2, and / or HCDR3 in Table 5. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VH domain comprising a HCDR1, HCDR2, and / or HCDR3 in Table 5. Table 5

[0249] Provided herein is an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) that includes a variable heavy (VH) domain that includes a HCDR1 that is an HCDR1 in any one of SEQ ID NOs:18-34, 139 or 140, a HCDR2 that is an HCDR2 in any one of SEQ ID NOs:18-34, 139 or 140, and a HCDR3 that is an HCDR3 in any one of SEQ ID NOs:18-34, 139 or 140. In some embodiments, the HCDR1, HCDR2, and HCDR3 are combined as they are arranged in each one of SEQ ID NOs:18-34, 139 or 140. In some embodiments, the antigen binding construct includes a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:18, a HCDR2 that is an HCDR2 in SEQ ID NO:18, and a HCDR3 that is an HCDR3 in SEQ ID NO:18. In some embodiments, the antigen binding construct includes a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:19 or 20, a HCDR2 that is an HCDR2 in SEQ ID NO:19 or 20, and a HCDR3 that is an HCDR3 in SEQ ID NO:19 or 20. In some embodiments, the antigen binding construct includes a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:21 or 22, a HCDR2 that is an HCDR2 in SEQ ID NO:21 or 22, and a HCDR3 that is an HCDR3 in SEQ ID NO:21 or 22. In some embodiments, the antigen binding construct includes a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:23 or 24, a HCDR2 that is an HCDR2 in SEQ ID NO:23 or 24, and a HCDR3 that is an HCDR3 in SEQ ID NO:23 or 24. In some embodiments, the antigen binding construct includes a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:25, a HCDR2 that is an HCDR2 in SEQ ID NO:25, and a HCDR3 that is an HCDR3 in SEQ ID NO:25. In some embodiments, the antigen binding construct includes a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:26, a HCDR2 that is an HCDR2 in SEQ ID NO:26, and a HCDR3 that is an HCDR3 in SEQ ID NO:26. In some embodiments, the antigen binding construct includes a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:27, a HCDR2 that is an HCDR2 in SEQ ID NO:27, and a HCDR3 that is an HCDR3 in SEQ ID NO:27. In some embodiments, the antigen binding construct includes a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:28, a HCDR2 that is an HCDR2 in SEQ ID NO:28, and a HCDR3 that is an HCDR3 in SEQ ID NO:28. In some embodiments, the antigen binding construct includes a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:29, a HCDR2 that is an HCDR2 in SEQ ID NO:29, and a HCDR3 that is an HCDR3 in SEQ ID NO:29. In someembodiments, the antigen binding construct includes a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:30, a HCDR2 that is an HCDR2 in SEQ ID NO:30, and a HCDR3 that is an HCDR3 in SEQ ID NO:30. In some embodiments, the antigen binding construct includes a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:31, a HCDR2 that is an HCDR2 in SEQ ID NO:31, and a HCDR3 that is an HCDR3 in SEQ ID NO:31. In some embodiments, the antigen binding construct includes a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:32, a HCDR2 that is an HCDR2 in SEQ ID NO:32, and a HCDR3 that is an HCDR3 in SEQ ID NO:32. In some embodiments, the antigen binding construct includes a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:33, a HCDR2 that is an HCDR2 in SEQ ID NO:33, and a HCDR3 that is an HCDR3 in SEQ ID NO:33. In some embodiments, the antigen binding construct includes a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:34, a HCDR2 that is an HCDR2 in SEQ ID NO:34, and a HCDR3 that is an HCDR3 in SEQ ID NO:34. In some embodiments, the antigen binding construct includes a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:139, a HCDR2 that is an HCDR2 in SEQ ID NO:139, and a HCDR3 that is an HCDR3 in SEQ ID NO:139. In some embodiments, the antigen binding construct includes a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:140, a HCDR2 that is an HCDR2 in SEQ ID NO:140, and a HCDR3 that is an HCDR3 in SEQ ID NO:140. In some embodiments, the antigen binding construct is a minibody. In some embodiments, the antigen binding construct is a scFv-Fc fusion.

[0250] Also provided is an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) that includes a variable light (VL) domain that includes a LCDR1 that is an LCDR1 in any one of SEQ ID NOs:1-17, a LCDR2 that is an LCDR2 in any one of SEQ ID NOs:1-17, and a LCDR3 that is an LCDR3 in any one of SEQ ID NOs:1-17; and a variable heavy (VH) domain that includes a HCDR1 that is an HCDR1 in any one of SEQ ID NOs:18-34, 139 or 140, a HCDR2 that is an HCDR2 in any one of SEQ ID NOs:18-34, 139 or 140, and a HCDR3 that is an HCDR3 in any one of SEQ ID NOs:18- 34, 139, or 140. In some embodiments, the LCDR1, LCDR2 and LCDR3 are combined as they are arranged in each one of SEQ ID NOs:1-17, and the HCDR1, HCDR2 and HCDR3 are combined as they are arranged in each one of SEQ ID NOs:18-34, 139, or 140. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 thatis an LCDR1 in SEQ ID NO:1, a LCDR2 that is an LCDR2 in SEQ ID NO:1, and a LCDR3 that is an LCDR3 in SEQ ID NO:1; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:18, a HCDR2 that is an HCDR2 in SEQ ID NO:18, and a HCDR3 that is an HCDR3 in SEQ ID NO:18. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:2 or 3, a LCDR2 that is an LCDR2 in SEQ ID NO:2 or 3, and a LCDR3 that is an LCDR3 in SEQ ID NO:2 or 3; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:19 or 20, a HCDR2 that is an HCDR2 in SEQ ID NO:19 or 20, and a HCDR3 that is an HCDR3 in SEQ ID NO:19 or 20. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:4 or 5, a LCDR2 that is an LCDR2 in SEQ ID NO:4 or 5, and a LCDR3 that is an LCDR3 in SEQ ID NO:4 or 5; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:21 or 22, a HCDR2 that is an HCDR2 in SEQ ID NO:21 or 22, and a HCDR3 that is an HCDR3 in SEQ ID NO:21 or 22. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:6 or 7, a LCDR2 that is an LCDR2 in SEQ ID NO:6 or 7, and a LCDR3 that is an LCDR3 in SEQ ID NO:6 or 7; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:23 or 24, a HCDR2 that is an HCDR2 in SEQ ID NO:23 or 24, and a HCDR3 that is an HCDR3 in SEQ ID NO:23 or 24. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:8, a LCDR2 that is an LCDR2 in SEQ ID NO:8, and a LCDR3 that is an LCDR3 in SEQ ID NO:8; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:25, a HCDR2 that is an HCDR2 in SEQ ID NO:25, and a HCDR3 that is an HCDR3 in SEQ ID NO:25. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:9, a LCDR2 that is an LCDR2 in SEQ ID NO:9, and a LCDR3 that is an LCDR3 in SEQ ID NO:9; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:26, a HCDR2 that is an HCDR2 in SEQ ID NO:26, and a HCDR3 that is an HCDR3 in SEQ ID NO:26. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:10, a LCDR2 that is an LCDR2 in SEQ ID NO:10, and a LCDR3 that is an LCDR3 in SEQ ID NO:10; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:27, a HCDR2 that is an HCDR2 in SEQ ID NO:27, and a HCDR3 that is an HCDR3 in SEQ IDNO:27. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:11, a LCDR2 that is an LCDR2 in SEQ ID NO:11, and a LCDR3 that is an LCDR3 in SEQ ID NO:11; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:28, a HCDR2 that is an HCDR2 in SEQ ID NO:28, and a HCDR3 that is an HCDR3 in SEQ ID NO:28. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:12, a LCDR2 that is an LCDR2 in SEQ ID NO:12, and a LCDR3 that is an LCDR3 in SEQ ID NO:12; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:29, a HCDR2 that is an HCDR2 in SEQ ID NO:29, and a HCDR3 that is an HCDR3 in SEQ ID NO:29. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:13, a LCDR2 that is an LCDR2 in SEQ ID NO:13, and a LCDR3 that is an LCDR3 in SEQ ID NO:13; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:30, a HCDR2 that is an HCDR2 in SEQ ID NO:30, and a HCDR3 that is an HCDR3 in SEQ ID NO:30. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:14, a LCDR2 that is an LCDR2 in SEQ ID NO:14, and a LCDR3 that is an LCDR3 in SEQ ID NO:14; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:31, a HCDR2 that is an HCDR2 in SEQ ID NO:31, and a HCDR3 that is an HCDR3 in SEQ ID NO:31. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:15, a LCDR2 that is an LCDR2 in SEQ ID NO:15, and a LCDR3 that is an LCDR3 in SEQ ID NO:15; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:32, a HCDR2 that is an HCDR2 in SEQ ID NO:32, and a HCDR3 that is an HCDR3 in SEQ ID NO:32. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:16, a LCDR2 that is an LCDR2 in SEQ ID NO:16, and a LCDR3 that is an LCDR3 in SEQ ID NO:16; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:33, a HCDR2 that is an HCDR2 in SEQ ID NO:33, and a HCDR3 that is an HCDR3 in SEQ ID NO:33. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:17, a LCDR2 that is an LCDR2 in SEQ ID NO:17, and a LCDR3 that is an LCDR3 in SEQ ID NO:17; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:34, a HCDR2 that is an HCDR2 in SEQ ID NO:34,and a HCDR3 that is an HCDR3 in SEQ ID NO:34. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:8, a LCDR2 that is an LCDR2 in SEQ ID NO:8, and a LCDR3 that is an LCDR3 in SEQ ID NO:8; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:139, a HCDR2 that is an HCDR2 in SEQ ID NO:139, and a HCDR3 that is an HCDR3 in SEQ ID NO:139. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:9, a LCDR2 that is an LCDR2 in SEQ ID NO:9, and a LCDR3 that is an LCDR3 in SEQ ID NO:9; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:28, a HCDR2 that is an HCDR2 in SEQ ID NO:28, and a HCDR3 that is an HCDR3 in SEQ ID NO:28. In some embodiments, the antigen binding construct includes a VL domain that includes a LCDR1 that is an LCDR1 in SEQ ID NO:11, a LCDR2 that is an LCDR2 in SEQ ID NO:11, and a LCDR3 that is an LCDR3 in SEQ ID NO:11; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:140, a HCDR2 that is an HCDR2 in SEQ ID NO:140, and a HCDR3 that is an HCDR3 in SEQ ID NO:140. In some embodiments, the antigen binding construct is a minibody. In some embodiments, the antigen binding construct is a scFv-Fc fusion.

[0251] Also provided is an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) that includes: a variable light (VL) domain that includes a LCDR1 that is an LCDR1 in an LCDR1 in SEQ ID NO:2 or 3, a LCDR2 that is an LCDR2 in SEQ ID NO:2 or 3, and a LCDR3 that is an LCDR3 in SEQ ID NO:2 or 3, wherein the VL domain includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:2 or 3; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:19 or 20, a HCDR2 that is an HCDR2 in SEQ ID NO:19 or 20, and a HCDR3 that is an HCDR3 in SEQ ID NO:19 or 20, wherein the VH domain includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:19 or 20. In some embodiments, the VL domain includes a LCDR1 of SEQ ID NO:35, a LCDR2 of SEQ ID NO:37, a LCDR3 of SEQ ID NO:38; andthe VH domain includes a HCDR1 of SEQ ID NO:43, a HCDR2 of SEQ ID NO:48, a HCDR3 of SEQ ID NO:49.

[0252] Provided herein is an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) that includes: a variable light (VL) domain that includes a LCDR1 that is an LCDR1 in an LCDR1 in SEQ ID NO:8, a LCDR2 that is an LCDR2 in SEQ ID NO:8, and a LCDR3 that is an LCDR3 in SEQ ID NO:8, wherein the VL domain includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:8; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:25, a HCDR2 that is an HCDR2 in SEQ ID NO:25, and a HCDR3 that is an HCDR3 in SEQ ID NO:25, wherein the VH domain includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:25. In some embodiments, the VL domain includes a LCDR1 of SEQ ID NO:35, a LCDR2 of SEQ ID NO:37, a LCDR3 of SEQ ID NO:38; and the VH domain includes a HCDR1 of SEQ ID NO:43, a HCDR2 of SEQ ID NO:48, a HCDR3 of SEQ ID NO:50.

[0253] Provided herein is an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) that includes: a variable light (VL) domain that includes a LCDR1 that is an LCDR1 in an LCDR1 in SEQ ID NO:9, a LCDR2 that is an LCDR2 in SEQ ID NO:9, and a LCDR3 that is an LCDR3 in SEQ ID NO:9, wherein the VL domain includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:9; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:26, a HCDR2 that is an HCDR2 in SEQ ID NO:26, and a HCDR3 that is an HCDR3 in SEQ ID NO:26, wherein the VH domain includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:26. In some embodiments, the VL domain includes a LCDR1 of SEQID NO:35, a LCDR2 of SEQ ID NO:37, a LCDR3 of SEQ ID NO:39; and the VH domain includes a HCDR1 of SEQ ID NO:43, a HCDR2 of SEQ ID NO:136, a HCDR3 of SEQ ID NO:50.

[0254] Provided herein is an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) that includes: a variable light (VL) domain that includes a LCDR1 that is an LCDR1 in an LCDR1 in SEQ ID NO:8, a LCDR2 that is an LCDR2 in SEQ ID NO:8, and a LCDR3 that is an LCDR3 in SEQ ID NO:8, wherein the VL domain includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:8; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:139, a HCDR2 that is an HCDR2 in SEQ ID NO:139, and a HCDR3 that is an HCDR3 in SEQ ID NO:139, wherein the VH domain includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:139. In some embodiments, the VL domain includes a LCDR1 of SEQ ID NO:35, a LCDR2 of SEQ ID NO:37, a LCDR3 of SEQ ID NO:38; and the VH domain includes a HCDR1 of SEQ ID NO:43, a HCDR2 of SEQ ID NO:136, a HCDR3 of SEQ ID NO:50.

[0255] Provided herein is an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) that includes: a variable light (VL) domain that includes a LCDR1 that is an LCDR1 in an LCDR1 in SEQ ID NO:11, a LCDR2 that is an LCDR2 in SEQ ID NO:11, and a LCDR3 that is an LCDR3 in SEQ ID NO:11, wherein the VL domain includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identical to SEQ ID NO:11; and a VH domain that includes a HCDR1 that is an HCDR1 in SEQ ID NO:28, a HCDR2 that is an HCDR2 in SEQ ID NO:28, and a HCDR3 that is an HCDR3 in SEQ ID NO:28, wherein the VH domain includes an amino acid sequence that is, is about, or is at least 70%, 71%, 72%, 73%, 74%, 75%, 75%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%identical to SEQ ID NO:28. In some embodiments, the VL domain includes a LCDR1 of SEQ ID NO:36, a LCDR2 of SEQ ID NO:37, a LCDR3 of SEQ ID NO:40; and the VH domain includes a HCDR1 of SEQ ID NO:43, a HCDR2 of SEQ ID NO:119, a HCDR3 of SEQ ID NO:50.

[0256] Also provided herein is an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) that includes a variable light (VL) domain that includes a LCDR1 that is an LCDR1 in Table 6, a LCDR2 that is an LCDR2 in Table 6, and a LCDR3 that is an LCDR3 in Table 6. In some embodiments, the LCDR1, LCDR2 and LCDR3 are combined as they are arranged in each construct in Table 6. In some embodiments, the VL domain includes a LCDR1 of SEQ ID NO:35, a LCDR2 of SEQ ID NO:37, a LCDR3 of SEQ ID NO:38. In some embodiments, the VL domain includes a LCDR1 of SEQ ID NO:35, a LCDR2 of SEQ ID NO:37, a LCDR3 of SEQ ID NO:39. In some embodiments, the VL domain includes a LCDR1 of SEQ ID NO:36, a LCDR2 of SEQ ID NO:37, a LCDR3 of SEQ ID NO:39. In some embodiments, the VL domain includes a LCDR1 of SEQ ID NO:36, a LCDR2 of SEQ ID NO:37, a LCDR3 of SEQ ID NO:40. In some embodiments, the antigen binding construct is a minibody. In some embodiments, the antigen binding construct is a scFv-Fc fusion. Table 6

[0257] Also provided herein is an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) that includes a variable heavy (VH) domain that includes a HCDR1 that is an HCDR1 in Table 6, a HCDR2 that is an HCDR2 in Table 6, and a HCDR3 that is an HCDR3 in Table 6. In some embodiments, the HCDR1, HCDR2 and HCDR3 are combined as they are arranged in each construct in Table 6. In some embodiments, the VH domain includes a HCDR1 of SEQ ID NO:43, a HCDR2 of SEQ ID NO:47, a HCDR3 of SEQ ID NO:49. In some embodiments, the VH domain includes a HCDR1 of SEQ ID NO:43, a HCDR2 of SEQ ID NO:48, a HCDR3 of SEQ ID NO:50. In some embodiments, the VH domain includes a HCDR1 of SEQ ID NO:44, a HCDR2 of SEQ ID NO:48, a HCDR3 of SEQ ID NO:50. In some embodiments, the VH domain includes a HCDR1 of SEQ ID NO:45, a HCDR2 of SEQ ID NO:48, a HCDR3 of SEQ ID NO:50. In some embodiments, the VH domain includes a HCDR1 of SEQ ID NO:43, a HCDR2 of SEQ ID NO:136, a HCDR3 of SEQ ID NO:50. In some embodiments, the VH domain includes a HCDR1 of SEQ ID NO:43, a HCDR2 of SEQ ID NO:119, a HCDR3 of SEQ ID NO:50. In some embodiments, the VH domain includes a HCDR1 of SEQ ID NO:46, a HCDR2 of SEQ ID NO:119, a HCDR3 of SEQ ID NO:50. In some embodiments, the antigen binding construct is a minibody. In some embodiments, the antigen binding construct is a scFv-Fc fusion.

[0258] Also provided herein is an antigen binding construct (e.g., an antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) that includes a variable light (VL) domain that includes a LCDR1 that is an LCDR1 in Table 6, a LCDR2 that is an LCDR2 in Table 6, and a LCDR3 that is an LCDR3 in Table 6; and a variable heavy (VH) domain that includes a HCDR1 that is an HCDR1 in Table 6, a HCDR2 that is an HCDR2 in Table 6, and a HCDR3 that is an HCDR3 in Table 6. In some embodiments, the LCDR1, LCDR2, and LCDR3 are combined as they are arranged in each construct in Table 6, and the HCDR1, HCDR2, and HCDR3 are combined as they are arranged in a corresponding construct in Table6. In some embodiments, the VL domain includes a LCDR1 of SEQ ID NO:35, a LCDR2 of SEQ ID NO:37, a LCDR3 of SEQ ID NO:38; and the VH domain includes a HCDR1 of SEQ ID NO:43, a HCDR2 of SEQ ID NO:47, a HCDR3 of SEQ ID NO:49. In some embodiments, the VL domain includes a LCDR1 of SEQ ID NO:35, a LCDR2 of SEQ ID NO:37, a LCDR3 of SEQ ID NO:38; and the VH domain includes a HCDR1 of SEQ ID NO:43, a HCDR2 of SEQ ID NO:48, a HCDR3 of SEQ ID NO:50. In some embodiments, the VL domain includes a LCDR1 of SEQ ID NO:35, a LCDR2 of SEQ ID NO:37, a LCDR3 of SEQ ID NO:38; and the VH domain includes a HCDR1 of SEQ ID NO:44, a HCDR2 of SEQ ID NO:48, a HCDR3 of SEQ ID NO:49. In some embodiments, the VL domain includes a LCDR1 of SEQ ID NO:35, a LCDR2 of SEQ ID NO:37, a LCDR3 of SEQ ID NO:38; and the VH domain includes a HCDR1 of SEQ ID NO:45, a HCDR2 of SEQ ID NO:48, a HCDR3 of SEQ ID NO:50. In some embodiments, the VL domain includes a LCDR1 of SEQ ID NO:35, a LCDR2 of SEQ ID NO:37, a LCDR3 of SEQ ID NO:39; and the VH domain includes a HCDR1 of SEQ ID NO:43, a HCDR2 of SEQ ID NO:136, a HCDR3 of SEQ ID NO:50. In some embodiments, the VL domain includes a LCDR1 of SEQ ID NO:36, a LCDR2 of SEQ ID NO:37, a LCDR3 of SEQ ID NO:40; and the VH domain includes a HCDR1 of SEQ ID NO:43, a HCDR2 of SEQ ID NO:119, a HCDR3 of SEQ ID NO:50. In some embodiments, the VL domain includes a LCDR1 of SEQ ID NO:36, a LCDR2 of SEQ ID NO:37, a LCDR3 of SEQ ID NO:40; and the VH domain includes a HCDR1 of SEQ ID NO:46, a HCDR2 of SEQ ID NO:119, a HCDR3 of SEQ ID NO:50. In some embodiments, the antigen binding construct is a minibody. In some embodiments, the antigen binding construct is a scFv-Fc fusion.

[0259] Provided herein is a minibody (e.g., a minibody that binds to ĮVȕ6 integrin) that includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to any one of the sequences set forth in SEQ ID NOs: 103-118 or 133-135, optionally the amino acid sequence has a sequence identity to any one of the sequences set forth in SEQ ID NOs: 103-118 or 133-135 in a range defined by any two of the preceding values (e.g., 70-100%, 80-100%, 90-100%, 85-98%, 80-95%, etc.). In some embodiments, the minibody includes any one of the sequences set forth in SEQ ID NOs: 103-118 or 133-135. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forthin SEQ ID NO: 103, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 103 in a range defined by any two of the preceding values (e.g., 70- 100%, 80-100%, 90-100%, 85-98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 103. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 104, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 104 in a range defined by any two of the preceding values (e.g., 70-100%, 80-100%, 90-100%, 85-98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 104. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 105, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 105 in a range defined by any two of the preceding values (e.g., 70-100%, 80-100%, 90-100%, 85- 98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 105. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 106, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 106 in a range defined by any two of the preceding values (e.g., 70- 100%, 80-100%, 90-100%, 85-98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 106. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 107, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 107 in a range defined by any two of the preceding values (e.g., 70-100%, 80-100%, 90-100%, 85-98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 107. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%,97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 108, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 108 in a range defined by any two of the preceding values (e.g., 70-100%, 80-100%, 90-100%, 85- 98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 108. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 109, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 109 in a range defined by any two of the preceding values (e.g., 70- 100%, 80-100%, 90-100%, 85-98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 109. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 110, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 110 in a range defined by any two of the preceding values (e.g., 70-100%, 80-100%, 90-100%, 85-98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 110. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 111, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 111 in a range defined by any two of the preceding values (e.g., 70-100%, 80-100%, 90-100%, 85- 98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 111. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 112, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 112 in a range defined by any two of the preceding values (e.g., 70- 100%, 80-100%, 90-100%, 85-98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 112. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is,is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 113, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 113 in a range defined by any two of the preceding values (e.g., 70-100%, 80-100%, 90-100%, 85-98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 113. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 114, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 114 in a range defined by any two of the preceding values (e.g., 70-100%, 80-100%, 90-100%, 85- 98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 114. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 115, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 115 in a range defined by any two of the preceding values (e.g., 70- 100%, 80-100%, 90-100%, 85-98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 115. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 116, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 116 in a range defined by any two of the preceding values (e.g., 70-100%, 80-100%, 90-100%, 85-98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 116. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 117, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 117 in a range defined by any two of the preceding values (e.g., 70-100%, 80-100%, 90-100%, 85- 98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 117. In some embodiments, a minibody of the present disclosure (e.g., that bindsto ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 118, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 118 in a range defined by any two of the preceding values (e.g., 70- 100%, 80-100%, 90-100%, 85-98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 118. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 133, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 133 in a range defined by any two of the preceding values (e.g., 70-100%, 80-100%, 90-100%, 85-98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 133. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 134, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 134 in a range defined by any two of the preceding values (e.g., 70-100%, 80-100%, 90-100%, 85- 98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 134. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 135, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 135 in a range defined by any two of the preceding values (e.g., 70- 100%, 80-100%, 90-100%, 85-98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 135. In some embodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 139, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 139 in a range defined by any two of the preceding values (e.g., 70-100%, 80-100%, 90-100%, 85-98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 139. In someembodiments, a minibody of the present disclosure (e.g., that binds to ĮVȕ6 integrin) includes an amino acid sequence that is, is about, or is at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the sequence set forth in SEQ ID NO: 140, optionally the amino acid sequence has a sequence identity to the sequence set forth in SEQ ID NO: 140 in a range defined by any two of the preceding values (e.g., 70-100%, 80-100%, 90-100%, 85- 98%, 80-95%, etc.). In some embodiments, the minibody includes the sequence set forth in SEQ ID NO: 140.

[0260] Table 7 lists some embodiments of substitutions in the VL, VH, CH2, CH3, and / or Fc domains of an antibody or antigen binding fragment thereof that, either alone, or in combination, may reduce healthy organ uptake of the antibody or antigen binding fragment thereof. In some embodiments, the VH domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises one or more mutations (e.g. any 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, or all of the non-overlapping mutations) in Table 7. For example, in some embodiments, the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%- 90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%- 93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VH domain in SEQ ID NO. 18, wherein the VH domain comprises one or more mutations in Table 7. In some embodiments, the VL domain comprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VL domain in SEQ ID NO. 1, wherein the VL domain comprises one or more mutations in Table 7. For example, in some embodiments, the VL domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%- 90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%- 90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%- 100%, 95%-97%, or 97%-100%, identity to a VL domain in SEQ ID NO. 1, wherein the VL domain comprises one or more mutations in Table 7. In some embodiments, the VL domaincomprises an amino acid sequence having, having about, or having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or a percentage in a range that is defined by any two of the preceding values, identity to a VL domain in SEQ ID NO. 1, and the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%-97%, or 97%-100%, identity to a VH domain in SEQ ID NO. 18, wherein the VL and / or the VH domain comprises one or more mutations in Table 7. For example, in some embodiments, the VL domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%-90%, 70%- 80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%-90%, 90%- 100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%-100%, 95%- 97%, or 97%-100%, identity to a VL domain in SEQ ID NO.1, and the VH domain comprises an amino acid sequence having between about, or in a range of, 70%-100%, 70%-95%, 70%- 90%, 70%-80%, 75%-100%, 75%-95%, 75%-90%, 75%-85%, 80%-100%, 80%-95%, 80%- 90%, 90%-100%, 90%-97%, 90%-95%, 90%-93%, 93%-100%, 93%-97%, 93%-95%, 95%- 100%, 95%-97%, or 97%-100%, identity to a VH domain in SEQ ID NO.18, wherein the VL and / or the VH domain comprises one or more mutations in Table 7. Table 7* X can be any amino acid other than K or R.

[0261] In some embodiments, the antigen binding construct (e.g., antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a hinge region comprising an upper hinge that includes the sequence EPKSSDKTHT (SEQ ID NO: 51). In some embodiments, the antigen binding construct (e.g., antibody, minibody, cys-diabody, scFv-Fc fusion, nanobody®-Fc) includes a hinge region comprising an upper hinge that includes the sequence GGSGGGSTHT (SEQ ID NO: 52). In some embodiments, the antigen binding construct includes a hinge region comprising EPKSSDKTHTCPPCPPCGGGSSGGGSG (SEQ ID NO: 137). In some embodiments, the antigen binding construct includes a hinge region comprising GGSGGGSTHTCPPCPPCGGGSSGGGSGG (SEQ ID NO: 138). In some embodiments, the antigen binding construct includes a hinge region comprising at least one of EPKSSDKTHT (SEQ ID NO: 51) and EPKSSDKTHTCPPCPPCGGGSSGGGSG (SEQ ID NO: 137).

[0262] In some embodiments, the antigen binding construct, minibody, or scFv-Fc fusion, accumulates to a detectable level in a subject’s blood, lungs, muscle, bone (bonemarrow), liver, kidneys, heart, stomach, small intestine, large intestine, pancreas, tumor, testis, ovaries, and / or any combination therein. In some embodiments, the subject is a mammal.

[0263] In some embodiments, the antigen binding construct, minibody, or scFv-Fc fusion, has a KD (e.g., KD of binding to an antigen, e.g., an ĮVȕ6 protein) of, or about, or of less than 10-5M, 10-6M, 10-7M, 10-8M, 10-9M, 10-10M, 10-11M, 10-12M, 10-13M, 10-14M, or 10-15M, or a KD in a range defined by any two of the preceding values. For example, in some embodiments, the antigen binding construct, minibody, or scFv-Fc fusion, has a KD (e.g., KD of binding to an antigen) of between about, or in a range of, 10-5M and 10-15M, 10-5M and 10-12M, 10-5M and 10-10M, 10-7M and 10-15M, 10-7M and 10-12M, 10-7M and 10-10M, 10-10M and 10-15M, or 10-10M and 10-12M. In some embodiments, the KD is determined for an antibody dissolved in buffer. In some embodiments, the buffer is a phosphate buffer. If desired, the KD can be determined using any suitable option, e.g., biolayer interferometry.

[0264] In some embodiments, the antigen binding construct, minibody, or scFv-Fc fusion, has an EC50(e.g., EC50of binding to a ĮVȕ6 protein, or a cell expressing a ĮVȕ63 protein) of, of about, of at most, or less than 0.001, 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20, 0.21, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29, 0.30, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38, 0.39, or 0.40 nM, or an EC50 in a range that is defined by any two of the preceding values. For example, in some embodiments, the antigen binding construct, minibody, or scFv-Fc fusion, has an EC50 (e.g., EC50 of binding to an antigen, or a cell expressing an antigen) of between about, or in a range of, 0.001 and 0.20 nM, 0.001 and 0.17 nM, 0.001 and 0.12 nM, 0.01 and 0.20 nM, 0.01 and 0.17 nM, 0.01 and 0.12 nM, 0.05 and 0.20 nM, 0.05 and 0.17 nM.0.05 and 0.15 nM, 0.05 and 0.12 nM, 0.08 and 0.20 nM, 0.08 and 0.17 nM, or 0.08 and 0.12 nM. In some embodiments, the antigen binding construct, minibody, or cys-diabody, has an EC50(e.g., EC50of binding to a ĮVȕ6 protein, or a cell expressing a ĮVȕ6 protein) of up to about 10 nM. In some embodiments, the antigen binding construct, minibody, or cys-diabody, has an EC50 (e.g., EC50 of binding to a ĮVȕ6 protein, or a cell expressing a ĮVȕ6 protein) of up to about 5 nM. If desired, the EC50 can be determined using any suitable option, e.g., using ELISA or flow cytometry.

[0265] In some embodiments, the antigen binding construct, minibody, or scFv-Fc fusion, includes at least one modification. Exemplary modifications include, but are not limitedto, antigen binding constructs that have been modified by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting / blocking groups, conjugation to metal chelator, fluorescence or infrared dye, or conjugation to a toxin, proteolytic cleavage, and linkage to a cellular ligand or other protein. Any of numerous chemical modifications may be carried out by known techniques, including, but not limited to, amine coupling to amine containing amino acids, cystine coupling, specific chemical cleavage, acetylation, and metabolic synthesis of tunicamycin. In some embodiments, the derivative can contain one or more non-natural amino acids.

[0266] In some embodiments, the antigen binding construct, minibody, or scFv-Fc fusion, is conjugated to another substance to form an anti-target conjugate. The conjugates described herein can be prepared by known methods of linking antigen binding constructs with lipids, carbohydrates, protein or other atoms and molecules. In some embodiments, the conjugate is formed by site-specific conjugation using a suitable linkage or bond. Site-specific conjugation is more likely to preserve the binding activity of an antigen binding construct. The substance may be conjugate or attached at the hinge region of a reduced antigen binding construct via thioether bond formation. In some embodiments, tyrosine conjugation can be employed. Other linkages or bonds used to form the conjugate can include, but are not limited to, a covalent bond, a non-covalent bond, a disulfide linkage, a hydrazone linkage, an ester linkage, an amido linkage, and amino linkage, an imino linkage, a thiosemicarbazone linkage, a semicarbazone linkage, an oxime linkage and a carbon-carbon linkage. In some embodiments, no cysteine or other linking aspect, need be included in the antigen binding construct.

[0267] In some embodiments, the antigen binding construct, minibody, or scFv-Fc fusion, is conjugated to a chemotherapeutic agent. Chemotherapeutic agents are often cytotoxic or cytostatic in nature and may include alkylating agents, antimetabolites, anti-tumor antibiotics, topoisomerase inhibitors, mitotic inhibitors hormone therapy, targeted therapeutics and immunotherapeutics. In some embodiments the chemotherapeutic agents that may be used as detectable markers in accordance with the embodiments of the disclosure are selected from among those chemotherapeutic agents defined elsewhere herein.

[0268] In some embodiments, the antigen binding construct, minibody, or scFv-Fc fusion, is conjugated to a toxin. Toxins that may be used in accordance with the embodimentsof the disclosure include, but are not limited to, Auristatin E, Auristatin F, Dolastatin 10, Dolastatin 15, combretastatin and their analogs, maytansinoid, calicheamicin, alpha-amanitin, pyrrolobenzodiazepine dimers, epothilones, duocarmycin and their analogs, tubulysin D, basillistatins, ricin, abrin, ribonuclease (RNase), DNase I, Staphylococcal enterotoxin-A, pokeweed antiviral protein, gelonin, diphtheria toxin, Pseudomonas exotoxin, and Pseudomonas endotoxin.

[0269] In some embodiments, the antigen binding construct, minibody, or scFv-Fc fusion, is conjugated to a radiolabel. In some embodiments, the radiolabel comprises Iodine- 131, a beta-emitter or alpha-emitter, such as, Yttrium-90, Copper-67, Terbium-149, Terbium- 161, Lutetium-177, Astatine-211, Lead-212, Bismuth-212, Actinium-225, Bismuth-213, or 227-Thorium, a positron emitter such as Zirconium-89, Copper-64, Gallium-68, Fluorine-18, Indium-111, Iodine-124. In some embodiments, the radiolabel comprises Terbium-149, Terbium-161, or Lead-212. In some embodiments, the antigen binding construct, minibody, and / or cys-diabody, are conjugated to a gamma emitter. In some embodiments the radio label comprises a gamma emitter.

[0270] In some embodiments, the antigen binding construct, minibody, or scFv-Fc fusion, is conjugated to a radioisotope, a fluorescent compound, a bioluminescent compound, chemiluminescent compound, a metal chelator or an enzyme. In some embodiments, the at least one radioisotope comprises18F,18F-FDG,32P,33P,45Ti,47Sc,52Fe,59Fe,62Cu,64Cu,67Cu,67Ga,68Ga,75Sc,77As,86Y,90Y,89Sr,89Zr,94Tc,94Tc,99mTc,99Mo,105Pd,105Rh,111Ag,111In,123I,124I,125I,131I,142Pr,143Pr,149Pm,149Tb,153Sm,154-158Gd,161Tb,166Dy,166Ho,169Er,175Lu,177Lu,186Re,188Re,189Re,194Ir,198Au,199Au,211At,211Pb,212Bi,212Pb,213Bi,223Ra,227Th and225Ac, or any combination thereof. In some embodiments, the at radioisotope comprises149Tb,161Tb, or212Pb. In some embodiments, the bioluminescence or fluorescent compound comprises fluorescein, fluorescein isothiocyanate (FITC), OREGON GREEN™, rhodamine, Texas red, tetrarhodimine isothiocynate (TRITC), Cy3, Cy5, and the like), fluorescent markers (e.g., green fluorescent protein (GFP), phycoerythrin, and the like), autoquenched fluorescent compounds that are activated by tumor-associated proteases, enzymes (e.g., luciferase, horseradish peroxidase, alkaline phosphatase, and the like), nanoparticles, biotin, digoxigenin or combination thereof. In some embodiments, the organic dye is IndoCyanine Green (ICG) or one of the dyes that fluoresces in the near infrared region, such as IR800. In someembodiments, the chelating ligand comprises ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), dodecane tetraacetic acid (DOTA), 1,4,7,10- tetraazacyclododececane,1-(glutaric acid)-4,7,10-triacetic acid (DOTAGA), 1,4,7- triazacyclononane-1,4,7-triacetic acid (NOTA), 1,4,7-triazacyclononane, 1-glutaric acid-5,7 acetic acid (NODAGA), 4-[2-(bis-carboxymethyl-amino)-ethyl]-7-carboxymethyl- [1,4,7]triazonan-1-yl-acetic acid (NETA), deferoxamine (Df, which may also be referred to as DFO), porphyrins, polyamines, crown ethers, bis-thiosemicarbazones, polyoximes, and like groups.

[0271] In some embodiments, the antigen binding construct is an antibody (~140- 170 kDa), a multi-specific antibody (~100 kDa and greater), a single-arm or one-arm antibody (~110 kDa and less), a modified antibody, or the like. In some embodiments, the antigen binding construct, minibody, or scFv-Fc fusion, binds specifically to ĮVȕ6. In some embodiments, the antigen binding construct, minibody, or scFv-Fc fusion, is bispecific. In some embodiments, the antigen binding construct, minibody, or scFv-Fc fusion, is bivalent. In some embodiments, the antigen binding construct, minibody, or scFv-Fc fusion, comprises a monovalent scFv. In some embodiments, the antigen binding construct is a scFv-Fc (e.g., a scFv fused to a Fc). In some embodiments, the antigen binding construct is a nanobody®-Fc (e.g., a nanobody® (or single domain fragment) fused to a Fc).

[0272] In some embodiments, the order of the variable domains, from N terminus to C terminus of the antigen binding construct, minibody, or scFv-Fc fusion, from N terminus to C terminus of the polypeptide is VL, VH. In some embodiments, the order of the variable domains, from N terminus to C terminus of the polypeptide is VH, VL. Therapeutic agents and Compositions

[0273] Some embodiments herein are directed to a therapeutic agent or composition comprising an antigen binding construct comprising: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO. 35 or 36; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 38 or 39; and a variable heavy (VH) domain comprising: a HCDR1 that is any one of SEQ ID NO.43-46; a HCDR2 that is any one of SEQ ID NO. 47- 48 or 119; and a HCDR3 that is SEQ ID NO.49 or 50. In some embodiments, the composition comprises a hinge-extension domain comprising a IgG1 hinge region; a IgG CH3 sequence; and a therapeutic agent, toxic payload, and / or a detectable marker.

[0274] Some embodiments herein are directed to a therapeutic agent or composition comprising an antigen binding construct comprising: a LCDR1 that is SEQ ID NO.35; a LCDR2 that is ...

Claims

WHAT IS CLAIMED IS:

1. An antigen binding construct that comprises: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO. 35 or 36; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is any one of SEQ ID NOs.38-40; and a variable heavy (VH) domain comprising: a HCDR1 that is any one of SEQ ID NO.43-46; a HCDR2 that is any one of SEQ ID NO.47-48, 119, or 136; and a HCDR3 that is SEQ ID NO.49 or 50.

2. An antigen binding construct that comprises: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO. 35; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 38; and a variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO.43; a HCDR2 that is SEQ ID NO.48; and a HCDR3 that is SEQ ID NO.

50.

3. An antigen binding construct that comprises: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO. 35; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 39; and a variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO.43; a HCDR2 that is SEQ ID NO.48; and a HCDR3 that is SEQ ID NO.

50.

4. An antigen binding construct that comprises: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO. 35;a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 39; and a variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO.43; a HCDR2 that is SEQ ID NO.136; and a HCDR3 that is SEQ ID NO.

50.

5. An antigen binding construct that comprises: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO. 36; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 40; and a variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO.43; a HCDR2 that is SEQ ID NO.119; and a HCDR3 that is SEQ ID NO.

50.

6. The antigen binding construct of any one of the preceding claims, further comprising an IgG CH2 sequence. The antigen binding construct of any one of the preceding claims, further comprising an IgG CH3 sequence.

8. The antigen binding construct of any one of the preceding claims, further comprising a Fc domain.

9. The antigen binding construct of any one of the preceding claims, wherein the antigen binding construct is or comprises a single-chain variable fragment-Fc fusion (scFv-Fc) that binds to ĮVȕ6, the scFv-Fc comprising the variable light (VL) domain linked to the variable heavy (VH) domain.

10. The antigen binding construct of any one of the preceding claims, wherein the antigen binding construct is or comprises a minibody that binds to integrin ĮVȕ6, the minibody comprising: a single-chain variable fragment (scFv) that binds to ĮVȕ6, the scFv comprising the variable light (VL) domain linked to the variable heavy (VH) domain.

11. A single-chain variable fragment-Fc fusion (scFv-Fc) that binds to ĮVȕ6, the scFv- Fc comprising a variable light (VL) domain linked to a variable heavy (VH) domain, the VL domain comprising: a LCDR1 that is SEQ ID NO. 35 or 36; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is any one of SEQ ID NO.38-40; and the variable heavy (VH) domain comprising: a HCDR1 that is any one of SEQ ID NO.43-46; a HCDR2 that is any one of SEQ ID NO.47-48, 119, or 136; and a HCDR3 that is SEQ ID NO.49 or 50.

12. A single-chain variable fragment-Fc fusion (scFv-Fc) that binds to ĮVȕ6, the scFv- Fc comprising a variable light (VL) domain linked to a variable heavy (VH) domain, the VL domain comprising: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO. 35; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 38; and a variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO.43; a HCDR2 that is SEQ ID NO.48; and a HCDR3 that is SEQ ID NO.

50.

13. A single-chain variable fragment-Fc fusion (scFv-Fc) that binds to ĮVȕ6, the scFv- Fc comprising a variable light (VL) domain linked to a variable heavy (VH) domain, the VLdomain comprising: a variable light (VL) domain comprising: a LCDR1 that is SEQ ID NO. 35; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 39; and a variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO.43; a HCDR2 that is SEQ ID NO.48; anda HCDR3 that is SEQ ID NO.

50.

14. The antigen binding construct or scFv-Fc fusion of any one of the preceding claims, further comprising a hinge-extension domain comprising a hinge region.

15. A minibody that binds to integrin ĮVȕ6, the minibody comprising: a single-chain variable fragment (scFv) that binds to ĮVȕ6, the scFv comprising a variable light (VL) domain linked to a variable heavy (VH) domain, the VLdomain comprising: a LCDR1 that is SEQ ID NO. 35 or 36; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is any one of SEQ ID NO.38-40; and the variable heavy (VH) domain comprising: a HCDR1 that is any one of SEQ ID NO.43-46; a HCDR2 that is any one of SEQ ID NO.47-48, 119, 136; and a HCDR3 that is SEQ ID NO.49 or 50; a hinge-extension domain comprising a hinge region; and a IgG CH3 sequence.

16. A minibody that binds to integrin ĮVȕ6, the minibody comprising: a single-chain variable fragment (scFv) that binds to ĮVȕ6, the scFv comprising a variable light (VL) domain linked to a variable heavy (VH) domain, the VL domain comprising: a LCDR1 that is SEQ ID NO. 35; a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 38; and the variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO.43; a HCDR2 that is SEQ ID NO.48; and a HCDR3 that is SEQ ID NO.50; a hinge-extension domain comprising a hinge region; and a IgG CH3 sequence.

17. A minibody that binds to integrin ĮVȕ6, the minibody comprising:a single-chain variable fragment (scFv) that binds to ĮVȕ6, the scFv comprising a variable light (VL) domain linked to a variable heavy (VH) domain, the VL domain comprising: a LCDR1 that is SEQ ID NO. 35; a LCDR2 that is any one of SEQ ID NO.37; a LCDR3 that is SEQ ID NO. 39; and the variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO.43; a HCDR2 that is SEQ ID NO.48; and a HCDR3 that is SEQ ID NO.50; a hinge-extension domain comprising a hinge region; and a IgG CH3 sequence.

18. The antigen binding construct, minibody, or scFv-Fc fusion, of any one of the preceding claims, further comprising a linker.

19. The antigen binding construct, minibody, or scFv-Fc fusion, of claim 14, wherein the linker comprises a linker that is SEQ ID NO.

55.

20. The antigen binding construct, minibody, or scFv-Fc fusion, of claim 14 or 15, wherein the linker comprises a linker comprising the sequence GSTSGGGSGGGSGGGGSS (SEQ ID NO. 55).

21. The antigen binding construct, minibody, or scFv-Fc fusion, of any one of the preceding claims, wherein the hinge region comprises an upper hinge region comprising SEQ ID NO.

51.

22. The antigen binding construct, minibody, or scFv-Fc fusion, of any one of the preceding claims, wherein the hinge region comprises an upper hinge region comprising SEQ ID NO.

52.

23. The antigen binding construct, minibody, or scFv-Fc fusion, of any one of the preceding claims, further comprising a signal peptide.

24. The antigen binding construct, minibody, or scFv-Fc fusion, of any one of the preceding claims, wherein administration of a therapeutic amount of the antigen binding construct, minibody, or scFv-Fc fusion, to a subject having a tumor reduces tumor size by up to about 2000 mm3.

25. The antigen binding construct, minibody, or scFv-Fc fusion, of any one of the preceding claims, wherein administration of a therapeutic amount of the antigen binding construct, minibody, or scFv-Fc fusion, to a subject having a tumor reduces tumor size by up to about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, or 100%, or by an amount in a range defined by any two of the preceding values.

26. The antigen binding construct, minibody, or scFv-Fc fusion, of any one of the preceding claims, wherein administration of a therapeutic amount of the antigen binding construct, minibody, or scFv-Fc fusion, to a subject having a tumor reduces tumor size by up to about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, or 100%, or by an amount in a range defined by any two of the preceding values.

27. The antigen binding construct, minibody, or scFv-Fc fusion, of any one of the preceding claims, wherein the antigen binding construct, minibody, or scFv-Fc fusion, has a KD of less than about 1x10-12M.

28. The antigen binding construct, minibody, or scFv-Fc fusion, of any one of the preceding claims, wherein the antigen binding construct, minibody, or scFv-Fc fusion, has a KD of less than about 1x10-10M.

29. The antigen binding construct, minibody, or scFv-Fc fusion, of any one of the preceding claims, wherein the antigen binding construct, minibody, or scFv-Fc fusion, has an EC50 of, of about, or of at most 0.001, 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20, 0.25, 0.3, 0.4, 0.5 nM, or an EC50in a range that is defined by any two of the preceding values.

30. The antigen binding construct, minibody, or scFv-Fc fusion, of any one of the preceding claims, wherein the antigen binding construct, minibody, or scFv-Fc fusion, has an EC50 of up to about 10 nM, or up to about 5 nM.

31. A minibody that binds to integrin ĮVȕ6, the minibody comprising: a single-chain variable fragment (scFv) that binds to ĮVȕ6, the scFv comprising a variable light (VL) domain linked to a variable heavy (VH) domain, the VL domain comprising: a LCDR1 that is SEQ ID NO. 36;a LCDR2 that is SEQ ID NO. 37; a LCDR3 that is SEQ ID NO. 40; and the variable heavy (VH) domain comprising: a HCDR1 that is SEQ ID NO.46; a HCDR2 that is SEQ ID NO.119; and a HCDR3 that is SEQ ID NO.50; a hinge-extension domain comprising a hinge region; and a IgG CH3 sequence.

32. A minibody, scFv or scFv-Fc fusion comprising: a variable light (VL) domain of SEQ ID NO.1 linked to a variable heavy (VH) domain of SEQ ID NO.

18.

33. The antigen binding construct, minibody, or scFv-Fc fusion of any one of the preceding claims, comprising a LCDR1 that is SEQ ID NO: 35 or 36; LCDR2 that is SEQ ID NO:37; and LCDR3 that is any one of SEQ ID NOs:38-40, wherein the LCDR1, LCDR2 and LCDR3 are combined as they are arranged in each construct in Table 6.

34. The antigen binding construct, minibody, or scFv-Fc fusion of any one of the preceding claims, comprising a HCDR1 that is any one of SEQ ID NOs:43-46, HCDR2 that is any one of SEQ ID NOs:47, 48, 119, or 136, and HCDR3 that is SEQ ID NO: 49 or 50, wherein the HCDR1, HCDR2 and HCDR3 are combined as they are arranged in each construct in Table 6.

35. The antigen binding construct, minibody, or scFv-Fc fusion of any one of the preceding claims, comprising: a LCDR1 that is SEQ ID NO: 35 or 36; LCDR2 that is SEQ ID NO:37; and LCDR3 that is any one of SEQ ID NOs:38-40, wherein the LCDR1, LCDR2 and LCDR3 are combined as they are arranged in any construct in Table 6; and a HCDR1 that is any one of SEQ ID NOs:43-46, HCDR2 that is any one of SEQ ID NOs:47, 48, 119, or 136, and HCDR3 that is SEQ ID NO: 49 or 50, wherein the HCDR1, HCDR2 and HCDR3 are combined as they are arranged in a corresponding construct in Table 6.

36. The antigen binding construct, minibody, or scFv-Fc fusion of any one of the preceding claims, comprising: a variable light (VL) domain comprising an amino acid sequence at least 80% identical to any one of SEQ ID NOs:1-17; and a variable heavy (VH) domain comprising an amino acid sequence at least 80% identical to any one of SEQ IDNOs:18-34, 139, and 140, wherein the VL and VH domains are combined as they are arranged in any construct in Table 6.

37. The antigen binding construct or minibody of any one of the preceding claims, comprising an amino acid sequence at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to any one of SEQ ID NO. 103-118 or 133-135, optionally wherein the antigen binding construct comprises the amino acid sequence set forth in any one of SEQ ID NO.103-118 or 133-135.

38. An antigen binding construct comprising: a variable light (VL) domain comprising: an LCDR1 that is an LCDR1 in SEQ ID NO:2 or 3: an LCDR2 that is an LCDR2 in SEQ ID NO:2 or 3; and an LCDR3 that is an LCDR3 in SEQ ID NO:2 or 3, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:2 or 3; a variable heavy (VH) domain comprising: an HCDR1 that is an HCDR1 in SEQ ID NO:19 or 20: an HCDR2 that is an HCDR2 in SEQ ID NO:19 or 20; and an HCDR3 that is an HCDR3 in SEQ ID NO:19 or 20, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:19 or 20.

39. An antigen binding construct comprising: a variable light (VL) domain comprising: an LCDR1 that is an LCDR1 in SEQ ID NO:8: an LCDR2 that is an LCDR2 in SEQ ID NO:8; and an LCDR3 that is an LCDR3 in SEQ ID NO:8, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:8; a variable heavy (VH) domain comprising: an HCDR1 that is an HCDR1 in SEQ ID NO:25: an HCDR2 that is an HCDR2 in SEQ ID NO:25; and an HCDR3 that is an HCDR3 in SEQ ID NO:25, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:

25.

40. An antigen binding construct comprising: a variable light (VL) domain comprising: an LCDR1 that is an LCDR1 in SEQ ID NO:9: an LCDR2 that is an LCDR2 in SEQ ID NO:9; and an LCDR3 that is an LCDR3 in SEQ ID NO:9, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:9;a variable heavy (VH) domain comprising: an HCDR1 that is an HCDR1 in SEQ ID NO:26: an HCDR2 that is an HCDR2 in SEQ ID NO:26; and an HCDR3 that is an HCDR3 in SEQ ID NO:26, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:

26.

41. An antigen binding construct comprising: a variable light (VL) domain comprising: an LCDR1 that is an LCDR1 in SEQ ID NO:8: an LCDR2 that is an LCDR2 in SEQ ID NO:8; and an LCDR3 that is an LCDR3 in SEQ ID NO:8, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:8; a variable heavy (VH) domain comprising: an HCDR1 that is an HCDR1 in SEQ ID NO:139: an HCDR2 that is an HCDR2 in SEQ ID NO:139; and an HCDR3 that is an HCDR3 in SEQ ID NO:139, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:

139.

42. An antigen binding construct comprising: a variable light (VL) domain comprising: an LCDR1 that is an LCDR1 in SEQ ID NO:11: an LCDR2 that is an LCDR2 in SEQ ID NO:11; and an LCDR3 that is an LCDR3 in SEQ ID NO:11, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:11; a variable heavy (VH) domain comprising: an HCDR1 that is an HCDR1 in SEQ ID NO:28: an HCDR2 that is an HCDR2 in SEQ ID NO:28; and an HCDR3 that is an HCDR3 in SEQ ID NO:28, wherein the VL domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:

28.

43. A pharmaceutical composition comprising the antigen binding construct, minibody, scFv, or scFv-Fc fusion of any one of the preceding claims; and a therapeutic agent, toxic payload, and / or a detectable marker.

44. The pharmaceutical composition of claim 43, wherein the therapeutic agent, toxic payload, and / or a detectable marker is selected from a cytotoxic agent, a radioisotope, a chemotherapeutic agent, an immunotherapeutic agent, a DNA damage or DNA repair specific agent, a fluorescent compound, a bioluminescent compound, chemiluminescent compound, a metal chelator or an enzyme.

45. A method of detecting the presence or absence of a target molecule, comprising:applying the antigen binding construct, minibody, scFv, or scFv-Fc fusion of any one of claims 1-42 or the pharmaceutical composition of claim 43 or 44 to a sample; and detecting binding (or lack thereof) of the antigen binding construct, minibody, scFv, or scFv-Fc fusion to the target molecule in the sample.

46. The method of claim 45, comprising applying the antigen binding construct, minibody, scFv, or scFv-Fc fusion to the sample in vivo.

47. The method of claim 45, comprising applying the antigen binding construct, minibody, scFv, or scFv-Fc fusion to the sample in vitro.

48. A method of treating cancer, comprising administering the antigen binding construct, minibody, scFv, or scFv-Fc fusion any one of claims 1-42 or the pharmaceutical composition of claim 43 or 44 to a subject in need thereof, optionally wherein the method comprises administering the minibody of any one of claims 1-42 or the pharmaceutical composition of claim 43 or 44 to the subject.

49. The antigen binding construct, minibody, scFv, or scFv-Fc fusion any one of claims 1-37, for treatment of cancer in a subject in need thereof.

50. The pharmaceutical composition of claim 43 or 44, for treatment of cancer in a subject in need thereof.

51. The antigen binding construct, minibody, scFv, scFv-Fc fusion, composition or method of any one of the preceding claims, wherein the antigen binding construct, minibody, scFv, or scFv-Fc fusion binds to a ȕ6 subunit of ĮVȕ6 integrin, optionally wherein the antigen binding construct, minibody, scFv, or scFv-Fc fusion does not bind to an ĮV subunit of ĮVȕ6 integrin.

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