Anti-CD3 binding domains and antibodies containing said domains, and methods of making and using them

Abstract

Anti-CD3 binding domains and antibodies containing the domains, as well as methods for making and using them, are provided. [Solution] Anti-CD3 binding domains and antibodies containing the same, including multispecific antibodies with particularly desirable T cell activation and (re)directed target cell killing capabilities and developability profiles, as well as methods for identifying, isolating, and producing them, as well as methods for preparing and using them, are provided. Reagents for identifying, isolating, selecting, generating, and characterizing CD3 binding domains and antibodies containing the domains are also provided.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 62 / 503,315, filed May 8, 2017, which is incorporated by reference in its entirety.

[0002] Sequence Listing This application contains a Sequence Listing that has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy was created on April 7, 2017, is named 2009186-0189_SL.TXT, and is 2,923,505 bytes in size.

[0003] The present invention particularly relates to anti-Cluster of Differentiation 3 (CD3) binding domains and antibodies containing such domains, including multispecific and bispecific antibodies, and functional fragments thereof, and to methods and reagents for their identification, isolation, preparation, and use. Reagents for identifying, isolating, selecting, generating, and characterizing CD3 binding domains and antibodies containing such domains are also provided. [Background technology]

[0004] All references cited herein, including but not limited to patents, patent applications, and non-patent references and publications referenced throughout the specification, are hereby expressly incorporated by reference in their entirety for all purposes.

[0005] The body's immune system serves as a defense against infection, injury, and cancer. Two separate but interrelated systems, the humoral immune system and the cellular immune system, work together to defend the body. The humoral immune system is mediated by soluble factors called antibodies. Antibodies neutralize products recognized by the body as foreign. In contrast, the cellular immune system includes cells such as T cells and macrophages, which remove and neutralize foreign invaders.

[0006] T cell activation is crucial for stimulating immune responses. T cells exhibit immune specificity and direct the majority of cell-mediated immune responses. T cells do not secrete antibodies, but they are essential for antibody secretion by B lymphocytes. T cell activation requires the engagement of the T cell receptor complex and many cell surface molecules, such as CD4 or CD8 molecules. Antigen-specific T cell receptors (TcRs) consist of disulfide-linked heterodimers of membrane glycoproteins with alpha and beta (α and β) or gamma and delta (γ and δ) chains. TcRs are noncovalently associated with a complex of an invariant protein designated CD3.

[0007] The TcR confers antigen specificity, and the CD3 structure transmits activation signals to the T cell. The CD3 complex contains four subunits: two zeta subunits, one epsilon subunit, and may contain either a gamma or delta subunit. Antigen binding results in cross-linking and activation of the TCR complex. T cell receptor signaling results in T cell activation and production of IL-2 and other cytokines in a complex process.

[0008] The ligands for TcRs are MHC-peptide complexes on the surface of target cells, such as virus-infected cells. After recognizing the MHC-peptide on a target cell, T cells can have a cytotoxic or apoptotic effect on the target cell. In particular, cytotoxic T cells (CD8+ T cells) can have a beneficial effect by directly eliminating virus-infected cells. This weapon in the cellular immune response is particularly useful and is important for fighting viral infections and eliminating tumor cells.

[0009] Cytotoxic T cell activation can occur through direct binding of the CD3 antigen without TcR recognition of the MHC-peptide complex. This alternative activation pathway can be achieved with anti-CD3 antibodies. Non-human monoclonal antibodies against portions (subunits) of the CD3 chain have been developed, including the murine antibodies OKT3, SP34, UCHT1, and 64.1. (See, e.g., June, et al., J. Immunol. 136:3945-3952 (1986); Yang, et al., J. Immunol. 137:1097-1100 (1986); and Hayward, et al., Immunol. 64:87-92 (1988)). Other CD3 antibodies have been disclosed, for example, in U.S. Patent Nos. 5,585,097, 5,929,212, 5,968,509, 6,706,265, 6,750,325, 7,381,803, and 7,728,114. Bispecific antibodies with CD3 binding specificity have also been disclosed, for example, in U.S. Patent Nos. 7,262,276, 7,635,472, and 7,862,813.

[0010] Many of these anti-CD3 antibodies bind to the epsilon chain and generate highly activated T cells. Conventional monoclonal antibody-based cancer immunotherapy often does not sufficiently activate T lymphocytes to elicit meaningful, targeted pharmacological activity against target cell types or tissues.

[0011] The development and use of multispecific antibodies, such as bispecific antibodies (bsAbs), which redirect effector T cells to target killing of tumor cells, has shown considerable promise in recent years in preclinical and clinical settings (see, e.g., Topp et al., 2012, Blood 120:5185-87; Bargou et al., 2008, Science 321:974-77). Many bispecific antibodies developed to date contain a first binding site specific for CD3 for T cell recruitment and activation, and a second binding site for a target disease-associated antigen, such as CD19 (Bassan, 2012, Blood 120:5094-95). Bispecific antibodies bind to CD3 + It is believed that direct contact between T cells and target disease cells induces cell-mediated cytotoxicity (Bassan, 2012). Anti-CD3x anti-CD19 bispecific antibodies are effective in reducing MRD. + It has been reported that very low concentrations of CD3 induce complete and durable molecular remission in approximately 70% of adult patients with ALL (Topp et al, 2012, Blood 120:5185-87). Bispecific antibodies that recognize the CD3 epitope on glioma and T cells have been successfully used to treat brain tumors in human patients (Nitta, et al. Lancet 1990; 355:368-371). Additionally, CD3xCD20 bispecific antibodies (US2015 / 0166661 and US2017 / 0202194) and CD3xCLL-1 (US2016 / 0368994) are being produced for clinical trials.

[0012] Leukocyte-redirecting bsAbs are not limited to T cells. Bispecific killer engagers (BiKEs) containing scFvs against the NK cell antigen CD16 and tumor-associated antigens (e.g., CD19, CD22, and CD33) have also shown potent anticancer activity (e.g., Miller, Hematology Soc Hematol Educ Program 2013:247-53). Other examples include trispecific killer engagers (TriKEs), such as anti-CD16xanti-CD19xanti-CD22 (Miller, 2013; Gleason et al., 2012, Mol Cancer Ther 11:2674-84). Anti-CD16xanti-CD33 BiKEs have been used to treat AML and myelodysplastic syndromes (Miller, 2013; Wiernik et al., 2013, Clin Cancer Res 19:3844-55). In refractory AML, CD16xCD33 BiTEs resulted in potent tumor cell killing and cytokine production by NK cells. Inhibition of ADAM17 enhanced the CD16xCD33 BiTE response (Miller, 2013). Other trispecific constructs, such as trivalent constructs directed against CD16 / CD19 / HLA-DR, have also been reported (Schubert et al., 2012, mAbs 4:45-56).

[0013] Many methods for producing bispecific antibodies are known (see, e.g., U.S. Pat. No. 7,405,320). Bispecific antibodies can be produced by the quadroma method, which involves the fusion of two different hybridomas, each producing a monoclonal antibody that recognizes a different antigenic site (Milstein and Cuello, Nature 1983; 305:537-540). The fused hybridomas are capable of synthesizing two different heavy chains and two different light chains, which randomly associate to produce a heterogeneous population of 10 different antibody structures, only one of which is bispecific, accounting for 1 / 8 of the total antibody molecules. Therefore, it is difficult to distinguish this antibody from other types. The hybridomas must be further purified. Fused hybridomas are often less cytogenetically stable than the parent hybridomas, and therefore generation of producer cell lines is often problematic.

[0014] Another method for generating bispecific antibodies involves using heterobifunctional cross-linkers to link two different monoclonal antibodies, resulting in a hybrid conjugate that binds to two different targets (Staerz, et al. Nature 1985; 314:628-631; Perez, et al. Nature 1985; 316:354-356). Bispecific antibodies generated in this manner are essentially heteroconjugates of two IgG molecules, which diffuse slowly into tissues and are rapidly cleared from the circulation. Bispecific antibodies can also be generated by reducing each of the two parent monoclonal antibodies to their half molecules, combining them, and reoxidizing them to obtain a hybrid structure (Staerz and Bevan. Proc Natl Acad Sci USA 1986; 83:1453-1457). Another method involves chemically cross-linking two or three separate purified Fab' fragments using an appropriate linker. All these chemical methods are undesirable for commercial development due to their high production costs, laboratory manufacturing processes, extensive purification steps, low yields (less than 20%), and heterogeneous products.

[0015] Separate VH and VL domains of antibodies produced by recombinant DNA techniques can be paired with each other to form binding-competent dimers (recombinant Fv fragments) (U.S. Patent No. 4,642,334). However, such noncovalently linked molecules are not very stable under physiological conditions and are not suitable for practical use. Cognate VH and VL domains can also be linked with a peptide linker of appropriate composition and length (usually consisting of more than 12 amino acid residues) to form binding-competent single-chain Fvs (scFvs). Methods for generating multivalent and multispecific scFv-based agents by varying the linker length are disclosed in U.S. Patent No. 5,844,094, U.S. Patent No. 5,837,242, and WO 98 / 44001. Common challenges are often associated with generating multivalent and multispecific scFv-based agents: low expression levels, heterogeneous products, instability in solution resulting in aggregate formation, instability in serum, and diminished affinity.

[0016] Several bispecific antibodies targeting CD3 and CD19 are in various stages of development and / or have been approved as therapeutics. The scFv-based bispecific antibody construct, known as BITE® (Bispecific T-cell Engager), employs a single polypeptide containing two antigen-binding specificities, each conferred by a cognate VH and VL domain linked in tandem via a flexible linker (see, e.g., Nagorsen et al., 2009, Leukemia & Lymphoma 50:886-91; Amann et al., 2009, J Immunother 32:453-64; Baeuerle and Reinhardt, 2009, Cancer Res 69:4941-44). Another bispecific antibody is called DART® (Dual-Affinity Re-Targeting) and utilizes a disulfide-stabilized diabody design (see, e.g., Moore et al., 2011, Blood 117:4542-51; Veri et al., 2010, Arthritis Rheum 62: 1933-43). Both BITE® and DART® exhibit rapid blood clearance due to their small size (approximately 55 kDa). Therefore, the bispecific antibody requires frequent administration to maintain therapeutic levels.

[0017] SCORPION Therapeutics (Emergent Biosolutions, Inc., Seattle, WA) is a platform technology that combines two antigen-binding domains in a single-chain protein: one binding domain on the C-terminus of the effector domain base on the immunoglobulin Fc region, and the second binding domain on the N-terminus.

[0018] A tetravalent and bispecific antibody-like protein is DVD-Ig, which was created from two monoclonal antibodies (Wu, C. et al., Nature Biotechnology, 25, pp. 1290-1297, 2007). To construct a DVD-Ig molecule, the V domains of two mAbs are fused in tandem with a short linker (TVAAP) (SEQ ID NO: 6714), with the variable domain of the light chain (VL) of the first antibody at the N-terminus, followed by the VL and Ck of the other antibody, to form the DVD-Ig protein light chain. Similarly, the variable regions of the heavy chains (VH) of two mAbs are fused in tandem with a short linker (ASTKGP) (SEQ ID NO: 6715), with the variable domain of the first antibody at the N-terminus, followed by the heavy chain constant domain of the other antibody, to form the DVD-Ig protein heavy chain (VH1 / VL1). All light and heavy chain constant domains are preserved in the DVD-Ig design. This is because they are important for the formation of a complete disulfide-linked IgG-like molecule. Co-transfection of mammalian cells with expression vectors encoding the light and heavy chains of DVD-Ig results in the secretion of a single IgG-like molecule with a molecular weight of approximately 200 kDa. This molecule contains four binding sites, two from each mAb.

[0019] Bispecific antibodies have shown significant advantages over monospecific antibodies for cancer treatment and detection. Widespread commercial application of bispecific antibodies has been hindered by the lack of efficient / low-cost production methods, the lack of stability of bispecific polypeptides, and their short half-life in humans. Many methods for generating bispecific monoclonal antibodies (BsMABs) have been developed over the past decades.

[0020] However, although many clinical candidate and therapeutic antibodies exhibit sufficient selectivity and high potency against many target targets during early-stage development, the majority of these antibodies have been found to suffer from undesirable properties throughout their subsequent downstream development and clinical efficacy activities, such as binding promiscuity, polyspecific binding (also referred to throughout this specification as "polyspecificity"), off-target binding; nonspecific binding; poor expression levels or profiles in eukaryotic host cells, e.g., mammalian host cells and yeast cells; poor chemical and physical properties, such as poor stability during storage (e.g., poor / low "shelf-life" stability), poor (low) solubility, poor (high) viscosity, aggregation tendency, and the like; and poor clinical and biophysical profiles, such as poor pharmacokinetic profiles, poor pharmacodynamic profiles, rapid or poor in vivo clearance rates, short circulating half-lives, and the like, which often result in the termination of further therapeutic development of the candidate antibodies. Furthermore, antibodies derived from display technologies have historically been a minority of all clinical and commercial antibodies, a trend that many believe is at least in part due to promiscuous binding, poor PK profiles, and poor CMC characteristics, and it has further been hypothesized that these obstacles are largely due to the lack of appropriate tools and methods that can detect and / or counterselect undeviating antibodies when screening antibodies using display technologies (see, e.g., Meninger 2012; available at hyper-text transfer protocol: proteins-congress.com / wordpress / wp-content / uploads / 2012 / 01 / Trends-in-Therapeutic-Monoclonal-Antibody-Discovery-Technology.pdf).

[0021] The art has developed techniques and assays for evaluating many of the aforementioned developability characteristics of antibodies developed during downstream development activities (post-discovery antibodies), such as CIC, SIC, BVP-ELISA, TMA, and other assays. However, such assays are often not amenable to integration into high-throughput early polypeptide and antibody development platforms, such as antibody display platforms. Furthermore, these characterizations often require milligram- to gram-scale quantities of protein, often imposing a practical limit on the number of leads that can be considered for development. This, in turn, reduces the likelihood of program success. As a result, significant resources are often expended attempting to correct poorly characterized lead candidates, leaving few backups available in later development stages.

[0022] Recognizing this bottleneck, many attempts have been made to develop assays with reduced material requirements and move developability assessment further upstream in the development process (Esfandiary et al., 2013, Protein Eng Des Sel 26 (10): 663-670, 2013; Sathish et al., 2013, Nat Rev Drug Discov. 12(4):306-24). Many such assays aim to predict the antibody solubility and aggregation properties of identified lead candidates. Self-interaction chromatography (SIC) and cross-interaction chromatography (CIC) are column-based, low- to medium-throughput assays that correlate and predict antibody solubility at relatively low concentrations (Ahamed et al., 2005, J Biol Chem 280(37):32090-100; Jacobs et al., Pharm Res 27:65-71, 2010; Spencer et al., Mabs 4(3)319-325, 2012). Longer retention times on the SIC or CIC column suggest interactions with the column-bound antibody and correlate with poor solubility (Jacobs et al., 2010). Sule et al. reported a medium-throughput gold nanoparticle assay that predicts solubility at very low concentrations and further extended the assay range to be compatible with complex cell culture media (Sule et al., Biophys J 101(7):1749-1757, 2011; Mol Pharmaceutics 10(4):1322-1331, 2013).

[0023] As mentioned above, polyspecificity is a highly undesirable property associated with poor antibody pharmacokinetics (Wu et al., J Mol Biol 368:652-665, 2007; Hotzel et al., 2012, MAbs 4(6):753-760). A polyspecific assay has been reported in the art that serves as a medium-throughput alternative to broad-panel tissue immunohistochemistry. Wardemann et al. reported an enzyme-linked immunosorbent assay (ELISA) using LPS, insulin, dsDNA, and ssDNA to test for polyreactivity in the natural antibody repertoire during B cell maturation (Wardemann et al., 2003, Science 301(5638):1374-7). Protein biochips for high-throughput ELISA, in which a diverse set of proteins is arranged on an array, are another type of screening tool. Protagen (Dortmund, Germany)'s chip containing approximately 400 different human proteins has been reported to be comparable to IHC staining assays (Lueking et al., 2008, Bio Techniques 45(4):Pi-Pv) and to measure off-target binding of clinically approved TNF-alpha inhibitors (Feyen et al., Anal Bioanal Chem 391:1713-1720, 2008). Recently, Frese et al. reported a 384-well assay measuring the polyreactivity of 32 test proteins, termed Protein Panel Profiling or 3P (Frese et al., 2013, MAbs 5:2, 279-287). Using this assay, the authors demonstrated that FDA-approved therapeutic antibodies exhibited a high specificity profile for the 32 test proteins and that this assay could be applied to screening candidates from the phage selection process. These specific polyreactive profiling assays have not yet been correlated with downstream development issues such as solubility, expression and stability.Recent progress in this area was reported in Hotzel et al. 2012 MAbs 4(6):753-760), which showed that baculovirus particle (BVP) ELISA predicted non-target-mediated early clearance of antibodies in vivo, whereas traditional biophysical properties, such as size exclusion chromatography retention time, hydrophobic interaction chromatography elution time, Fv charge, and pI, did not (Hotzel, et al., 2012 MAbs 4(6):753-760). Recently, reagents, methods, and tools have been disclosed for evaluating, predicting, selecting, and enriching developable therapeutic antibodies from, for example, antibody libraries, and ultimately generating and obtaining developable antibodies (see, for example, WO2014 / 179363). CD3-targeting approaches have shown great promise. However, a common side effect of certain T cell immunostimulatory therapies is the production of related cytokines, which often leads to toxic cytokine release syndrome (CRS), also known as cytokine storm or cytokine release crisis. The anti-CD3 binding domain of bispecific antibodies engages all T cells, resulting in the recruitment of a highly cytokine-producing CD4+ T cell subset, which further includes regulatory T cells. The recruitment and expansion of regulatory T cells can potentially lead to immunosuppression, which may have a negative impact on long-term tumor suppression. Cell proliferative disorders, such as cancer, are characterized by the uncontrolled growth of a cell subpopulation. Cell proliferative disorders are the leading cause of death in developed countries and the second leading cause of death in developing countries. Each year, more than 12 million new cases of cancer are diagnosed, and 7 million people die from cancer. The National Cancer Institute estimates that more than 500,000 Americans died from cancer in 2013, accounting for nearly one in four deaths in the country. As the aging population grows, the incidence of cancer also rises simultaneously; the likelihood of developing cancer more than doubles after age 70. Cancer treatment, therefore, represents a significant and ever-increasing societal burden. Thus, there is a need to provide CD3-binding domains and antibodies (including multispecific antibodies) comprising the same that exhibit a desirable developability profile and / or CRS risk profile, e.g., in terms of binding specificity for CD3 expressed on T cells, T cell activation, (re)targeting of activated T cells, and target cell killing, while attenuating the risk of inducing cytokine release syndrome (also known as cytokine storm or cytokine release crisis), and that are safe and effective. [Prior art documents] [Patent documents]

[0024] [Patent Document 1] U.S. Patent No. 5,585,097 [Patent Document 2] U.S. Patent No. 5,929,212 [Patent Document 3] U.S. Patent No. 5,968,509 [Patent Document 4] U.S. Patent No. 6,706,265 [Patent Document 5] U.S. Patent No. 6,750,325 [Patent Document 6] U.S. Patent No. 7,381,803 [Patent Document 7] U.S. Patent No. 7,728,114 [Patent Document 8] U.S. Patent No. 7,262,276 [Patent Document 9] U.S. Patent No. 7,635,472 [Patent Document 10] U.S. Patent No. 7,862,813 [Patent Document 11] US Patent Application Publication No. 2015 / 0166661 [Patent Document 12] US Patent Application Publication No. 2017 / 0202194 [Patent Document 13] US Patent Application Publication No. 2016 / 0368994 [Patent Document 14] U.S. Patent No. 7,405,320 [Patent Document 15] U.S. Patent No. 4,642,334 [Patent Document 16] U.S. Patent No. 5,844,094 [Patent Document 17] U.S. Patent No. 5,837,242 [Patent Document 18] International Publication No. 98 / 44001 [Non-patent literature]

[0025] [Non-Patent Document 1] NJune, et al., J. Immunol. 136:3945-3952 (1986) [Non-patent document 2] Yang, et al., J. Immunol. 137:1097-1100 (1986) [Non-patent document 3] Hayward, et al., Immunol. 64:87-92 (1988) [Non-patent document 4] Topp et al., 2012, Blood 120:5185-87 [Non-Patent Document 5] Bargou et al, 2008, Science 321:974-77

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Summary of the Invention

[0026] A series of CD3 binding domains and antibodies containing the same, as well as methods of making and using them, have been discovered and are disclosed throughout the present specification. Members of this series of CD3 binding domains collectively exhibit a wide range of desirable properties, such as a wide range of affinities for CD3 epsilon, cross-reactivity to both human CD3 (Hu CD3) and cynomolgus monkey CD3 (Cy CD3), and a desirable developability profile and / or cytokine release syndrome (CRS) risk profile. These desirable properties have been found for many of the CD3 binding domains disclosed herein to be superior to the developability profiles of other anti-CD3 antibodies (e.g., I2C; SP34; 38E4; CAB21609_A01, CAB21609_B01, CAB21609_C01, CAB21609_D01, as disclosed herein and in Yang et al., J Immunol, Vol. 137, pp. 1097-1100 (August 4, 1986); US 2014 / 008295; and WO2015 / 095392).

[0027] Advantageously, CD3 binding domains can be incorporated into essentially all antibody formats, including immunoglobulin formats (e.g., IgG, IgM, IgA, IgE and their isotypes) and multispecific formats (including bispecific). Examples of multispecific formats suitable for incorporation into the CD3 binding domain of the present invention include: Fab-Fc-scFv (Bottle-Opener) (XENCOR), Mab-scFv (XENCOR), Mab-Fv (XENCOR), Dual scFv (XENCOR), central Fv (XENCOR), central scFv (XENCOR), one-arm central scFv (XENCOR), Fab-Fab (XENCOR), Fab-Fv (XENCOR), mAb-Fv (XENCOR), mAb-Fab (XENCOR), DART (MACROGENICS), BiTE (Amgen / Micromet), KiTE, common light chain-IgG (Genentech), TandAb (Affimed), Cross-Mab (Roche), SEED (EMD Serono), BEAT (Glenmark), TrioMab (Trion Pharma / FresEnius Biotech), DuetMab (MedImmune), and others, as described, for example, in (WO 95 / 09917; WO 2008 / 119566; WO 2008 / 119567; WO2011 / 121110; WO 2010 / 037835; WO 2007 / 042261; WO 2007 / 110205; WO 2011 / 121110; WO 2012 / 055961; WO 2012 / 16067; WO 2016 / 086189; WO 2016 / 182751; WO 2015 / 006749; WO 2014 / 049003; WO 2013 / 177101; WO 2015 / 128509; US 7,951,917; US 2009 / 0252729; US 2014 / 0348839; US 7,183,076; al., Mabs, Vol. 7, pages 377-389 (2015); Muda et al., Protein Engineering, Design, & Selection, Vol. 24, pp. 447-454 (2011); and Del Bano et al., Antibodies, Vol. 5, pp. 1-23 (2016).

[0028] The CD3-binding domains of the invention and antibodies comprising them collectively possess a wide range of affinities for cell-surface-expressed CD3, a wide range of T cell activation and (re)directed target cell killing capacities, and are amenable to essentially any multispecific (including bispecific) antibody format of any valency of interest. As such, the CD3 antibodies of the invention can be selected for and incorporated into therapeutic molecules designed to target almost any cell type, tissue type, and physiological compartment, and thus can serve as components of therapeutic molecules designed to address almost any disease type or disease state.

[0029] In certain embodiments, the present invention provides CD3 binding domains and antibodies comprising the same that bind to CD3 (eg, CD3ε and / or CD3γ).

[0030] In certain embodiments, CD3 binding domains of the invention and antibodies comprising the same are provided that exhibit an enhanced developability profile compared to other CD3 binding domains (or antibodies comprising the same). In certain embodiments, CD3 binding domains of the invention and antibodies comprising the same are provided that exhibit an enhanced developability profile compared to trastuzumab (Herceptin®), lintuzumab, blinatumomab (Blincyto®), and one or more of Mab 364, Mab 366, Mab 367, Mab 368, Mab 369, Mab 370, or Mab 22, as presented in Table 2.

[0031] In specific embodiments, the CD3-binding domains of the present invention and antibodies comprising the same have an affinity for an antibody having an affinity of about 0 MFI to about 500 MFI, about 0 MFI to about 450 MFI, about 0 MFI to about 400 MFI, about 0 MFI to about 350 MFI, about 0 MFI to about 300 MFI, about 0 MFI to about 250 MFI, about 0 MFI to about 200 MFI, about 0 MFI to about 150 MFI, about 0 MFI to about 100 MFI, about 0 MFI to about 50 MFI, about 200 MFI and exhibits a developability score of 500MFI, about 200MFI to about 450MFI, about 200MFI to about 400MFI, about 200MFI to about 350MFI, about 200MFI to about 300MFI, about 200MFI to about 250MFI, about 100MFI to about 450MFI, about 100MFI to about 400MFI, about 100MFI to about 350MFI, about 100MFI to about 300MFI, about 100MFI to about 250MFI, about 100MFI to about 200MFI, or about 100MFI to about 150MFI.

[0032] In other embodiments, the CD3-binding substances of the present invention and antibodies comprising the same have a CpG of about 0.0 to about 0.6, about 0.0 to about 0.57, about 0.0 to about 0.55, about 0.0 to about 0.53, about 0.0 to about 0.51, about 0.0 to about 0.49, about 0.0 to about 0.47, about 0.0 to about 0.45, about 0.0 to about 0.43, about 0.0 to about 0.41, about 0.0 to about 0.39, about 0.0 to about 0.37, about 0.0 to about 0.35, about 0.0 to about 0.4 exhibits a normalized developability score of about 0.33, about 0.0 to about 0.31, about 0.0 to about 0.29, about 0.0 to about 0.27, about 0.0 to about 0.25, about 0.0 to about 0.23, about 0.0 to about 0.21, about 0.0 to about 0.19, about 0.0 to about 0.17, about 0.0 to about 0.15, about 0.0 to about 0.13, about 0.0 to about 0.11, about 0.0 to about 0.09, about 0.0 to about 0.07, or about 0.0 to about 0.05.

[0033] In certain embodiments, the developability profile and / or developability score for the CD3 binding agents of the present invention and antibodies comprising the same are obtained by performing a PSR assay, an SCP assay, an AS-CINS, a BVP assay, an ELISA, a DSF assay, a Tm assay, an HIC assay, a CIC assay, or a combination thereof.

[0034] In other embodiments, the CD3-binding domains of the invention and antibodies comprising same elicit potent T cell activation or killing while exhibiting a reduced tendency to elicit cytokine production to levels capable of inducing cytokine release syndrome. In certain embodiments, cytokine production levels are measured to assess the tendency to elicit cytokine production levels capable of inducing cytokine release syndrome, and the at least one cytokine is selected from the group consisting of interleukin-6 (IL-6), interleukin-12 (IL-12), tumor necrosis factor alpha (TNFα), (TGFβ), interleukin-2 (IL-2), and interferon-gamma (IFNγ).

[0035] In certain embodiments, the CD3 binding domains of the invention and antibodies comprising same, while eliciting T cell activation or T cell killing, exhibit a reduced tendency to elicit cytokine production to levels capable of inducing cytokine release compared to the tendency observed with trastuzumab (Herceptin®), lintuzumab, blinatumomab (Blincyto®), and one or more of Mab 364, Mab 366, Mab 367, Mab 368, Mab 369, Mab 370, or Mab 22, as presented in Table 2. In certain embodiments, the level of cytokine production is measured to assess the propensity for at least one cytokine to elicit cytokine production levels capable of inducing cytokine release syndrome, wherein the at least one cytokine is selected from the group consisting of interleukin 6 (IL-6), interleukin 12 (IL-12), tumor necrosis factor alpha (TNFa), (TGFb), interleukin 2 (IL-2), and interferon gamma (IFNg).

[0036] In certain embodiments, the CD3-binding domains of the present invention and antibodies comprising the same exhibit a cytokine release syndrome risk profile indicative of a reduced risk of inducing cytokine release syndrome (CRS). In other embodiments, the CD3-binding domains of the present invention and antibodies comprising the same exhibit a cytokine release syndrome risk profile indicative of a reduced risk of inducing cytokine release syndrome (CRS) compared to the cytokine release syndrome risk profiles assessed with trastuzumab (Herceptin®), lintuzumab, blinatumomab (Blincyto®), and one or more of Mab 364, Mab 366, Mab 367, Mab 368, Mab 369, Mab 370, or Mab 22, as presented in Table 2.

[0037] In certain embodiments, the present invention provides antibodies containing a CDRH3 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRH3 selected from the group consisting of the CDRH3s of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2, provided that the CDRH3 is not 100% identical to the CDRH3 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370 as presented in Table 2.

[0038] In certain embodiments, the present invention provides antibodies containing a CDRH2 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRH2 selected from the group consisting of CDRH2 of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2, provided that the CDRH2 is not 100% identical to the CDRH2 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370 as presented in Table 2.

[0039] In certain embodiments, the present invention provides antibodies containing a CDRH1 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRH1 selected from the group consisting of CDRH1 of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2, provided that the CDRH1 is not 100% identical to the CDRH1 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370 as presented in Table 2.

[0040] In certain embodiments, the present invention provides antibodies containing a CDRL3 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRL3 selected from the group consisting of CDRL3s of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2, provided that the CDRL3 is not 100% identical to the CDRL3 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370 as presented in Table 2.

[0041] In certain embodiments, the present invention provides antibodies containing a CDRL2 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRL2 selected from the group consisting of CDRL2 of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2, provided that the CDRL2 is not 100% identical to the CDRL2 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370 as presented in Table 2.

[0042] In certain embodiments, the present invention provides antibodies containing a CDRL1 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRL1 selected from the group consisting of CDRL1 of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2, provided that the CDRL1 is not 100% identical to the CDRL1 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370 as presented in Table 2.

[0043] In certain embodiments, the present invention provides antibodies containing a heavy chain (HC) that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a HC selected from the group consisting of the HC of Mab 1-21, 23-222, 224-363, and 371-405 as presented in Table 2, provided that the HC is not 100% identical to the HC of any of Mab 223, 364, 365, 366, 367, 368, 369, or 370 as presented in Table 2.

[0044] In certain embodiments, the present invention provides antibodies containing a light chain (LC) that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a LC selected from the group consisting of the LC of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2, provided that the LC is not 100% identical to the LC of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370 as presented in Table 2.

[0045] In certain embodiments, the present invention provides a CDRH3 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRH3 selected from the group consisting of CDRH3s of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2. and a CDRH2 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRH2 selected from the group consisting of CDRH2s 1-21, 23-222, 224-363, and 371-405, as well as a Mab as set forth in Table 2.100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 89% identical, at least 9 ... Antibodies are provided that contain a CDRH1 that is 7% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical, provided that none of the CDRH3, CDRH2, and CHRH1 are 100% identical to the CHRH3, CDRH2, or CDRH1 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370, respectively, as set forth in Table 2.

[0046] In certain embodiments, the present invention provides a CDRL3 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRL3 selected from the group consisting of CDRL3s of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2. and a CDRL2 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRL2 selected from the group consisting of CDRL2s 1-21, 23-222, 224-363, and 371-405, as well as a Mab as set forth in Table 2.100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 89% identical, at least 9 ... Antibodies are provided that contain a CDRL1 that is 7% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical, provided that none of the CDRL3, CDRL2, and CHRL1 are 100% identical to the CHRL3, CDRL2, or CDRL1 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370, respectively, as set forth in Table 2.

[0047] In certain embodiments, the present invention provides a CDRH3 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRH3 selected from the group consisting of CDRH3s of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2. a CDRH2 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRH2 selected from the group consisting of CDRH2s 1-21, 23-222, 224-363, and 371-405, as presented in Table 2;a CDRH1 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRH1 selected from the group consisting of CDRH1s 1-21, 23-222, 224-363, and 371-405, as presented in Table 2; a CDRL3 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRL3 selected from the group consisting of CDRL3s 1-21, 23-222, 224-363, and 371-405, as presented in Table 2;and a CDRL2 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRL2 selected from the group consisting of CDRL2s 1-21, 23-222, 224-363, and 371-405, as well as a Mab as set forth in Table 2. 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, or at least 86% identical to a CDRL1 selected from the group consisting of CDRL1s 1 to 21, 23 to 222, 224 to 363, and 371 to 405. or at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical, provided that none of the CDRH3, CDRH2, CDRH1, CDRL3, CDHL2, and CHRL1 is 100% identical to the CDRH3, CDRH2, CDRH1, CDRL3, CDHL2, or CHRL1 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370, respectively, as set forth in Table 2.

[0048] In certain embodiments, the present invention provides antibodies containing a CD3 binding domain selected from the group consisting of the CD3 binding domains of ADI-15512, ADI-15516, and ADI-16513, as presented in Table 2.

[0049] In certain embodiments, the present invention provides a method for the treatment of a rheumatoid arthritis (RA) comprising administering to a patient a therapeutically effective amount ... Antibodies are provided that contain a CD3 binding domain selected from the group consisting of the CD3 binding domains of ADI-18590; ADI-18591; ADI-18593; ADI-18594; ADI-18595; ADI-18596; ADI-18597; ADI-18592; ADI-18587; ADI-18586, and ADI-16606.

[0050] In certain embodiments, the present invention provides a method for the preparation of a compound selected from the group consisting of: ADI-18576; ADI-20820; ADI-20578; ADI-20571; ADI-21097; ADI-20577; ADI-20576; ADI-20568; ADI-20582; ADI-20575; ADI-20567; ADI-20574; ADI-20573; ADI-20579; ADI-18565; ADI-20818; ADI-20587; ADI-20588; ADI-20589; ADI-20590; ADI-20594; ADI-20596; ADI-20599; ADI-20605; ADI-20607; An antibody containing a CD3 binding domain selected from the group consisting of the CD3 binding domains of ADI-20608 and ADI-20609 is provided.

[0051] In certain embodiments, the present invention is ADI-16606; ADI-20587; ADI-20607; ADI-20590; ADI-28708; ADI-28709; ADI-28710; ADI-21943; ADI-28711; ADI-28712; ADI-28713; ADI-28714; ADI-28715; ADI-21944; ADI-28716; ADI-21945; ADI-21946; ADI-28717; ADI-21947; ADI-28718; ADI-28719; ADI-28720; ADI-28721; ADI-28722; ADI-28723; ADI-28724; ADI-28725; ADI-28726; ADI-28727; ADI-28728; ADI-28729; ADI-28730; ADI-28731; ADI-28732; ADI-28733; ADI-28734; ADI-28735; ADI-28736; ADI-28737; ADI-28738; ADI-28739; ADI-28740; ADI-28741; ADI-28742; ADI-28743; ADI-21948; ADI-21949; ADI-28744; ADI-21950; ADI-28745; ADI-28746; ADI-28747; ADI-28748; ADI-21951; ADI-21952; ADI-28749; ADI-28750; ADI-28751; ADI-21953; ADI-28752; ADI-21954; ADI-28753; ADI-28754; ADI-28755; ADI-28756; ADI-28757; ADI-28758; ADI-28759; ADI-28760; ADI-28761; ADI-28762; ADI-28763; ADI-28764; ADI-28765; ADI-28766; ADI-28767; ADI-28768; ADI-21955; ADI-28769; ADI-28770; ADI-21956; ADI-28771; ADI-28772; ADI-28773 as presented in Table 2.An antibody containing a CD3-binding domain selected from the group consisting of the CD3-binding domains of ADI-28774 and ADI-28775 is provided.

[0052] In certain embodiments, the present invention provides a method for the treatment of rhodopsin-related ... Antibodies are provided that contain a CD3 binding domain selected from the group consisting of the CD3 binding domains of ADI-21954; ADI-21955; and ADI-21956.

[0053] In certain embodiments, the present invention provides a method for the treatment of a pulmonary artery disease comprising administering to a patient a therapeutically effective amount of a pulmonary artery disease (PAD) comprising administering to a patient a therapeutically effective amount of a pulmonary artery disease (PAH ... ADI-24443; ADI-24444; ADI-24445; ADI-24446; ADI-24449; ADI-24388; ADI-24389; ADI-24390; ADI-24391; ADI-24392; ADI-24393; ADI-24396; ADI-24397; ADI-24398; ADI-24399; ADI-24400; ADI-24401; ADI-24402; ADI-24419; ADI-24420; ADI-24421; ADI-24422; ADI-24425; Antibodies are provided that contain a CD3 binding domain selected from the group consisting of the CD3 binding domains of ADI-24426; ADI-24427; ADI-24428; ADI-24429; ADI-24430; ADI-24431; ADI-24432; ADI-24433; ADI-24447, and ADI-24448.

[0054] In certain embodiments, the present invention provides a method for the preparation of a compound selected from the group consisting of: ADI-22523; ADI-23652; ADI-23653; ADI-23654; ADI-23655; ADI-23656; ADI-23657; ADI-23658; ADI-23651; ADI-23644; ADI-23645; ADI-23646; ADI-23647; ADI-23648; ADI-23649; ADI-23650; ADI-23667; ADI-23668; ADI-23669; ADI-23670; ADI-23671; ADI-23672; ADI-23673; ADI-23659; ADI-23660; ADI-23661; Antibodies are provided that contain a CD3 binding domain selected from the group consisting of the CD3 binding domains of ADI-23663; ADI-23664; ADI-23639; ADI-23641; ADI-23642; ADI-23640; ADI-23643; ADI-21952; ADI-23633; ADI-23634; ADI-23635; ADI-23636; ADI-23637; ADI-23638; ADI-23632, and ADI-23629.

[0055] In certain embodiments, the present invention provides a method for the treatment of rhodium-containing ... ADI-26941; ADI-26942; ADI-26943; ADI-26944; ADI-26945; ADI-26950; ADI-26954; ADI-23672; ADI-23673; ADI-23664; ADI-26955; ADI-26958; ADI-26959; ADI-26960; ADI-26962; ADI-26963; ADI-26964; ADI-26965; ADI-26966; ADI-26974; Antibodies are provided that contain a CD3 binding domain selected from the group consisting of the CD3 binding domains of ADI-26975; ADI-26976; ADI-26977; ADI-26978; ADI-26979; ADI-26980; ADI-26981; ADI-26982; ADI-26983; ADI-26984; ADI-26985; ADI-26986; ADI-26987; ADI-26988; ADI-26989; ADI-26990; ADI-26991; ADI-26992; ADI-26993; ADI-26994, and ADI-26995.

[0056] In certain embodiments, the present invention provides antibodies containing a CD3 binding domain selected from the group consisting of the CD3 binding domains of ADI-22523; ADI-26906; ADI-26907; ADI-26908; ADI-26910; ADI-26913; ADI-26915; ADI-26919; ADI-26920; ADI-26921; ADI-26943; ADI-26954; ADI-21952; ADI-26955; ADI-26956; ADI-26962; ADI-26978; ADI-26983, and ADI-26994, as presented in Table 2.

[0057] In certain embodiments, the present invention provides an antibody containing a CD3 binding domain selected from the group consisting of: ADI-15512, ADI-16513, ADI-15516, ADI-18565, ADI-18589, ADI-18585, ADI-18590, ADI-18576, ADI-20568, ADI-20580, ADI-21978, ADI-22523, ADI-25133, and ADI-26906.

[0058] In certain embodiments, the present invention provides an antibody containing a CD3 binding domain selected from the group consisting of: ADI-16606, ADI-29601, ADI-29602, ADI-29603, ADI-20587, ADI-20607, ADI-20590, ADI-21952, ADI-23633, ADI-26955, ADI-26956, ADI-26957, ADI-26958, ADI-26959, ADI-26960, ADI-26961, ADI-26962, ADI-26963, ADI-26964, ADI-26965, ADI-26966, ADI-26967, ADI-26968, ADI-26969, ADI-26970, ADI-26971, ADI-26972, ADI-26973, ADI-26974, ADI-26975, ADI-26976, ADI-26977, ADI-26978, ADI-26979, ADI-26980, ADI-26981, ADI-26982, ADI-26983, ADI-26984, ADI-26985, ADI-26986, ADI-26987, ADI-26988, ADI-26989, ADI-26990, ADI-26991, ADI-26992, ADI-26993, ADI-26994, ADI-26995, ADI-26996 DI-26968, ADI-26969, ADI-26970, ADI-26971, ADI-26972, ADI-26973, ADI-26974 , ADI-26975, ADI-26976, ADI-26977, ADI-26978, ADI-26979, ADI-26980, ADI-269 81, ADI-26982, ADI-26983, ADI-26984, ADI-26985, ADI-26986, ADI-26987, ADI-2 6988, ADI-26989, ADI-26990, ADI-26991, ADI-26992, ADI-26993, and ADI-26994.

[0059] In certain embodiments, either alone or in combination with other embodiments of the invention, the CD3 binding domains of the invention and antibodies comprising same exhibit a reduced tendency to degradation compared to trastuzumab (Herceptin®), lintuzumab, blinatumomab (Blincyto®), and one or more of Mab 364, Mab 366, Mab 367, Mab 368, Mab 369, Mab 370 or Mab 22, as presented in Table 2.

[0060] In certain embodiments, either alone or in combination with other embodiments of the invention, the CD3 binding domains of the invention and antibodies comprising same exhibit a reduced CRS risk profile compared to trastuzumab (Herceptin®), lintuzumab, blinatumomab (Blincyto®), and one or more of Mab 364, Mab 366, Mab 367, Mab 368, Mab 369, Mab 370 or Mab 22, as presented in Table 2.

[0061] In certain embodiments, either alone or in combination with other embodiments of the invention, CD3 binding domains and antibodies comprising the same are provided that exhibit a reduced tendency to degradation compared to one or more of trastuzumab (Herceptin®), lintuzumab, blinatumomab (Blincyto®), and Mab 364, Mab 366, Mab 367, Mab 368, Mab 369, Mab 370, or Mab 22, as set out in Table 2.

[0062] In certain embodiments, and / or in combination with any of the embodiments disclosed throughout this specification, CD3 binding domains and humanized antibodies comprising the same are provided. In certain embodiments, such CD3 binding domains comprise the CDRs of such other embodiments and further comprise an acceptor human framework, such as a human immunoglobulin framework or a human consensus framework.

[0063] In particular embodiments, and / or in combination with any of the embodiments disclosed throughout the specification, there is provided an antibody comprising a CD3 binding domain, and comprising the same, a VH as in any of the embodiments presented throughout the specification, and a VL as in any of the embodiments presented throughout the specification, wherein one or both of the variable domain sequences comprises a post-translational modification.

[0064] In a further aspect of the present invention, there are provided CD3 binding domains and antibodies comprising same that bind to the same epitope as the CD3 binding domains provided in other embodiments disclosed throughout this specification.

[0065] In certain embodiments, the CD3 binding domain and / or antibodies comprising same have a cytotoxicity of ≦1 μM, ≦100 nM, ≦10 nM, ≦1 nM, ≦0.1 nM, ≦0.01 nM, or ≦0.001 nM (e.g., 10 -8 M or less, e.g. 10 -8 M~10 -13 M, e.g. 10 -9 M~10 -13 It has a CD3 dissociation constant (Kd) of 1 M.

[0066] In a further aspect of the invention, the CD3 binding domain and antibodies comprising the same comprise multispecific antibodies. In a further aspect of the invention, the CD3 binding domain and antibodies comprising the same comprise bispecific antibodies.

[0067] In further embodiments of the invention, the CD3 binding domain and antibodies comprising the same contain at least a second antigen-binding domain that specifically binds to an oncological target, an immuno-oncological target, a neurodegenerative disease target, an autoimmune disorder target, an infectious disease target, a metabolic disease target, a cognitive disorder target, a blood-brain barrier target, or a hematological disorder target.

[0068] In certain embodiments, the CD3 binding domains of the invention and antibodies comprising same comprise multispecific antibodies, e.g., bispecific antibodies, wherein the multispecific antibodies comprise a second binding domain having specificity for a second antigen selected from the group consisting of: 0772P (CA125, MUC16; Genbank Accession No. AF36148); adipophilin (perilipin-2, Adipose differentiation-related protein, ADRP, ADFP, MGC10598; NCBI Reference Sequence: NP-001113.2); AIM-2 (Absent In Melanoma 2, PYHIN4, Interferon-Inducible Protein AIM2; NCBI Reference Sequence: NP-004824.1); ALDH1 A1 (Aldehyde Dehydrogenase 1 Family, Member A1, ALDH1, PUMB1, Retinaldehyde Dehydrogenase 1, ALDC, ALDH-E1, ALHDII, RALDH 1, EC 1.2.1.36, ALDH11, HEL-9, HEL-S-53e, HEL12, RALDH1, Acetaldehyde Dehydrogenase 1, Aldehyde Dehydrogenase 1, Soluble, Aldehyde Dehydrogenase, liver cytosolic, ALDH Class 1, Epididymis Luminal Protein 12, Epididymis Luminal Protein 9, Epididymis Secretory Sperm Binding Protein Li 53e, Retinal Dehydrogenase 1, RaIDH1, Aldehyde Dehydrogenase Family 1 Member A1, Aldehyde Dehydrogenase, Cytosolic, EC 1.2.1; NCBI Reference Sequence: NP-000680.2); alpha-actinin-4 (ACTN4, Actinin, Alpha 4, FSGS1, Focal Segmental Glomerulosclerosis 1, Non-Muscle Alpha-Actinin 4, F-Actin Cross-Linking Protein, FSGS, ACTININ-4, Actinin Alpha4 Isoform, alpha-actinin-4; NCBI Reference Sequence: NP-004915.2); alpha-fetoprotein (AFP, HPAFP, FETA, alpha-1-fetoprotein, alpha-fetoglobulin, Alpha-1-fetoprotein, Alpha-fetoglobulin, HP; GenBank: AAB58754.1); Amphiregulin (AREG, SDGF, Schwannoma-Derived Growth Factor, Colorectum Cell-Derived Growth Factor, AR, CRDGF; GenBank: AAA51781.1); ARTC1 (ART1, ADP-Ribosyltransferase 1, Mono(ADP-Ribosyl)Transferase 1, ADP-Ribosyltransferase C2 And C3 Toxin-Like 1, ART2, CD296, RT6, ADP-Ribosyltransferase 2, GPI-Linked NAD(P)(+)-Arginine ADP-Ribosyltransferase 1, EC 2.4.2.31, CD296 Antigen; NP); ASLG659; ASPHD1 (Aspartate Beta-Hydroxylase Domain Containing 1, Aspartate Beta-Hydroxylase Domain-Containing Protein 1, EC 1.14.11., GenBank: AAI44153.1); B7-H4 (VTCN1, V-Set Domain Containing T Cell Activation Inhibitor 1, B7H4, B7 Superfamily Member 1, Immune Costimulatory Protein B7-H4, B7h.5, T-Cell Costimulatory Molecule B7x, B7S1, B7X, VCTN1, H4, B7 Family Member, PRO1291, B7 Family Member, H4, T Cell Costimulatory Molecule B7x, V-Set Domain-Containing T-Cell Activation Inhibitor 1, Protein B7S1; GenBank: AAZ17406.1); BAFF-R (TNFRSF13C, Tumor Necrosis Factor Receptor Superfamily, Member 13C, BAFFR, B-Cell-Activating Factor Receptor, BAFF Receptor, BLyS Receptor 3, CVID4, BROMIX, CD268, B Cell-Activating Factor Receptor, prolixin, Tumor Necrosis Factor Receptor Superfamily Member 13C, BR3, CD268 Antigen; NCBI reference sequence: NP-443177.1); BAGE-1; BCLX (L); BCR-ABL fusion protein (b3a2); beta-catenin (CTNNB1, Catenin (Cadherin-Associated Protein), Beta 1, 88 kDa, CTNNB, MRD19, Catenin (Cadherin-Associated Protein), Beta 1 (88kD), armadillo, Catenin Beta-1; GenBank: CAA61107.1); BING-4 (WDR46, WD Repeat Domain 46, C6orf11, BING4, WD Repeat-Containing Protein BING4, Chromosome 6 Open Reading Frame 11, FP221, UTP7, WD Repeat-Containing Protein 46; NP); BMPR1 B (bone morphogenetic protein receptor-type IB, Genbank accession number NM-00120; NP); B-RAF (Brevican (BCAN, BEHAB, Genbank accession number AF22905); Brevican (BCAN, Chondroitin Sulfate Proteoglycan 7, Brain-Enriched Hyaluronan-Binding Protein, BEHAB, CSPG7, Brevican Proteoglycan, Brevican Core Protein, Chondroitin Sulfate Proteoglycan BEHAB; GenBank: AAH27971.1); CALCA (Calcitonin-Related Polypeptide Alpha, CALC1, Calcitonin 1, calcitonin, Alpha-Type CGRP, Calcitonin Gene-Related Peptide I, CGRP-I, CGRP, CGRP1, CT, KC, Calcitonin / Calcitonin-Related Polypeptide, Alpha, katacalcin; NP); CASP-5 (CASP5, Caspase 5, Apoptosis-Related Cysteine Peptidase, Caspase 5, Apoptosis-Related Cysteine Protease, Protease ICH-3, Protease TY, ICE(rel)-111, ICE(rel)III, ICEREL-III, ICH-3, caspase-5, TY Protease, EC 3.4.22.58, ICH3, EC 3.4.22; NP); CASP-8; CD19 (CD19 - B-lymphocyte antigen CD19 isoform 2 precursor, B4, CVID3 [Homo sapiens], NCBI Reference Sequence: NP-001761.3); CD20 (CD20 - B-lymphocyte antigen CD20, membrane-spanning 4-domains, subfamily A, member 1, B1, Bp35, CD20, CVID5, LEU-16, MS4A2, S7; NCBI Reference Sequence: NP-690605.1); CD21 (CD21 (CR2 (Complement receptor) or C3DR (C3d / Epstein-Barr virus receptor) or Hs.73792 Genbank Accession Number M2600); (CD22 (B-cell receptor CD22-B isoform, BL-CAM, Lyb-8, LybB, SIGLEC-2, FLJ22814, GenBank accession number AK02646); CD22; CD33 (CD33 molecule, CD33 antigen (Gp67), Sialic Acid Binding Ig-Like Lectin 3, Sialic Acid-Binding Ig-Like Lectin 3, SIGLEC3, gp67, SIGLEC-3, Myeloid Cell Surface Antigen CD33, p67, Siglec-3, CD33 antigen; GenBank: AAH28152.1); CD45; CD70 (CD70-tumor necrosis factor (ligand) superfamily, member 7; surface antigen CD70; Ki-24 antigen; CD27 ligand; CD27-L; tumor necrosis factor ligand superfamily member 7; NCBI reference sequence for Homo sapiens species: NP-001243.1); CD72 (CD72 (B-cell differentiation antigen CD72, Lyb-; 359 aa, μl: 8.66, MW: 40225, TM: 1 [P] gene chromosome: 9p13.3, Genbank accession number NP-001773.); CD79a (CD79a (CD79A, CD79a, immunoglobulin-associated alpha, a B cell-specific protein that covalently interacts with Ig beta (CD79B) to form a complex on the surface with IgM molecules and transmits signals involved in B cell differentiation), μl: 4.84, MW: 25028 TM: 2 [P] gene chromosome: 19q13.2, Genbank accession number NP-001774.1); CD79b (CD79b (CD79B, CD79b, IGb (immunoglobulin-associated beta), B29, Genbank accession number NM-000626 or 1103867); Cdc27 (Cell Division Cycle 27, D0S1430E, D17S978E, Anaphase Promoting Complex Subunit 3, Anaphase-Promoting Complex Subunit 3, ANAPC3, APC3, CDC27Hs, H-NUC, CDC27 Homolog, Cell Division Cycle 27 Homolog (S.Cerevisiae), HNUC, NUC2, Anaphase-Promoting Complex, Protein 3, Cell Division Cycle 27 Homolog, Cell Division Cycle Protein 27 Homolog, Nuc2 Homolog; GenBank: AAH11656.1); CDK4 (Cyclin-Dependent Kinase 4, Cell Division Protein Kinase 4, PSK-J3, EC 2.7.11.22, CMM3, EC 2.7.11); NCBI reference sequence: NP-000066.1); CDKN2A (Cyclin-Dependent Kinase Inhibitor 2A, MLM, CDKN2, MTS1, Cyclin-Dependent Kinase Inhibitor 2A (Melanoma, P16, Inhibits CDK4), Cyclin-Dependent Kinase 4 Inhibitor A, Multiple Tumor Suppressor 1, CDK4I, MTS-1, CMM2, P16, ARF, INK4, INK4A, P14, P14ARF, P16-INK4A, P16INK4, P16INK4A, P19, P19ARF, TP16, CDK4 inhibitor P16-INK4, Cell Cycle Negative Regulator Beta, p14ARF, p16-INK4, p16-INK4a, p16INK4A, p19ARF; NP); CEA; CLL1 (CLL-1), CLEC12A, MICL, and DCAL, are members of the C-type lectin / C-type lectin-like domain (CTL / CTLD) superfamily. Members of this family share a common protein fold and have diverse functions, including roles in adhesion, cell-cell signaling, glycoprotein turnover, and inflammation and immune responses. The protein encoded by this gene is a negative regulator of granulocyte and monocyte function. Several alternatively spliced ​​transcript variants of this gene have been reported, but the full-length nature of some of these variants is unknown. This gene is closely related to other CTL / CTLD superfamily members in the natural killer gene complex region on chromosome 12p13 (Drickamer, K. Curr. Opin. Struct. Biol. 9:585-90

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[2006] ; Bakker AB, et al Cancer Res.64:8443-50

[2004] ; Marshall AS, et al J. Biol. Chem. 279:14792-80, 2004. CLL-1 has been shown to be a type II transmembrane receptor containing a single C-type lectin-like domain (not predicted to bind either calcium or sugars), a stalk region, a transmembrane domain, and a short cytoplasmic tail containing ITIM motifs; CLPP (Caseinolytic Mitochondrial Matrix Peptidase Proteolytic Subunit, Endopeptidase Clp, EC 3.4.21.92, PRLTS3, ATP-Dependent Protease ClpAP (E. coli), ClpP (Caseinolytic Protease, ATP-Dependent, Proteolytic Subunit, E. coli) Homolog, ClpP Caseinolytic Peptidase, ATP-Dependent, Proteolytic Subunit Homolog (E. coli), ClpP Caseinolytic Protease, ATP-Dependent, Proteolytic Subunit Homolog (E. coli)coli), human, Proteolytic Subunit, ATP-Dependent Protease ClpAP, Proteolytic Subunit, Human, ClpP Caseinolytic Peptidase ATP-Dependent, Proteolytic Subunit, ClpP Caseinolytic Peptidase, ATP-Dependent, Proteolytic Subunit Homolog, ClpP Caseinolytic Protease, ATP-Dependent, Proteolytic Subunit Homolog, Putative ATP-Dependent Clp Protease Proteolytic Subunit, Mitochondrial; NP); COA-1; CPSF; CRIPTO CXCR5 (Burkitt's lymphoma receptor CXORF61 (CXORF61-chromosome X open reading frame 61 [Homo sapiens], NCBI reference sequence: NP-001017978).1); cyclin D1 (CCND1, BCL1, PRAD1, D11S287E, B-Cell CLL / Lymphoma 1, B-Cell Lymphoma 1 Protein, BCL-1 Oncogene, PRAD1 Oncogene, Cyclin D1 (PRAD1: Parathyroid Adenomatosis 1), G1 / S-Specific Cyclin D1, Parathyroid Adenomatosis 1, U21B31, G1 / S-Specific Cyclin-D1, BCL-1; NCBI reference sequence: NP-444284.1); Cyclin-A1 (CCNA1, CT146, Cyclin A1; GenBank: AAH36346.1); dek-can fusion protein; DKK1 (Dickkopf WNT Signaling Pathway Inhibitor 1, SK, hDkk-1, Dickkopf (Xenopus Laevis) Homolog 1, Dickkopf 1 Homolog (Xenopus Laevis), DKK-1, Dickkopf 1 Homolog, Dickkopf Related Protein-1, Dickkopf-1 Like, Dickkopf-Like Protein 1, Dickkopf-Rel. Dickkopf-1, Dkk-1; GenBank: AAQ89364.1); DR1 (Down-Regulator Of Transcription 1, TBP-Binding (Negative Cofactor 2), Negative Cofactor 2-Beta, TATA-Binding Protein-Associated Phosphoprotein, NC2, NC2-BETA, Protein Dr1, NC2-beta, Down-Regulator Of Transcription 1; NCBI reference sequence: NP-001929.1);DR13 (Major Histocompatibility Complex, Class II, DR Beta 1, HLA-DR1B, DRw10, DW2.2 / DR2.2, SS1, DRB1, HLA-DRB, HLA Class II Histocompatibility Antigen, DR-1 Beta Chain, Human Leucocyte Antigen DRB1, Lymphocyte Antigen DRB1, MHC Class II Antigen, MHC Class II HLA-DR Beta 1 Chain, MHC Class II HLA-DR-Beta Cell Surface Glycoprotein, MHC Class II HLA-DRw10-Beta, DR-1, DR-12, DR-13, DR-14, DR-16, DR-4, DR-5, DR-7, DR-8, DR-9, DR1, DR12, DR13, DR14, DR16, DR4, DR5, DR7, DRB, DR9, DRw11, DRw8, HLA-DRB2, Clone P2-Beta-3, MHC Class II Antigen DRB1*1, MHC Class II Antigen DRB1*10, MHC Class II Antigen DRB1*11, MHC Class II Antigen DRB1*12, MHC Class II Antigen DRB1*13, MHC Class II Antigen DRB1*14, MHC Class II Antigen DRB1*15, MHC Class II Antigen DRB1*16, MHC Class II Antigen DRB1*3, MHC Class II Antigen DRB1*4, MHC Class II Antigen DRB1*7, MHC Class II Antigen DRB1*8, MHC Class II Antigen DRB1*9; NP); E16 (E16 (LAT1, SLC7A5, Genbank accession number NM-00348); EDAR (EDAR - tumor necrosis factor receptor superfamily member EDAR precursor, EDA-A1 receptor;downless homolog; ectodysplasin-A receptor; ectodermal dysplasia receptor; anhydrotic ectodysplasin receptor 1、DL; ECTD10A; ECTD10B; ED1R; ED3; ED5; EDA-A1R; EDA1R; EDA3; HRM1 [Homo sapiens]; NCBI Publication Number: NP-071731.1); EFTUD2 (Elongation Factor Tu GTP Binding Domain Containing 2、Elongation Factor Tu GTP-Binding Domain-Containing Protein 2、hSNU114、SNU114 Homolog、U5 SnRNP-Specific Protein、116 KDa、MFDGA、KIAA0031、116 KD、U5 SnRNP Specific Protein、116 KDa U5 Small Nuclear Ribonucleoprotein Component, MFDM, SNRNP116, Snrp116, Snu114, U5-116KD, SNRP116, U5-116 KDa;EGFR (Epidermal Growth Factor Receptor、ERBB、Proto-Oncogene C-ErbB-1、Tyrosine-Protein Kinase Receptor ErbB-1、ERBB1、HER1、EC 2.7.10.1、Epidermal Growth Factor Receptor (Avian Erythroblastic Leukemia Viral (V-Erb-B) Oncogene Homolog)、Viral Erythroblastic Leukemia (V-Erb-B) Oncogene Homolog (Avian)、PlG61、Avian Erythroblastic Leukemia Viral (V-Erb-B) Oncogene Homolog、Cell Growth Inhibiting Protein 40、Cell Proliferation-Inducing Protein 61、mENA、EC 2.7.10; EGFR-G719A; EGFR-G719C; EGFR-G719S; EGFR-L858R; EGFR-L861Q; EGFR-57681; EGFR-T790M; Elongation factor 2 (EEF2、Eukaryotic Translation Elongation Factor 2、EF2、Polypeptidyl-TRNA Translocase、EF-2、SCA26、EEF-2; NCBI National Park Service: NP-001952.1); ENAH (hMena) (Enabled Homolog (Drosophila), MENA, Mammalian Enabled, ENA, NDPP1, Enabled Protein Homolog; GenBank: AAH95481.1)-“ENAH” means “ENAH (hMena)”;EpCAM (Epithelial Cell Adhesion Molecule、M4S1、MIC18、Tumor-Associated Calcium Signal Transducer 1、TACSTD1、TROP1、Adenocarcinoma-Associated Antigen、Cell Surface Glycoprotein Trop-1、Epithelial Glycoprotein 314、Major Gastrointestinal Tumor-Associated Protein GA733-2、EGP314、KSA、DIAR5、HNPCC8、Antigen Identified By Monoclonal Antibody AUA1、EGP-2、EGP40、ESA、KS1 / 4、MK-1、Human Epithelial Glycoprotein-2, Membrane Component, Chromosome 4, Surface Marker (35kD Glycoprotein)、EGP、Ep-CAM、GA733-2、M1S2、CD326 Antigen、Epithelial Cell Surface Antigen、hEGP314、KS 1 / 4 Antigen、ACSTD1; GenBank: AAH14785.1); EphA3 (EPH Receptor A3、ETK1、ETK、TYRO4、HEK、Eph-Like Tyrosine Kinase 1、Tyrosine-Protein Kinase Receptor ETK1、EK4、EPH-Like Kinase 4、EC 2.7.10.1、EPHA3、HEK4、Ephrin Type-A Receptor 3、Human Embryo Kinase 1、TYRO4 Protein Tyrosine Kinase、hEK4、Human Embryo Kinase、Tyrosine-Protein Kinase TYRO4、EC 2.7.10; GenBank: AAH63282.1); EphB2R; Epiregulin (EREG、ER、proepiregulin; GenBank: AAI36405.1);ETBR (EDNRB, Endothelin Receptor Type B, HSCR2, HSCR, Endothelin Receptor Non-Selective Type, ET-B, ET-BR, ETRB, ABCDS, WS4A, ETB, Endothelin B Receptor; NP); ETV6-AML1 fusion protein; EZH2 (Enhancer Of Zeste Homolog 2 (Drosophila), Lysine N-Methyltransferase 6, ENX-1, KMT6 EC 2.1.1.43, EZH1, WVS, Enhancer Of Zeste (Drosophila) Homolog 2, ENX1, EZH2b, KMT6A, WVS2, Histone-Lysine N-Methyltransferase EZH2, Enhancer Of Zeste Homolog 2, EC 2.1.1; GenBank: AAH10858.1); FcRH1 (FCRL1, Fc Receptor-Like 1, FCRH1, Fc Receptor Homolog 1, FcR-Like Protein 1, Immune Receptor Translocation-Associated Protein 5, IFGP1, IRTA5, hIFGP1, IFGP Family Protein 1, CD307a, Fc Receptor-Like Protein 1, Immunoglobulin Superfamily Fc Receptor, Gp42, FcRL1, CD307a Antigen; GenBank: AAH33690.1);FcRH2 (FCRL2、Fc Receptor-Like 2、SPAP1、SH2 Domain-Containing Phosphatase Anchor Protein 1、Fc Receptor Homolog 2、FcR-Like Protein 2、Immunoglobulin Receptor Translocation-Associated Protein 4、FCRH2、IFGP4、IRTA4、IFGP Family Protein 4、SPAP1A、SPAP1 B、SPAP1C、CD307b、Fc Receptor-Like Protein 2、Immune Receptor Translocation-Associated Protein 4、Immunoglobulin Superfamily Fc Receptor、Gp42、SH2 Domain Containing Phosphatase Anchor Protein 1、FcRL2、CD307b Antigen; GenBank: AAQ88497.1); FcRH5 (FCRL5、Fc Receptor-Like 5、IRTA2、Fc Receptor Homolog 5、FcR-Like Protein 5、Immune Receptor Translocation-Associated Protein 2、BXMAS1、FCRH5、CD307、CD307e、PRO820、Fc Receptor-Like Protein 5、Immunoglobulin Superfamily Receptor Translocation Associated 2 (IRTA2)、FCRL5、CD307e Antigen; GenBank: AAI01070.1); FLT3-ITD; FN1(Fibronectin 1、Cold-Insoluble Globulin、FN、Migration-Stimulating Factor、CIG、FNZ、GFND2、LETS、ED-B、FINC、GFND、MSF、fibronectin; GenBank: AAI43764.1);G250 (MN、CAIX、Carbonic Anhydrase IX、Carbonic Dehydratase、RCC-Associated Protein G250、Carbonate Dehydratase IX、Membrane Antigen MN、Renal Cell Carcinoma-Associated Antigen G250, CA-IX, P54 / 58N, pMW1, RCC-Associated Antigen G250, Carbonic Anhydrase 9; NP); GAGE-1,2,8; GAGE-3,4,5,6,7; GDNF-Ra1 (GDNF family receptor alpha 1; GFRA1; GDNFR; GDNFRA; RETL1; TRNR1; RET1 L; GDNFR-alpha1; GFR-ALPHA-; U95847; BC014962; NM-145793 NM-005264); GEDA (Genbank database AY26076); GFRA1-GDNF family receptor alpha-1; GDNF receptor alpha-1; GDNFR-alpha-1; GFR-alpha-1; RET ligand 1; TGF-beta-related neurotrophic factor receptor 1 [Homo sapiens]; ProtKB / Swiss-Prot: P56159.2; glypican-3 (GPC3, Glypican 3, SDYS, Glypican Proteoglycan 3, Intestinal Protein OCI-5, GTR2-2, MXR7, SGBS1, DGSX, OCI-5. SGB, SGBS, Heparan Sulphate Proteoglycan, Secreted glypican-3、OCI5;GenBank:AAH35972.1); GnTVf;gp100 (PMEL、Premelanosome Protein、SILV、D12S53E、PMEL17、SIL、Melanocyte Protein Pmel 17、Melanocytes Lineage-Specific Antigen GP100、Melanoma-Associated ME20 Antigen、Silver Locus Protein Homolog、ME20-M、ME20M、P1、P100、Silver (Mouse Homolog) Like、Silver Homolog (マウス)、ME20、SI、Melanocyte Protein Mel 17、Melanocyte Protein PMEL、Melanosomal Matrix Protein17, Silver, Mouse, Homolog Of; GenBank: AAC60634.1); GPC; GPNMB (Glycoprotein (Transmembrane) Nmb、Glycoprotein NMB、Glycoprotein Nmb-Like Protein、osteoactivin、Transmembrane Glycoprotein HGFIN、HGFIN、NMB、Transmembrane Glycoprotein、Transmembrane Glycoprotein NMB; GenBank: AAH32783.1); GPR172A (G protein-coupled receptor 172A; GPCR41; FLJ11856; D15Ertd747e); NP-078807.1; NM-024531.3); GPR19 (G protein-coupled receptor 19; Mm.478; NP-006134.1; NM-006143.2); GPR54 (KISS1 receptor; KISS1R; GPR54; HOT7T175; AXOR1; NP-115940.2; NM-032551.4);HAVCR1 (Hepatitis A Virus Cellular Receptor 1、T-Cell Immunoglobulin Mucin Family Member 1、Kidney Injury Molecule 1、KIM-1、KIM1、TIM、TIM-1、TIM1、TIMD-1、TIMD1、T-Cell Immunoglobulin Mucin Receptor 1、T-Cell Membrane Protein 1、HAVCR、HAVCR-1、T Cell Immunoglobin Domain And Mucin Domain Protein 1、HAVcr-1、T-Cell Immunoglobulin And Mucin Domain-Containing Protein 1; GenBank: AAH13325.1);HER2 (ERBB2, V-Erb-B2 Avian Erythroblastic Leukemia Viral Oncogene Homolog 2, NGL, NEU, Neuro / Glioblastoma Derived Oncogene Homolog, Metastatic Lymph Node Gene 19 Protein, Proto-Oncogene C-ErbB-2, Proto-Oncogene Neu, Tyrosine Kinase-Type Cell Surface Receptor HER2, MLN 19, p185erbB2, EC 2.7.10.1, V-Erb-B2 Avian Erythroblastic Leukemia Viral Oncogene Homolog 2 (Neuro / Glioblastoma Derived Oncogene Homolog), CD340, HER-2, HER-2 / neu, TKR1, C-Erb B2 / Neu Protein, herstatin, Neuroblastoma / Glioblastoma Derived Oncogene Homolog, Receptor Tyrosine-Protein Kinase ErbB-2, V-Erb-B2 Erythroblastic Leukemia Viral Oncogene Homolog 2, Neuro / Glioblastoma Derived Oncogene Homolog, MLN19, CD340 Antigen, EC 2.7.10; NP); HER-2 / neu - synonyms of the above; HERV-K-MEL; HLA-DOB (beta subunit (la antigen) of MHC class II molecule that binds peptides and presents them to CD4+ T lymphocytes); 273 aa, μl: 6.56, MW: 30820. TM: 1 [P] gene chromosome: 6p21.3, GenBank accession number NP-002111);hsp70-2 (HSPA2, Heat Shock 70 kDa Protein 2, Heat Shock 70kD Protein 2, HSP70-3, Heat Shock-Related 70 KDa Protein 2, Heat Shock 70 KDa Protein 2; GenBank: AAD21815.1); IDO1 (Indoleamine 2,3-Dioxygenase 1, IDO, INDO, Indoleamine-Pyrrole 2,3-Dioxygenase, IDO-1, Indoleamine-Pyrrole 2,3 Dioxygenase, Indolamine 2,3 Dioxygenase, Indole 2,3 Dioxygenase, EC 1.13.11.52; NCBI reference sequence: NP-002155.1); IGF2B3; IL13Ralpha2 (IL13RA2, Interleukin 13 Receptor, Alpha 2, Cancer / Testis Antigen 19, Interleukin-13-Binding Protein, IL-13R-alpha-2, IL-13RA2, IL-13 Receptor Subunit Alpha-2, IL-13R Subunit Alpha-2, CD213A2, CT19, IL-; 13R、IL13BP、Interleukin 13 Binding Protein、Interleukin 13 Receptor Alpha 2 Chain、Interleukin-13 Receptor Subunit Alpha-2、IL13R、CD213a2 Antigen; NP); IL20Rα; Intestinal carboxyl esterase; IRTA2 (alias of FcRH5); Kallikrein 4 (KLK4、Kallikrein-Related Peptidase 4、PRSS17、EMSP1、Enamel Matrix Serine Proteinase 1、Kallikrein-Like Protein 1、Serine Protease 17、KLK-L1、PSTS、AI2A1、Kallikrein 4 (Prostase、Enamel Matrix、Prostate)、ARM1、EMSP、Androgen-Regulated Message 1、Enamel Matrix Serine Protease 1、kallikrein、kallikrein-4、prostase、EC 3.4.21.-、Prostase、EC 3.4.21; GenBank: AAX30051.1); KIF20A (Kinesin Family Member 20A、RAB6KIFL、RAB6 Interacting、Kinesin-Like (Rabkinesin6)、Mitotic a; LAGE-1; LDLR-fucosyltransferase AS fusion protein;Lens Glutamine Synthase-Like; GenBank: AAF61255.1); LGR5 (leucine-rich repeat-containing G protein-coupled receptor 5; GPR49, GPR6; NP-003658.1; NM-003667.2; LY64 (Lymphocyte antigen 64) (RP10, a type I membrane protein of the leucine-rich repeat (LRR) family that regulates B cell activation and apoptosis. Loss of its function has been associated with increased disease activity in patients with systemic lupus erythematosus); 661 aa, μl: 6.20, MW: 74147 TM: 1 [P] Gene chromosome: 5q12, GenBank accession number NP-005573.; Ly6E (lymphocyte antigen 6 complex, locus E; Ly67, RIG-E, SCA-2, TSA-; NP-002337.1; NM-002346.2); Ly6G6D (lymphocyte antigen 6 complex, locus G6D; Ly6-D, MEGT; NP-067079.2; NM-021246.2); LY6K (lymphocyte antigen 6 complex, locus K; LY6K; HSJ001348; FLJ3522; NP-059997.3; NM-017527.3); LyPD1-LY6 / PLAUR domain containing 1, PHTS [Homo sapiens], GenBank: AAH17318.1);MAGE-A1 (Melanoma Antigen Family A, 1 (Directs Expression Of Antigen MZ2-E, MAGE1, Melanoma Antigen Family A 1, MAGEA1, Melanoma Antigen MAGE-1, Melanoma-Associated Antigen 1, Melanoma-Associated Antigen MZ2-E, Antigen MZ2-E, Cancer / Testis Antigen 1.1, CT1.1, MAGE-1 Antigen, Cancer / Testis Antigen Family 1, Member 1, Cancer / Testis Antigen Family 1, Member 1, MAGE1A; NCBI reference sequence: NP-004979.3); MAGE-A10 (MAGEA10, Melanoma Antigen Family A, 10, MAGE10, MAGE-10 Antigen, Melanoma-Associated Antigen 10, Cancer / Testis Antigen 1.10, CT1.10, Cancer / Testis Antigen Family 1, Member 10, Cancer / Testis Antigen Family 1, Member 10; NCBI reference sequence: NP-001238757.1); MAGE-A12 (MAGEA12, Melanoma Antigen Family A, 12, MAGE12, Cancer / Testis Antigen 1.12, CT1.12, MAGE12F Antigen, Cancer / Testis Antigen Family 1, Member 12, Cancer / Testis Antigen Family 1, Member 12, Melanoma-Associated Antigen 12, MAGE-12 Antigen; NCBI reference sequence: NP-001159859.1);MAGE-A2 (MAGEA2, Melanoma Antigen Family A, 2, MAGE2, Cancer / Testis Antigen 1.2, CT1.2, MAGEA2A, MAGE-2 Antigen, Cancer / Testis Antigen Family 1, Member 2, Cancer / Testis Antigen Family 1, Member 2, Melanoma Antigen 2, Melanoma-Associated Antigen 2; NCBI Reference Sequence: NP-001269434.1); MAGE-A3 (MAGEA3, Melanoma Antigen Family A, 3, MAGE3, MAGE-3 Antigen, Antigen MZ2-D, Melanoma-Associated Antigen 3, Cancer / Testis Antigen 1.3, CT1.3, Cancer / Testis Antigen Family 1, Member 3, HIPS, HYPD, MAGEA6, Cancer / Testis Antigen Family 1, Member 3; NCBI Reference Sequence: NP-005353.1); MAGE-A4 (MAGEA4, Melanoma Antigen Family A, 4, MAGE4, Melanoma-Associated Antigen 4, Cancer / Testis Antigen 1.4, CT1.4, MAGE-4 Antigen, MAGE-41 Antigen, MAGE-X2 Antigen, MAGE4A, MAGE4B, Cancer / Testis Antigen Family 1, Member 4, MAGE-41, MAGE-X2, Cancer / Testis Antigen Family 1, Member 4; NCBI Reference Sequence: NP-001011550.1);MAGE-A6 (MAGEA6, Melanoma Antigen Family A, 6, MAGE6, MAGE-6 Antigen, Melanoma-Associated Antigen 6, Cancer / Testis Antigen 1.6, CT1.6, MAGE3B Antigen, Cancer / Testis Antigen Family 1, Melanoma Antigen Family A 6, Member 6, MAGE-3b, MAGE3B, Cancer / Testis Antigen Family 1, Member 6; NCBI reference sequence: NP-787064.1); MAGE-A9 (MAGEA9, Melanoma Antigen Family A, 9, MAGE9, MAGE-9 Antigen, Melanoma-Associated Antigen 9, Cancer / Testis Antigen 1.9, CT1.9, Cancer / Testis Antigen Family 1, Member 9, Cancer / Testis Antigen Family 1, Member 9, MAGEA9A; NCBI reference sequence: NP-005356.1); MAGE-C1 (MAGEC1, Melanoma Antigen Family C, 1, Cancer / Testis Antigen 7.1, CT7.1, MAGE-C1 Antigen, Cancer / Testis Antigen Family 7, Member 1, CT7, Cancer / Testis Antigen Family 7, Member 1, Melanoma-Associated Antigen C1; NCBI reference sequence: NP-005453.2);MAGE-C2 (MAGEC2, Melanoma Antigen Family C, 2, MAGEE1, Cancer / Testis Antigen 10, CT10, HCA587, Melanoma Antigen, Family E, 1, Cancer / Testis Specific, Hepatocellular Carcinoma-Associated Antigen 587, MAGE-C2 Antigen, MAGE-E1 Antigen, Hepatocellular Cancer Antigen 587, Melanoma-Associated Antigen C2; NCBI Reference Sequence: NP-057333.1); mammaglobin-A (SCGB2A2, Secretoglobin, Family 2A, Member 2, MGB1, Mammaglobin 1, UGB2, Mammaglobin A, mammaglobin-A, Mammaglobin-1, Secretoglobin Family 2A Member 2; NP); MART2 (H HAT, Hedgehog Acyltransferase, SKI1, Melanoma Antigen Recognized By T-Cells 2, Skinny Hedgehog Protein 1, Skn, Melanoma Antigen Recognized By T Cells 2, Protein-Cysteine N-Palmitoyltransferase HHAT, EC 2.3.1.-; GenBank: AAH39071.1); M-CSF (CSF1, Colony Stimulating Factor 1 (Macrophage), MCSF, CSF-1, lanimostim, Macrophage Colony-Stimulating Factor 1, Lanimostim; GenBank: AAH21117.1);MCSP (SMCP, Sperm Mitochondria - Associated Cysteine - Rich Protein, MCS, Mitochondrial Capsule Selenoprotein, HSMCSGEN1, Sperm Mitochondrial - Associated Cysteine - Rich Protein; NCBI reference sequence: NP - 109588.2); XAGE - 1b / GAGED2a; WT1 (Wilms Tumor 1, WAGR, GUD, WIT - 2, WT33, Amino - Terminal Domain Of EWS, NPHS4, Last Three Zinc Fingers Of The DNA - Binding Domain Of WT1, AWT1, Wilms Tumor Protein, EWS - WT1; GenBank: AAB33443.1); VEGF; Tyrosinase (TYR; OCAIA; OCA1A; tyrosinase; SHEP; NP - 000363.1; NM - 000372.4; GenBank: AAB60319.1); TrpM4 (BR22450, FLJ20041, TRPM4, TRPM4B, transient receptor potential cation channel, subfamily M, member 4, Genbank accession number NM - 01763); TRP2 - INT2; TRP -Triosephosphate isomerase (Triosephosphate isomerase 1、TPID、Triose-Phosphate Isomerase、HEL-S-49、TIM、Epididymis Secretory Protein Li 49、TPI、Triosephosphate Isomerase、EC 5.3.1.1; TRAG-3 (CSAG Family Member 2、Cancer / Testis Antigen Family 24、CSAG3B, Member 2、CSAG Family Member 3B、Cancer / Testis Antigen Family 24 Member 2、Cancer / Testis Antigen 24.2、Chondrosarcoma-Associated Gene 2 / 3 Protein、Taxol-Resistant-Associated Gene 3 Protein、Chondrosarcoma-Associated Gene 2 / 3 Protein-Like、CT24.2、Taxol Resistance Associated Gene 3、TRAG-3、CSAG3A、TRAG3;); TMEM46 (shisa homolog 2 (Xenopus Laevis); SHISA; NP-001007539.1; NM-001007538.1; TMEM118 (ring finger protein、transmembrane2; RNFT2; FLJ1462; NP-001103373.1; NM-001109903.1; TMEFF1 (transmembrane protein with EGF-like and two follistatin-like domains 1; Tomoregulin-; H7365; C9orf2; C9ORF2; U19878; X83961; NM-080655; NM-003692;TGF-betaRII (TGFBR2, Transforming Growth Factor, Beta Receptor II (70 / 80 kDa), TGFbeta-RII, MFS2, tbetaR-II, TGFR-2, TGF-Beta Receptor Type IIB, TGF-Beta Type II Receptor, TGF-Beta Receptor Type-2, EC 2.7.11.30, Transforming Growth Factor Beta Receptor Type TENB2 (TMEFF2, tomoregulin, TPEF, HPP1, TR, putative transmembrane proteoglycan, related to the EGF / heregulin family of growth factors and follistatin); 374 aa, NCBI accession numbers: AAD55776, AAF91397, AAG49451, NCBI reference sequence: NP-057276; NCBI Gene: 23671; OMIM: 605734; SwissProt Q9UIK5; Genbank accession numbers AF179274; AY358907, CAF85723, CQ782436; TAG-2; TAG-1 (Contactin 2 (Axonal), TAG-1, AXT, Axonin-1 Cell Adhesion Molecule, TAX, Contactin 2 (transiently PRF: 444868);SYT-SSX1 or -SSX2 fusion protein; survivin; STEAP2 (HGNC 8639, IPCA-1, PCANAP1, STAMP1, STEAP2, STMP, prostate cancer associated gene 1, prostate cancer associated protein 1, six transmembrane epithelial antigen of prostate 2, six transmembrane prostate protein, Genbank accession number AF45513; STEAP1 (six transmembrane epithelial antigen of prostate, Genbank accession number NM-01244); SSX-4; SSX-2 (SSX2, Synovial Sarcoma, X Breakpoint2, X Breakpoint 2, SSX, X Breakpoint 2B, Cancer / Testis Antigen 5.2, X-Chromosome-Related 2, Tumor Antigen HOM-MEL-40, CT5.2, HD21, Cancer / Testis Antigen Family 5, HOM-MEL-40, Isoform B, Cancer / Testis Antigen Family 5 member 2a, member 2a, Protein SSX2, Sarcoma, Sarcoma, Synovial, X-Chromosome-Related 2, synovial, Synovial Sarcoma, X Breakpoint 2B, Synovial Sarcomam, SSX2A; Sp17; SOX10 (SRY (Sex Determining Region Y)-Box 10, mouse, PCWH, DOM, WS4, WS2E, WS4C, Dominant Megacolon, mouse, Human Homolog Of, Dominant Megacolon, SRY-Related HMG-Box Gene 10, Human Homolog Of, transcription Factor SOX-10;GenBank: CAG30470.1); SNRPD1 (Small Nuclear Ribonucleoprotein D1、Sma; ll Nuclear Ribonucleoprotein D1、Polypeptide 16 kDa、Polypeptide (16kD)、SNRPD、HsT2456、Sm-D1、SMD1、Sm-D Autoantigen、Small Nuclear Ribonucleoprotein D1 Polypeptide 16 kDa Pseudogene、SnRNP Core Protein D1、Small Nuclear Ribonucleoprotein Sm D1; SLC35D3 (Solute Carrier Family 35, Member D3、FRCL1、Fringe Connection-Like Protein 1、bA55K22.3、Frc、Fringe-Like 1、Solute Carrier Family 35 Member D3; NCBI GenBank: NC-000006.11 NC-018917.2 NT-025741.16); SIRT2 (Sirtuin 2、NAD-Dependent Deacetylase Sirtuin-2、SIRL2、Silent Information Regulator 2、Regulatory Protein SIR2 Homolog 2、Sir2-Related Protein Type 2、SIR2-Like Protein 2、Sirtuin Type 2、Sirtuin (Silent Mating Type Information Regulation 2 Homolog) 2 (S. cerevisiae)、Sirtuin-2、Sirtuin (Silent Mating Type Information Regulation 2, S. cerevisiae, Homolog) 2、EC 3.5.1.、SIR2; GenBank: AAK51133.1); Sema 5b (FLJ10372、KIAA1445、Mm.42015, SEMA5B, SEMAG, Semaphorin 5b Hlog, sema domain, seven thrombospondin repeats (type 1 and type 1-like), Transmembrane Domain (TM) and short cytoplasmic domain, (semaphorin) 5B, Genbank accession number AB04087; secernin 1 (SCRN1, SES1, KIAA0193, secerin-1; GenBank: EAL24458.1); SAGE (SAGE1, Sarcoma Antigen 1, Cancer / Testis Antigen 14, CT14, Putative Tumor Antigen; NCBI reference sequence: NP-061136.2); RU2AS (KAAG1, Kidney Associated Antigen 1, RU2AS, RU2 Antisense Gene Protein, Kidney-Associated Antigen 1; GenBank: AAF23613.1); RNF43 - E3 ubiquitin-protein ligase RNF43 precursor [Homo sapiens], RNF124; URCC; NCBI reference sequence: NP-060233.3; RhoC (RGS5 (Regulator Of G-Protein Signaling 5, MSTP032, Regulator Of G-Protein Signalling 5, MSTP092, MST092, MSTP106, MST106, MSTP129, MST129; GenBank: AAB84001.1); RET (ret proto-oncogene; MEN2A; HSCR1; MEN2B; MTC1; PTC; CDHF12; Hs.168114; RET51; RET-ELE; NP-066124.1; NM-020975.4); RBAF600 (UBR4、Ubiquitin Protein Ligase E3 Component N-Recognin 4、Zinc Finger、UBR1 Type 1、ZUBR1、E3 Ubiquitin-Protein Ligase UBR4、RBAF600、600 KDa Retinoblastoma Protein-Associated Factor、Zinc Finger UBR1-Type Protein 1、EC 6.3.2.、N-recognin-4、KIAA0462、p600、EC 6.3.2、KIAA1307; GenBank: AAL83880.1); RAGE-1 (MOK、MOK Protein Kinase、Renal Tumor Antigen、RAGE、MAPK / MAK / MRK Overlapping Kinase、Renal Tumor Antigen 1、Renal Cell Carcinoma Antigen、RAGE-1、EC 2.7.11.22、RAGE1; UniProtKB / Swiss-Prot: Q9UQ07.1); RAB38 / NY-MEL-1 (RAB38、NY-MEL-1、RAB38、Member RAS Oncogene Family、Melanoma Antigen NY-MEL-1、Rab-Related GTP-Binding Protein、Ras-Related Protein Rab-38、rrGTPbp; GenBank: AAH15808.1); PTPRK (DJ480J14.2.1 (Protein Tyrosine Phosphatase, Receptor Type, K R-PTP-KAPPA, Protein Tyrosine Phosphatase Kappa, Protein Tyrosine Phosphatase Kappa), Protein Tyrosine Phosphatase, Receptor Type, K, Protein-Tyrosine Phosphatase Kappa, Protein-Tyrosine Phosphatase, Receptor Type, Kappa, R-PTP-kappa, Receptor-Type Tyrosine-Protein Phosphatase Kappa, EC 3.1.3.48, PTPK; GenBank: AAI44514.1); PSMA; PSCA hIg(2700050C12Rik, C530008016Rik, RIKEN cDNA 2700050C12, RIKEN cDNA 2700050C12 gene, Genbank accession number AY358628); PSCA (Prostate stem cell antigen precursor, Genbank accession number AJ29743; PRDX5 (Peroxiredoxin 5, EC 1.11.1.15, TPx Type VI, B166, Antioxidant Enzyme B166, HEL-S-55, Liver Tissue 2D-Page Spot 71 B, PMP20, Peroxisomal Antioxidant Enzyme, PRDX6, Thioredoxin Peroxidase PMP20, PRXV, AOEB166, Epididymis Secretory Protein Li 55, Alu Co-Repressor 1, Peroxiredoxin-5, Mitochondrial, Peroxiredoxin V, prx-V, Thioredoxin Reductase, Prx-V, ACR1, Alu Corepressor, PLP; GenBank: CAG33484.1); PRAME (Preferentially Expressed Antigen In Melanoma, Preferentially Expressed Antigen Of Melanoma, MAPE, 01P-4, OIPA, CT130, Cancer / Testis Antigen 130, Melanoma Antigen Preferentially Expressed In Tumors, Opa-Interacting Protein 4, Opa-Interacting Protein 01P4; GenBank: CAG30435.1); pml-RARalpha fusion protein; PMEL17 (silver homolog; SILV; D12S53E; PMEL17; SI; SIL); ME20; gp10 BC001414; BT007202; M32295; M77348; NM-006928; PBF (ZNF395, Zinc Finger Protein 395, PRF-1, Huntington disease regulatory, HD Gene Regulatory Region-Binding Protein, Region-Binding Protein 2, Protein 2, Papillomavirus Regulatory Factor 1, HD-Regulating Factor 2, Papillomavirus-Regulatory Factor, PRF1, HDBP-2, Si-1-8-14, HDBP2, Huntington'S Disease Gene Regulatory Region-Binding Protein 2, HDRF-2, Papillomavirus Regulatory Factor PRF-1, PBF; GenBank: AAH01237.1); PAX5 (Paired Box 5、Paired Box Homeotic Gene 5、BSAP、Paired Box Protein Pax-5、B-Cell Lineage Specific Activator、Paired Domain Gene 5、Paired Box Gene 5 (B-Cell Lineage Specific Activator Protein)、B-Cell-Specific Transcription Factor、Paired Box Gene 5 (B-Cell Lineage Specific Activator); PAP (REG3A、Regenerating Islet-Derived 3 Alpha、INGAP、PAP-H、Hepatointestinal Pancreatic Protein、PBBCGF. , Human Proislet Peptide, REG-Ill, Pancreatitis-Associated Protein 1, Regi, Reg III-Alpha, hepatocarcinoma-intestine-pancreas, Regenerating Islet-Derived Protein III-Alpha, Pancreatic Beta Cell Growth Factor, HIP, PAP Homologous Protein, HIP / PAP, Proliferation-Inducing Protein 34, PAP1, Proliferation-Inducing Protein 42, REG-3-alpha, Regenerating Islet-Derived Protein 3-Alpha, Pancreatitis-Associated Protein; GenBank: AAH36776.1); p53 (TP53, Tumor Protein P53, TPR53, P53, Cellular Tumor Antigen P53, Antigen NY-CO-13, Mutant Tumor Protein 53, Phosphoprotein P53, P53 Tumor Suppressor, BCC7, Transformation-Related Protein 53, LFS1, tumor Protein 53, Li-Fraumeni Syndrome, Tumor Suppressor P53; P2X5 (Purinergic receptor P2X ligand-gated ion channel 5, an ion channel gated by extracellular ATP that may be involved in synaptic transmission and neurogenesis. Its deficiency may contribute to the pathophysiology of idiopathic detrusor instability); 422 aa), μl: 7.63, MW: 47206 TM: 1 [P] Gene chromosome: 17p13.3, Genbank accession number NP-002552.OGT (0-Linked N-Acetylglucosamine (GlcNAc) Transferase、O-GlcNAc Transferase P110 Subunit、0-Linked N-Acetylglucosamine (GlcNAc) Transferase (UDP-N-Acetylglucosamine:Polypeptide-N-Acetylglucosaminyl Transferase、UDP-N-Acetylglucosamine-Peptide N-Acetylglucosaminyltransferase 110 KDa Subunit、UDP-N-Acetylglucosamine:Polypeptide-N-Acetylglucosaminyl Transferase、Uridinediphospho-N-Acetylglucosamine:Polypeptide Beta-N-Acetylglucosaminyl Transferase、O-GlcNAc Transferase Subunit P110、EC 2.4.1.255、0-Linked N-Acetylglucosamine Transferase 110 KDa Subunit、EC 2.4.1、HRNT1、EC 2.4.1.186、0-GLCNAC; GenBank: AAH38180.1); 0A1 (Osteoarthritis QTL 1、OASD; GenBank: CAA88742.1); NY-ESO-1 / LAGE-2 (Cancer / Testis Antigen 1 B、CTAG1 B、NY-ESO-1、LAGE-2、ESO1、CTAG1、CTAG、LAGE2B、Cancer / Testis Antigen 1、Autoimmunogenic Cancer / Testis Antigen NY-ESO-1、Ancer Antigen 3、Cancer / Testis Antigen 6.1、New York Esophageal Squamous Cell Carcinoma 1、L Antigen Family Member 2、LAGE2、CT6.1、LAGE2A; GenBank: AAI30365.1);NY-BR-1 (ANKRD30A、Ankyrin Repeat Domain 30A、Breast Cancer Antigen NY-BR-1、Serologically Defined Breast Cancer Antigen NY-BR-1、Ankyrin Repeat Domain-Containing Protein 30A; NCBI Reference Sequence: NP-443723.2); N-ras (NRAS、Neuroblastoma RAS Viral (V-Ras) Oncogene Homolog、NRAS1、Transforming Protein N-Ras、GTPase NRas、ALPS4、N-Ras Protein Part 4、NS6、Oncogene Homolog、HRAS1; GenBank: AAH05219.1); NFYC (Nuclear Transcription Factor Y、Gamma、HAP5、HSM、Nuclear Transcription Factor Y Subunit C、Transactivator HSM-1 / 2、CCAAT Binding Factor Subunit C、NF-YC、CCAAT Transcription Binding Factor Subunit Gamma、CAAT Box DNA-Binding Protein Subunit C、Histone H1 Transcription Factor Large Subunit 2A、CBFC、Nuclear Transcription Factor Y Subunit Gamma、CBF-C、Transactivator HSM-1、H1TF2A、Transcription Factor NF-Y、C Subunit;neo-PAP (PAPOLG, Poly(A) Polymerase Gamma, Neo-Poly(A) Polymerase, Nuclear Poly(A) Polymerase Gamma, Polynucleotide Adenylyltransferase Gamma, SRP RNA 3′ Adenylating Enzyme / Pap2, PAP-gamma, Neo-PAP, SRP RNA 3′-Adenylating Enzyme, PAP2, EC 2.7.7.19, PAPG; NCBI Reference Sequence: NP-075045.2); NCA (CEACAM6, Genbank accession number M1872); Napi3b (NAPI-3B, NPTIIb, SLC34A2, solute carrier family 34 (sodium phosphate), member 2, type II sodium-dependent phosphate transporter 3b, Genbank accession number NM-00642); Myosin class I; MUM-3; MUM-2 (TRAPPC1, Trafficking Protein Particle Complex 1, BETS, BETS Homolog, MUM2, Melanoma Ubiquitous Mutated 2, Multiple Myeloma Protein 2, Trafficking Protein Particle Complex Subunit 1; MUM-1f;Mucin (MUC1、Mucin 1、Cell Surface Associated、PEMT、PUM、CA 15-3、MCKD1、ADMCKD、Medullary Cystic Kidney Disease 1 (autosomal dominant)、ADMCKD1、Mucin 1、Transmembrane、CD227、Breast Carcinoma-Associated Antigen DF3、MAM6、Cancer Antigen 15-3、MCD、Carcinoma-Associated Mucin、MCKD、Krebs Von Den Lungen-6、MUC-1 / SEC、Peanut-Reactive Urinary Mucin、MUC1 / ZD、Tumor-Associated Epithelial Membrane Antigen、DF3 Antigen、Tumor-Associated Mucin, episialin、EMA、H23 Antigen、H23AG、Mucin-1、KL-6、Tumor Associated Epithelial Mucin、MUC-1、Episialin、PEM、CD227 Antigen; UniProtKB / Swiss-Prot: P15941.3); MUCSAC (Mucin SAC、Oligomeric Mucus / Gel-Forming、Tracheobronchial Mucin′ MUC5、TBM、Mucin 5、Subtypes A And C、Tracheobronchial / Gastric、leB、Gastric Mucin、Mucin SAC、Oligomeric Mucus / Gel-Forming Pseudogene、Lewis B Blood Group Antigen、LeB、Major Airway Glycoprotein、MUC-SAC、Mucin-5 Subtype AC、Tracheobronchial;MUC1 (Mucin 1, Cell Surface Associated, PEMT, PUM, CA 15-3, MCKD1, ADMCKD, Medullary Cystic Kidney Disease 1 (Autosomal Dominant), ADMCKD1, Mucin 1, Transmembrane, CD227, Breast Carcinoma-Associated Antigen DF3, MAM6, Cancer Antigen 15-3, MCD, Carcinoma-Associated Mucin, MCKD, Krebs Von Den Lungen-6, MUC-1 / SEC, Peanut-Reactive Urinary Mucin, MUC-1 / X, Polymorphic Epithelial Mucin, MUC1 / ZD, Tumor-Associated Epithelial Membrane Antigen, DF3 Antigen, Tumor-Associated Mucin, episialin, EMA, h23 Antigen, H23AG, mucin-1, KL-6, Tumor Associated Epithelial Mucin, MUC-1, Episialin, PEM, CD227 Antigen; MSG783 (RNF124, hypothetical protein FLJ20315, Genbank accession number NM-01776; MRP4 - multidrug resistance-associated protein 4 isoform 3, MOAT-B; MOATB [Homo sapiens]; NCBI reference sequence: NP-001288758.1; MPF (MPF, MSLN, SMR, megakaryocyte potentiating factor, mesothelin, Genbank accession number NM-00582;MMP-7 (MMP7、matrilysin、MPSL1、matrin、Matrix Metalloproteinase 7 (Matrilysin、Uterine)、Uterine Matrilysin、Matrix Metalloproteinase-7、EC 3.4.24.23、Pump-1 Protease、Matrin、Uterine Metalloproteinase、PUMP1、MMP-7、EC 3.4.24、PUMP-1; GenBank: AAC37543.1); MMP-2 (MMP2、Matrix Metallopeptidase 2 (Gelatinase A、72 kDa Gelatinase、72 kDa Type IV Collagenase)、MONA、CLG4A、Matrix Metalloproteinase 2 (Gelatinase A、72kD Gelatinase、72kD Type IV Collagenase)、CLG4、72 kDa Gelatinase、72 kDa Type IV Collagenase)、Matrix Metalloproteinase-2、MMP-II、72 KDa Gelatinase、Collagenase Type IV-A、MMP-2、Matrix Metalloproteinase-II、TBE-1、Neutrophil Gelatinase、EC 3.4.24.24、EC 3.4.24; GenBank: AAH02576.1);およびMeloe。;

[0069] In certain embodiments, the CD3 binding domains of the invention and antibodies comprising same comprise multispecific antibodies, e.g., bispecific antibodies, wherein the multispecific antibody comprises a second binding domain having specificity for a second antigen selected from the group consisting of: 17-IA, 4-1BB, 4Dc, 6-keto-PGF1a, 8-iso-PGF2a, 8-oxo-dG, Al adenosine receptor, A33, ACE, ACE-2, activin, activin A, activin AB, activin B, activin C, activin RIA, activin RIA ALK-2, activin RIB ALK-4, activin RIIA, activin RUB, ADAM, ADAM10, ADAM12, ADAM15, ADAM17 / TACE, ADAM8, ADAM9, ADAMTS, ADAMTS4, ADAMTS5, addressin, aFGF, ALCAM, ALK, ALK-1, ALK-7, alpha-l-antitrypsin, alpha-V / beta-1 antagonist, ANG, Ang, APAF-1, APE, APJ, APP, APRIL, AR, ARC, ART, Artemin, anti-Id, ASPARTIC, atrial natriuretic factor, av / b3 integrin, Axl, b2M, B7-1, B7-2, B7-H, B-lymphocyte stimulating factor (BlyS), BACE, BACE-1, Bad, BAFF, BAFF-R, Bag-1, BAK, Bax, BCA-1, BCAM, Bel, BCMA, BDNF, b-ECGF, bFGF, BID, Bik, BIM, BLC, BL-CAM, BLK, BMP, BMP-2 BMP-2a, BMP-3 osteogenin, BMP-4 BMP-2b, BMP-5, BMP-6 Vgr-1, BMP-7 (OP-1), BMP-8 (BMP-8a, OP-2), BMPR, BMPR-IA (ALK-3), BMPR-IB (ALK-6), BRK-2, RPK-1, BMPR-II (BRK-3), BMPs, b-NGF, BOK, bombesin, bone-derived neurotrophic factor, BPDE, BPDE-DNA, BTC, complement factor 3(C3), C3a, C4, C5, C5a, CIO, CA125, CAD-8, calcitonin, cAMP, carcinoembryonic antigen (CEA), tumor-associated antigen, cathepsin A, cathepsin B, cathepsin C / DPPI, cathepsin D, cathepsin E, cathepsin H, cathepsin L, cathepsin O, cathepsin S, cathepsin V, cathepsin X / Z / P, CBL, CCI, CCK2, CCL, CCLl, CCLll, CCL12, CCL13, CCL 14, CCL15, CCL16, CCLl 7, CCL18, CCL19, CCL2, CCL20, CCL21, CCL22, CCL23, CCL24, CCL25, CCL26, CCL27, CCL28, CCL3, CCL4, CCL5, CCL6, CCL7, CCL8, CCL9 / 10, CCR, CCR1, CCR10, CCR10, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CD1, CD2, CD4, CD5, CD6, CD7, CD8, CD10, CDlla, CDllb, CDllc, CD13, CD14, CD 15, CD16, CD18, CD19, CD20, CD21, CD22, CD23, CD25, CD27L, CD28, CD29, CD30, CD30L, CD32, CD33 (p67 protein), CD34, CD38, CD40, CD40L, CD44, CD45, CD46, CD49a, CD52, CD54, CD55, CD56, CD61, CD64, CD66e, CD74, CD80 (B7-1), CD89, CD95, CD123, CD137, CD138, CD140a, CD146, CD147, CD148, CD152, CD164, CEACAM5, CFTR, cGMP, CINC, botulinum toxin, Clostridium perfringens toxin, CKb8-l, CLC, CMV, CMVUL, CNTF, CNTN-1, COX, C-Ret, CRG-2, CT-1, CTACK, CTGF, CTLA-4, CX3CL1, CX3CR1, CXCL, CXCLl, CXCL2, CXCL3, CXCL4, CXCL5, CXCL6, CXCL7, CXCL8, CXCL9, CXCL10, CX CL11, CXCL12, CXCL13, CXCL14, CXCL15, CXCL16, CXCR, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6, cytokeratin tumor-associated antigen, DAN, DCC, DcR3, DC-SIGN, decay-promoting factor, des(l-3)-IGF-I (brain IGF-1), Dhh, digoxin, DNAM-1, Dnase, Dpp, DPPIV / CD26, Dtk, ECAD, EDA, EDA-A1, EDA-A2, EDAR, EGF, EGFR (ErbB-1), EMA, EMMPRIN, EN A, endothelin receptor, enkephalinase, eNOS, Eot, eotaxinl, EpCAM, ephrin B2 / EphB4, EPO, ERCC, E-selectin, ET-1, Factor Ila, Factor VII, Factor VIIIc, Factor IX, fibroblast activation protein (FAP), Fas, FcRl, FEN-1, ferritin, FGF, FGF-19, FGF-2, FGF3, FGF-8, FGFR, FGFR-3, Fibrin, FL, FLIP, Flt-3, Flt-4, follicle-stimulating hormone, fractalkine, FZD1, FZD2, FZD3, FZD4, FZD5, FZD6, FZD7, FZD8, FZD9, FZD10, G250, Gas 6, GCP-2, GCSF, GD2, GD3, GDF, GDF-1, GDF-3 (Vgr-2), GDF-5 (BMP-14, CDMP- 1), GDF-6 (BMP-13, CDMP-2), GDF-7 (BMP-12, CDMP-3), GDF-8 (myostatin), GDF-9, GDF- 15 (MIC-1), GDNF, GDNF, GFAP, GFRa-1, GFR-alphal, GFR-alpha2, GFR-alpha3, GITR, glucagon, Glut 4, glycoprotein Ilb / IIIa (GPIlb / IIIa), GM-CSF, gpl30, gp72, GRO, growth hormone-releasing factor, hapten (NP-cap or NIP-cap), HB-EGF, HCC, HCMV gB envelope glycoprotein, HCMV gH envelope glycoprotein, HCMV UL, hemopoietic growth factor (HGF), Hep B gpl20, heparanase, Her2, Her2 / neu (ErbB-2), Her3 (ErbB-3), Her4 (ErbB-4), herpes simplex virus (HSV) gB glycoprotein, HSV gD glycoprotein, HGFA, high molecular weight melanoma-associated antigen (HMW-MAA), HIV gpl20, HIV IIIB gp 120 V3 loop, HLA, HLA-DR, HM1.24, HMFG PEM, HRG, Hrk, human cardiac myosin, human cytomegalovirus (HCMV), human growth hormone (HGH), HVEM, 1-309, IAP, ICAM, ICAM-1, ICAM-3, ICE, ICOS, IFNg, Ig, IgA receptor, IgE, IGF, IGF binding protein, IGF-1R, IGFBP, IGF-I, IGF-II, IL, IL-1, IL-1R, IL-2, IL-2R, IL-4, IL-4R, IL-5, IL-5R, IL-6, IL-6R, IL-8, IL-9, IL-10, IL-12, IL-13, IL-15, IL-18, IL-18R, IL-23, interferon (INF)-alpha, INF-beta, INF-gamma, inhibin, iNOS, insulin A-chain, insulin B-chain, insulin-like growth factor 1, integrin alpha 2, integrin alpha 3, integrin alpha 4, integrin alpha 4 / beta l, integrin alpha 4 / beta 7, integrin alpha 5(alpha V), integrin alpha 5 / beta l, integrin alpha 5 / beta 3, integrin alpha 6, integrin beta l, integrin beta 2, interferon gamma, IP-10, 1-TAC, JE, kallikrein 2, kallikrein 5, kallikrein 6, kallikrein 11, kallikrein 12, kallikrein 14, kallikrein 15, kallikrein LI, kallikrein L2, kallikrein L3, kallikrein L4, KC, KDR, keratinocyte growth factor (KGF), laminin 5, LAMP, LAP, LAP (TGF-1), latent TGF-1, latent TGF-1 bpl, LBP, LDGF, LECT2, Lefty, Lewis-Y antigen, Lewis-Y related antigen, LFA-1, LFA-3, Lfo, LIF, LIGHT, lipoprotein, LIX, LKN, Lptn, L-selectin, LT-a, LT-b, LTB4, LTBP-1, pulmonary surfactant, progesterone, lymphotoxin beta receptor, Mac-1, MAdCAM, MAG, MAP2, MARC, MCAM, MCAM, MCK-2, MCP, M-CSF, MDC, Mer, metalloproteinase, MGDF receptor, MGMT, MHC (HLA-DR), MIF, MIG, MIP, MIP-1-alpha, MK, MMAC1, MMP, MMP-1, MMP-10, MMP-11, MMP-12, MMP-13, MMP-14, MMP-15, MMP-2, MMP-24, MMP-3, MMP-7, MMP-8, MMP-9, MPIF, Mpo, MSK, MSP, mucin (Mucl), MUC18, Müllerian inhibitory factor, Mug, MuSK, NAIP, NAP, NCAD, N-cadherin, NCA 90, NCAM, NCAM, neprilysin, neurotrophin-3, -4, or -6, neurturin, nerve growth factor (NGF), NGFR, NGF-beta, nNOS, NO, NOS, Npn, NRG-3, NT, NTN, OB, OGG1, OPG, OPN, OSM, OX40L, OX40R, pl50, p95, PADPr, Parathyroidhormone, PARC, PARP, PBR, PBSF, PCAD, P-cadherin, PCNA, PDGF, PDGF, PDK-1, PECAM, PEM, PF4, PGE, PGF, PGI2, PGJ2, PIN, PLA2, placental alkaline phosphatase (PLAP), P1GF, PLP, PP14, proinsulin, prorelaxin, protein C, PS, PSA, PSCA, prostate-specific membrane antigen (PSMA), PTEN, PTHrp, Ptk, PTN, R51, RANK, RANKL, RANTES, relaxin A-chain, relaxin B-chain, renin, respiratory syncytial virus (RSV) F, RSV Fgp, Ret, rheumatoid factor, RLIP76, RPA2, RSK, S100, SCF / KL, SDF-1, SERINE, serum albumin, sFRP-3, Shh, SIGIRR, SK-1, SLAM, SLPI, SMAC, SMDF, SMOH, SOD, SPARC, Stat, STEAP, STEAP-II, TACE, TACI, TAG-72 (tumor-associated glycoprotein-72), TARC, TCA-3, T cell receptor (e.g., T cell receptor alpha / beta), TdT, TECK, TEM1, TEM5, TEM7, TEM8, TERT, testicular PLAP-like alkaline phosphatase, TfR, TGF, TGF-alpha, TGF-beta, TGF-beta pan-specific, TGF-beta RI (ALK-5), TGF-beta RII, TGF-beta Rllb, TGF-beta RIII, TGF-beta l, TGF-beta 2, TGF-beta 3, TGF-beta 4, TGF-beta 5, thrombin, thymus Ck-1, thyroid stimulating hormone, Tie, TIMP, TIQ, tissue factor, TMEFF2, Tmpo, TMPRSS2, TNF, TNF-alpha, TNF-alpha beta, TNF-beta 2, TNFc, TNF-RI, TNF-RII, TNFRSF10A (TRAIL Rl Apo-2, DR4), TNFRSFIOB (TRAIL R2 DR5, KILLER, TRICK-2A, TRICK-B), TNFRSF10C (TRAIL R3 DcRl, LIT, TRID), TNFRSF10D (TRAIL R4 DcR2, TRUNDD), TNFRSF11A (RANK ODF R, TRANCER)、TNFRSFllB (OPG OCIF、TR1)、TNFRSF12 (TWEAK R FN14)、TNFRSF13B (TACI)、TNFRSF13C (BAFF R)、TNFRSF14 (WHEM ATAR、HveA、LIGHT R、TR2)、TNFRSF16 (NGFR p75NTR), TNFRSF17 (BCMA), TNFRSF18 (GITR AITR), TNFRSF19 (TROY CROWN), TNFRSF19L (RELT), TNFRSFIA (TNF RI CD120a, p55-60), TNFRSFIB (TNF RII). CD120b, p75-80, TNFRSF26 (TNFRH3), TNFRSF3 (LTbR TNF RIII, TNFC R), TNFRSF4 (OX40 ACT35, TXGP1 R), TNFRSF5 (CD40 p50), TNFRSF6 (Fas). Apo-1, APT1, CD95, TNFRSF6B (DcR3 M68, TR6), TNFRSF7 (CD27), TNFRSF8 (CD30), TNFRSF9 (4-lBB CD137, ILA), TNFRSF21 (DR6), TNFRSF22 (DcTRAIL R2). TNFRH2), TNFRST23 (DcTRAIL Rl TNFRH1), TNFRSF25 (DR3 Apo-3, LARD, TR-3, TRAMP, WSL-1), TNFSF10 (TRAIL Apo-2 dilution, TL2), TNFSF11 (TRANCE / RANK ODF (OPG) TNFSF12 (TWEAK Apo-3 ODF) TNFSF13 (APRIL TALL2) TNFSF13B (BAFF). BLYS、TALL1、THANK、TNFSF20)、TNFSF14 (LIGHT WHIME LINES、LTg) TNFSF15 (TL1A / VEGI) TNFSF18 (GITRリンドAITRリガド、TL6), TNFSFIA (TNF-a Connectin, DIF, TNFSF2), TNFSF1B (TNF-b LTa, TNFSF1), TNFSF3 (LTb TNFC, p33), TNFSF4 (OX40ダツgp34, TXGP1, TNFSF5 (CD40 substrate).CD154, gp39, HIGM1, IMD3, TRAP), TNFSF6 (Fas ligand, Apo-1 ligand, APT1 ligand), TNFSF7 (CD27 ligand, CD70), TNFSF8 (CD30 ligand, CD153), TNFSF9 (4-lBB ligand, CD137 ligand), TP-1, t-PA, Tpo, TRAIL, TRAIL R, TRAIL-R1, TRAIL-R2, TRANCE, transfer receptor, TRF, Trk, TROP-2, TSG, TSLP, tumor-associated antigen CA 125, tumor-associated antigen expressing Lewis Y-related glycoprotein, TWEAK, TXB2, Ung, uPAR, uPAR-1, urokinase, VCAM, VCAM-1, VECAD, VE-cadherin, VE-cadherin-2, VEFGR-1 (flt-1), VEGF, VEGFR, VEGFR-3 (flt-4), VEGI, VIM, viral antigen, VLA, VLA-1, VLA-4, VNR integrin, von Willebrand factor, WIF-1, WNT1, WNT2, WNT2B / 13, WNT3, WNT3A, WNT4, WNT5A, WNT5B, WNT6, WNT7A, WNT7B, WNT8A, WNT8B, WNT9A, WNT9A, WNT9B, WNT10A, WNT10B, WNT11, WNT16, XCL1, XCL2, XCR1, XCR1, XEDAR, XIAP, XPD, CTLA4 (cytotoxic T lymphocyte antigen-4), PD1 (programmed cell death protein 1), PD-L1 (programmed cell death ligand 1), LAG-3 (lymphocyte activation gene-3), TIM-3 (T cell immunoglobulin and mucin protein-3), receptors for hormones, and growth factors.

[0070] In certain embodiments, the CD3 binding domains of the invention and antibodies comprising same comprise multispecific antibodies, e.g., bispecific antibodies, wherein the multispecific antibodies comprise a second binding domain having specificity for a second antigen selected from the group consisting of BCMA, CTLA4 (cytotoxic T lymphocyte antigen-4), PD1 (programmed cell death protein 1), PD-L1 (programmed cell death ligand 1), LAG-3 (lymphocyte activation gene-3), TIM-3, CD20, CD2, CD19, Her2, EGFR, EpCAM, FcyRIIIa (CD16), FcyRIIa (CD32a), FcyRIIb (CD32b), FcyRI. (CD64), Toll-like receptors (TLRs), TLR4, TLR9, cytokines, IL-2, IL-5, IL-13, IL-6, IL-17, IL-12, IL-23, TNFα, TGFβ, cytokine receptors, IL-2R, chemokines, chemokine receptors, growth factors, VEGF, and HGF.

[0071] In certain embodiments, the CD3 binding domains of the invention and antibodies comprising same comprise at least a second antigen binding domain that specifically binds to an antigen, wherein the antibody comprises a multispecific format selected from the group consisting of: Fab-Fc-scFv, "bottle-opener", Mab-scFv, Mab-Fv, dual-scFv, core Fv, core scFv, one-arm core scFv, Fab-Fab, Fab-Fv, mAb-Fv, mAb-Fab, DART, BiTE, common light chain-IgG, TandAb, Cross-Mab, SEED, BEAT, TrioMab, and DuetMab.

[0072] In certain embodiments, isolated nucleic acid sequences encoding the CD3 binding domains of the invention and antibodies comprising same are provided.

[0073] In certain embodiments, expression vectors comprising nucleic acid sequences encoding the CD3 binding domains of the invention and antibodies comprising same are provided.

[0074] In certain embodiments, host cells are provided that are transfected, transformed, or transduced with nucleic acid sequences encoding the CD3 binding domains of the invention and antibodies comprising same.

[0075] In certain embodiments, host cells are provided that are transfected, transformed, or transduced with vectors encoding the CD3 binding domains of the invention and antibodies comprising same.

[0076] In certain embodiments, pharmaceutical compositions comprising one or more of the CD3 binding domains of the invention and antibodies comprising same are provided.

[0077] In certain embodiments, pharmaceutical compositions are provided comprising one or more of the CD3 binding domains of the invention and antibodies comprising same, and a pharmaceutically acceptable carrier and / or excipient.

[0078] In certain embodiments, pharmaceutical compositions comprising one or more nucleic acids encoding one or more of the CD3 binding domains of the invention and antibodies comprising same are provided.

[0079] In certain embodiments, pharmaceutical compositions are provided comprising one or more vectors encoding one or more of the CD3 binding domains of the invention and antibodies comprising the same, and a pharmaceutically acceptable carrier and / or excipient.

[0080] In certain embodiments, methods are provided for treating a disorder or delaying the progression of a disorder in a mammal in need of such treatment, comprising administering one or more of the CD3 binding domains of the invention and antibodies comprising the same, wherein the disorder is reduced or ameliorated as a result of the administration. In certain embodiments, methods are provided for preventing a disorder or reducing the risk of developing a disorder in a mammal by administering one or more of the CD3 binding domains of the invention and antibodies comprising the same, wherein the disorder is prevented as a result of the administration. In certain embodiments, methods are provided for treating a disorder in a mammal in need of such treatment, wherein the disorder comprises a proliferative disorder, a cancer disorder, an immune-cancer disorder, a neurological disorder, a neurodegenerative disorder, or an autoimmune disorder, comprising administering one or more of the CD3 binding domains of the invention and antibodies comprising the same, wherein the disorder is reduced or ameliorated as a result of the administration. In certain embodiments, the method further comprises administering an additional therapeutic agent to the mammal. In certain embodiments, the mammal is a human. In certain embodiments, heterodimeric CD3 fusion proteins are provided comprising a first polypeptide chain comprising a CD3 epsilon polypeptide fused to a first Fc region and a second polypeptide chain comprising a CD3 delta polypeptide fused to a second Fc region. In certain embodiments, the CD3 epsilon polypeptide comprises a human CD3 epsilon polypeptide and the CD3 delta polypeptide comprises a human CD3 delta polypeptide. In certain embodiments, the CD3 epsilon polypeptide is a human CD3 epsilon polypeptide and the CD3 delta polypeptide is a human CD3 delta polypeptide. In certain embodiments, the CD3 epsilon polypeptide comprises a cynomolgus monkey CD3 epsilon polypeptide and the CD3 delta polypeptide comprises a cynomolgus monkey CD3 delta polypeptide. In certain embodiments, the CD3 epsilon polypeptide is a cynomolgus monkey CD3 epsilon polypeptide and the CD3 delta polypeptide is a cynomolgus monkey CD3 delta polypeptide. In certain embodiments, the first polypeptide of the heterodimeric CD3 fusion protein comprises the following amino acid sequence: QDGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMDGGSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (sequence number 6716).The second polypeptide chain comprises the following amino acid sequence: FKIPIEELEDRVFVNCNTSITWVEGTVGTLLSDITRLDLGKRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDGGSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 6717). In certain embodiments, the first polypeptide of the heterodimeric CD3 fusion protein comprises the following amino acid sequence: QDGNEEMGSITQTPYQVSISGTTVILTCSQHLGSEAQWQHNGKNKEDSGDRLFLPEFSEMEQSGYYVCYPRGSNPEDASHHLYLKARVCENCMEMDGGSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (sequence number 6718).The second polypeptide chain comprises the following amino acid sequence: FKIPVEELEDRVFVKCNTSVTWVEGTVGTLLTNNTRLDLGKRILDPRGIYRCNGTDIYKDKESAVQVHYRMCQNCVELDPGSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 6719). In certain embodiments, the present invention provides the use of such heterodimeric CD3 fusion proteins in identifying, isolating, selecting, producing and / or characterizing CD3 binding domains and antibodies comprising the same. In certain embodiments, for example, the following are provided: (Item 1) 1. An antibody or antigen-binding polypeptide comprising a CDRH1 comprising the amino acid sequence of FNIKDYYMH (SEQ ID NO: 6720), YTFTSYTIH (SEQ ID NO: 6721), or FTFX1TYAMN (SEQ ID NO: 6722), wherein X1 is any amino acid. (Item 2) 2. The antibody or antigen-binding polypeptide of item 1, wherein X1 is N or D. (Item 3) An antibody or antigen-binding polypeptide comprising a CDRH2 comprising the amino acid sequence of WIDLENX1NTX2YDX3KFOG (SEQ ID NO: 6723), wherein X1, X2, and X3 are independently any amino acid. (Item 4) 4. The antibody or antigen-binding polypeptide of item 3, wherein X1 is not G. (Item 5) 5. The antibody or antigen-binding polypeptide of any one of items 3 to 4, wherein X2 is V or I. (Item 6) 6. The antibody or antigen-binding polypeptide of any one of items 3 to 5, wherein X3 is not G. (Item 7) 1. An antibody or antigen-binding polypeptide comprising a CDRH3 comprising the amino acid sequence of X1QSYSX2RT (SEQ ID NO: 6724) or X3RDX4YGX5YFYDV (SEQ ID NO: 6725), wherein X1, X2, X3, X4, and X5 are independently any amino acid. (Item 8) 8. The antibody or antigen-binding polypeptide of item 7, wherein X1 is K or T. (Item 9) 8. The antibody or antigen-binding polypeptide of item 7, wherein X1 is T. (Item 10) 10. The antibody or antigen-binding polypeptide of any one of items 7 to 9, wherein X2 is R or L. (Item 11) 10. The antibody or antigen-binding polypeptide of any one of items 7 to 9, wherein X3 is A or G. (Item 12) 12. The antibody or antigen-binding polypeptide of any one of items 7 to 11, wherein X4 is not G. (Item 13) 13. The antibody or antigen-binding polypeptide of any one of items 7 to 12, wherein X5 is R, A, G, or L. (Item 14) An antibody or antigen-binding polypeptide comprising a CDRL1 comprising the amino acid sequence of X1KSSQX2LLX3X4RTGKX5YLA (SEQ ID NO: 6726), wherein X1, X2, X3, X4 and X5 are independently any amino acid. (Item 15) 15. The antibody or antigen-binding polypeptide of item 14, wherein X1 is K or R. (Item 16) 15. The antibody or antigen-binding polypeptide of item 14, wherein X2 is S or N. (Item 17) 15. The antibody or antigen-binding polypeptide of item 14, wherein X3 is E or N. (Item 18) 15. The antibody or antigen-binding polypeptide of item 14, wherein X4 is A or S. (Item 19) 15. The antibody or antigen-binding polypeptide of item 14, wherein X5 is N or S. (Item 20) An antibody or antigen-binding polypeptide comprising a CDRL2 comprising the amino acid sequence of WASTRES (SEQ ID NO: 6727) or GTX1KRAP (SEQ ID NO: 6728), wherein X1 is any amino acid. (Item 21) 21. The antibody or antigen-binding polypeptide of item 20, wherein X1 is N or D. (Item 22) An antibody or antigen-binding polypeptide comprising a CDRL3 comprising the amino acid sequence of KQSYSX1RT (SEQ ID NO: 6729), wherein X1 is any amino acid. (Item 23) 23. The antibody or antigen-binding polypeptide of item 22, wherein X1 is R. (Item 24) 23. The antibody or antigen-binding polypeptide of item 22, wherein X1 is L or I. (Item 25) An antibody or antigen-binding polypeptide comprising one or more of the CDRH1 according to any one of items 1 to 2, the CDRH2 according to any one of items 3 to 6, and the CDRH3 according to any one of items 7 to 13. (Item 26) 2. An antibody or antigen-binding polypeptide comprising one or more of CDRL1 according to any one of items 14 to 17, CDRL2 according to any one of items 20 to 21, and CDRL3 according to any one of items 22 to 24. (Item 27) An antibody or antigen-binding polypeptide comprising a CDRH1 according to any one of items 1 to 2, a CDRH2 according to any one of items 3 to 6, a CDRH3 according to any one of items 7 to 13, and a CDRL1 according to any one of items 14 to 17, a CDRL2 according to any one of items 20 to 21, and a CDRL3 according to any one of items 22 to 24. [Brief explanation of the drawings]

[0081] [Figure 1] Not specified [Figure 2] Not specified [Figure 3] Not specified [Figure 4A] Not specified [Figure 4B] Not specified [Figure 5] Not specified [Figure 6-1] Not specified [Figure 6-2] Not specified [Figure 7] Not specified [Figure 8] Not specified [Figure 9] Not specified [Figure 10A-1] Not specified [Figure 10A-2] Not specified [Figure 10B] Not specified [Figure 10C] Not specified [Figure 10D] Not specified [Figure 11A] Not specified [Figure 11B] Not specified [Figure 11C] Not specified [Figure 12A] Not specified [Figure 12B] Not specified [Figure 13-1] Not specified [Figure 13-2] Not specified [Figure 14] Not specified [Figure 15] Not specified [Figure 16] Not specified [Figure 17] Not specified [Figure 18] Not specified [Figure 19] Not specified [Figure 20] Not specified [Figure 21] Not specified [Figure 22] Not specified [Figure 23A] Not specified [Figure 23B] Not specified [Figure 24] Not specified [Figure 25A] Not specified [Figure 25B] Not specified [Figure 25C] Not specified [Figure 25D] Not specified [Figure 26A] Not specified [Figure 26B] Not specified [Figure 26C] Not specified [Figure 26D] Not specified [Figure 26E] Not specified [Figure 26F] Not specified [Figure 26G] Not specified [Figure 27-1] Not specified [Figure 27-2] Not specified [Figure 28A] Not specified [Figure 28B-1] Not specified [Figure 28B-2] Not specified [Figure 28C] Not specified [Figure 28D-1] Not specified [Figure 28D-2] Not specified [Figure 29A] Not specified [Figure 29B] Not specified [Figure 30] Not specified [Figure 31] Not specified [Figure 32] Not specified [Figure 33A] Not specified [Figure 33B] Not specified [Figure 33C] Not specified [Figure 34A] Not specified [Figure 34B] Not specified [Figure 34C] Not specified [Figure 34D] Not specified [Figure 35A-1] Not specified [Figure 35A-2] Not specified [Figure 35B] Not specified [Figure 36A-1] Not specified [Figure 36A-2] Not specified [Figure 36A-3] Not specified [Figure 36B-1] Not specified [Figure 36B-2] Not specified [Figure 36B-3] Not specified [Figure 36C] Not specified [Figure 37-1] Not specified [Figure 37-2] Not specified [Figure 38A] Not specified [Figure 38B] Not specified DETAILED DESCRIPTION OF THE INVENTION

[0082] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs.As used herein, the term "about" when used in connection with a specific recited numerical value means that the value can vary from the recited value by 1% or less.For example, as used herein, the expression "about 100" includes 99 and 101, and all values ​​therebetween (for example, 99.1, 99.2, 99.3, 99.4, etc.).

[0083] It will be understood that aspects and embodiments of the invention described herein include "comprising," "consisting of," and "consisting essentially of" aspects and embodiments.

[0084] "Cluster of Differentiation 3" or "CD3" generally refers to any native CD3 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats), and includes, for example, the CD3ε chain, CD3γ chain, CD3α chain, and CD3β chain, unless otherwise indicated. The term encompasses "full-length," unprocessed CD3 (e.g., unprocessed or unmodified CD3ε or CD3γ) as well as any form of CD3 that results from processing in cells. The term also encompasses naturally occurring variants of CD3, such as splice variants or allelic variants. CD3 includes, for example, the human CD3ε protein (NCBI Reference SEQ ID NO: NP _ 000724), which is a protein of 207 amino acids in length and is similar to the human CD3γ protein (NCBI reference sequence number NP _000064), which protein is 182 amino acids in length. The term further refers to either human or cynomolgus monkey CD3 epsilon protein, the amino acid sequences of which are shown, for example, in Figure 18 herein. "CD3εN27" and "CD3εN13" refer to the N-terminal 27 amino acids and the N-terminal 13 amino acids of CD3, respectively, including any chemical modifications or linkages thereto.

[0085] The "class" of an antibody refers to the type of constant domain or constant region possessed by its heavy chain. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, some of which may be further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

[0086] The term "cytotoxic agent," as used herein, refers to a substance that inhibits or prevents the function of cells and / or leads to the death or destruction of cells. Cytotoxic agents include, but are not limited to, radioisotopes (e.g., At 211 , I 131 , I 125 , Y 90 ,Re 186 ,Re 188 , Sm 153 , Bi 212 , P 32 , Pb 212 , and radioactive isotopes of Lu); chemotherapeutic agents or drugs (e.g., methotrexate, adriamycin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents); growth inhibitory agents; enzymes, such as nucleases and fragments thereof; antibiotics; toxins, including fragments and / or variants thereof, such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin; and various anti-tumor or anti-cancer agents disclosed below.

[0087] "Disorder" refers to any condition or disease that would benefit from treatment, including, but not limited to, chronic and acute disorders or diseases, including pathological conditions that predispose the mammal to the disorder in question.

[0088] The terms "cell proliferative disorder" and "proliferative disorder" refer to disorders associated with some degree of abnormal cell proliferation. In one embodiment, a cell proliferative disorder comprises cancer. In one embodiment, a cell proliferative disorder comprises a tumor.

[0089] 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 specific examples of such cancers include, but are not limited to, squamous cell carcinoma (e.g., epithelial squamous cell carcinoma), lung cancer including small cell lung cancer, non-small cell lung cancer, lung adenoma, squamous cell carcinoma of the lung, cancer of the peritoneum, hepatocellular carcinoma, gastric cancer including gastrointestinal cancer and gastrointestinal stromal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, cancer of the urinary tract, liver cancer (hepatoma), breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial or uterine cancer, salivary gland cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, anal cancer, penile cancer, melanoma, superficial spreading melanoma, lentigo maligna melanoma, acral lentigo melanoma, nodular melanoma, multiple myeloma, and B-cell lymphoma (low grade / follicular non-Hodgkin's lymphoma (NH)). L), small lymphocytic (SL) NHL, intermediate-grade / follicular NHL, intermediate-grade diffuse NHL, high-grade immunoblastic NHL, high-grade lymphoblastic NHL, high-grade small noncleaved cell NHL, bulky disease NHL, mantle cell lymphoma, AIDS-related lymphoma, and Waldenstrom's macroglobulinemia), chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), hairy cell leukemia, chronic myeloblastic leukemia, and post-transplant lymphoproliferative disorder (PTLD), as well as abnormal blood vessel growth associated with phacomatosis, edema (e.g., edema associated with brain tumors), Meige syndrome, brain tumors and head and neck cancers, and associated metastases. In certain embodiments, cancers suitable for treatment with the antibodies of the invention include breast cancer, colon cancer, rectal cancer, non-small cell lung cancer, glioblastoma, non-Hodgkin's lymphoma (NHL), renal cell carcinoma, prostate cancer, liver cancer, pancreatic cancer, soft tissue sarcoma, Kaposi's sarcoma, carcinoid carcinoma, head and neck cancer, ovarian cancer, mesothelioma, and multiple myeloma. In some embodiments, the cancer is selected from small cell lung cancer, glioblastoma, neuroblastoma, melanoma, breast cancer, gastric cancer, colorectal cancer (CRC), and hepatocellular carcinoma. However, in some embodiments, the cancer is selected from non-small cell lung cancer, colon cancer, glioblastoma, and breast cancer, including metastatic forms of these cancers.In other embodiments, the cancer excludes Hodgkin's lymphoma, but is selected from the group consisting of germinal center B-cell-like (GCB) DLBCL, activated B-cell-like (ABC) DLBCL, follicular lymphoma (FL), mantle cell lymphoma (MCL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), marginal zone lymphoma (MZL), small lymphocytic leukemia (SLL), lymphoplasmacytic lymphoma (LL), Waldenstrom's macroglobulinemia (WM), central nervous system (CNS) lymphoma (CNS), and leukemia-associated lymphoma (CLM). Neoplastic lymphoma (CNSL), Burkitt's lymphoma (BL), B-cell prolymphocytic leukemia, splenic marginal zone lymphoma, hairy cell leukemia, splenic lymphoma / leukemia, unclassifiable, splenic diffuse red pulp small B-cell lymphoma, hairy cell leukemia variant, Waldenstrom's macroglobulinemia, heavy chain disease, alpha heavy chain disease, gamma heavy chain disease, mucous membrane-associated lymphoid tissue disease, plasma cell myeloma, isolated plasmacytoma of bone, extraskeletal plasmacytoma, extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue MALT lymphoma, nodal marginal zone lymphoma, pediatric nodal marginal zone lymphoma, pediatric follicular lymphoma, primary cutaneous follicle-centered lymphoma, T-cell / histiocytic-rich large B-cell lymphoma, primary DLBCL of the CNS, primary cutaneous DLBCL, foot type, EBV-positive DLBCL in the elderly, DLBCL associated with chronic inflammation, lymphomatoid granulomatosis, primary mediastinal (thymic) large B-cell lymphoma, intravascular large B-cell lymphoma, ALK-positive large cell Selected from certain cancers of mature B cells, including diffuse large B-cell lymphoma, plasmablastic lymphoma, large B-cell lymphoma occurring in HHV8-associated multicentric Castleman disease, primary effusion lymphoma: unclassified B-cell lymphoma with features intermediate between diffuse large B-cell lymphoma and Burkitt lymphoma, and unclassified B-cell lymphoma with features intermediate between diffuse large B-cell lymphoma and classical Hodgkin lymphoma.

[0090] As used herein, the term "tumor" 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 when referred to herein.

[0091] As used herein, the term "tumor antigen" can be understood as an antigen presented on tumor cells. These antigens can be presented on the cell surface by their extracellular portion, which is often combined with the transmembrane and cytoplasmic portions of the molecule. These antigens are sometimes presented only by tumor cells and never by normal cells. Tumor antigens may be expressed only on tumor cells or may exhibit tumor-specific mutations compared to normal cells. In this case, they are called tumor-specific antigens. Tumor antigens presented by both tumor and normal cells are more common and are called tumor-associated antigens. These tumor-associated antigens may be overexpressed in tumor cells compared to normal cells, allowing antibodies to access and bind to tumor cells. This is because the structure of tumor tissue is not less dense than that of normal tissue.

[0092] "EC 50 The term "median effective concentration" refers to the "median effective concentration." This value measures the effectiveness of a compound (e.g., an anti-CD3 binding domain-containing antibody) relative to its biological or biochemical utility. This quantitative measure indicates the amount or concentration of a particular compound or antibody required to elicit half of the maximum response of a given biological process.

[0093] "Effector function" refers to the biological activities attributable to the Fc region of an antibody and varies depending on the antibody isotype. Examples of antibody effector functions include: C1q binding and complement-dependent cytotoxicity (CDC), Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, down-regulation of cell surface receptors (e.g., B cell receptors), and B cell activation.

[0094] An "effective amount" of a compound, e.g., an anti-CD3 antibody of the invention, or a composition thereof (e.g., a pharmaceutical composition), is at least the minimum amount required to achieve a desired therapeutic or prophylactic result, such as a measurable improvement or prevention of a particular disorder (e.g., a cell proliferative disorder such as cancer). As used herein, an effective amount may vary according to factors such as the disease state, the patient's age, sex, and weight, and the ability of the antibody to elicit a desired response in an individual. An effective amount is also one in which any toxic or detrimental effects of the treatment are outweighed by the therapeutic beneficial effects. With respect to prophylactic use, beneficial or desired results include, for example, elimination or reduction of risk, reduction in severity, or delay in the onset of disease, including the biochemical, histological, and / or behavioral symptoms of the disease, its complications, and intermediate pathological phenotypes manifesting during the development of the disease. With respect to therapeutic use, beneficial or desired results include, for example, a reduction in one or more symptoms resulting from the disease, an increase in the quality of life of an individual afflicted with the disease, a reduction in the dosage of other pharmaceutical agents required to treat the disease, or an enhancement of the effect of another pharmaceutical agent, for example, through targeting the disease, delaying disease progression, and / or extending survival. In the case of cancer or tumors, an effective amount of a pharmaceutical agent may have the effect of reducing the number of cancer cells, reducing tumor size, inhibiting (i.e., slowing or, preferably, stopping) the infiltration of cancer cells into peripheral organs, inhibiting tumor growth to some extent, and / or alleviating one or more symptoms associated with the disorder to some extent. An effective amount can be administered in one or more administrations. For purposes of this invention, an effective amount of a drug, compound, or pharmaceutical composition is an amount sufficient to directly or indirectly achieve prophylactic or therapeutic treatment. As will be understood in clinical practice, an effective amount of a drug, compound, or pharmaceutical composition may or may not be achieved in combination with another drug, compound, or pharmaceutical composition. Thus, an "effective amount" may be considered in the context of administering one or more therapeutic agents, which may be considered to be administered in an effective amount when combined with one or more other agents to produce a desired result, or when a single agent is administered in an effective amount when a desired result is achieved.

[0095] The term "Fc region" is used herein to define the C-terminal region of an immunoglobulin heavy chain containing at least a portion of the constant region. This term includes native-sequence Fc regions and variant Fc regions. In one embodiment, the Fc region of a human IgG heavy chain extends from Cys226 or from Pro230 to the carboxyl terminus of the heavy chain. However, the C-terminal lysine (Lys447) of the Fc region may or may not be present. Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or constant region follows the EU numbering system, also known as the EU index. This system is described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991.

[0096] "Framework" or "FR" refers to variable domain residues other than complementarity-determining region (CDR) residues. The FR of a variable domain generally consists of four FR domains: FR1, FR2, FR3, and FR4. Thus, the CDR and FR sequences generally occur in the following order in VH (or VL): FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

[0097] The terms "full length antibody," "complete antibody," and "whole antibody" are used interchangeably herein and refer to an antibody having a structure substantially similar to that of a native antibody, or an antibody having a heavy chain containing an Fc region as described herein.

[0098] As used herein, a "growth inhibitory agent" refers to a compound or composition that inhibits cell proliferation either in vitro or in vivo. In one embodiment, the growth inhibitory agent is a growth inhibitory antibody that prevents or reduces proliferation of cells expressing the antigen to which the antibody binds. In another embodiment, the growth inhibitory agent may be an agent that significantly reduces the proportion of cells in S phase. Examples of growth inhibitory agents include agents that disrupt cell cycle progression (at a stage other than S phase), such as agents that induce G1 arrest or M-phase arrest. Classical M-phase blockers include vincas (vincristine and vinblastine), taxanes, and topoisomerase II inhibitors, such as doxorubicin, epirubicin, daunorubicin, etoposide, and bleomycin. Agents that arrest G1 are also effective in arresting S phase, such as DNA alkylating agents, such as tamoxifen, prednisone, dacarbazine, mechlorethamine, cisplatin, methotrexate, 5-fluorouracil, and ara-C. Further details can be found, for example, on page 13, in Mendelsohn and Israel, eds., The Molecular Basis of Cancer, Chapter 1, entitled "Cell cycle regulation, oncogenes, and antitineoplastic drugs" by Murakami et al. (WB Saunders, Philadelphia, 1995). Taxanes (paclitaxel and docetaxel) are anticancer drugs both derived from the yew tree. Docetaxel (TAXOTERE®, Rhone-Poulenc Rorer) is derived from the European yew and is a semisynthetic analog of paclitaxel (TAXOL®, Bristol-Myers Squibb). Paclitaxel and docetaxel promote the assembly of microtubules from tubulin dimers and stabilize microtubules by inhibiting depolymerization, which inhibits cell mitosis.

[0099] Phrases including an antigen and the term "positive," e.g., "HER2-positive," mean that the disease, condition, or disorder is characterized by elevated above normal levels of the specifically referenced antigen or target. In certain embodiments, the antigen or target includes a moiety that is expressed at a higher level on cancer cells than on normal cells, or is otherwise present at a higher level on cancer cells than on normal cells.

[0100] The terms "host cell," "host cell line," and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells. Host cells include "transformants" and "transformed cells," which include the primary transformed cell and its progeny, regardless of the number of transfers. Progeny cells may not be completely identical in nucleic acid content to the parent cell and may contain mutations. Mutant progeny that have the same function or biological activity as screened or selected for in the originally transformed cell are also included herein.

[0101] A "human antibody" is an antibody that possesses an amino acid sequence corresponding to that of an antibody produced by a human or human cell, or an antibody derived from a non-human source utilizing the human antibody repertoire or other human antibody coding sequence. This definition of a human antibody specifically excludes humanized antibodies, which contain non-human antigen-binding residues. Human antibodies can be produced using various techniques known in the art, including phage display library methods. Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991). Further methods available for producing human monoclonal antibodies are described in Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boerner et al., J. Immunol. 147(1):86-95 (1991). See also van Dijk and van de Winkel, Curr. Opin. Pharmacol., 5: 368-74 (2001). Human antibodies can be produced by administering antigen to transgenic animals, such as immunized xenomouse, that have been engineered to produce the antibody in response to antigen challenge but whose endogenous loci are disabled (see, e.g., U.S. Patent Nos. 6,075,181 and 6,150,584 for a description of xenomouse technology). See also Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006) for a description of human antibodies produced, e.g., via human B cell hybridomas. As used herein, the term "human antibody" is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences.The human mAbs of the invention may include amino acid residues, for example in the CDRs, particularly CDR3, that are not encoded in human germline immunoglobulin sequences (e.g., mutations introduced by random or site-directed mutagenesis in vitro or by somatic mutation in vivo). However, as used herein, the term "human antibody" is not intended to include mAbs in which CDR sequences derived from the germline of another mammalian species (e.g., a mouse) have been grafted onto human FR sequences.

[0102] A "human consensus framework" is a framework that represents the most commonly occurring amino acid residues in a selection of human immunoglobulin VL or VH framework sequences. Generally, the selection of human immunoglobulin VL or VH sequences is from a subgroup of variable domain sequences. Generally, the subgroup of sequences is a subgroup as set forth in Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, NIH Publication 91-3242, Bethesda, Md. (1991), vols. 1-3. In one embodiment, for the VL, the subgroup is subgroup kappa I as set forth in Kabat et al., supra. In one embodiment, for the VH, the subgroup is subgroup III as set forth in Kabat et al., supra.

[0103] A "humanized antibody" refers to an antibody containing amino acid residues derived from non-human CDRs and amino acid residues derived from human FRs. In certain embodiments, a humanized antibody contains substantially all of at least one, and typically two, variable domains, in which all or substantially all of the (e.g., CDRs) correspond to those of a non-human antibody and all or substantially all of the FRs correspond to those of a human antibody. A humanized antibody can optionally contain at least a portion of an antibody constant region derived from a human antibody. A humanized form of an antibody, e.g., a non-human antibody, refers to an antibody that has undergone humanization.

[0104] An "immunoconjugate" is an antibody conjugated to one or more heterologous molecules, including but not limited to, cytotoxic agents.

[0105] A "subject" or "individual" is a mammal. Mammals include, but are not limited to, domesticated animals (e.g., cows, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats). In certain embodiments, the subject or individual is a human.

[0106] An "isolated antibody" is an antibody that has been separated from a component of its natural environment. In some embodiments, the antibody is purified to greater than 95% or 99% purity, as determined, for example, by electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatography (e.g., ion exchange or reverse-phase HPLC). For a review of methods for assessing antibody purity, see, e.g., Flatman et al., J. Chromatogr. B 848:79-87 (2007).

[0107] An "isolated nucleic acid" refers to a nucleic acid molecule that has been separated from a component of its natural environment. Isolated nucleic acid includes a nucleic acid molecule contained in a cell that normally contains the nucleic acid molecule, but where the nucleic acid molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location.

[0108] The terms "monoclonal antibody" or "mAb," as used herein, refer to an antibody obtained from a population of substantially homogeneous antibodies; i.e., the individual antibodies comprising the population are identical and / or bind to the same epitope, excluding potential variant antibodies containing, for example, natural mutations or generated during production of the monoclonal antibody preparation. Such variant antibodies are generally present in low amounts. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen. Thus, the modifier "monoclonal" indicates the character of the antibody as obtained from a substantially homogeneous population of antibodies, but is not intended to require production of the antibody by any particular method. For example, monoclonal antibodies for use in accordance with the present invention may be produced by a variety of techniques, including but not limited to hybridoma methods, recombinant DNA methods, phage display methods, and methods utilizing transgenic animals containing all or part of the human immunoglobulin loci; such methods and other exemplary methods for producing monoclonal antibodies are described herein.

[0109] The term "pharmaceutical formulation" or "pharmaceutical composition" refers to a preparation in a form that allows the biological activity of the active ingredient contained therein, such as the CD3-binding domain of the present invention and an antibody comprising the same, to be effective, and that does not contain additional ingredients that are unacceptably toxic to the subject to which the preparation is administered.

[0110] A "pharmaceutically acceptable carrier" refers to an ingredient in a pharmaceutical formulation, other than an active ingredient, that is not toxic to a subject. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

[0111] The phrase "therapeutically effective amount" means the amount for which it is administered that produces the desired effect. The precise amount will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lloyd (1999) The Art, Science and Technology of Pharmaceutical Compounding).

[0112] As used herein, "immune response" or "immunological response" refers to the development in a subject of a humoral immune response, a cellular immune response, or a humoral and cellular immune response to an antigen / immunogen. A "humoral immune response" refers to a response that is mediated at least in part by antibodies. A "cellular immune response" is a response mediated by T lymphocytes or other white blood cells, or both, and includes the production of cytokines, chemokines, and similar molecules produced by activated T cells, white blood cells, or both. Immune responses can be determined by standard immunoassays and neutralization assays, which are known in the art.

[0113] "Immunogenicity," as used herein, refers to the ability of a protein or polypeptide to elicit an immune response that is specifically directed against a particular disease-causing bacterium or virus.

[0114] As used herein, "treatment" (and grammatical variations such as "treat" or "treating") refers to clinical intervention in an attempt to alter the natural course of the individual being treated and can be performed either prophylactically or during the course of clinical pathology. Desirable effects of treatment include, but are not limited to, prevention of disease onset or recurrence, alleviation of symptoms, diminishment of any direct or indirect pathological consequences of the disease, prevention of metastasis, reduction in the rate of disease progression, improvement or palliation of the disease state, and remission or improved prognosis. In some embodiments, the antibodies of the invention are used to delay the onset of a disorder or disease or slow the progression of a disorder or disease.

[0115] As used herein, "delaying the progression" of a disorder or disease means suspending, preventing, slowing, fixing, stabilizing, and / or postponing the onset of the disorder or disease (e.g., a cell proliferative disorder, e.g., cancer). This delay can be for varying lengths of time, depending on the history of the disease and / or the individual being treated. As will be apparent to one of skill in the art, a sufficient or significant delay effectively encompasses prevention, in that the individual does not develop the disease. For example, the onset of, e.g., metastasis, of a late-stage cancer may be delayed.

[0116] "Reduce" or "inhibit" refers to the ability to cause an overall decrease, for example, of 20% or more, 50% or more, or 75%, 85%, 90%, 95% or more. In certain embodiments, reduction or inhibition may refer to antibody effector functions mediated by the antibody Fc region, including complement-dependent cytotoxicity (CDC), antibody-dependent cellular cytotoxicity (ADCC), and antibody-dependent cellular phagocytosis (ADCP), among others.

[0117] As used herein, the terms "prevent," "preventing," and "prevention" refer to preventing or inhibiting the onset or occurrence of a disorder or disease.

[0118] As used herein, the terms "amelioration" and "alleviation" refer to a decrease or lessening in the severity of a condition or any of its symptoms.

[0119] The term "antibody" as used herein is used in the broadest sense and encompasses a variety of antibody structures, including, but not limited to, monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments so long as they exhibit the desired antigen-binding activity. With regard to multispecific antibodies, such antibodies contain at least two different antigen-binding domains that recognize and specifically bind to at least two different antigens. With regard to bispecific antibodies, such antibodies contain two different antigen-binding domains that recognize and specifically bind to at least two different antigens. "Different antigens" may refer to different and / or distinct proteins, polypeptides, or molecules, as well as different and / or distinct epitopes, which may be contained within a single protein, polypeptide, or other molecule. Unless otherwise indicated, the term "antibody," as used herein, in addition to the above, should also be understood to encompass antibody molecules comprising two immunoglobulin heavy chains and two immunoglobulin light chains (i.e., "intact antibody molecules"), as well as antigen-binding fragments thereof. The terms "antigen-binding portion" of an antibody, "antigen-binding fragment" of an antibody, and the like, as used herein, include any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds to an antigen to form a complex.

[0120] "Antibody" can refer to an immunoglobulin molecule (i.e., "intact antibody molecule") composed of four polypeptide chains, two heavy (H) and two light (L) chains inter-connected by disulfide bonds, as well as multimers thereof (e.g., IgM) or antigen-binding fragments thereof. Each heavy chain contains a heavy chain variable region (VH) and a heavy chain constant region (domain C). H 1. C H 2 and C H Each light chain is composed of a light chain variable region (VL) and a light chain constant region (CL). H Area and V LThe regions can be further divided into more conserved regions termed framework regions (FR) and intervening regions of hypervariability termed complementarity determining regions (CDR). H and V L Each CDR is composed of three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In certain embodiments of the invention, the FRs of an antibody (or antigen-binding fragment thereof) may be identical to human germline sequences, or may be naturally occurring or artificially modified. A consensus sequence of amino acids may be defined based on the alignment of two or more CDRs. Thus, the heavy chain CDRs are designated "CHRH1," "CDRH2," and "CDRH3," respectively, and the light chain CDRs are designated "CDRL1," "CDRL2," and "CDRL3," respectively.

[0121] "Antibody fragment" refers to a molecule other than an intact antibody that contains a portion of the intact antibody that binds to the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2, diabodies, linear antibodies, single-chain antibody molecules (scFv), and multispecific antibodies formed from antibody fragments.

[0122] "Binding domain" or "antigen-binding domain" refers to the portion of an antibody, compound, or molecule that specifically binds to a target epitope, antigen, ligand, or receptor. Binding domains include, but are not limited to, antibodies (e.g., monoclonal, polyclonal, recombinant, humanized, and chimeric antibodies), antibody fragments or portions thereof (e.g., Fab fragments, Fab2, scFv antibodies, Fv fragments, SMIPs, domain antibodies, diabodies, minibodies, scFv-Fc, affibodies, nanobodies, and antibody VH and / or VL domains), receptors, ligands, aptamers, and other molecules with identified binding partners.

[0123] An "affinity matured antibody" refers to an antibody with one or more modifications in one or more complementarity-determining regions (CDRs), which result in improved affinity of the antibody for its antigen compared to a parent antibody lacking such modifications. Substitution of one or more CDR residues or omission of one or more CDRs is also possible. Antibodies have been described in the scientific literature in which one or two CDRs are not required for binding. Padlan et al. (1995 FASEB J. 9:133-139) analyzed the contact areas between an antibody and its antigen based on published crystal structures and concluded that only approximately one-fifth to one-third of the CDR residues actually contact the antigen. Padlan also found that in many antibodies, one or two CDRs do not contain any amino acids that contact the antigen (see also Vajdos et al. 2002 J Mol Biol 320:415-428). CDR residues that do not contact antigen can be identified based on previous studies, using molecular modeling and / or empirically from regions of the Kabat CDRs outside the Chothia CDRs (e.g., residues H60-H65 of CDRH2 are often not required). When a CDR or its residues are omitted, they are typically replaced with an amino acid at the corresponding position in another human antibody sequence or a consensus of such a sequence. The position of substitution within the CDR and the substituting amino acid can also be selected empirically.

[0124] The fully human monoclonal antibodies disclosed herein may contain one or more amino acid substitutions, insertions, and / or deletions in the framework and / or CDR regions of the heavy and light chain variable domains compared to the corresponding germline sequences. Such mutations can be readily ascertained by comparing the amino acid sequences disclosed herein to germline sequences available, for example, from public antibody sequence databases. The present invention includes antibodies, and antigen-binding fragments thereof, derived from any of the amino acid sequences disclosed herein, in which one or more amino acids in one or more framework and / or CDR regions have been mutated to the corresponding residue in the germline sequence from which the antibody is derived, or to the corresponding residue in another human germline sequence, or to a conservative amino acid substitution of the corresponding residue in the corresponding germline sequence (such sequence changes are collectively referred to herein as "germline mutations"). Starting with the heavy and light chain variable region sequences disclosed herein, one of skill in the art can readily generate numerous antibodies and antigen-binding fragments containing one or more individual germline mutations, or combinations thereof. In certain embodiments, V H and / or V LFramework and / or CDR residues within the domains are mutated back to residues present in the original germline sequence from which the antibody is derived. In other embodiments, only specific residues are mutated back to the original germline sequence. For example, only mutated residues present within the first 8 amino acids of FR1, or the last 8 amino acids of FR4, or only mutated residues present within CDR1, CDR2, or CDR3 are mutated back. In other embodiments, one or more framework and / or CDR residues are mutated to the corresponding residue in a different germline sequence (i.e., a germline sequence that differs from the germline sequence from which the antibody was originally derived). Furthermore, antibodies of the present invention may contain any combination of two or more germline mutations within the framework and / or CDR regions, e.g., certain individual residues are mutated to the corresponding residue in a particular germline sequence, while certain other residues that differ from the original germline sequence are maintained or mutated to the corresponding residue in a different germline sequence. Once obtained, antibodies and antigen-binding fragments containing one or more germline mutations can be readily tested for one or more desired properties. Desired properties include, for example, improved binding specificity, increased binding affinity, improved or enhanced antagonistic or agonistic biological properties (as the case may be), reduced immunogenicity, etc. Antibodies and antigen-binding fragments obtained by this general method are encompassed within the scope of the present invention.

[0125] The present invention also includes fully monoclonal antibodies containing variants of any of the CDR amino acid sequences disclosed herein with one or more conservative substitutions, for example, the present invention includes antibodies having CDR amino acid sequences with, e.g., 10 or fewer, 8 or fewer, 6 or fewer, 4 or fewer, etc., conservative amino acid substitutions compared to any of the CDR amino acid sequences disclosed herein.

[0126] The terms "anti-CD3 antibody," "anti-CD3 binding domain-containing antibody," "antibody that binds to CD3," "CD3 binding domain," "antibody containing a CD3 binding domain," and the like refer to an antibody (or CD3 binding domain) that can bind to CD3 with sufficient affinity and / or specificity to make the antibody useful as a CD3-targeted diagnostic and / or therapeutic agent. In one embodiment, the extent to which an anti-CD3 antibody binds to an unrelated, non-CD3 protein is less than about 10% of the binding of the antibody to CD3, as measured, for example, by one of a variety of solution- and surface-based affinity measurement techniques and devices. Such techniques and devices include, for example, biolayer interferometry (BLI), surface plasmon resonance (SPR), solution equilibrium based kinetic exclusion assay, KinExA direct association assay using ForteBio instruments and reagents, such as Octet RED 384 and HTX BLI-based instruments, enzyme-linked immunosorbent assay (ELISA), and radioimmunoassay (RIA) (see, e.g., Estep et al, MAbs, Vol. 5, pages 270-278 (2013); Yu et al., J Biomol Screen, Vol. 21, pages 88-95 (2016)). In certain embodiments, antibodies that bind to CD3 have a cytotoxicity of <1 mM, <100 nM, <10 nM, <1 nM, <0.1 nM, <0.01 nM, or <0.001 nM (e.g., 10 -8 M or less, e.g. 10 -8 M~10 -13 M, e.g. 10 -9 M~10 -13 In certain embodiments, the anti-CD3 antibody binds to an epitope of CD3 that is conserved among CD3s of different species.

[0127] The term "developable" refers to the extent to which one or more polypeptides in a plurality of polypeptides possess desirable characteristics, such as, for example, desirable expression in mammalian cells, solubility, viscosity, aggregation, chemical and / or physical stability, a desirable "shelf-life," melting point, pharmacokinetic profile, circulating half-life, and clearance characteristics. Such characteristics may be treated individually, as a combination of a subset of such characteristics, or collectively as indicia regarding the likelihood that the one or more polypeptides will be successfully developed as therapeutic candidates and ultimately become approved drugs. Thus, as understood in the art, polypeptides having desirable developability characteristics generally possess, for example, relatively high solubility, relatively low viscosity, relatively low tendency to aggregate, relatively high chemical stability, relatively high physical stability, a relatively long "shelf-life," a relatively high melting point, a relatively long circulating half-life, a relatively long clearance time, etc. Polypeptides with undesirable developability characteristics possess, for example, relatively low solubility, relatively high viscosity, relatively high tendency to aggregate, relatively poor chemical stability, relatively poor physical stability, relatively short "shelf life," relatively low melting point, relatively short circulatory half-life, relatively long clearance time, etc.

[0128] Methods and assays that can be employed to ascertain the extent to which polypeptides, such as, for example, the CD3 binding domains of the invention and antibodies comprising the same, possess such desirable (or, in some cases, undesirable) developability characteristics are available in the art, and include, for example, the methods and assays disclosed in WO 2014 / 179363 and Xu et al., Protein Eng Des Sol, Vol. 26, pp. 663-670 (2013); cross-interaction chromatography (CIC), such as SMP and SCP assays; self-interaction chromatography (SIC); dynamic light scattering; size-exclusion chromatography (SEC), dynamic light scattering (DLS) spectroscopy; photon correlation spectroscopy; quasi-elastic light scattering, circular dichroism (CD), viscometry; whole cell binding; tissue microarray techniques; BVP ELISA assay; AC-SINS assay (Liu et al; MAbs, Vol. 6, pages 483-492 (2014)); differential scanning calorimetry (see, for example, He et al., J. Pharm. Sci., Vol. 100(4), pp. 1330-1340 (2011); Wagner et al., Pharm. Develop. & Technol (posted online 2012; hyper-text transfer protocol: informahealthcare.com / doi / abs / 10.3109 / 10837450.2011.649851); Hotzel et al., mAbs, Vol. 4(6), pages 753-7601 (2012); Weiqiang et al., J. Pharm. Sci., Vol. 101(5), pp. 1701-1720 (2012); Banks et al., J. Pharm. Sci., Vol. 101(8), pp. 2720-2732 (2012); Lie et al., J. Pharm. Sci., Vol. 94(9), pp. 1928-1948 (2005); and Payne et al., Biopolymers, Vol. 85(5), pp. 527-533 (2006).

[0129] Additionally, polypeptides selected or identified as having enhanced developability, such as the CD3-binding domains of the invention and antibodies comprising the same, are referred to as "developable." Polypeptides detected according to the disclosed and claimed methods as having reduced developability are so detected by reason of their interaction with the disclosed and claimed PSR, and are therefore referred to as "multispecific" polypeptides. Such multispecific polypeptides are further referred to as relatively "non-developable" or relatively "non-developable" polypeptides.

[0130] Any means for detecting an interaction between one or more polypeptides and the interacting moieties of the one or more polypeptides may be employed in accordance with the disclosed and claimed methods. Examples of such means include, for example, flow cytometry; magnetic-activated cell sorting (MAGS); fluorescence-assisted cell sorting (FACS); immunohistochemistry; column and / or affinity chromatography or separation; precipitation techniques (e.g., centrifugation); immunoprecipitation; two-hybrid assays, such as mammalian two-hybrid and yeast two-hybrid assays; fluorescence resonance energy transfer (FRET) assays; affinity chromatography, etc. In certain embodiments, detecting such an interaction involves the use of magnetic-activated cell sorting (MAGS); fluorescence-assisted cell sorting (FACS); and / or a combination of magnetic-activated cell sorting (MAGS) and fluorescence-assisted cell sorting (FACS).

[0131] A "developability score," also known as a "developability profile," refers to an index that can be assigned to a CD3-binding domain of the present invention and / or an antibody comprising the same when evaluating its developability as described herein, and for example, in WO 2014 / 179363 and Xu et al., Protein Eng Des Sol, Vol. 26, pages 663-670 (2013). Thus, a developability score is a scale or criterion by which the developability of a CD3-binding agent and / or an antibody comprising the same can be evaluated, compared, and / or ranked. Such a developability score serves as a measure of the degree of interaction between the CD3-binding agent and an antibody comprising the same. The degree of interaction may be assessed by any number of means available in the art that provide an output value that correlates with the strength or affinity of the polypeptide for the moiety to be bound. Examples of such means include flow cytometry methods, such as FACS; ELISA; quantitative immunoaffinity or immunoprecipitation assays; mammalian two-hybrid assays; or yeast two-hybrid assays. In the case of FACS, as shown in the examples, the degree of interaction between a polypeptide in a plurality of polypeptides and PSR may be determined by generating a mean fluorescence intensity (MFI) for each detected polypeptide-PSR interaction, then ordering the MFIs in either ascending or descending order, thereby ranking the polypeptides in the plurality of polypeptides according to the relative degree of interaction between each detected polypeptide and PSR. By performing such a ranking on a plurality of polypeptides, polypeptides with high developability are readily identified, as well as polypeptides with low developability. In certain embodiments, an MFI of 500 or less is an indicator of a polypeptide with high developability. In certain embodiments, an MFI of 400 or less is an indicator of a polypeptide with high developability. In certain embodiments, an MFI of 300 or less is an indicator of a polypeptide with high developability. In certain embodiments, an MFI of 200 or less is an indicator of a polypeptide with high developability.In certain embodiments, an MFI of 100 or less is indicative of a polypeptide possessing high developability. In certain embodiments, an MFI of 1000 or more is indicative of a polypeptide possessing low developability. In certain embodiments, an MFI of 900 or more is indicative of a polypeptide possessing low developability. In certain embodiments, an MFI of 800 or more is indicative of a polypeptide possessing low developability. In certain embodiments, an MFI of 700 or more is indicative of a polypeptide possessing low developability. In certain embodiments, an MFI of 600 or less is indicative of a polypeptide possessing low developability.

[0132] The developability score may also take the form of a normalized score, e.g., on a scale of 0.0 to 1.0, where the score of one or more test CD3 binding agents and / or antibodies comprising the same is normalized to a developability score determined by performing an assay with a standard or control polypeptide or antibody. Examples of such standard or control antibodies include, for example, hen egg lysozyme-binding (HEL) antibodies, e.g., ADI-03847.

[0133] The terms "cytokine release syndrome," "cytokine release crisis," or "cytokine storm" refer to a pro-inflammatory positive feedback loop between cytokines and immune cells, which results in excessive or uncontrolled release of pro-inflammatory cytokines by cells within the immune system (e.g., T cells) (see, e.g., Lee et al., Blood, Vol. 124, pages 188-195 (2014) and Tisoncik et al., Microbiol Mol Biol Rev, Vol. 76, pages 16-32 (2012)). Without wishing to be bound by any theory, it is believed that upon stimulation and activation, immune cells (e.g., T cells) release a series of cytokines to a level and extent that adverse biological / physiological effects occur, or to a degree and severity that includes, for example, redness (redness), swelling or edema, fever (fever), pain (ache), and "loss of function" (loss of function): when localized to the skin or other tissues: increased blood flow, allowing blood leukocytes and plasma proteins to reach the extravascular site of injury, increased local temperature and pain, tissue edema and extravascular pressure, and decreased tissue perfusion; organ and systemic dysfunction, including, for example, cardiac dysfunction, adult respiratory distress syndrome, neurotoxicity, renal and / or hepatic failure, and acute inflammation characterized by disseminated intravascular coagulation. Although many cytokines appear to be associated with the development of CRS, elevated levels of IFNγ, IL-6, TNFα, TGFβ, IL-2, granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-10, IL-8, IL-5, and / or fractalkine may be involved as predictors and / or causative agents of CRS, or may indicate a propensity for CRS to develop upon T cell stimulation. In certain embodiments of the present invention, elevated levels of interleukin-6 (IL-6), interleukin-12 (IL-12), tumor necrosis factor alpha (TNFα), (TGFβ), interleukin-2 (IL-2), and / or interferon gamma (IFNγ) production by T cells incubated with CD3-binding domains and antibodies comprising the same predict a propensity for CRS to develop.In certain embodiments of the invention, elevated levels of interferon gamma (IFNg) production by T cells incubated with the CD3 binding domain and antibodies comprising same predicts a propensity to develop CRS.

[0134] The "cytokine release syndrome risk score," also known as the "cytokine release syndrome risk profile," "CRS risk score," or "CRS risk profile," refers to an index that can be assigned to the CD3-binding domain of the present invention and / or an antibody comprising the same when assessing its propensity to generate cytokine production levels from T cells incubated with or contacted with the CD3-binding domain of the present invention (and / or an antibody comprising the same) at a level that is predicted, expected, and / or actually shown to be sufficient to induce cytokine release syndrome in a subject. Thus, the developability score is a scale or criterion by which the CRS risk of CD3-binding agents and / or antibodies comprising the same can be assessed, compared, and / or ranked. Such CRS risk scores serve as an index or indicator of the propensity of CD3-binding agents and antibodies comprising the same to elicit cytokine release when assessed in cytokine release assays either in vitro (e.g., in cell lines) or in vivo (e.g., in test subjects or patients). Such assays are available to those skilled in the art and, in certain embodiments of the invention, include, for example, the use of PBMCs, either cultured cell lines or primary cells obtained from living donors, to assess the ability of, for example, CD3-binding domains and antibodies comprising the same to induce cytokine production / secretion in cell-based cytokine release assays performed in fixed or, preferably, soluble antibody formats (see, e.g., Vessillier et al., J Immunol Methods, Vol. 424, pages 43-52 (2015)). Generally, cytokine production levels measured as an indicator or predictor of CRS risk are indicative of a low risk of CRS induction by a test molecule lower than the levels measured with a known inducer of cytokine release syndrome or cytokine storm (i.e., a positive control), as will be understood by those skilled in the art and / or disclosed in the literature.

[0135] The term "recombinant" generally refers to any protein, polypeptide, or cell expressing a gene of interest that has been produced by genetic engineering methods. When used in connection with a protein or polypeptide, the term "recombinant" means a polypeptide that has been produced by expression of a recombinant polynucleotide. Proteins used in the immunogenic compositions of the invention may be isolated from natural sources or may be produced by genetic engineering methods.

[0136] In some embodiments, antibodies of the present invention may be recombinant human antibodies. The term "recombinant human antibody," as used herein, is intended to include all antibodies, including human or humanized antibodies prepared, expressed, generated, or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell (described in more detail below), antibodies isolated from a recombinant combinatorial human antibody library (described in more detail below), antibodies isolated from an animal (e.g., a mouse) transgenic for human immunoglobulin genes (see, e.g., Taylor et al. (1992) Nucl. Acids Res. 20:6287-6295), or antibodies prepared, expressed, generated, or isolated by any other means, including splicing human immunoglobulin gene sequences into other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or, when animals transgenic for human Ig sequences are used, in vivo somatic mutagenesis) to thereby modify the V H and V L The amino acid sequence of the region is similar to that of the human germline V H and V L While derived from and related to sequences, they may not naturally occur within the human antibody germline repertoire in vivo.

[0137] The term "specifically binds" or "specifically binds" means that an antibody or antigen-binding fragment thereof forms a complex with an antigen that is relatively stable under physiological conditions. Specific binding is defined as binding to an antigen with a specific binding affinity of at least about 1 x 10 -6 can be characterized by an equilibrium dissociation constant less than or equal to M (e.g., K D (A smaller value indicates tighter binding.) Methods for determining whether two molecules specifically bind are known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. As disclosed herein, antibodies that specifically bind to CD3 are identified by surface plasmon resonance, such as BIACORE™ biolayer interferometry using, for example, a FORTEBIO Octet HTX instrument (Pall Life Sciences). Furthermore, multispecific antibodies that bind to CD3 and one or more additional antigens, or bispecific antibodies that bind to two different regions of CD3 and / or additional antigens, are also considered "specifically binding" antibodies as used herein. In certain embodiments, the antibodies disclosed herein have a specific binding affinity of about 1x10 -6 M, about 1x10 -7 M, about 1x10 -8 M, about 1x10 -9 M, about 1x10 -10 M, about 1x10 -6 M ~ approx. 1x10 -7 M, about 1x10 -7 M ~ approx. 1x10 -8 M, about 1x10 -8 M ~ approx. 1x10 -9 M, or approximately 1x10 -9 M ~ approx. 1x10 -10 The equilibrium dissociation constant (and hence specificity) of M is given.

[0138] The term "high affinity" antibody refers to an antibody with an affinity of at least 10 as measured by surface plasmon resonance, e.g., BIACORE, Biolayer Interferometry, using, e.g., a FORTEBIO Octet HTX instrument (Pall Life Sciences), or solution affinity ELISA. -9 M, preferably 10 -10M, more preferably 10 -11 M, more preferably 10 -12 K of M D This refers to a mAb with binding affinity for CD3 expressed as

[0139] The term "slow off rate" refers to a slow off rate of 1x10 as determined by surface plasmon resonance, for example, on a BIACORE™ or FORTEBIO Octet HTX instrument (Pall Life Sciences). -3 s "1 Less than or equal to 1x10 -4 s "1 The following rate constants (K off or K D ) refers to an antibody that dissociates from CD3.

[0140] Further, with regard to antibody fragments, such antibody fragments can include Fab fragments, F(ab')2 fragments, Fv fragments, dAb fragments, fragments containing CDRs, or isolated CDRs. Antigen-binding fragments of antibodies can be derived from intact antibody molecules using any suitable standard technique, such as, for example, proteolytic digestion or recombinant genetic engineering methods involving the manipulation and expression of DNA encoding the variable and (optionally) constant domains of the antibody. Such DNA is known and / or readily available, for example, from commercial sources, DNA libraries (including, for example, phage-antibody libraries), or can be synthesized. The DNA can be sequenced and manipulated chemically or using molecular biology techniques, for example, to arrange one or more variable and / or constant domains into the appropriate configuration, or to introduce codons, generate or modify cysteine ​​residues, or add or delete amino acids.

[0141] Non-limiting examples of antigen-binding fragments include (i) Fab fragments, (ii) F(ab')2 fragments, (iii) Fd fragments, (iv) Fv fragments, (v) single-chain Fv (scFv) molecules, (vi) dAb fragments, and (vii) minimal recognition units consisting of amino acid residues mimicking the hypervariable regions of an antibody (e.g., isolated complementarity-determining regions (CDRs), such as CDR3 peptides) or constrained FR3-CDR3-FR4 peptides. Other engineered molecules, such as domain-specific antibodies, single-domain antibodies, domain-deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains, are also encompassed within the term "antigen-binding fragment" as used herein.

[0142] Antigen-binding fragments of antibodies will often contain at least one variable domain, which may be of any size or amino acid composition and will generally contain at least one CDR, either contiguous or in-frame with one or more framework sequences. L V associated with the domain H In an antigen-binding fragment having a domain, V H Domains and V L The domains may be positioned in any suitable arrangement relative to each other. For example, the variable region may be a dimer, with the V H -V H , V H -V L or V L -V L Alternatively, the antigen-binding fragment of an antibody may contain a dimer of monomeric V H or V L It may contain domains.

[0143] In certain embodiments, an antigen-binding fragment of an antibody may contain at least one variable domain covalently linked to at least one constant domain. Non-limiting examples of variable and constant domain configurations that may be present in an antigen-binding fragment of an antibody of the invention include: (i) V H -C H 1 ; (ii) V H -C H 2; (iii) V H -C H 3; (iv) V H -C h 1 -C h 2; (V) V H -C h 1-C h 2-C h 3; (vi) V H -C H 2-C H 3; (vii) V H -C L (viii) V L -C H 1 ; (ix) V L -C H 2; (x) V L -C H 3; (xi) V L -C H 1-C H 2; (xii) V L -C H 1-C H 2-C H 3; (xiii) V L -C H 2-C H 3; and (xiv) V L -C LIn any configuration of variable and constant domains, including any of the exemplary configurations above, the variable and constant domains may be directly linked to each other or may be linked by a complete or partial hinge or linker region. The hinge region may consist of at least two (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids, which provides a flexible or semi-flexible linkage between adjacent variable and / or constant domains in a single polypeptide molecule. Furthermore, antigen-binding fragments of antibodies of the present invention may be non-covalently linked to each other and / or to one or more monomeric V H or V L It may comprise homodimers or heterodimers (or other multimers) of any of the above variable and constant domain configurations, with the domains non-covalently linked (eg, by disulfide bonds).

[0144] Like intact antibody molecules, antigen-binding fragments may be monospecific or multispecific (e.g., bispecific). Multispecific antigen-binding fragments of antibodies often contain at least two different variable domains, where each variable domain can specifically bind to a distinct antigen or to different epitopes on the same antigen. Any multispecific antibody format, including the exemplary bispecific antibody formats disclosed herein, may be adapted for use in the context of antigen-binding fragments of antibodies of the invention using routine techniques available in the art. Multispecific antibody fragments suitable for incorporation of the anti-CD3 binding domain of the invention may be used in the context of antigen-binding fragments of antibodies of the invention. Non-limiting examples of multispecific and bispecific formats include, for example, Fab-Fc-scFv (Bottle-Opener) (XENCOR), Mab-scFv (XENCOR), Mab-Fv (XENCOR), Dual scFv (XENCOR), central Fv (XENCOR), central scFv (XENCOR), one-arm central scFv (XENCOR), Fab-Fab (XENCOR), Fab-Fv (XENCOR), mAb-Fv (XENCOR), mAb-Fab (XENCOR), DART (MacroGenics), BiTE (Amgen / Micromet), KiTE, common light chain-IgG (Genentech), TandAb (SFFIMED), Cross-Mab (Roche), SEED (EMD Serono), BEAT (Glenmark), TrioMab (Trion Pharma / FresEnius Biotech), DuetMab (MedImmune), and others, including, for example, (WO 95 / 09917; WO 2008 / 119566; WO 2008 / 119567; WO2011 / 121110; WO 2010 / 037835; WO 2007 / 042261; WO 2007 / 110205; WO 2011 / 121110; WO 2012 / 055961; WO 2012 / 16067; WO 2016 / 086189; WO 2016 / 182751; WO 2015 / 006749; WO 2014 / 049003; WO 2013 / 177101; WO 2015 / 128509; US 7,951,917; US 2009 / 0252729; US 2014 / 0348839; US 7,183,076; Mazor et al., Mabs, Vol. 7, pp. 377-389 (2015); Muda et al., Protein Engineering, Design, & Selection, Vol. 24, pp. 447-454 (2011); and Del Bano et al., Antibodies, Vol.5, pp. 1-23 (2016).

[0145] In certain embodiments, the antibodies or antibody fragments of the invention may be conjugated to a therapeutic moiety (immunoconjugate), such as, for example, an antibiotic, a second anti-CD3 antibody, a vaccine or toxoid, or any other useful therapeutic moiety.

[0146] As used herein, an "isolated antibody" is intended to refer to an antibody that is substantially free of other antibodies (Abs) having different antigen specificities (e.g., an isolated antibody that specifically binds CD3, or a fragment thereof, is substantially free of Abs that specifically bind to antigens other than CD3).

[0147] As used herein, "inhibitory antibody" or "neutralizing antibody" (or an antibody that neutralizes CD3 activity) is intended to refer to an antibody whose binding to CD3 results in the inhibition of at least one biological activity of CD3, as the case may be, as disclosed herein.

[0148] As used herein, the term "surface plasmon resonance" refers to an optical phenomenon that allows the analysis of biomolecular interactions in real time by detecting changes in protein concentration within a biosensor matrix, for example using the BIACORE™ system (Pharmacia Biosensor AB, Uppsala, Sweden and Piscataway, New Jersey).

[0149] As used herein, "K D The term "antibody-antigen interaction" is intended to refer to the equilibrium dissociation constant of a particular antibody-antigen interaction.

[0150] The term "epitope" refers to an antigenic determinant that interacts with a specific antigen-binding site in the variable region of an antibody molecule, known as the paratope. A single antigen may have multiple epitopes. Thus, different antibodies may bind to different regions on an antigen and have different biological effects. The term "epitope" also refers to the site on an antigen to which B cells and / or T cells respond. It also refers to the region of an antigen that is bound by an antibody. Epitopes can be defined structurally or functionally. Functional epitopes are generally a subset of structural epitopes and contain residues that directly contribute to the affinity of the interaction. Epitopes can also be conformational; that is, they may be composed of nonlinear amino acids. In certain embodiments, epitopes may include determinants that are chemically active surface groupings of molecules, such as amino acids, sugar side chains, phosphoryl groups, or sulfonyl groups. In certain embodiments, epitopes may have specific three-dimensional structural characteristics and / or specific charge characteristics.

[0151] The term "substantial identity" or "substantially identical," when referring to a nucleic acid or fragment thereof, or an amino acid sequence or fragment thereof, indicates that there is at least about 100%, at least about 99%, at least about 98%, at least about 97%, at least about 96%, at least about 95%, at least about 94%, at least about 93%, at least about 92%, at least about 91%, at least about 90%, at least about 89%, at least about 88%, at least about 87%, at least about 86%, at least about 85%, at least about 84%, at least about 83%, at least about 82%, at least about 80%, etc., sequence identity when optimally aligned with another nucleic acid or amino acid sequence, as the case may be, with appropriate insertions or deletions (or their complements), and as measured by any known algorithm for sequence identity, such as, for example, FASTA, BLAST, or GAP, as discussed below. Thus, sequences exhibiting a particular percentage of "identity" share that percentage of identity and / or are "identical" to each other to that degree.

[0152] In certain embodiments, the disclosed CD3 binding domains, and / or individual heavy chain (HC) sequences, light chain (LC) sequences, CDRH3 sequences, CDRH2 sequences, CHRH1 sequences, CDRL3 sequences, CDRL2 sequences, CDRL1 sequences, and / or framework sequences are independently at least about 100%, at least about 99%, at least about 98%, at least about 97%, at least about 96%, at least about 95%, at least about 94%, at least about 93%, at least about 92%, at least about 91%, at least about 90%, at least about 89%, at least about 88%, at least about 87%, at least about 86%, at least about 85%, at least about 84%, at least about 83%, at least about 82%, at least about 80% identical to other sequences, and / or all percentages therebetween, and / or share such percentages of identity with each other (or with specific subsets of the antibody sequences disclosed herein).

[0153] In certain embodiments, the amino acid sequences of the antibodies of the present disclosure are, for example, at least 70% identical, at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, and / or all percentages of identity therebetween to other sequences and / or share such percentages of identity with each other (or with particular subsets of the antibody sequences disclosed herein).

[0154] Preferably, residue positions that are not identical differ by conservative amino acid substitutions. A "conservative amino acid substitution" is one in which an amino acid residue is replaced with another amino acid residue having a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity). Generally, conservative amino acid substitutions do not substantially change the functional properties of a protein. When two or more amino acid sequences differ from each other by conservative substitutions, the percentage or degree of similarity may be adjusted upward to correct for the conservative nature of the substitution. Means for making this adjustment are known to those of skill in the art. (See, e.g., Pearson (1994) Methods Mol. Biol. 24: 307-331.) Examples of groups of amino acids with side chains with similar chemical properties include: 1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; 2) aliphatic-hydroxyl side chains: serine and threonine; 3) amide-containing side chains: asparagine and glutamine; 4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; 5) basic side chains: lysine, arginine, and histidine; 6) acidic side chains: aspartic acid and glutamic acid; and 7) sulfur-containing side chains: cysteine ​​and methionine. Preferred conservative amino acid substitution groups are valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamic acid-aspartic acid, and asparagine-glutamine. Alternatively, a conservative substitution is any change that has a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet et al. (1992) Science 256: 1443 45. A "moderately conservative" substitution is any change that has a non-negative value in the PAM250 log-likelihood matrix.

[0155] Polypeptide sequence similarity is often measured using sequence analysis software. Protein analysis software matches similar sequences using similarity measures assigned to various substitutions, deletions, and other modifications, including conservative amino acid substitutions. For example, GCG software contains programs such as GAP and BESTFIT, which can be used with default parameters to determine sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different species or homologous polypeptides between wild-type proteins and their mutants. See, for example, GCG version 6.1. Polypeptide sequences can also be compared using FASTA with default or recommended parameters; the GCG version 6.1 program. FASTA (FASTA2 and FASTA3) provides alignments and percentage sequence identity of the regions of best overlap between the query and search sequences (Pearson (2000) supra). Another preferred algorithm for comparing the sequences of the present invention to a database containing a large number of sequences from different organisms is the computer program BLAST, particularly BLASTP or TBLASTN with default parameters. (See, e.g., Altschul et al. (1990) J. Mol. Biol. 215: 403 410 and (1997) Nucleic Acids Res. 25:3389 402).

[0156] In certain embodiments, antibodies or antibody fragments for use in the methods of the invention may be monospecific, bispecific, or multispecific. Multispecific antibodies may be specific for different epitopes of a single target polypeptide, or may contain antigen-binding domains specific for epitopes of multiple target polypeptides. Examples of bispecific antibody formats that can be used in the context of the present invention include a first immunoglobulin (Ig) C H 3 domain and second IgC H 3 domains, in which the first and second IgC HThe three domains differ from each other by at least one amino acid, and wherein the at least one amino acid difference reduces binding of the bispecific antibody to Protein A compared to a bispecific antibody without that amino acid difference. H The 3 domain binds to protein A and the second IgC H The C3 domain contains a mutation that reduces or eliminates Protein A binding, for example, the H95R (according to IMGT exon numbering; H435R in EU numbering) modification. H 3 may further comprise a Y96F modification (according to IMGT; Y436F in EU). H Additional modifications that may be present within 3 include: D16E, L18M, N44S, K52N, V57M, and V82I for IgG1 mAbs (according to IMGT; D356E, L358M, N384S, K392N, V397M, and V422I for EU); N44S, K52N, and V82I for IgG2 mAbs (according to IMGT; N384S, K392N, and V422I for EU); and Q15R, N44S, K52N, V57M, R69K, E79Q, and V82I for IgG4 mAbs (according to IMGT; Q355R, N384S, K392N, V397M, R409K, E419Q, and V422I for EU). Variations on the bispecific antibody format described above are also contemplated within the scope of the present invention.

[0157] Examples of anti-CD3 binding domains and antibodies containing them

[0158] As described above, certain embodiments of the present invention provide a series of CD3-binding domains and antibodies comprising the same, as well as methods for producing and using them. Surprisingly, this series of CD3-binding domains collectively exhibit desirable properties, such as broad affinity for CD3 epsilon, cross-reactivity with both human CD3 (Hu CD3) and cynomolgus monkey CD3 (Cy CD3), and a desirable developability profile and / or cytokine release syndrome (CRS) risk profile. In certain embodiments, the CD3-binding domains of the present invention and antibodies comprising the same exhibit a developability profile and / or CRS risk profile that is superior to the developability profile and / or CRS risk profile of other anti-CD3 antibodies. In certain embodiments, the CD3 binding domains of the invention and antibodies comprising same exhibit a developability profile and / or CRS risk profile that is superior to the developability profile and / or CRS risk profile of I2C, SP34, 38E4, CAB21609_A01, CAB21609_B01, CAB21609_C01, and / or CAB21609_D01, e.g., as disclosed herein and as disclosed in Yang et al., J Immunol, Vol 137, pp. 1097-1100 (August 4, 1986), US 2014 / 008295, and WO 2015 / 095392.

[0159] In certain embodiments, the present invention provides CD3 binding domains and antibodies comprising the same that bind to CD3 (eg, CD3ε and / or CD3γ).

[0160] In certain embodiments, CD3 binding domains of the invention and antibodies comprising the same are provided that exhibit an enhanced developability profile compared to other CD3 binding domains (or antibodies comprising the same). In certain embodiments, CD3 binding domains of the invention and antibodies comprising the same are provided that exhibit an enhanced developability profile compared to trastuzumab (Herceptin®), lintuzumab, blinatumomab (Blincyto®), and one or more of Mab 364, Mab 366, Mab 367, Mab 368, Mab 369, Mab 370, or Mab 22, as presented in Table 2.

[0161] In specific embodiments, the CD3-binding domains of the present invention and antibodies comprising the same have an affinity for an antibody having an affinity of about 0 MFI to about 500 MFI, about 0 MFI to about 450 MFI, about 0 MFI to about 400 MFI, about 0 MFI to about 350 MFI, about 0 MFI to about 300 MFI, about 0 MFI to about 250 MFI, about 0 MFI to about 200 MFI, about 0 MFI to about 150 MFI, about 0 MFI to about 100 MFI, about 0 MFI to about 50 MFI, about 200 MFI and exhibits a developability score of 500MFI, about 200MFI to about 450MFI, about 200MFI to about 400MFI, about 200MFI to about 350MFI, about 200MFI to about 300MFI, about 200MFI to about 250MFI, about 100MFI to about 450MFI, about 100MFI to about 400MFI, about 100MFI to about 350MFI, about 100MFI to about 300MFI, about 100MFI to about 250MFI, about 100MFI to about 200MFI, or about 100MFI to about 150MFI.

[0162] In other embodiments, the CD3-binding substances of the present invention and antibodies comprising the same have a CpG of about 0.0 to about 0.6, about 0.0 to about 0.57, about 0.0 to about 0.55, about 0.0 to about 0.53, about 0.0 to about 0.51, about 0.0 to about 0.49, about 0.0 to about 0.47, about 0.0 to about 0.45, about 0.0 to about 0.43, about 0.0 to about 0.41, about 0.0 to about 0.39, about 0.0 to about 0.37, about 0.0 to about 0.35, about 0.0 to about 0.4 exhibits a normalized developability score of about 0.33, about 0.0 to about 0.31, about 0.0 to about 0.29, about 0.0 to about 0.27, about 0.0 to about 0.25, about 0.0 to about 0.23, about 0.0 to about 0.21, about 0.0 to about 0.19, about 0.0 to about 0.17, about 0.0 to about 0.15, about 0.0 to about 0.13, about 0.0 to about 0.11, about 0.0 to about 0.09, about 0.0 to about 0.07, or about 0.0 to about 0.05.

[0163] In certain embodiments, the developability profile and / or developability score for the CD3 binding agents of the present invention and antibodies comprising the same are obtained by performing a PSR assay, an SCP assay, an AS-CINS, a BVP assay, an ELISA, a DSF assay, a Tm assay, an HIC assay, a CIC assay, or a combination thereof.

[0164] In other embodiments, the CD3-binding domains of the invention and antibodies comprising same elicit potent T cell activation or killing while exhibiting a reduced tendency to elicit cytokine production to levels capable of inducing cytokine release syndrome. In certain embodiments, cytokine production levels are measured to assess the tendency to elicit cytokine production levels capable of inducing cytokine release syndrome, and the at least one cytokine is selected from the group consisting of interleukin-6 (IL-6), interleukin-12 (IL-12), tumor necrosis factor alpha (TNFα), (TGFβ), interleukin-2 (IL-2), and interferon-gamma (IFNγ).

[0165] In certain embodiments, the CD3 binding domains of the invention and antibodies comprising same, while eliciting T cell activation or T cell killing, exhibit a reduced tendency to elicit cytokine production to levels capable of inducing cytokine release compared to the tendency observed with trastuzumab (Herceptin®), lintuzumab, blinatumomab (Blincyto®), and one or more of Mab 364, Mab 366, Mab 367, Mab 368, Mab 369, Mab 370, or Mab 22, as presented in Table 2. In certain embodiments, the level of cytokine production is measured to assess the propensity for at least one cytokine to elicit cytokine production levels capable of inducing cytokine release syndrome, wherein the at least one cytokine is selected from the group consisting of interleukin 6 (IL-6), interleukin 12 (IL-12), tumor necrosis factor alpha (TNFa), (TGFb), interleukin 2 (IL-2), and interferon gamma (IFNg).

[0166] In certain embodiments, the CD3-binding domains of the present invention and antibodies comprising the same exhibit a cytokine release syndrome risk profile indicative of a reduced risk of inducing cytokine release syndrome (CRS). In other embodiments, the CD3-binding domains of the present invention and antibodies comprising the same exhibit a cytokine release syndrome risk profile indicative of a reduced risk of inducing cytokine release syndrome (CRS) compared to the cytokine release syndrome risk profiles assessed with trastuzumab (Herceptin®), lintuzumab, blinatumomab (Blincyto®), and one or more of Mab 364, Mab 366, Mab 367, Mab 368, Mab 369, Mab 370, or Mab 22, as presented in Table 2.

[0167] In certain embodiments, the present invention provides antibodies containing a CDRH3 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRH3 selected from the group consisting of the CDRH3s of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2, provided that the CDRH3 is not 100% identical to the CDRH3 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370 as presented in Table 2.

[0168] In certain embodiments, the present invention provides antibodies containing a CDRH2 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRH2 selected from the group consisting of CDRH2 of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2, provided that the CDRH2 is not 100% identical to the CDRH2 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370 as presented in Table 2.

[0169] In certain embodiments, the present invention provides antibodies containing a CDRH1 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRH1 selected from the group consisting of CDRH1 of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2, provided that the CDRH1 is not 100% identical to the CDRH1 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370 as presented in Table 2.

[0170] In certain embodiments, the present invention provides antibodies containing a CDRL3 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRL3 selected from the group consisting of CDRL3s of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2, provided that the CDRL3 is not 100% identical to the CDRL3 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370 as presented in Table 2.

[0171] In certain embodiments, the present invention provides antibodies containing a CDRL2 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRL2 selected from the group consisting of CDRL2 of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2, provided that the CDRL2 is not 100% identical to the CDRL2 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370 as presented in Table 2.

[0172] In certain embodiments, the present invention provides antibodies containing a CDRL1 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRL1 selected from the group consisting of CDRL1 of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2, provided that the CDRL1 is not 100% identical to the CDRL1 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370 as presented in Table 2.

[0173] In certain embodiments, the present invention provides Mabs as presented in Table 2. Provided are antibodies containing a heavy chain (HC) variable region that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a HC variable region selected from the group consisting of HC variable regions 1-21, 23-222, 224-363, and 371-405, provided that the HC variable region is not 100% identical to the HC variable region of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370 as set forth in Table 2.

[0174] In certain embodiments, the present invention provides Mabs as presented in Table 2. and 371-405, provided are antibodies containing a light chain (LC) variable region that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a LC variable region selected from the group consisting of LC variable regions 1-21, 23-222, 224-363, and 371-405, provided that the LC variable region is not 100% identical to any of LCs Mab223, 364, 365, 366, 367, 368, 369, or 370 as presented in Table 2.

[0175] In certain embodiments, the present invention provides a CDRH3 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRH3 selected from the group consisting of CDRH3s of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2. and a CDRH2 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRH2 selected from the group consisting of CDRH2s 1-21, 23-222, 224-363, and 371-405, as well as a Mab as set forth in Table 2.100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 89% identical, at least 9 ... Antibodies are provided that contain a CDRH1 that is 7% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical, provided that none of the CDRH3, CDRH2, and CHRH1 are 100% identical to the CHRH3, CDRH2, or CDRH1 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370, respectively, as set forth in Table 2.

[0176] In certain embodiments, the present invention provides a CDRL3 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRL3 selected from the group consisting of CDRL3s of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2. and a CDRL2 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRL2 selected from the group consisting of CDRL2s 1-21, 23-222, 224-363, and 371-405, as well as a Mab as set forth in Table 2.100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 89% identical, at least 9 ... Antibodies are provided that contain a CDRL1 that is 7% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical, provided that none of the CDRL3, CDRL2, and CHRL1 are 100% identical to the CHRL3, CDRL2, or CDRL1 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370, respectively, as set forth in Table 2.

[0177] In certain embodiments, the present invention provides a CDRH3 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRH3 selected from the group consisting of CDRH3s of Mabs 1-21, 23-222, 224-363, and 371-405 as presented in Table 2. a CDRH2 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRH2 selected from the group consisting of CDRH2s 1-21, 23-222, 224-363, and 371-405, as presented in Table 2;a CDRH1 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRH1 selected from the group consisting of CDRH1s 1-21, 23-222, 224-363, and 371-405, as presented in Table 2; a CDRL3 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRL3 selected from the group consisting of CDRL3s 1-21, 23-222, 224-363, and 371-405, as presented in Table 2;and a CDRL2 that is 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, at least 86% identical, at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical to a CDRL2 selected from the group consisting of CDRL2s 1-21, 23-222, 224-363, and 371-405, as well as a Mab as set forth in Table 2. 100% identical, at least 99% identical, at least 98% identical, at least 97% identical, at least 96% identical, at least 95% identical, at least 94% identical, at least 93% identical, at least 92% identical, at least 91% identical, at least 90% identical, at least 89% identical, at least 88% identical, at least 87% identical, or at least 86% identical to a CDRL1 selected from the group consisting of CDRL1s 1 to 21, 23 to 222, 224 to 363, and 371 to 405. or at least 85% identical, at least 84% identical, at least 83% identical, at least 82% identical, or at least 80% identical, provided that none of the CDRH3, CDRH2, CDRH1, CDRL3, CDHL2, and CHRL1 is 100% identical to the CDRH3, CDRH2, CDRH1, CDRL3, CDHL2, or CHRL1 of any of Mabs 223, 364, 365, 366, 367, 368, 369, or 370, respectively, as set forth in Table 2.

[0178] In certain embodiments, the present invention provides antibodies containing a CD3 binding domain selected from the group consisting of the CD3 binding domains of ADI-15512, ADI-15516, and ADI-16513, as presented in Table 2.

[0179] In certain embodiments, the present invention provides a method for the treatment of a rheumatoid arthritis (RA) comprising administering to a patient a therapeutically effective amount ... Antibodies are provided that contain a CD3 binding domain selected from the group consisting of the CD3 binding domains of ADI-18590; ADI-18591; ADI-18593; ADI-18594; ADI-18595; ADI-18596; ADI-18597; ADI-18592; ADI-18587; ADI-18586; and ADI-16606.

[0180] In certain embodiments, the present invention provides a method for the preparation of a compound selected from the group consisting of: ADI-18576; ADI-20820; ADI-20578; ADI-20571; ADI-21097; ADI-20577; ADI-20576; ADI-20568; ADI-20582; ADI-20575; ADI-20567; ADI-20574; ADI-20573; ADI-20579; ADI-18565; ADI-20818; ADI-20587; ADI-20588; ADI-20589; ADI-20590; ADI-20594; ADI-20596; ADI-20599; ADI-20605; ADI-20607; An antibody containing a CD3 binding domain selected from the group consisting of the CD3 binding domains of ADI-20608 and ADI-20609 is provided.

[0181] In certain embodiments, the present invention provides, as presented in Table 2, ADI-16606; ADI-20587; ADI-20607; ADI-20590; ADI-28708; ADI-28709; ADI-28710; ADI-21943; ADI-28711; ADI-28712; ADI-28713; ADI-28714; ADI-28715; ADI-21944; ADI-28716; ADI-21945; ADI-21946; ADI-28717; ADI-21947; ADI-28718; ADI-28719; ADI-28720; ADI-28721; ADI-28722; ADI-28723; ADI-28724; ADI-28725; ADI-28726; ADI-28727; ADI-28728; ADI-28729; ADI-28730; ADI-28731; ADI-28732; ADI-28733; ADI-28734; ADI-28735; ADI-28736; ADI-28737; ADI-28738; ADI-28739; ADI-28740; ADI-28741; ADI-28742; ADI-28743; ADI-21948; ADI-21949; ADI-28744; ADI-21950; ADI-28745; ADI-28746; ADI-28747; ADI-28748; ADI-21951; ADI-21952; ADI-28749; ADI-28750; ADI-28751; ADI-21953; ADI-28752; ADI-21954; ADI-28753; ADI-28754; ADI-28755; ADI-28756; ADI-28757; ADI-28758; ADI-28759; ADI-28760; ADI-28761; ADI-28762; ADI-28763; ADI-28764; ADI-28765; ADI-28766; ADI-28767; ADI-28768; ADI-21955; ADI-28769; ADI-28770; ADI-21956; ADI-28771; ADI-28772; ADI-28773An antibody containing a CD3-binding domain selected from the group consisting of the CD3-binding domains of ADI-28774 and ADI-28775 is provided.

[0182] In certain embodiments, the present invention provides a method for the treatment of a pulmonary artery disease comprising administering to a patient a therapeutically effective amount of a pulmonary artery disease (PAD) ... An antibody containing a CD3 binding domain selected from the group consisting of the CD3 binding domains of ADI-21955 and ADI-21956 is provided.

[0183] In certain embodiments, the present invention provides a method for the treatment of a pulmonary artery disease comprising administering to a patient a therapeutically effective amount of a pulmonary artery disease (PAD) comprising administering to a patient a therapeutically effective amount of a pulmonary artery disease (PAH ... ADI-24443; ADI-24444; ADI-24445; ADI-24446; ADI-24449; ADI-24388; ADI-24389; ADI-24390; ADI-24391; ADI-24392; ADI-24393; ADI-24396; ADI-24397; ADI-24398; ADI-24399; ADI-24400; ADI-24401; ADI-24402; ADI-24419; ADI-24420; ADI-24421; ADI-24422; ADI-24425; Antibodies are provided that contain a CD3 binding domain selected from the group consisting of the CD3 binding domains of ADI-24426; ADI-24427; ADI-24428; ADI-24429; ADI-24430; ADI-24431; ADI-24432; ADI-24433; ADI-24447, and ADI-24448.

[0184] In certain embodiments, the present invention provides a method for the preparation of a compound selected from the group consisting of: ADI-22523; ADI-23652; ADI-23653; ADI-23654; ADI-23655; ADI-23656; ADI-23657; ADI-23658; ADI-23651; ADI-23644; ADI-23645; ADI-23646; ADI-23647; ADI-23648; ADI-23649; ADI-23650; ADI-23667; ADI-23668; ADI-23669; ADI-23670; ADI-23671; ADI-23672; ADI-23673; ADI-23659; ADI-23660; ADI-23661; Antibodies are provided that contain a CD3 binding domain selected from the group consisting of the CD3 binding domains of ADI-23663; ADI-23664; ADI-23639; ADI-23641; ADI-23642; ADI-23640; ADI-23643; ADI-21952; ADI-23633; ADI-23634; ADI-23635; ADI-23636; ADI-23637; ADI-23638; ADI-23632, and ADI-23629.

[0185] In certain embodiments, the present invention provides a method for the treatment of rhodium-containing ... ADI-26941; ADI-26942; ADI-26943; ADI-26944; ADI-26945; ADI-26950; ADI-26954; ADI-23672; ADI-23673; ADI-23664; ADI-26955; ADI-26958; ADI-26959; ADI-26960; ADI-26962; ADI-26963; ADI-26964; ADI-26965; ADI-26966; ADI-26974; Antibodies are provided that contain a CD3 binding domain selected from the group consisting of the CD3 binding domains of ADI-26975; ADI-26976; ADI-26977; ADI-26978; ADI-26979; ADI-26980; ADI-26981; ADI-26982; ADI-26983; ADI-26984; ADI-26985; ADI-26986; ADI-26987; ADI-26988; ADI-26989; ADI-26990; ADI-26991; ADI-26992; ADI-26993; ADI-26994; and ADI-26995.

[0186] In certain embodiments, the present invention provides antibodies containing a CD3 binding domain selected from the group consisting of the CD3 binding domains of ADI-22523, ADI-26906, ADI-26907, ADI-26908, ADI-26910, ADI-26913, ADI-26915, ADI-26919, ADI-26920, ADI-26921, ADI-26943, ADI-26954, ADI-21952, ADI-26955, ADI-26956, ADI-26962, ADI-26978, ADI-26983, and ADI-26994, as presented in Table 2.

[0187] In certain embodiments, the present invention provides an antibody containing a CD3 binding domain selected from the group consisting of: ADI-15512, ADI-16513, ADI-15516, ADI-18565, ADI-18589, ADI-18585, ADI-18590, ADI-18576, ADI-20568, ADI-20580, ADI-21978, ADI-22523, ADI-25133, and ADI-26906.

[0188] In certain embodiments, the invention provides the following methods: ADI-16606; ADI-29601; ADI-29602; ADI-29603; ADI-20587; ADI-20607; ADI-20590; ADI-26957; ADI-26958; ADI-26959; ADI-26960; ADI-26961; ADI-26962; ADI-26963; ADI-26964; ADI-26965; ADI-26966; ADI-26967; ADI-26970; ADI-26971; Provided are antibodies containing a CD3 binding domain selected from the group consisting of: ADI-26972; ADI-26973; ADI-26974; ADI-26975; ADI-26976; ADI-26977; ADI-26978; ADI-26979; ADI-26980; ADI-26981; ADI-26982; ADI-26983; ADI-26984; ADI-26985; ADI-26986; ADI-26987; ADI-26988; ADI-26989; ADI-26990; ADI-26991; ADI-26992; ADI-26993; and ADI-26994:

[0189] In certain embodiments, the present invention provides antibodies containing a CD3-binding domain selected from the group consisting of ADI-32238, ADI-32241, ADI-32244, ADI-32247, ADI-32250, ADI-32253, ADI-32256, ADI-32259, ADI-32239, ADI-32242, ADI-32245, ADI-32248, ADI-32251, ADI-32254, ADI-32257, ADI-32260, ADI-32240, ADI-32243, ADI-32246, ADI-32249, ADI-32252, ADI-32255, ADI-32258, and ADI-32261:

[0190] In certain embodiments, the present invention provides an antibody containing a CD3 binding domain selected from the group consisting of: ADI-29295, ADI-32249, ADI-29298, ADI-29300, and ADI-26915.

[0191] In certain embodiments, either alone or in combination with other embodiments of the invention, the CD3 binding domains of the invention and antibodies comprising same exhibit a reduced tendency to degradation compared to trastuzumab (Herceptin®), lintuzumab, blinatumomab (Blincyto®), and one or more of Mab 364, Mab 366, Mab 367, Mab 368, Mab 369, Mab 370 or Mab 22, as presented in Table 2.

[0192] In certain embodiments, either alone or in combination with other embodiments of the invention, the CD3 binding domains of the invention and antibodies comprising same exhibit a reduced CRS risk profile compared to trastuzumab (Herceptin®), lintuzumab, blinatumomab (Blincyto®), and one or more of Mab 364, Mab 366, Mab 367, Mab 368, Mab 369, Mab 370 or Mab 22, as presented in Table 2.

[0193] In certain embodiments, and / or in combination with any of the embodiments disclosed throughout this specification, CD3 binding domains and humanized antibodies comprising the same are provided. In certain embodiments, such CD3 binding domains comprise the CDRs of such other embodiments and further comprise an acceptor human framework, such as a human immunoglobulin framework or a human consensus framework.

[0194] In particular embodiments, and / or in combination with any of the embodiments disclosed throughout the specification, there is provided an antibody comprising a CD3 binding domain, and comprising the same, a VH as in any of the embodiments presented throughout the specification, and a VL as in any of the embodiments presented throughout the specification, wherein one or both of the variable domain sequences comprises a post-translational modification.

[0195] In a further aspect of the present invention, there are provided CD3 binding domains and antibodies comprising same that bind to the same epitope as the CD3 binding domains provided in other embodiments disclosed throughout this specification.

[0196] In certain embodiments, the CD3 binding domain and / or antibodies comprising same have a cytotoxicity of ≦1 μM, ≦100 nM, ≦10 nM, ≦1 nM, ≦0.1 nM, ≦0.01 nM, or ≦0.001 nM (e.g., 10 -8 M or less, e.g. 10 -8 M~10 -13 M, e.g. 10 -9 M~10 -13 It has a CD3 dissociation constant (Kd) of 1 M.

[0197] In certain embodiments, Kd is measured by biolayer interferometry (BLI), surface plasmon resonance (SPR), solution equilibrium-based kinetic exclusion assay, KinExA direct association assay using ForteBio instruments and reagents, such as Octet RED 384 and HTX BLI-based instruments, enzyme-linked immunosorbent assay (ELISA), and radioimmunoassay (RIA). In certain embodiments, Kd measurements are performed using a Fab form of the CD3-binding domain of the present invention or an antibody comprising the same. For example, the solution binding affinity of a Fab for an antigen is measured by equilibrating the Fab with a minimal concentration of (I)-labeled antigen in the presence of a titration series of unlabeled antigen, followed by capturing the bound antigen with a plate coated with an anti-Fab antibody (see, e.g., Chen et al., J. Mol. Biol. 293:865-881 (1999)). To establish assay conditions, MICROTITER® multiwell plates (Thermo Scientific) were coated overnight with 5 μg / ml of capture anti-Fab antibody (Cappel Labs) in 50 mM sodium carbonate (pH 9.6). They were then blocked with 2% (w / v) bovine serum albumin in PBS for 2-5 hours at room temperature (approximately 23°C). Serial dilutions of the Fab of interest were mixed with 100 pM or 26 pM [I]-antigen in a non-adsorbent plate (Nunc #269620) (e.g., consistent with the evaluation of the anti-VEGF antibody Fab-12 in Presta et al., Cancer Res. 57:4593-4599 (1997)). The Fab of interest was then incubated overnight. However, incubation could be continued for a longer period (e.g., approximately 65 hours) to ensure equilibrium was reached. The mixture was then transferred to the capture plate for incubation at room temperature (e.g., 1 hour). The solution was then removed and the plates were washed eight times with 0.1% polysorbate 20 (TWEEN-20®) in PBS.When the plates are dry, 150 μl / well of scintillant (MICROSCINT-20™; Packard) is added and the plates are counted for 10 minutes on a TOPCOUNT™ gamma counter (Packard). Concentrations of each Fab that result in 20% or less of maximal binding are selected for use in competitive binding assays.

[0198] In another embodiment, Kd is measured using a BIACORE® surface plasmon resonance assay. For example, assays using a BIACORE®-2000 or BIACORE®-3000 (BIACORE, Inc., Piscataway, NJ) are performed at 25°C using approximately 10 response units (RU) of immobilized antigen on a CM5 chip. In one embodiment, a carboxymethylated dextran biosensor chip (CM5, BIACORE, Inc.) is activated with N-ethyl-N'-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) according to the manufacturer's instructions. The antigen is diluted to 5 μg / ml (approximately 0.2 μM) with 10 mM sodium acetate, pH 4.8, and then injected at a flow rate of 5 μl / min, resulting in approximately 10 response units (RU) of bound protein. Following injection of antigen, 1 M ethanolamine was injected to block unreacted groups. For kinetic measurements, two-fold serial dilutions of Fab (0.78 nM to 500 nM) were injected in PBS containing 0.05% polysorbate 20 (TWEEN-20™) surfactant (PBST) at a flow rate of approximately 25 μl / min at 25°C. The association rate (K onThe equilibrium dissociation constant (Kd) and dissociation rate (K) are calculated by simultaneously fitting the association and dissociation sensorgrams using a one-to-one Langmuir binding model (BIACORE® Evaluation Software version 3.2). The equilibrium dissociation constant (Kd) is calculated as the ratio of k / k. See, e.g., Chen et al., J. Mol. Biol. 293:865-881 (1999). If the association rate by the surface plasmon resonance assay described above exceeds 106 M-1s-1, the association rate can be determined using fluorescence quenching techniques. The technique involves measuring the increase or decrease in fluorescence emission intensity of a 20 nM anti-antigen antibody (Fab type) in PBS, pH 7.2, at 25°C with increasing antigen concentrations using a stirred cuvette in a spectrometer, e.g., a stop-flow equipped spectrophotometer (Aviv Instrument) or an 8000-series SLM-AMINC spectrophotometer (ThermoSpectronic) (excitation = 295 nm, emission = 340 nm, 16 nm bandpass).

[0199] In certain embodiments, the CD3-binding domains of the present invention and antibodies comprising the same comprise antibody fragments. Antibody fragments include, but are not limited to, Fab, Fab', Fab'-SH, F(ab')2, Fv, and scFv fragments, as well as other fragments described throughout the specification. For a review of specific antibody fragments, see, e.g., Hudson et al. Nat. Med. 9:129-134 (2003). For a review of scFv fragments, see, e.g., Pluckthun, in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., (Springer-Verlag, New York), pp. 269-315 (1994). See also WO 93 / 16185 and U.S. Patent Nos. 5,571,894 and 5,587,458. See US Pat. No. 5,869,046 for Fab and F(ab')2 fragments containing salvage receptor binding epitope residues and having extended in vivo half-lives.

[0200] Diabodies are antibody fragments with two antigen-binding sites that may be bivalent or bispecific. See, e.g., EP 404,097, WO 1993 / 01161, Hudson et al. Nat. Med. 9:129-134 (2003), and Hollinger et al. Proc. Natl. Acad. Sci. USA 90: 6444-6448 (1993). Triabodies and tetrabodies are also described in Hudson et al. Nat. Med. 9:129-134 (2003).

[0201] Single-domain antibodies are antibody fragments that contain all or part of the heavy chain variable domain or all or part of the light chain variable domain of an antibody. In certain embodiments, single-domain antibodies are human single-domain antibodies (Domantis, Inc., Waltham, Mass.; see, e.g., U.S. Patent No. 6,248,516 B1).

[0202] Antibody fragments can be produced by various techniques, including, but not limited to, proteolytic digestion of whole antibodies and production by recombinant host cells (e.g., E. coli or phage), as described herein.

[0203] In certain embodiments, the CD3-binding domains of the present invention and antibodies comprising the same comprise chimeric antibodies. Specific chimeric antibodies are described, for example, in U.S. Pat. No. 4,816,567; and Morrison et al. Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984). In one example, a chimeric antibody comprises a non-human variable region (e.g., a variable region derived from a mouse, rat, hamster, rabbit, or a non-human primate, such as a monkey) and a human constant region. In a further example, a chimeric antibody is a "class-switched" antibody whose class or subclass has been changed from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0204] In certain embodiments, the CD3-binding domains of the present invention and antibodies comprising the same include chimeric antibodies, which are humanized antibodies. Typically, non-human antibodies are humanized to reduce immunogenicity in humans while retaining the specificity and affinity of the parent non-human antibody. Generally, a humanized antibody comprises one or more variable domains, in which the CDRs (or portions thereof) are derived from a non-human antibody and the FRs (or portions thereof) are derived from a human antibody sequence. Optionally, a humanized antibody also contains at least a portion of a human constant region. In some embodiments, some FR residues in a humanized antibody are substituted with corresponding residues from the non-human antibody (e.g., the antibody from which the CDR residues are derived) to restore or improve, for example, the antibody's specificity, affinity, lack of immunogenicity, stability, developability profile, CRS risk, etc.

[0205] Humanized antibodies and methods for making humanized antibodies are generally described, e.g., in Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008), and in, e.g., Riechmann et al., Nature 332:323-329 (1988); Queen et al., Proc. Nat'l Acad. Sci. USA 86:10029-10033 (1989); U.S. Pat. Nos. 5,821,337, 7,527,791, 6,982,321, and 7,087,409; Kashmiri et al., Methods 36:25-34 (2005) (describing specificity-determining region (SDR) grafting); Padlan, Mol. Immunol. 28:489-498 (1991). (describing "resurfacing"); Dall'Acqua et al., Methods 36:43-60 (2005) (describing "FR shuffling"); and Osbourn et al., Methods 36:61-68 (2005) and Klimka et al., Br. J. Cancer, 83:252-260 (2000) (describing a "guided selection" method for FR shuffling).

[0206] Human framework regions that can be used for humanization include, but are not limited to, framework regions selected using the "best-fit" method (see, e.g., Sims et al. J. Immunol. 151:2296 (1993)); framework regions derived from human antibody consensus sequences of light or heavy chain variable regions of specific subgroups (see, e.g., Carter et al. Proc. Natl. Acad. Sci. USA, 89:4285 (1992); and Presta et al. J. Immunol., 151:2623 (1993)); human mature (somatically matured) framework regions or human germline framework regions (see, e.g., Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)); and framework regions derived by screening FR libraries (see, e.g., Baca et al., J. Biol. Chem. 272:10678-10684 (1997) and Rosok et al., J. Biol. Chem. 271:22611-22618 (1996).

[0207] In certain embodiments, the CD3-binding domains of the present invention and antibodies comprising the same comprise human antibodies. Human antibodies can be produced using various antibody techniques known in the art. Human antibodies are reviewed in van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5: 368-74 (2001) and Lonberg, Curr. Opin. Immunol. 20: 450-459 (2008).

[0208] Human antibodies may be produced by administering immunogens to transgenic animals that have been engineered to produce fully human antibodies or to produce fully human antibodies with human variable regions in response to antigen challenge. Such animals often contain all or part of a human immunoglobulin locus that replaces an endogenous immunoglobulin locus, or that is present extrachromosomally or randomly integrated into the animal's chromosomes. In such transgenic mice, the endogenous immunoglobulin locus is generally inactivated. For a review of methods for obtaining human antibodies from transgenic animals, see Lonberg, Nat. Biotech. 23:1117-1125 (2005). See, e.g., U.S. Patent Nos. 6,075,181 and 6,150,584, which describe Xenomouse, U.S. Patent No. 5,770,429, which describes HUMAB®, U.S. Patent No. 7,041,870, which describes KM MOUSE® technology, and U.S. Patent Application Publication No. US 2007 / 0061900, which describes VELOCIMOUSE® technology. The human variable regions derived from intact antibodies made by such animals may be further modified, for example, by combining with different human constant regions.

[0209] Human antibodies can also be produced using hybridoma-based methods. Human myeloma and mouse-human heteromyeloma cell lines for producing human monoclonal antibodies have been described. (See, for example, Kozbor J. Immunol., 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al., J. Immunol., 147: 86 (1991)). Human antibodies produced via human B cell hybridoma technology are described in Li et al., Proc. Natl. Acad. Sci. LISA. 103; 3557-3562 (2006). Additional methods include those described in, for example, U.S. Patent No. 7,189,826 (describing the production of monoclonal human IgM antibodies from hybridoma cell lines) and Ni, Xiandai Mianyixue, 26(4):265-268 (2006) (describing human-human hybridomas). Human hybridoma technology (trioma technology) is also described in Vollmers and Brandlein, Histology and Histopathology, 20(3):927-937 (2005) and Vollmers and Brandlein, Methods and Findings in Experimental and Clinical Pharmacology, 27(3):185-91 (2005).

[0210] Human antibodies can also be generated by isolating Fv clone variable domain sequences selected from human-derived phage display libraries. These variable domain sequences can then be combined with desired human constant domains. Techniques for selecting human antibodies from antibody libraries are described throughout this specification.

[0211] Antibodies of the invention can be isolated by screening combinatorial libraries for antibodies with the desired activity. For example, various methods are known in the art for generating phage display libraries and screening such libraries for antibodies possessing the desired binding characteristics. Such methods are reviewed, for example, by Hoogenboom et al. in Methods in Molecular Biology 178:1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, 2001), and also by, for example, McCafferty et al., Nature 348:552-554; Clackson et al., Nature 352: 624-628 (1991); Marks et al., J. Mol. Biol. 222: 581-597 (1992); Marks and Bradbury, in Methods in Molecular Biology 248:161-175 (Lo, ed., Human Press, Totowa, NJ, 2003); Sidhu et al., J. Mol. Biol. 338(2): 299-310 (2004); Lee et al., J. Mol. Biol. 340(5): 1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34): 12467-12472 (2004); and Lee et al., J. Immunol. Methods 284(1-2): 119-132(2004).

[0212] In certain embodiments, the CD3-binding domains of the invention and antibodies comprising the same may be selected from yeast-based antibody libraries, such as the libraries disclosed in WO 2009 / 0363379, WO 2010 / 105256, and WO 2012 / 009568. In certain embodiments, the CD3-binding domains of the invention and antibodies comprising the same may be optimized (also known as "affinity matured") as disclosed throughout this specification and in WO 2009 / 0363379, WO 2010 / 105256, and WO 2012 / 009568, and through other techniques available in the art.

[0213] In a specific phage display method, repertoires of VH and VL genes are cloned separately by polymerase chain reaction (PCR) and randomly recombined into a phage library. This library can then be screened for antigen-binding phage as described in Winter et al., Ann. Rev. Immunol., 12: 433-455 (1994). Typically, the phage display antibody fragments as single-chain Fv (scFv) fragments or Fab fragments. Libraries from immunized sources provide high-affinity antibodies to the immunogen of interest without the need for hybridoma construction. Alternatively, naive repertoires can be cloned (e.g., from humans) to provide a single source of antibodies against a wide range of non-self and also self antigens without any immunization, as described in Griffiths et al., EMBO J, 12: 725-734 (1993). Finally, naive libraries can be synthetically generated by cloning unrearranged V gene segments from stem cells and using PCR primers containing random sequences to encode the hypervariable CDR3 regions, allowing for in vitro rearrangement, as described in Winter, J. Mol. Biol., 227:381-388 (1992). Patent publications describing human antibody phage libraries include, for example, U.S. Patent No. 5,750,373, and U.S. Patent Application Publications 2005 / 0079574, 2005 / 0119455, 2005 / 0266000, 2007 / 0117126, 2007 / 0160598, 2007 / 0237764, 2007 / 0292936, and 2009 / 0002360.

[0214] Antibodies or antibody fragments isolated from a human antibody library are considered human antibodies or human antibody fragments herein.

[0215] In certain embodiments, the CD3-binding domains of the present invention and antibodies comprising the same comprise multispecific antibodies, e.g., bispecific antibodies. Multispecific antibodies are monoclonal antibodies that have binding specificities for at least two different antigens or at least two different epitopes on the same antigen. In certain embodiments, bispecific antibodies may bind to two different epitopes of CD3 (e.g., CD3ε or CD3γ). In certain embodiments, one binding specificity is specific for CD3 (e.g., CD3ε or CD3γ), and the other is specific for any other antigen (e.g., a second biological molecule, e.g., a cell surface antigen, e.g., a tumor antigen). Thus, a bispecific anti-CD3 antibody may have binding specificities for CD3 and a second biological molecule, such as a second biological molecule (e.g., a tumor antigen) listed in Table 1 and U.S. Patent Application Publication No. 2010 / 0111856. In certain embodiments, the CD3 binding domains of the invention and antibodies comprising same comprise multispecific antibodies, e.g., bispecific antibodies, in which the multispecific antibody comprises a second binding domain with specificity for a second antigen, e.g., a human leukocyte antigen (HLA)-peptide complex presented on the cell surface by MHC.

[0216] In certain embodiments, the CD3 binding domains of the invention and antibodies comprising same comprise multispecific antibodies, e.g., bispecific antibodies, wherein the multispecific antibodies comprise a second binding domain having specificity for a second antigen selected from the group consisting of: 0772P (CA125, MUC16; Genbank Accession No. AF36148); adipophilin (perilipin-2, Adipose differentiation-related protein, ADRP, ADFP, MGC10598; NCBI Reference Sequence: NP-001113.2); AIM-2 (Absent In Melanoma 2, PYHIN4, Interferon-Inducible Protein AIM2; NCBI Reference Sequence: NP-004824.1); ALDH1 A1 (Aldehyde Dehydrogenase 1 Family, Member A1, ALDH1, PUMB1, Retinaldehyde Dehydrogenase 1, ALDC, ALDH-E1, ALHDII, RALDH 1, EC 1.2.1.36, ALDH11, HEL-9, HEL-S-53e, HEL12, RALDH1, Acetaldehyde Dehydrogenase 1, Aldehyde Dehydrogenase 1, Soluble, Aldehyde Dehydrogenase, liver cytosolic, ALDH Class 1, Epididymis Luminal Protein 12, Epididymis Luminal Protein 9, Epididymis Secretory Sperm Binding Protein Li 53e, Retinal Dehydrogenase 1, RaIDH1, Aldehyde Dehydrogenase Family 1 Member A1, Aldehyde Dehydrogenase, Cytosolic, EC 1.2.1; NCBI Reference Sequence: NP-000680.2); alpha-actinin-4 (ACTN4, Actinin, Alpha 4, FSGS1, Focal Segmental Glomerulosclerosis 1, Non-Muscle Alpha-Actinin 4, F-Actin Cross-Linking Protein, FSGS, ACTININ-4, Actinin Alpha4 Isoform, alpha-actinin-4; NCBI reference sequence: NP-004915.2); alpha-fetoprotein (AFP, HPAFP, FETA, alpha-1-fetoprotein, alpha-fetoglobulin, Alpha-1-fetoprotein, Alpha-fetoglobulin, HP; GenBank: AAB58754.1); Amphiregulin (AREG, SDGF, Schwannoma-Derived Growth Factor, Colorectum Cell-Derived Growth Factor, AR, CRDGF; GenBank: AAA51781.1); ARTC1 (ART1, ADP-Ribosyltransferase 1, Mono(ADP-Ribosyl)Transferase 1, ADP-Ribosyltransferase C2 And C3 Toxin-Like 1, ART2, CD296, RT6, ADP-Ribosyltransferase 2, GPI-Linked NAD(P)(+)-Arginine ADP-Ribosyltransferase 1, EC 2.4.2.31, CD296 Antigen; NP); ASLG659; ASPHD1 (Aspartate Beta-Hydroxylase Domain Containing 1, Aspartate Beta-Hydroxylase Domain-Containing Protein 1, EC 1.14.11.-, EC 1.14.11., GenBank: AAI44153.1); B7-H4 (VTCN1, V-Set Domain Containing T Cell Activation Inhibitor 1, B7H4, B7 Superfamily Member 1, Immune Costimulatory Protein B7-H4, B7h.5, T-Cell Costimulatory Molecule B7x, B7S1, B7X, VCTN1, H4, B7 Family Member, PRO1291, B7 Family Member, H4, T Cell Costimulatory Molecule B7x, V-Set Domain-Containing T-Cell Activation Inhibitor 1, Protein B7S1; GenBank: AAZ17406.1); BAFF-R (TNFRSF13C, Tumor Necrosis Factor Receptor Superfamily, Member 13C, BAFFR, B-Cell-Activating Factor Receptor, BAFF Receptor, BLyS Receptor 3, CVID4, BROMIX, CD268, B Cell-Activating Factor Receptor, prolixin, Tumor Necrosis Factor Receptor Superfamily Member 13C, BR3, CD268 Antigen; NCBI reference sequence: NP-443177.1); BAGE-1; BCLX (L); BCR-ABL fusion protein (b3a2); beta-catenin (CTNNB1, Catenin (Cadherin-Associated Protein), Beta 1, 88 kDa, CTNNB, MRD19, Catenin (Cadherin-Associated Protein), Beta 1 (88kD), armadillo, Catenin Beta-1; GenBank: CAA61107.1); BING-4 (WDR46, WD Repeat Domain 46, C6orf11, BING4, WD Repeat-Containing Protein BING4, Chromosome 6 Open Reading Frame 11, FP221, UTP7, WD Repeat-Containing Protein 46; NP); BMPR1 B (bone morphogenetic protein receptor-type IB, Genbank accession number NM-00120; NP); B-RAF (Brevican (BCAN, BEHAB, Genbank accession number AF22905); Brevican (BCAN, Chondroitin Sulfate Proteoglycan 7, Brain-Enriched Hyaluronan-Binding Protein, BEHAB, CSPG7, Brevican Proteoglycan, Brevican Core Protein, Chondroitin Sulfate Proteoglycan BEHAB; GenBank: AAH27971.1); CALCA (Calcitonin-Related Polypeptide Alpha, CALC1, Calcitonin 1, calcitonin, Alpha-Type CGRP, Calcitonin Gene-Related Peptide I, CGRP-I, CGRP, CGRP1, CT, KC, Calcitonin / Calcitonin-Related Polypeptide, Alpha, katacalcin; NP); CASP-5 (CASP5, Caspase 5, Apoptosis-Related Cysteine Peptidase, Caspase 5, Apoptosis-Related Cysteine Protease, Protease ICH-3, Protease TY, ICE(rel)-111, ICE(rel)III, ICEREL-III, ICH-3, caspase-5, TY Protease, EC 3.4.22.58, ICH3, EC 3.4.22; NP); CASP-8; CD19 (CD19 - B-lymphocyte antigen CD19 isoform 2 precursor, B4, CVID3 [Homo sapiens], NCBI Reference Sequence: NP-001761.3); CD20 (CD20 - B-lymphocyte antigen CD20, membrane-spanning 4-domains, subfamily A, member 1, B1, Bp35, CD20, CVID5, LEU-16, MS4A2, S7; NCBI Reference Sequence: NP-690605.1); CD21 (CD21 (CR2 (Complement receptor) or C3DR (C3d / Epstein-Barr virus receptor) or Hs.73792 Genbank Accession Number M2600); (CD22 (B-cell receptor CD22-B isoform, BL-CAM, Lyb-8, LybB, SIGLEC-2, FLJ22814, GenBank accession number AK02646); CD22; CD33 (CD33 molecule, CD33 antigen (Gp67), Sialic Acid Binding Ig-Like Lectin 3, Sialic Acid-Binding Ig-Like Lectin 3, SIGLEC3, gp67, SIGLEC-3, Myeloid Cell Surface Antigen CD33, p67, Siglec-3, CD33 antigen; GenBank: AAH28152.1); CD45; CD70 (CD70-tumor necrosis factor (ligand) superfamily, member 7; surface antigen CD70; Ki-24 antigen; CD27 ligand; CD27-L; tumor necrosis factor ligand superfamily member 7; NCBI reference sequence for Homo sapiens species: NP-001243.1); CD72 (CD72 (B-cell differentiation antigen CD72, Lyb-; 359 aa, μl: 8.66, MW: 40225, TM: 1 [P] gene chromosome: 9p13.3, Genbank accession number NP-001773.); CD79a (CD79a (CD79A, CD79a, immunoglobulin-associated alpha, a B cell-specific protein that covalently interacts with Ig beta (CD79B) to form a complex on the surface with IgM molecules and transmits signals involved in B cell differentiation), μl: 4.84, MW: 25028 TM: 2 [P] gene chromosome: 19q13.2, Genbank accession number NP-001774.1); CD79b (CD79b (CD79B, CD79b, IGb (immunoglobulin-associated beta), B29, Genbank accession number NM-000626 or 1103867); Cdc27 (Cell Division Cycle 27, D0S1430E, D17S978E, Anaphase Promoting Complex Subunit 3, Anaphase-Promoting Complex Subunit 3, ANAPC3, APC3, CDC27Hs, H-NUC, CDC27 Homolog, Cell Division Cycle 27 Homolog (S. Cerevisiae), HNUC, NUC2, Anaphase-Promoting Complex, Protein 3, Cell Division Cycle 27 Homolog, Cell Division Cycle Protein 27 Homolog, Nuc2 Homolog; GenBank: AAH11656.1); CDK4 (Cyclin-Dependent Kinase 4, Cell Division Protein Kinase 4, PSK-J3, EC 2.7.11.22, CMM3, EC 2.7.11); NCBI reference sequence: NP-000066.1); CDKN2A (Cyclin-Dependent Kinase Inhibitor 2A, MLM, CDKN2, MTS1, Cyclin-Dependent Kinase Inhibitor 2A (Melanoma, P16, Inhibits CDK4), Cyclin-Dependent Kinase 4 Inhibitor A, Multiple Tumor Suppressor 1, CDK4I, MTS-1, CMM2, P16, ARF, INK4, INK4A, P14, P14ARF, P16-INK4A, P16INK4, P16INK4A, P19, P19ARF, TP16, CDK4 inhibitor P16-INK4, Cell Cycle Negative Regulator Beta, p14ARF, p16-INK4, p16-INK4a, p16INK4A, p19ARF; NP); CEA; CLL1 (CLL-1 (CLEC12A, MICL, and DCAL, many members of the C-type lectin / C-type lectin-like domain (CTL / CTLD) superfamily. Members of this family share a common protein fold and have diverse functions, including roles in adhesion, cell-cell signaling, glycoprotein turnover, and inflammation and immune responses. The protein encoded by this gene is a negative regulator of granulocyte and monocyte function. Several alternatively spliced ​​transcript variants of this gene have been reported, but the full-length nature of some of these variants is unknown. This gene is closely related to other CTL / CTLD superfamily members in the natural killer gene complex region on chromosome 12p13 (Drickamer, K Curr. Opin. Struct. Biol. 9):585-90; van Rhenen, A, et al., Blood 110):2659-66; Chen CH, et al. Blood 107):1459-67; Marshall AS, et al. Eur. J. Immunol. 36):2159-69; Bakker AB, et al Cancer Res. 64:8443-50; Marshall AS, et al J. Biol. Chem.279:14792-80. CLL-1 has been shown to be a type II transmembrane receptor containing a single C-type lectin-like domain (not predicted to bind either calcium or sugar), a stalk region, a transmembrane domain, and a short cytoplasmic tail containing ITIM motifs); CLPP (Caseinolytic Mitochondrial Matrix Peptidase Proteolytic Subunit, Endopeptidase Clp, EC 3.4.21.92, PRLTS3, ATP-Dependent Protease ClpAP (E. coli), ClpP (Caseinolytic Protease, ATP-Dependent, Proteolytic Subunit, E. coli) Homolog, ClpP Caseinolytic Peptidase, ATP-Dependent, Proteolytic Subunit Homolog (E. coli), ClpP Caseinolytic Protease, ATP-Dependent, Proteolytic Subunit Homolog (E. coli).coli), Human, Proteolytic Subunit, ATP-Dependent Protease ClpAP, Proteolytic Subunit, Human, ClpP Caseinolytic Peptidase ATP-Dependent, Proteolytic Subunit, ClpP Caseinolytic Peptidase, ATP-Dependent, Proteolytic Subunit Homolog, ClpP Caseinolytic Protease, ATP-Dependent, Proteolytic Subunit Homolog, Putative ATP-Dependent Clp Protease Proteolytic Subunit, Mitochondrial; NP); COA-1; CPSF; CRIPTO (CR, CR1, CRGF, CRIPTO, TDGF1, teratocarcinoma-derived growth factor, Genbank accession number NP-003203 or NM-0321); Cw6; (Burkitt's lymphoma receptor 1, a G protein-coupled receptor activated by the CXCL13 chemokine, functions in lymphocyte migration and humoral defense, and may contribute to HIV-2 infection and possibly other pathological conditions. involved in the development of AIDS, lymphoma, myeloma, and leukemia); 372 aa, μl: 8.54 MW: 41959 TM: 7 [P] gene chromosome: 11q23.3, GenBank accession number NP-1707.); CXORF61 CXORF61-chromosome X open reading frame 61 [Homo sapiens], NCBI reference sequence: NP-01017978.1); cyclin D1 (CCND1, BCL1, PRAD1, D11S287E, B-Cell CLL / Lymphoma 1, B-Cell Lymphoma 1 Protein, BCL-1 Oncogene, PRAD1 Oncogene, Cyclin D1 (PRAD1: Parathyroid Adenomatosis 1), G1 / S-Specific Cyclin D1, Parathyroid Adenomatosis 1, U21B31, G1 / S-Specific Cyclin-D1, BCL-1; NCBI reference sequence: NP-44284.1); Cyclin-A1 (CCNA1, CT146, Cyclin A1; GenBank: AAH36346.1); dek-can fusion protein; DKK1 (Dickkopf WNT Signaling Pathway Inhibitor 1, SK, hDkk-1, Dickkopf (Xenopus Laevis) Homolog 1, Dickkopf 1 Homolog (Xenopus Laevis), DKK-1, Dickkopf 1 Homolog, Dickkopf Related Protein-1, Dickkopf-1 Like, Dickkopf-Like Protein 1, Dickkopf-Related Protein 1, Dickkopf-1, Dkk-1; GenBank: AAQ89364.1);DR1 (Down-Regulator Of Transcription 1, TBP-Binding (Negative Cofactor 2), Negative Cofactor 2-Beta, TATA-Binding Protein-Associated Phosphoprotein, NC2, NC2-BETA, Protein Dr1, NC2-beta, Down-Regulator Of Transcription 1; NCBI Reference Sequence: NP-01929.1);DR13 (Major Histocompatibility Complex, Class II, DR Beta 1, HLA-DR1B, DRw10, DW2.2 / DR2.2, SS1, DRB1, HLA-DRB, HLA Class II Histocompatibility Antigen, DR-1 Beta Chain, Human Leucocyte Antigen DRB1, Lymphocyte Antigen DRB1, MHC Class II Antigen, MHC Class II HLA-DR Beta 1 Chain, MHC Class II HLA-DR-Beta Cell Surface Glycoprotein, MHC Class II HLA-DRw10-Beta, DR-1, DR-12, DR-13, DR-14, DR-16, DR-4, DR-5, DR-7, DR-8, DR-9, DR1, DR12, DR13, DR14, DR16, DR4, DR5, DR7, DRB, DR9, DRw11, DRw8, HLA-DRB2, Clone P2-Beta-3, MHC Class II Antigen DRB1*1, MHC Class II Antigen DRB1*10, MHC Class II Antigen DRB1*11, MHC Class II Antigen DRB1*12, MHC Class II Antigen DRB1*13, MHC Class II Antigen DRB1*14, MHC Class II Antigen DRB1*15, MHC Class II Antigen DRB1*16, MHC Class II Antigen DRB1*3, MHC Class II Antigen DRB1*4, MHC Class II Antigen DRB1*7, MHC Class II Antigen DRB1*8, MHC Class II Antigen DRB1*9; NP); E16 (E16 (LAT1, SLC7A5, Genbank accession number NM-00348); EDAR (EDAR - tumor necrosis factor receptor superfamily member EDAR precursor, EDA-A1 receptor;downless homolog; ectodysplasin-A receptor; ectodermal dysplasia receptor; anhydrotic ectodysplasin receptor 1、DL; ECTD10A; ECTD10B; ED1R; ED3; ED5; EDA-A1R; EDA1R; EDA3; HRM1 [Homo sapiens]; NCBI Publication Number: NP-071731.1); EFTUD2 (Elongation Factor Tu GTP Binding Domain Containing 2、Elongation Factor Tu GTP-Binding Domain-Containing Protein 2、hSNU114、SNU114 Homolog、U5 SnRNP-Specific Protein、116 KDa、MFDGA、KIAA0031、116 KD、U5 SnRNP Specific Protein、116 KDa U5 Small Nuclear Ribonucleoprotein Component, MFDM, SNRNP116, Snrp116, Snu114, U5-116KD, SNRP116, U5-116 KDa;EGFR (Epidermal Growth Factor Receptor、ERBB、Proto-Oncogene C-ErbB-1、Tyrosine-Protein Kinase Receptor ErbB-1、ERBB1、HER1、EC 2.7.10.1、Epidermal Growth Factor Receptor (Avian Erythroblastic Leukemia Viral (V-Erb-B) Oncogene Homolog)、Viral Erythroblastic Leukemia (V-Erb-B) Oncogene Homolog (Avian)、PlG61、Avian Erythroblastic Leukemia Viral (V-Erb-B) Oncogene Homolog、Cell Growth Inhibiting Protein 40、Cell Proliferation-Inducing Protein 61、mENA、EC 2.7.10; EGFR-G719A; EGFR-G719C; EGFR-G719S; EGFR-L858R; EGFR-L861Q; EGFR-57681; EGFR-T790M; Elongation factor 2 (EEF2、Eukaryotic Translation Elongation Factor 2、EF2、Polypeptidyl-TRNA Translocase、EF-2、SCA26、EEF-2; NCBI National Park Service: NP-001952.1); ENAH (hMena) (Enabled Homolog (Drosophila), MENA, Mammalian Enabled, ENA, NDPP1, Enabled Protein Homolog; GenBank: AAH95481.1)-“ENAH” means “ENAH (hMena)”;EpCAM (Epithelial Cell Adhesion Molecule、M4S1、MIC18、Tumor-Associated Calcium Signal Transducer 1、TACSTD1、TROP1、Adenocarcinoma-Associated Antigen、Cell Surface Glycoprotein Trop-1、Epithelial Glycoprotein 314、Major Gastrointestinal Tumor-Associated Protein GA733-2、EGP314、KSA、DIAR5、HNPCC8、Antigen Identified By Monoclonal Antibody AUA1、EGP-2、EGP40、ESA、KS1 / 4、MK-1、Human Epithelial Glycoprotein-2, Membrane Component, Chromosome 4, Surface Marker (35kD Glycoprotein)、EGP、Ep-CAM、GA733-2、M1S2、CD326 Antigen、Epithelial Cell Surface Antigen、hEGP314、KS 1 / 4 Antigen、ACSTD1; GenBank: AAH14785.1); EphA3 (EPH Receptor A3、ETK1、ETK、TYRO4、HEK、Eph-Like Tyrosine Kinase 1、Tyrosine-Protein Kinase Receptor ETK1、EK4、EPH-Like Kinase 4、EC 2.7.10.1、EPHA3、HEK4、Ephrin Type-A Receptor 3、Human Embryo Kinase 1、TYRO4 Protein Tyrosine Kinase、hEK4、Human Embryo Kinase、Tyrosine-Protein Kinase TYRO4、EC 2.7.10; GenBank: AAH63282.1); EphB2R; Epiregulin (EREG、ER、proepiregulin; GenBank: AAI36405.1);ETBR (EDNRB, Endothelin Receptor Type B, HSCR2, HSCR, Endothelin Receptor Non-Selective Type, ET-B, ET-BR, ETRB, ABCDS, WS4A, ETB, Endothelin B Receptor; NP); ETV6-AML1 fusion protein; EZH2 (Enhancer Of Zeste Homolog 2 (Drosophila), Lysine N-Methyltransferase 6, ENX-1, KMT6 EC 2.1.1.43, EZH1, WVS, Enhancer Of Zeste (Drosophila) Homolog 2, ENX1, EZH2b, KMT6A, WVS2, Histone-Lysine N-Methyltransferase EZH2, Enhancer Of Zeste Homolog 2, EC 2.1.1; GenBank: AAH10858.1); FcRH1 (FCRL1, Fc Receptor-Like 1, FCRH1, Fc Receptor Homolog 1, FcR-Like Protein 1, Immune Receptor Translocation-Associated Protein 5, IFGP, IRTA5, hIFGP1, IFGP Family Protein 1, CD307a, Fc Receptor-Like Protein 1, Immunoglobulin Superfamily Fc Receptor, Gp42, FcRL1, CD307a Antigen; GenBank: AAH33690.1);FcRH2 (FCRL2、Fc Receptor-Like 2、SPAP1、SH2 Domain-Containing Phosphatase Anchor Protein 1、Fc Receptor Homolog 2、FcR-Like Protein 2、Immunoglobulin Receptor Translocation-Associated Protein 4、FCRH2、IFGP4、IRTA4、IFGP Family Protein 4、SPAP1A、SPAP1 B、SPAP1C、CD307b、Fc Receptor-Like Protein 2、Immune Receptor Translocation-Associated Protein 4、Immunoglobulin Superfamily Fc Receptor、Gp42、SH2 Domain Containing Phosphatase Anchor Protein 1、FcRL2、CD307b Antigen; GenBank: AAQ88497.1); FcRH5 (FCRL5、Fc Receptor-Like 5、IRTA2、Fc Receptor Homolog 5、FcR-Like Protein 5、Immune Receptor Translocation-Associated Protein 2、BXMAS1、FCRH5、CD307、CD307e、PRO820、Fc Receptor-Like Protein 5、Immunoglobulin Superfamily Receptor Translocation Associated 2 (IRTA2)、FCRL5、CD307e Antigen; GenBank: AAI01070.1); FLT3-ITD; FN1(Fibronectin 1、Cold-Insoluble Globulin、FN、Migration-Stimulating Factor、CIG、FNZ、GFND2、LETS、ED-B、FINC、GFND、MSF、fibronectin; GenBank: AAI43764.1);G250 (MN、CAIX、Carbonic Anhydrase IX、Carbonic Dehydratase、RCC-Associated Protein G250、Carbonate Dehydratase IX、Membrane Antigen MN、Renal Cell Carcinoma-Associated Antigen G250, CA-IX, P54 / 58N, pMW1, RCC-Associated Antigen G250, Carbonic Anhydrase 9; NP); GAGE-1,2,8; GAGE-3,4,5,6,7; GDNF-Ra1 (GDNF family receptor alpha 1; GFRA1; GDNFR; GDNFRA; RETL1; TRNR1; RET1 L; GDNFR-alpha1; GFR-ALPHA-; U95847; BC014962; NM-145793 NM-005264); GEDA (Genbank database AY26076); GFRA1-GDNF family receptor alpha-1; GDNF receptor alpha-1; GDNFR-alpha-1; GFR-alpha-1; RET ligand 1; TGF-beta-related neurotrophic factor receptor 1 [Homo sapiens]; ProtKB / Swiss-Prot: P56159.2; glypican-3 (GPC3, Glypican 3, SDYS, Glypican Proteoglycan 3, Intestinal Protein OCI-5, GTR2-2, MXR7, SGBS1, DGSX, OCI-5. SGB, SGBS, Heparan Sulphate Proteoglycan, Secreted glypican-3、OCI5;GenBank:AAH35972.1); GnTVf;gp100 (PMEL、Premelanosome Protein、SILV、D12S53E、PMEL17、SIL、Melanocyte Protein Pmel 17、Melanocytes Lineage-Specific Antigen GP100、Melanoma-Associated ME20 Antigen、Silver Locus Protein Homolog、ME20-M、ME20M、P1、P100、Silver (Mouse Homolog) Like、Silver Homolog (マウス)、ME20、SI、Melanocyte Protein Mel 17、Melanocyte Protein PMEL、Melanosomal Matrix Protein17, Silver, Mouse, Homolog Of; GenBank: AAC60634.1); GPC; GPNMB (Glycoprotein (Transmembrane) Nmb、Glycoprotein NMB、Glycoprotein Nmb-Like Protein、osteoactivin、Transmembrane Glycoprotein HGFIN、HGFIN、NMB、Transmembrane Glycoprotein、Transmembrane Glycoprotein NMB; GenBank: AAH32783.1); GPR172A (G protein-coupled receptor 172A; GPCR41; FLJ11856; D15Ertd747e); NP-078807.1; NM-024531.3); GPR19 (G protein-coupled receptor 19; Mm.478; NP-006134.1; NM-006143.2); GPR54 (KISS1 receptor; KISS1R; GPR54; HOT7T175; AXOR1; NP-115940.2; NM-032551.4);HAVCR1 (Hepatitis A Virus Cellular Receptor 1、T-Cell Immunoglobulin Mucin Family Member 1、Kidney Injury Molecule 1、KIM-1、KIM1、TIM、TIM-1、TIM1、TIMD-1、TIMD1、T-Cell Immunoglobulin Mucin Receptor 1、T-Cell Membrane Protein 1、HAVCR、HAVCR-1、T Cell Immunoglobin Domain And Mucin Domain Protein 1、HAVcr-1、T-Cell Immunoglobulin And Mucin Domain-Containing Protein 1; GenBank: AAH13325.1);HER2 (ERBB2、V-Erb-B2 Avian Erythroblastic Leukemia Viral Oncogene Homolog 2、NGL、NEU、Neuro / Glioblastoma Derived Oncogene Homolog、Metastatic Lymph Node Gene 19 Protein、Proto-Oncogene C-ErbB-2、Proto-Oncogene Neu、Tyrosine Kinase-Type Cell Surface Receptor HER2、MLN 19、p185erbB2、EC 2.7.10.1、V-Erb-B2 Avian Erythroblastic Leukemia Viral Oncogene Homolog 2 (Neuro / Glioblastoma Derived Oncogene Homolog)、CD340、HER-2、HER-2 / neu、TKR1、C-Erb B2 / Neu Protein、herstatin、Neuroblastoma / Glioblastoma Derived Oncogene Homolog、Receptor Tyrosine-Protein Kinase ErbB-2、V-Erb-B2 Erythroblastic Leukemia Viral Oncogene Homolog 2、Neuro / Glioblastoma Derived Oncogene Homolog、MLN19、CD340 Antigen、EC; 2.7.10; NP); HER-2 / neu-also known as; HERV-K-MEL; HLA-DOB (beta subunit (la antigen) of MHC class II molecule that binds peptides and presents them to CD4+ T lymphocytes); 273 aa, μl: 6.56, MW: 30820.TM: 1 [P] gene chromosome: 6p21.3, GenBank accession number NP-002111); hsp70-2 (HSPA2, Heat Shock 70 kDa Protein 2, Heat Shock 70kD Protein 2, HSP70-3, Heat Shock-Related 70 KDa Protein 2, Heat Shock 70 KDa Protein 2; GenBank: AAD21815.1); IDO1 (Indoleamine 2,3-Dioxygenase 1, IDO, INDO, Indoleamine-Pyrrole 2,3-Dioxygenase, IDO-1, Indoleamine-Pyrrole 2,3 Dioxygenase, Indolamine 2,3 Dioxygenase, Indole 2,3 Dioxygenase, EC 1.13.11.52; NCBI reference sequence: NP-002155.1); IGF2B3; IL13Ralpha2 (IL13RA2、Interleukin 13 Receptor、Alpha 2、Cancer / Testis Antigen 19、Interleukin-13-Binding Protein、IL-13R-alpha-2、IL-13RA2、IL-13 Receptor Subunit Alpha-2、IL-13R Subunit Alpha-2、CD213A2、CT19、IL-13R、IL13BP、Interleukin 13 Binding Protein、Interleukin 13 Receptor Alpha 2 Chain、Interleukin-13 Receptor Subunit Alpha-2、IL13R、CD213a2 Antigen; NP); IL20Rα; Intestinal carboxyl esterase; IRTA2 (alias of FcRH5); Kallikrein 4 (KLK4、Kallikrein-Related Peptidase 4、PRSS17、EMSP1、Enamel Matrix Serine Proteinase 1、Kallikrein-Like Protein 1、Serine Protease 17、KLK-L1、PSTS、AI2A1、Kallikrein 4 (Prostase、Enamel Matrix、Prostate)、ARM1、EMSP、Androgen-Regulated Message 1、Enamel Matrix Serine Protease 1、kallikrein、kallikrein-4、prostase、EC 3.4.21.-、Prostase、EC 3.4.21; GenBank: AAX30051.1); KIF20A (Kinesin Family Member 20A, RAB6KIFL, RAB6 Interacting, Kinesin-Like (Rabkinesin6), Mitotic a; LAGE-1; LDLR-fucosyltransferase AS fusion protein; Lengsin (LGSN, Lengsin, Lens Protein With Glutamine Synthetase Domain, GLULD1, Glutamate-Ammonia Ligase Domain-Containing Protein 1, LGS, Glutamate-Ammonia Ligase (Glutamine Synthetase) Domain Containing 1, Lens Glutamine Synthase-Like; GenBank: AAF61255.1); LGR5 (leucine-rich repeat-containing G protein-coupled receptor 5; GPR49, GPR6; NP-003658.1; NM-003667.2; LY64 (Lymphocyte antigen 64 (RP10, a type I membrane protein of the leucine-rich repeat (LRR) family that regulates B cell activation and apoptosis. Loss of its function has been associated with increased disease activity in patients with systemic lupus erythematosus); 661 aa, μl: 6.20, MW: 74147 TM: 1 [P] Gene chromosome: 5q12, GenBank accession number NP-005573.; Ly6E (lymphocyte antigen 6 complex, locus E; Ly67, RIG-E, SCA-2, TSA-; NP-002337.1; NM-002346.2); Ly6G6D (lymphocyte antigen 6 complex, locus G6D; Ly6-D, MEGT; NP-067079.2; NM-021246.2); LY6K (lymphocyte antigen 6 complex, locus K; LY6K; HSJ001348; FLJ3522; NP-059997.3; NM-017527.3); LyPD1-LY6 / PLAUR domain containing 1, PHTS [Homo sapiens], GenBank: AAH17318.1); MAGE-A1 (Melanoma Antigen Family A, 1 (Directs Expression Of Antigen MZ2-E, MAGE1, Melanoma Antigen Family A 1, MAGEA1, Melanoma Antigen MAGE-1, Melanoma-Associated Antigen 1, Melanoma-Associated Antigen MZ2-E, Antigen MZ2-E, Cancer / Testis Antigen 1.1, CT1.1, MAGE-1 Antigen, Cancer / Testis Antigen Family 1, Member 1, Cancer / Testis Antigen Family 1, Member 1, MAGE1A; NCBI reference sequence: NP-004979.3); MAGE-A10 (MAGEA10, Melanoma Antigen Family A, 10, MAGE10, MAGE-10 Antigen, Melanoma-Associated Antigen 10, Cancer / Testis Antigen 1.10, CT1.10, Cancer / Testis Antigen Family 1, Member 10, Cancer / Testis Antigen Family 1, Member 10; NCBI reference sequence: NP-001238757.1); MAGE-A12 (MAGEA12, Melanoma Antigen Family A, 12, MAGE12, Cancer / Testis Antigen 1.12, CT1.12. MAGE12F Antigen, Cancer / Testis Antigen Family 1, Member 12, Cancer / Testis Antigen Family 1, Member 12, Melanoma-Associated Antigen 12, MAGE-12 Antigen; NCBI reference sequence: NP-001159859.1); MAGE-A2 (MAGEA2, Melanoma Antigen Family A, 2, MAGE2, Cancer / Testis Antigen 1.2, CT1.2, MAGEA2A, MAGE-2 Antigen, Cancer / Testis Antigen Family 1, Member 2, Cancer / Testis Antigen Family 1, Member 2, Melanoma Antigen 2, Melanoma-Associated Antigen 2; NCBI reference sequence: NP-001269434.1); MAGE-A3 (MAGEA3, Melanoma Antigen Family A, 3, MAGE3, MAGE-3 Antigen, Antigen MZ2-D, Melanoma-Associated Antigen 3, Cancer / Testis Antigen 1.3, CT1.3, Cancer / Testis Antigen Family 1, Member 3, HIPS, HYPD, MAGEA6, Cancer / Testis Antigen Family 1, Member 3; NCBI reference sequence: NP-005353.1); MAGE-A4 (MAGEA4, Melanoma Antigen Family A, 4, MAGE4, Melanoma-Associated Antigen 4, Cancer / Testis Antigen 1.4, CT1.4. MAGE-4 Antigen, MAGE-41 Antigen, MAGE-X2 Antigen, MAGE4A, MAGE4B, Cancer / Testis Antigen Family 1, Member 4, MAGE-41, MAGE-X2, Cancer / Testis Antigen Family 1, Member 4; NCBI Reference Sequence: NP-001011550.1); MAGE-A6 (MAGEA6, Melanoma Antigen Family A, 6, MAGE6, MAGE-6 Antigen, Melanoma-Associated Antigen 6, Cancer / Testis Antigen 1.6, CT1.6, MAGE3B Antigen, Cancer / Testis Antigen Family 1, Melanoma Antigen Family A 6, Member 6, MAGE-3b, MAGE3B, Cancer / Testis Antigen Family 1, Member 6; NCBI Reference Sequence: NP-787064.1); MAGE-A9 (MAGEA9, Melanoma Antigen Family A, 9, MAGE9, MAGE-9 Antigen, Melanoma-Associated Antigen 9, Cancer / Testis Antigen 1.9, CT1.9, Cancer / Testis Antigen Family 1, Member 9, Cancer / Testis Antigen Family 1, Member 9, MAGEA9A; NCBI Reference Sequence: NP-005356.1); MAGE-C1 (MAGEC1, Melanoma Antigen Family C, 1, Cancer / Testis Antigen 7.1, CT7.1, MAGE-C1 Antigen, Cancer / Testis Antigen Family 7, Member 1, CT7, Cancer / Testis Antigen Family 7, Member 1, Melanoma-Associated Antigen C1; NCBI Reference Sequence: NP-005453.2); MAGE-C2 (MAGEC2, Melanoma Antigen Family C, 2, MAGEE1, Cancer / Testis Antigen 10, CT10, HCA587, Melanoma Antigen, Family E, 1, Cancer / Testis Specific, Hepatocellular Carcinoma-Associated Antigen 587, MAGE-C2 Antigen, MAGE-E1 Antigen, Hepatocellular Cancer Antigen 587, Melanoma-Associated Antigen C2; NCBI Reference Sequence: NP-057333.1); mammaglobin-A (SCGB2A2, Secretoglobin, Family 2A, Member 2, MGB1, Mammaglobin 1, UGB2, Mammaglobin A, mammaglobin-A, Mammaglobin-1, Secretoglobin Family 2A Member 2; NP); MART2 (H HAT, Hedgehog Acyltransferase, SKI1, Melanoma Antigen Recognized By T-Cells 2, Skinny Hedgehog Protein 1, Skn, Melanoma Antigen Recognized By T Cells 2, Protein-Cysteine N-Palmitoyltransferase HHAT, EC 2.3.1.-; GenBank: AAH39071.1); M-CSF (CSF1, Colony Stimulating Factor 1 (Macrophage), MCSF, CSF-1, lanimostim, Macrophage Colony-Stimulating Factor 1, Lanimostim; GenBank: AAH21117.1); MCSP (SMCP, Sperm Mitochondria - Associated Cysteine - Rich Protein, MCS, Mitochondrial Capsule Selenoprotein, HSMCSGEN1, Sperm Mitochondrial - Associated Cysteine - Rich Protein; NCBI reference sequence: NP - 109588.2); XAGE - 1b / GAGED2a; WT1 (Wilms Tumor 1, WAGR, GUD, WIT - 2, WT33, Amino - Terminal Domain Of EWS, NPHS4, Last Three Zinc Fingers Of The DNA - Binding Domain Of WT1, AWT1, Wilms Tumor Protein, EWS - WT1; GenBank: AAB33443.1); VEGF; Tyrosinase (TYR; OCAIA; OCA1A; tyrosinase; SHEP; NP - 000363.1; NM - 000372.4; GenBank: AAB60319.1); TrpM4 (BR22450, FLJ20041, TRPM4, TRPM4B, transient receptor potential cation channel, subfamily M, member 4, Genbank accession number NM - 01763); TRP2 - INT2; TRP - 2; TRP - 1 / gp75 (Tyrosinase - Related Protein 1, 5,6 - Dihydroxyindole - 2 - Carboxylic Acid Oxidase, CAS2, CATB, TYRP, OCAS, Catalase B, b - PROTEIN, Glycoprotein 75, EC 1.14.18., Melanoma Antigen Gp75, TYRP1, TRP, TYRRP, TRP1, SHEP11, DHICA Oxidase, EC 1.14.18, GP75, EC 1.14.18.1; Triosephosphate isomerase (Triosephosphate isomerase 1、TPID、Triose-Phosphate Isomerase、HEL-S-49、TIM、Epididymis Secretory Protein Li 49、TPI、Triosephosphate Isomerase、EC 5.3.1.1; TRAG-3 (CSAG Family Member 2、Cancer / Testis Antigen Family 24、CSAG3B, Member 2、CSAG Family Member 3B、Cancer / Testis Antigen Family 24 Member 2、Cancer / Testis Antigen 24.2、Chondrosarcoma-Associated Gene 2 / 3 Protein、Taxol-Resistant-Associated Gene 3 Protein、Chondrosarcoma-Associated Gene 2 / 3 Protein-Like、CT24.2、Taxol Resistance Associated Gene 3、TRAG-3、CSAG3A、TRAG3;); TMEM46 (shisa homolog 2 (Xenopus Laevis); SHISA; NP-001007539.1; NM-001007538.1; TMEM118 (ring finger protein、transmembrane2; RNFT2; FLJ1462; NP-001103373.1; NM-001109903.1; TMEFF1 (transmembrane protein with EGF-like and two follistatin-like domains 1; Tomoregulin-; H7365; C9orf2; C9ORF2; U19878; X83961; NM-080655; NM-003692; TGF-betaRII (TGFBR2、Transforming Growth Factor、Beta Receptor II (70 / 80 kDa)、TGFbeta-RII、MFS2、tbetaR-II、TGFR-2、TGF-Beta Receptor Type IIB、TGF-Beta Type II Receptor、TGF-Beta Receptor Type-2、EC 2.7.11.30、Transforming Growth Factor Beta Receptor Type IIC、AAT3、TbetaR-II、Transforming Growth Factor、Beta Receptor II (70-80kD)、TGF-Beta Receptor Type II、FAA3、Transforming Growth Factor-Beta Receptor Type II、LDS1 B、HNPCC6、LDS2B、LDS2、RITC、EC 2.7.11, TAAD2; TENB2 (TMEFF2, tomoregulin, TPEF, HPP1, TR, putative transmembrane proteoglycan, related to the EGF / heregulin family of growth factors and follistatin); 374 aa, NCBI accession numbers: AAD55776, AAF91397, AAG49451, NCBI reference sequence: NP-057276; NCBI Gene: 23671; OMIM: 605734; SwissProt Q9UIK5; Genbank accession numbers AF179274; AY358907, CAF85723, CQ782436; TAG-2; TAG-1 (Contactin 2 (Axonal), TAG-1, AXT, Axonin-1 Cell Adhesion Molecule, TAX, Contactin 2 (transiently SYT-SSX1 or -SSX2 fusion protein; survivin; STEAP2 (HGNC 8639, IPCA-1, PCANAP1, STAMP1, STEAP2, STMP, prostate cancer associated gene 1, prostate cancer associated protein 1, six transmembrane epithelial antigen of prostate 2, six transmembrane prost. ate protein, Genbank accession number AF45513; STEAP1 (six transmembrane epithelial antigen of prostate, Genbank accession number NM-01244; SSX-4; SSX-2 (SSX2, Synovial Sarcoma, X Breakpoint2, X Breakpoint 2, SSX, X Breakpoint 2B, Cancer / Testis Antigen 5.2, X-Chromosome-Related 2, Tumor Antigen HOM-MEL-40, CT5.2, HD21, Cancer / Testis Antigen Family 5, HOM-MEL-40, Isoform B, Cancer / Testis Antigen Family 5 member 2a, member 2a, Protein SSX2, Sarcoma, Sarcoma, Synovial, X-Chromosome-Related 2, synovial, Synovial Sarcoma, X Breakpoint 2B, Synovial Sarcomam, SSX2A; Sp17; SOX10 (SRY (Sex Determining Region Y)-Box 10, mouse, PCWH, DOM, WS4, WS2E, WS4C, Dominant Megacolon, mouse, Human Homolog Of, Dominant Megacolon, SRY-Related HMG-Box Gene 10, Human Homolog Of, transcription Factor SOX-10; GenBank: CAG30470.1); SNRPD1 (Small Nuclear Ribonucleoprotein D1、Small Nuclear Ribonucleoprotein D1、Polypeptide 16 kDa、Polypeptide (16kD)、SNRPD、HsT2456、Sm-D1、SMD1、Sm-D Autoantigen、Small Nuclear Ribonucleoprotein D1 Polypeptide 16 kDa Pseudogene、SnRNP Core Protein D1、Small Nuclear Ribonucleoprotein Sm D1; SLC35D3 (Solute Carrier Family 35, Member D3、FRCL1、Fringe Connection-Like Protein 1、bA55K22.3、Frc、Fringe-Like 1、Solute Carrier Family 35 Member D3; NCBI GenBank: NC-000006.11 NC-018917.2 NT-025741.16); SIRT2 (Sirtuin 2、NAD-Dependent Deacetylase Sirtuin-2、SIRL2、Silent Information Regulator 2、Regulatory Protein SIR2 Homolog 2、Sir2-Related Protein Type 2、SIR2-Like Protein 2、Sirtuin Type 2、Sirtuin (Silent Mating Type Information Regulation 2 Homolog) 2 (S. cerevisiae)、Sirtuin-2、Sirtuin (Silent Mating Type Information Regulation 2, S. cerevisiae, Homolog) 2、EC 3.5.1.、SIR2; GenBank: AAK51133.1); Sema 5b (FLJ10372、KIAA1445、Mm.42015, SEMA5B, SEMAG, Semaphorin 5b Hlog, sema domain, seven thrombospondin repeats (type 1 and type 1-like), Transmembrane Domain (TM) and short cytoplasmic domain, (semaphorin) 5B, Genbank accession number AB04087; secernin 1 (SCRN1, SES1, KIAA0193, secerin-1; GenBank: EAL24458.1); SAGE (SAGE1, Sarcoma Antigen 1, Cancer / Testis Antigen 14, CT14, Putative Tumor Antigen; NCBI reference sequence: NP-061136.2); RU2AS (KAAG1, Kidney Associated Antigen 1, RU2AS, RU2 Antisense Gene Protein, Kidney-Associated Antigen 1; GenBank: AAF23613.1); RNF43 - E3 ubiquitin-protein ligase RNF43 precursor [Homo sapiens], RNF124; URCC; NCBI reference sequence: NP-060233.3; RhoC (RGS5 (Regulator Of G-Protein Signaling 5, MSTP032, Regulator Of G-Protein Signalling 5, MSTP092, MST092, MSTP106, MST106, MSTP129, MST129; GenBank: AAB84001.1); RET (ret proto-oncogene; MEN2A; HSCR1; MEN2B; MTC1; PTC; CDHF12; Hs.168114; RET51; RET-ELE; NP-066124.1; NM-020975.4); RBAF600 (UBR4、Ubiquitin Protein Ligase E3 Component N-Recognin 4、Zinc Finger、UBR1 Type 1、ZUBR1、E3 Ubiquitin-Protein Ligase UBR4、RBAF600、600 KDa Retinoblastoma Protein-Associated Factor、Zinc Finger UBR1-Type Protein 1、EC 6.3.2.、N-recognin-4、KIAA0462、p600、EC 6.3.2、KIAA1307; GenBank: AAL83880.1); RAGE-1 (MOK、MOK Protein Kinase、Renal Tumor Antigen、RAGE、MAPK / MAK / MRK Overlapping Kinase、Renal Tumor Antigen 1、Renal Cell Carcinoma Antigen、RAGE-1、EC 2.7.11.22、RAGE1; UniProtKB / Swiss-Prot: Q9UQ07.1); RAB38 / NY-MEL-1 (RAB38、NY-MEL-1、RAB38、Member RAS Oncogene Family、Melanoma Antigen NY-MEL-1、Rab-Related GTP-Binding Protein、Ras-Related Protein Rab-38、rrGTPbp; GenBank: AAH15808.1); PTPRK (DJ480J14.2.1 (Protein Tyrosine Phosphatase, Receptor Type, K R-PTP-KAPPA, Protein Tyrosine Phosphatase Kappa, Protein Tyrosine Phosphatase Kappa), Protein Tyrosine Phosphatase, Receptor Type, K, Protein-Tyrosine Phosphatase Kappa, Protein-Tyrosine Phosphatase, Receptor Type, Kappa, R-PTP-kappa, Receptor-Type Tyrosine-Protein Phosphatase Kappa, EC 3.1.3.48, PTPK; GenBank: AAI44514.1); PSMA; PSCA hIg(2700050C12Rik, C530008016Rik, RIKEN cDNA 2700050C12, RIKEN cDNA 2700050C12 gene, Genbank accession number AY358628); PSCA (Prostate stem cell antigen precursor, Genbank accession number AJ29743; PRDX5 (Peroxiredoxin 5, EC 1.11.1.15, TPx Type VI, B166, Antioxidant Enzyme B166, HEL-S-55, Liver Tissue 2D-Page Spot 71 B, PMP20, Peroxisomal Antioxidant Enzyme, PRDX6, Thioredoxin Peroxidase PMP20, PRXV, AOEB166, Epididymis Secretory Protein Li 55, Alu Co-Repressor 1, Peroxiredoxin-5, Mitochondrial, Peroxiredoxin V, prx-V, Thioredoxin Reductase, Prx-V, ACR1, Alu Corepressor, PLP; GenBank: CAG33484.1); PRAME (Preferentially Expressed Antigen In Melanoma, Preferentially Expressed Antigen Of Melanoma, MAPE, 01P-4, OIPA, CT130, Cancer / Testis Antigen 130, Melanoma Antigen Preferentially Expressed In Tumors, Opa-Interacting Protein 4, Opa-Interacting Protein 01P4; GenBank: CAG30435.1); pml-RARalpha fusion protein; PMEL17 (silver homolog; SILV; D12S53E; PMEL17; SI; SIL); ME20; gp10 BC001414; BT007202; M32295; M77348; NM-006928; PBF (ZNF395, Zinc Finger Protein 395, PRF-1, Huntington disease regulatory, HD Gene Regulatory Region-Binding Protein, Region-Binding Protein 2, Protein 2, Papillomavirus Regulatory Factor 1, HD-Regulating Factor 2, Papillomavirus-Regulatory Factor, PRF1, HDBP-2, Si-1-8-14, HDBP2, Huntington'S Disease Gene Regulatory Region-Binding Protein 2, HDRF-2, Papillomavirus Regulatory Factor PRF-1, PBF; GenBank: AAH01237.1); PAX5 (Paired Box 5、Paired Box Homeotic Gene 5、BSAP、Paired Box Protein Pax-5、B-Cell Lineage Specific Activator、Paired Domain Gene 5、Paired Box Gene 5 (B-Cell Lineage Specific Activator Protein)、B-Cell-Specific Transcription Factor、Paired Box Gene 5 (B-Cell Lineage Specific Activator); PAP (REG3A、Regenerating Islet-Derived 3 Alpha、INGAP、PAP-H、Hepatointestinal Pancreatic Protein、PBBCGF、Human Proislet Peptide、REG-Ill、Pancreatitis-Associated Protein 1、Regi、Reg III-Alpha、hepatocarcinoma-intestine-pancreas、Regenerating Islet-Derived Protein III-Alpha、Pancreatic Beta Cell Growth Factor、HIP、PAP Homologous Protein、HIP / PAP、Proliferation-Inducing Protein 34、PAP1、Proliferation-Inducing Protein 42、REG-3-alpha、Regenerating Islet-Derived Protein 3-Alpha、Pancreatitis-Associated Protein; GenBank: AAH36776.1); p53 (TP53, Tumor Protein P53, TPR53, P53, Cellular Tumor Antigen P53, Antigen NY-CO-13, Mutant Tumor Protein 53, Phosphoprotein P53, P53 Tumor Suppressor, BCC7, Transformation-Related Protein 53, LFS1, tumor Protein 53, Li-Fraumeni Syndrome, Tumor Suppressor P53; P2X5 (Purinergic receptor P2X ligand-gated ion channel 5, an ion channel gated by extracellular ATP that may be involved in synaptic transmission and neurogenesis. Its deficiency may contribute to the pathophysiology of idiopathic detrusor instability); 422 aa), μl: 7.63, MW: 47206 TM: 1 [P] Gene Chromosome: 17p13.3, GenBank accession number NP-002552. OGT (0-Linked N-Acetylglucosamine (GlcNAc) Transferase, O-GlcNAc Transferase P110 Subunit, 0-Linked N-Acetylglucosamine (GlcNAc) Transferase (UDP-N-Acetylglucosamine:Polypeptide-N-Acetylglucosaminyl Transferase, UDP-N-Acetylglucosamine-Peptide N-Acetylglucosaminyltransferase 110 KDa Subunit, UDP-N-Acetylglucosamine:Polypeptide-N-Acetylglucosaminyl Transferase, Uridinediphospho-N-Acetylglucosamine:Polypeptide Beta-N-Acetylglucosaminyl Transferase, O-GlcNAc Transferase Subunit P110, EC 2.4.1.255、0-Linked N-Acetylglucosamine Transferase 110 KDa Subunit、EC 2.4.1、HRNT1、EC 2.4.1.186、0-GLCNAC; GenBank: AAH38180.1); 0A1 (Osteoarthritis QTL 1、OASD; GenBank: CAA88742.1); NY-ESO-1 / LAGE-2 (Cancer / Testis Antigen 1 B、CTAG1 B、NY-ESO-1、LAGE-2、ESO1、CTAG1、CTAG、LAGE2B、Cancer / Testis Antigen 1、Autoimmunogenic Cancer / Testis Antigen NY-ESO-1、Ancer Antigen 3、Cancer / Testis Antigen 6.1、New York Esophageal Squamous Cell Carcinoma 1、L Antigen Family Member 2、LAGE2、CT6.1、LAGE2A; GenBank: AAI30365.1); NY-BR-1 (ANKRD30A、Ankyrin Repeat Domain 30A、Breast Cancer Antigen NY-BR-1、Serologically Defined Breast Cancer Antigen NY-BR-1、Ankyrin Repeat Domain-Containing Protein 30A; NCBI Reference Sequence: NP-443723.2); N-ras (NRAS、Neuroblastoma RAS Viral (V-Ras) Oncogene Homolog、NRAS1、Transforming Protein N-Ras、GTPase NRas、ALPS4、N-Ras Protein Part 4、NS6、Oncogene Homolog、HRAS1; GenBank: AAH05219.1); NFYC (Nuclear Transcription Factor Y、Gamma、HAP5、HSM、Nuclear Transcription Factor Y Subunit C、Transactivator HSM-1 / 2、CCAAT Binding Factor Subunit C、NF-YC、CCAAT Transcription Binding Factor Subunit Gamma、CAAT Box DNA-Binding Protein Subunit C、Histone H1 Transcription Factor Large Subunit 2A、CBFC、Nuclear Transcription Factor Y Subunit Gamma、CBF-C、Transactivator HSM-1、H1TF2A、Transcription Factor NF-Y、C Subunit; neo-PAP (PAPOLG、Poly(A) Polymerase Gamma、Neo-Poly(A) Polymerase、Nuclear Poly(A) Polymerase Gamma、Polynucleotide Adenylyltransferase Gamma、SRP RNA 3′ Adenylating Enzyme / Pap2、PAP-gamma、Neo-PAP、SRP RNA 3′-Adenylating Enzyme、PAP2、EC 2.7.7.19、PAPG; NCBI Reference Sequence: NP-075045.2); NCA (CEACAM6, Genbank accession number M1872); Napi3b (NAPI-3B, NPTIIb, SLC34A2, solute carrier family 34 (sodium phosphate), member 2, type II sodium-dependent phosphate transporter 3b, Genbank accession number NM-00642); Myosin class I; MUM-3; MUM-2 (TRAPPC1, Trafficking Protein Particle Complex 1, BETS, BETS Homolog, MUM2, Melanoma Ubiquitous Mutated 2, Multiple Myeloma Protein 2, Trafficking Protein Particle Complex Subunit 1; MUM-1f; Mucin (MUC1, Mucin 1, Cell Surface Associated, PEMT, PUM, CA 15-3, MCKD1, ADMCKD, Medullary Cystic Kidney Disease 1 (autosomal dominant), ADMCKD1, Mucin 1, Transmembrane, CD227, Breast Carcinoma-Associated Antigen DF3, MAM6, Cancer Antig. en 15-3、MCD、Carcinoma-Associated Mucin、MCKD、Krebs Von Den Lungen-6、MUC-1 / SEC、Peanut-Reactive Urinary Mucin、MUC1 / ZD、Tumor-Associated Epithelial Membrane Antigen、DF3 Antigen、Tumor-Associated Mucin, episialin、EMA、H23 Antigen、H23AG、Mucin-1、KL-6、Tumor Associated Epithelial Mucin、MUC-1、Episialin、PEM、CD227 Antigen; UniProtKB / Swiss-Prot: P15941.3); MUCSAC (Mucin SAC、Oligomeric Mucus / Gel-Forming、Tracheobronchial Mucin′ MUC5、TBM、Mucin 5、Subtypes A And C、Tracheobronchial / Gastric、leB、Gastric Mucin、Mucin SAC、Oligomeric Mucus / Gel-Forming Pseudogene、Lewis B Blood Group Antigen、LeB、Major Airway Glycoprotein、MUC-SAC、Mucin-5 Subtype AC、Tracheobronchial;MUC1 (Mucin 1, Cell Surface Associated, PEMT, PUM, CA 15-3, MCKD1, ADMCKD, Medullary Cystic Kidney Disease 1 (Autosomal Dominant), ADMCKD1, Mucin 1, Transmembrane, CD227, Breast Carcinoma-Associated Antigen DF3, MAM6, Cancer Antigen 15-3, MCD, Carcinoma-Associated Mucin, MCKD, Krebs Von Den Lungen-6, MUC-1 / SEC, Peanut-Reactive Urinary Mucin, MUC-1 / X, Polymorphic Epithelial Mucin, MUC1 / ZD, Tumor-Associated Epithelial Membrane Antigen, DF3 Antigen, Tumor-Associated Mucin, episialin, EMA, h23 Antigen, H23AG, mucin-1, KL-6, Tumor Associated Epithelial Mucin, MUC-1, Episialin, PEM, CD227 Antigen; MSG783 (RNF124, hypothetical protein FLJ20315, Genbank accession number NM-01776;MRP4-multidrug resistance-associated protein 4 isoform 3, MOAT-B; MOATB [Homo sapiens]; NCBI reference sequence: NP-001288758.1; MPF (MPF, MSLN, SMR, megakaryocyte potentiating factor, mesothelin, Genbank accession number NM-00582;MMP-7 (MMP7、matrilysin、MPSL1、matrin、Matrix Metalloproteinase 7 (Matrilysin、Uterine)、Uterine Matrilysin、Matrix Metalloproteinase-7、EC 3.4.24.23、Pump-1 Protease、Matrin、Uterine Metalloproteinase、PUMP1、MMP-7、EC 3.4.24、PUMP-1; GenBank: AAC37543.1); MMP-2 (MMP2、Matrix Metallopeptidase 2 (Gelatinase A、72 kDa Gelatinase、72 kDa Type IV Collagenase)、MONA、CLG4A、Matrix Metalloproteinase 2 (Gelatinase A、72kD Gelatinase、72kD Type IV Collagenase)、CLG4、72 kDa Gelatinase、72 kDa Type IV Collagenase)、Matrix Metalloproteinase-2、MMP-II、72 KDa Gelatinase、Collagenase Type IV-A、MMP-2、Matrix Metalloproteinase-II、TBE-1、Neutrophil Gelatinase、EC 3.4.24.24、EC 3.4.24; GenBank: AAH02576.1);およびMeloe。;

[0217] In certain embodiments, the CD3 binding domains of the invention and antibodies comprising same comprise multispecific antibodies, e.g., bispecific antibodies, wherein the multispecific antibody comprises a second binding domain having specificity for a second antigen selected from the group consisting of: 17-IA, 4-1BB, 4Dc, 6-keto-PGF1a, 8-iso-PGF2a, 8-oxo-dG, Al adenosine receptor, A33, ACE, ACE-2, activin, activin A, activin AB, activin B, activin C, activin RIA, activin RIA ALK-2, activin RIB ALK-4, activin RIIA, activin RUB, ADAM, ADAM10, ADAM12, ADAM15, ADAM17 / TACE, ADAM8, ADAM9, ADAMTS, ADAMTS4, ADAMTS5, addressin, aFGF, ALCAM, ALK, ALK-1, ALK-7, alpha-l-antitrypsin, alpha-V / beta-1 antagonist, ANG, Ang, APAF-1, APE, APJ, APP, APRIL, AR, ARC, ART, Artemin, anti-Id, ASPARTIC, atrial natriuretic factor, av / b3 integrin, Axl, b2M, B7-1, B7-2, B7-H, B-lymphocyte stimulating factor (BlyS), BACE, BACE-1, Bad, BAFF, BAFF-R, Bag-1, BAK, Bax, BCA-1, BCAM, Bel, BCMA, BDNF, b-ECGF, bFGF, BID, Bik, BIM, BLC, BL-CAM, BLK, BMP, BMP-2 BMP-2a, BMP-3 osteogenin, BMP-4 BMP-2b, BMP-5, BMP-6 Vgr-1, BMP-7 (OP-1), BMP-8 (BMP-8a, OP-2), BMPR, BMPR-IA (ALK-3), BMPR-IB (ALK-6), BRK-2, RPK-1, BMPR-II (BRK-3), BMPs, b-NGF, BOK, bombesin, bone-derived neurotrophic factor, BPDE, BPDE-DNA, BTC, complement factor 3(C3), C3a, C4, C5, C5a, CIO, CA125, CAD-8, calcitonin, cAMP, carcinoembryonic antigen (CEA), tumor-associated antigen, cathepsin A, cathepsin B, cathepsin C / DPPI, cathepsin D, cathepsin E, cathepsin H, cathepsin L, cathepsin O, cathepsin S, cathepsin V, cathepsin X / Z / P, CBL, CCI, CCK2, CCL, CCLl, CCLll, CCL12, CCL13, CCL 14, CCL15, CCL16, CCLl 7, CCL18, CCL19, CCL2, CCL20, CCL21, CCL22, CCL23, CCL24, CCL25, CCL26, CCL27, CCL28, CCL3, CCL4, CCL5, CCL6, CCL7, CCL8, CCL9 / 10, CCR, CCR1, CCR10, CCR10, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CD1, CD2, CD4, CD5, CD6, CD7, CD8, CD10, CDlla, CDllb, CDllc, CD13, CD14, CD 15, CD16, CD18, CD19, CD20, CD21, CD22, CD23, CD25, CD27L, CD28, CD29, CD30, CD30L, CD32, CD33 (p67 protein), CD34, CD38, CD40, CD40L, CD44, CD45, CD46, CD49a, CD52, CD54, CD55, CD56, CD61, CD64, CD66e, CD74, CD80 (B7-1), CD89, CD95, CD123, CD137, CD138, CD140a, CD146, CD147, CD148, CD152, CD164, CEACAM5, CFTR, cGMP, CINC, botulinum toxin, Clostridium perfringens toxin, CKb8-l, CLC, CMV, CMVUL, CNTF, CNTN-1, COX, C-Ret, CRG-2, CT-1, CTACK, CTGF, CTLA-4, CX3CL1, CX3CR1, CXCL, CXCLl, CXCL2, CXCL3, CXCL4, CXCL5, CXCL6, CXCL7, CXCL8, CXCL9, CXCL10, CX CL11, CXCL12, CXCL13, CXCL14, CXCL15, CXCL16, CXCR, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6, cytokeratin tumor-associated antigen, DAN, DCC, DcR3, DC-SIGN, decay-promoting factor, des(l-3)-IGF-I (brain IGF-1), Dhh, digoxin, DNAM-1, Dnase, Dpp, DPPIV / CD26, Dtk, ECAD, EDA, EDA-A1, EDA-A2, EDAR, EGF, EGFR (ErbB-1), EMA, EMMPRIN, EN A, endothelin receptor, enkephalinase, eNOS, Eot, eotaxinl, EpCAM, ephrin B2 / EphB4, EPO, ERCC, E-selectin, ET-1, Factor Ila, Factor VII, Factor VIIIc, Factor IX, fibroblast activation protein (FAP), Fas, FcRl, FEN-1, ferritin, FGF, FGF-19, FGF-2, FGF3, FGF-8, FGFR, FGFR-3, Fibrin, FL, FLIP, Flt-3, Flt-4, follicle-stimulating hormone, fractalkine, FZD1, FZD2, FZD3, FZD4, FZD5, FZD6, FZD7, FZD8, FZD9, FZD10, G250, Gas 6, GCP-2, GCSF, GD2, GD3, GDF, GDF-1, GDF-3 (Vgr-2), GDF-5 (BMP-14, CDMP- 1), GDF-6 (BMP-13, CDMP-2), GDF-7 (BMP-12, CDMP-3), GDF-8 (myostatin), GDF-9, GDF- 15 (MIC-1), GDNF, GDNF, GFAP, GFRa-1, GFR-alphal, GFR-alpha2, GFR-alpha3, GITR, glucagon, Glut 4, glycoprotein Ilb / IIIa (GPIlb / IIIa), GM-CSF, gpl30, gp72, GRO, growth hormone-releasing factor, hapten (NP-cap or NIP-cap), HB-EGF, HCC, HCMV gB envelope glycoprotein, HCMV gH envelope glycoprotein, HCMV UL, hemopoietic growth factor (HGF), Hep B gpl20, heparanase, Her2, Her2 / neu (ErbB-2), Her3 (ErbB-3), Her4 (ErbB-4), herpes simplex virus (HSV) gB glycoprotein, HSV gD glycoprotein, HGFA, high molecular weight melanoma-associated antigen (HMW-MAA), HIV gpl20, HIV IIIB gp 120 V3 loop, HLA, HLA-DR, HM1.24, HMFG PEM, HRG, Hrk, human cardiac myosin, human cytomegalovirus (HCMV), human growth hormone (HGH), HVEM, 1-309, IAP, ICAM, ICAM-1, ICAM-3, ICE, ICOS, IFNg, Ig, IgA receptor, IgE, IGF, IGF binding protein, IGF-1R, IGFBP, IGF-I, IGF-II, IL, IL-1, IL-1R, IL-2, IL-2R, IL-4, IL-4R, IL-5, IL-5R, IL-6, IL-6R, IL-8, IL-9, IL-10, IL-12, IL-13, IL-15, IL-18, IL-18R, IL-23, interferon (INF)-alpha, INF-beta, INF-gamma, inhibin, iNOS, insulin A-chain, insulin B-chain, insulin-like growth factor 1, integrin alpha 2, integrin alpha 3, integrin alpha 4, integrin alpha 4 / beta l, integrin alpha 4 / beta 7, integrin alpha 5(alpha V), integrin alpha 5 / beta l, integrin alpha 5 / beta 3, integrin alpha 6, integrin beta l, integrin beta 2, interferon gamma, IP-10, 1-TAC, JE, kallikrein 2, kallikrein 5, kallikrein 6, kallikrein 11, kallikrein 12, kallikrein 14, kallikrein 15, kallikrein LI, kallikrein L2, kallikrein L3, kallikrein L4, KC, KDR, keratinocyte growth factor (KGF), laminin 5, LAMP, LAP, LAP (TGF-1), latent TGF-1, latent TGF-1 bpl, LBP, LDGF, LECT2, Lefty, Lewis-Y antigen, Lewis-Y related antigen, LFA-1, LFA-3, Lfo, LIF, LIGHT, lipoprotein, LIX, LKN, Lptn, L-selectin, LT-a, LT-b, LTB4, LTBP-1, pulmonary surfactant, progesterone, lymphotoxin beta receptor, Mac-1, MAdCAM, MAG, MAP2, MARC, MCAM, MCAM, MCK-2, MCP, M-CSF, MDC, Mer, metalloproteinase, MGDF receptor, MGMT, MHC (HLA-DR), MIF, MIG, MIP, MIP-1-alpha, MK, MMAC1, MMP, MMP-1, MMP-10, MMP-11, MMP-12, MMP-13, MMP-14, MMP-15, MMP-2, MMP-24, MMP-3, MMP-7, MMP-8, MMP-9, MPIF, Mpo, MSK, MSP, mucin (Mucl), MUC18, Müllerian inhibitory factor, Mug, MuSK, NAIP, NAP, NCAD, N-cadherin, NCA 90, NCAM, NCAM, neprilysin, neurotrophin-3, -4, or -6, neurturin, nerve growth factor (NGF), NGFR, NGF-beta, nNOS, NO, NOS, Npn, NRG-3, NT, NTN, OB, OGG1, OPG, OPN, OSM, OX40L, OX40R, pl50, p95, PADPr, Parathyroidhormone, PARC, PARP, PBR, PBSF, PCAD, P-cadherin, PCNA, PDGF, PDGF, PDK-1, PECAM, PEM, PF4, PGE, PGF, PGI2, PGJ2, PIN, PLA2, placental alkaline phosphatase (PLAP), P1GF, PLP, PP14, proinsulin, prorelaxin, protein C, PS, PSA, PSCA, prostate-specific membrane antigen (PSMA), PTEN, PTHrp, Ptk, PTN, R51, RANK, RANKL, RANTES, relaxin A-chain, relaxin B-chain, renin, respiratory syncytial virus (RSV) F, RSV Fgp, Ret, rheumatoid factor, RLIP76, RPA2, RSK, S100, SCF / KL, SDF-1, SERINE, serum albumin, sFRP-3, Shh, SIGIRR, SK-1, SLAM, SLPI, SMAC, SMDF, SMOH, SOD, SPARC, Stat, STEAP, STEAP-II, TACE, TACI, TAG-72 (tumor-associated glycoprotein-72), TARC, TCA-3, T cell receptor (e.g., T cell receptor alpha / beta), TdT, TECK, TEM1, TEM5, TEM7, TEM8, TERT, testicular PLAP-like alkaline phosphatase, TfR, TGF, TGF-alpha, TGF-beta, TGF-beta pan-specific, TGF-beta RI (ALK-5), TGF-beta RII, TGF-beta Rllb, TGF-beta RIII, TGF-beta l, TGF-beta 2, TGF-beta 3, TGF-beta 4, TGF-beta 5, thrombin, thymus Ck-1, thyroid stimulating hormone, Tie, TIMP, TIQ, tissue factor, TMEFF2, Tmpo, TMPRSS2, TNF, TNF-alpha, TNF-alpha beta, TNF-beta 2, TNFc, TNF-RI, TNF-RII, TNFRSF10A (TRAIL Rl Apo-2, DR4), TNFRSFIOB (TRAIL R2 DR5, KILLER, TRICK-2A, TRICK-B), TNFRSF10C (TRAIL R3 DcRl, LIT, TRID), TNFRSF10D (TRAIL R4 DcR2, TRUNDD), TNFRSF11A (RANK ODF R, TRANCER)、TNFRSFllB (OPG OCIF、TR1)、TNFRSF12 (TWEAK R FN14)、TNFRSF13B (TACI)、TNFRSF13C (BAFF R)、TNFRSF14 (WHEM ATAR、HveA、LIGHT R、TR2)、TNFRSF16 (NGFR p75NTR), TNFRSF17 (BCMA), TNFRSF18 (GITR AITR), TNFRSF19 (TROY CROWN), TNFRSF19L (RELT), TNFRSFIA (TNF RI CD120a, p55-60), TNFRSFIB (TNF RII). CD120b, p75-80, TNFRSF26 (TNFRH3), TNFRSF3 (LTbR TNF RIII, TNFC R), TNFRSF4 (OX40 ACT35, TXGP1 R), TNFRSF5 (CD40 p50), TNFRSF6 (Fas). Apo-1, APT1, CD95, TNFRSF6B (DcR3 M68, TR6), TNFRSF7 (CD27), TNFRSF8 (CD30), TNFRSF9 (4-lBB CD137, ILA), TNFRSF21 (DR6), TNFRSF22 (DcTRAIL R2). TNFRH2), TNFRST23 (DcTRAIL Rl TNFRH1), TNFRSF25 (DR3 Apo-3, LARD, TR-3, TRAMP, WSL-1), TNFSF10 (TRAIL Apo-2 dilution, TL2), TNFSF11 (TRANCE / RANK ODF (OPG) TNFSF12 (TWEAK Apo-3 ODF) TNFSF13 (APRIL TALL2) TNFSF13B (BAFF). BLYS、TALL1、THANK、TNFSF20)、TNFSF14 (LIGHT WHIME LINES、LTg) TNFSF15 (TL1A / VEGI) TNFSF18 (GITRリンドAITRリガド、TL6), TNFSFIA (TNF-a Connectin, DIF, TNFSF2), TNFSF1B (TNF-b LTa, TNFSF1), TNFSF3 (LTb TNFC, p33), TNFSF4 (OX40ダツgp34, TXGP1, TNFSF5 (CD40 substrate).CD154, gp39, HIGM1, IMD3, TRAP), TNFSF6 (Fas ligand, Apo-1 ligand, APT1 ligand), TNFSF7 (CD27 ligand, CD70), TNFSF8 (CD30 ligand, CD153), TNFSF9 (4-lBB ligand, CD137 ligand), TP-1, t-PA, Tpo, TRAIL, TRAIL R, TRAIL-R1, TRAIL-R2, TRANCE, transfer receptor, TRF, Trk, TROP-2, TSG, TSLP, tumor-associated antigen CA 125, tumor-associated antigen expressing Lewis Y-related glycoprotein, TWEAK, TXB2, Ung, uPAR, uPAR-1, urokinase, VCAM, VCAM-1, VECAD, VE-cadherin, VE-cadherin-2, VEFGR-1 (flt-1), VEGF, VEGFR, VEGFR-3 (flt-4), VEGI, VIM, viral antigen, VLA, VLA-1, VLA-4, VNR integrin, von Willebrand factor, WIF-1, WNT1, WNT2, WNT2B / 13, WNT3, WNT3A, WNT4, WNT5A, WNT5B, WNT6, WNT7A, WNT7B, WNT8A, WNT8B, WNT9A, WNT9A, WNT9B, WNT10A, WNT10B, WNT11, WNT16, XCL1, XCL2, XCR1, XCR1, XEDAR, XIAP, XPD, CTLA4 (cytotoxic T lymphocyte antigen-4), PD1 (programmed cell death protein 1), PD-L1 (programmed cell death ligand 1), LAG-3 (lymphocyte activation gene-3), TIM-3 (T cell immunoglobulin and mucin protein-3), receptors for hormones, and growth factors.

[0218] In certain embodiments, the CD3 binding domains of the invention and antibodies comprising same comprise multispecific antibodies, e.g., bispecific antibodies, wherein the multispecific antibodies comprise a second binding domain having specificity for a second antigen selected from the group consisting of BCMA, CTLA4 (cytotoxic T lymphocyte antigen-4), PD1 (programmed cell death protein 1), PD-L1 (programmed cell death ligand 1), LAG-3 (lymphocyte activation gene-3), TIM-3, CD20, CD2, CD19, Her2, EGFR, EpCAM, FcyRIIIa (CD16), FcyRIIa (CD32a), FcyRIIb (CD32b), FcyRI. (CD64), Toll-like receptors (TLRs), TLR4, TLR9, cytokines, IL-2, IL-5, IL-13, IL-6, IL-17, IL-12, IL-23, TNFα, TGFβ, cytokine receptors, IL-2R, chemokines, chemokine receptors, growth factors, VEGF, and HGF.

[0219] In some embodiments, the present invention provides bispecific antibodies having affinity for CD3 and HER2. In some embodiments, the present invention provides bispecific antibodies comprising an antibody fragment (e.g., Fab) of trastuzumab. Exemplary bispecific CD3 antibodies include ADI-29291, ADI-29292, ADI-29293, ADI-29294, ADI-29295, ADI-29297, ADI-29298, ADI-29300, ADI-29301, ADI-29302, ADI-29303, ADI-29304, ADI-29305, and ADI-29306.

[0220] In specific embodiments, the CD3-binding domains of the present invention and antibodies comprising the same may be prepared according to various techniques available to those skilled in the art, including, but not limited to, recombinant coexpression of two immunoglobulin heavy-light chain pairs with different specificities (see Milstein and Cuello, Nature 305: 537 (1983)), WO 93 / 08829 and Traunecker et al., EMBO J. 10: 3655 (1991)), and "knob-in-hole" engineering (see, e.g., U.S. Pat. No. 5,731,168). "Knob-in-hole" engineering of multispecific antibodies may be used to create a first arm containing a knob and a second arm containing a hole to which the knob of the first arm can be bound. In one embodiment, the knob of a multispecific antibody of the present invention may be an anti-CD3 arm. Alternatively, in one embodiment, the knob of a multispecific antibody of the present invention may be an anti-target / antigen arm. In one embodiment, the hole of a multispecific antibody of the present invention may be an anti-CD3 arm. Alternatively, the holes of the multispecific antibodies of the present invention may be anti-target / antigen arms in one embodiment. Multispecific antibodies may be engineered using immunoglobulin crossover technology (also known as Fab domain exchange or CrossMab format) (see, e.g., WO2009 / 080253; Schaefer et al., Proc. Natl. Acad. Sci. USA, 108:11187-11192 (2011)).Multispecific antibodies can be generated by manipulating electrostatic steering effects to create antibody Fc-heterodimeric molecules (WO 2009 / 089004A1); cross-linking two or more antibodies or fragments (see, e.g., U.S. Pat. No. 4,676,980 and Brennan et al., Science, 229: 81 (1985)); using leucine zippers to create bispecific antibodies (see, e.g., Kostelny et al., J. Immunol, 148(5):1547-1553 (1992)); using "diabody" technology to create bispecific antibody fragments (see, e.g., Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993)); and using single-chain Fv (sFv) dimers (see, e.g., Gruber et al., J. Immunol, 152:5368 (1994)); and by preparing trispecific antibodies as described, for example, in Tutt et al. J. ImmunoL 147:60 (1991).Additional non-limiting examples of multispecific and bispecific formats suitable for incorporation of the anti-CD3 binding domains of the present invention include, for example, Fab-Fc-scFv (Bottle-Opener) (XENCOR), Mab-scFv (XENCOR), Mab-Fv (XENCOR), Dual scFv (XENCOR), central Fv (XENCOR), central scFv (XENCOR), one-arm central scFv (XENCOR), Fab-Fab (XENCOR), Fab-Fv (XENCOR), mAb-Fv (XENCOR), mAb-Fab (XENCOR), DART (MacroGenics), BiTE (Amgen / Micromet), KiTE, common light chain-IgG (Genentech), and TandAb (Affimed). These include Cross-Mab (Roche), SEED (EMD Serono), BEAT (Glenmark), TrioMab (Trion Pharma / FresEnius Biotech), DuetMab (MedImmune), and others.Methods for producing such multispecific antibodies are disclosed, for example, in WO 95 / 09917, WO 2008 / 119566, WO 2008 / 119567, WO2011 / 121110, WO 2010 / 037835, WO 2007 / 042261, WO 2007 / 110205, WO 2011 / 121110, WO 2012 / 055961, WO 2012 / 16067, WO 2016 / 086189, WO 2016 / 182751, WO 2015 / 006749, WO 2014 / 049003, WO 2013 / 177101, WO 2015 / 128509, US 7,951,917, US 2009 / 0252729, US 2014 / 0348839, US 7,183,076, Mazor et al., Mabs, Vol. 7, pp. 377-389 (2015), Muda et al., Protein Engineering, Design, & Selection, Vol. 24, pp. 447-454 (2011), and Del Bano et al., Antibodies, Vol. 5, pp. 1-23 (2016).

[0221] Further examples of bispecific antibody formats that can be used in the context of the present invention include the use of a first immunoglobulin (Ig) CH3 domain and a second Ig CH3 domain, where the first and second Ig CH3 domains differ from each other by at least one amino acid, and where the at least one amino acid difference reduces binding of the bispecific antibody to Protein A compared to a bispecific antibody without that amino acid difference. In one embodiment, the first Ig CH3 domain binds to Protein A and the second Ig CH3 domain contains a mutation that reduces or eliminates Protein A binding, such as, for example, an H95R (according to IMGT exon numbering; H435R in EU numbering) modification. The second CH3 may further comprise a Y96F modification (according to IMGT; Y436F in EU). Additional modifications that may be present within the second CH3 include: D16E, L18M, N44S, K52N, V57M, and V82I in the case of IgG1 antibodies (according to IMGT; D356E, L358M, N384S, K392N, V397M, and V422I in EU); N44S, K52N, and V82I in the case of IgG2 antibodies (according to IMGT; N384S, K392N, and V422I in EU); and Q15R, N44S, K52N, V57M, R69K, E79Q, and V82I in the case of IgG4 antibodies (according to IMGT; Q355R, N384S, K392N, V397M, R409K, E419Q, and V422I in EU).

[0222] In certain embodiments, the CD3 binding domains of the invention and antibodies comprising same may be prepared as engineered antibodies with three or more functional antigen binding sites, including, for example, the "Octopus antibodies" disclosed in US 2006 / 0025576A1.

[0223] In certain embodiments, the CD3-binding domains of the present invention and antibodies comprising the same may be prepared as "dual acting FAbs" or "DABs" that contain antigen-binding sites that bind to CD3 as well as another distinct antigen, as disclosed, for example, in US 2008 / 0069820.

[0224] As disclosed throughout this specification, the CD3-binding domains of the present invention and antibodies comprising the same can include monospecific, bispecific, or multispecific antibodies. Multispecific antibodies may be specific for different epitopes of a single target polypeptide or may contain antigen-binding domains specific for multiple target polypeptides. See, for example, Tutt et al., 1991, J. Immunol. 147:60-69; Kufer et al., 2004, Trends Biotechnol. 22:238-244. The antibodies of the present invention can be linked to or co-expressed with another functional molecule, such as another peptide or protein. For example, an antibody or fragment thereof can be functionally linked (e.g., by chemical conjugation, genetic fusion, noncovalent bonding, or other means) to one or more other molecular entities, such as another antibody or antibody fragment, to generate a bispecific or multispecific antibody with a second binding specificity.

[0225] In certain embodiments, amino acid sequence variants of the CD3-binding domain of the present invention and antibodies comprising the same are contemplated. For example, it may be desirable to improve the binding affinity and / or other biological properties of the antibody. Amino acid sequence variants of antibodies may be generated by introducing appropriate modifications into the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, deletion and / or insertion and / or substitution of residues within the amino acid sequence of the antibody. Any combination of deletion, insertion, and substitution can be made in the final construct, provided that the final construct retains the desired characteristics, such as antigen binding. In certain embodiments, such variants have one or more amino acid substitutions. Target sites for substitutional mutagenesis include the CDRs and FRs. Amino acid substitutions may be introduced into the subject antibody and the products screened for desired activity, such as retained / improved antigen binding, reduced immunogenicity, or improved ADCC or CDC.

[0226] One type of substitutional variant involves substituting one or more hypervariable region residues of a parent antibody (e.g., a humanized or human antibody). The resulting variants are typically selected for further testing and will have alterations (e.g., improvements) in certain biological properties (e.g., increased affinity, decreased immunogenicity, etc.) compared to the parent antibody and / or will substantially retain certain biological properties of the parent antibody. An exemplary substitutional variant is an affinity-matured antibody, which can be conveniently generated using affinity maturation techniques, e.g., based on phage display. Briefly, one or more CDR residues are mutated, and the variant antibodies are displayed on phage and screened for a particular biological activity (e.g., binding affinity, etc.).

[0227] Alterations (e.g., substitutions) may be made in the CDRs, for example, to improve antibody affinity. Such alterations may be made in CDR "hot spots," i.e., residues encoded by codons that undergo frequent mutation during the somatic maturation process (see, e.g., Chowdhury, Methods Mol. Biol. 207:179-196 (2008)), and / or in residues that contact the antigen, and the resulting variant VH or VL are tested for binding affinity. Affinity maturation by construction and reselection from a secondary library is reported, for example, by Hoogenboom et al. in Methods in Molecular Biology 178:1-37 (O'Brien et al., ed., Human Press, Totowa, NJ, (2001)). In some affinity maturation embodiments, diversity is introduced into the variable genes selected for mutation by any of a variety of methods (e.g., error-prone PCR, chain shuffling, or oligonucleotide-directed mutagenesis). A second library is then generated. The library is then screened to identify any antibody variants with the desired affinity. Another method for introducing diversity includes CDR-directed methods, in which several CDR residues (e.g., 4-6 residues at a time) are randomized. CDR residues involved in antigen binding may be specifically identified using, for example, alanine scanning mutagenesis or modeling methods. CDR-H3 and CDR-L3 in particular are often targeted.

[0228] In certain embodiments, substitutions, insertions, or deletions may occur within one or more CDRs of the CD3-binding domains of the present invention and antibodies comprising the same, so long as such changes do not substantially reduce the ability of the antibody to bind to the antigen. For example, conservative changes (e.g., conservative substitutions provided herein) that do not substantially reduce binding affinity may be made in the CDRs. Such changes may, for example, be outside the antigen-contacting residues in the CDRs. In certain embodiments of the variant VH and VL sequences provided above, each CDR is unchanged or contains no more than one, two, or three amino acid substitutions.

[0229] A useful method for identifying antibody residues or regions that can be targeted for mutagenesis is called "alanine scanning mutagenesis," described in Cunningham and Wells (1989) Science, 244:1081-1085. In this method, a residue or group of target residues (e.g., charged residues such as arg, asp, his, lys, and glu) is identified and substituted with neutral or negatively charged amino acids (e.g., alanine or polyalanine), and it is determined whether the antibody-antigen interaction is affected. Further substitutions can be introduced at amino acid positions that show functional sensitivity to the initial substitution. Alternatively, or in addition, crystal structures of antigen-antibody complexes can be used to identify contact points between the antibody and antigen. Such contact residues and adjacent residues can be targeted or excluded as substitution candidates. Variants can be screened to determine whether they contain desired properties.

[0230] Amino acid sequence insertions include amino- and / or carboxyl-terminal fusions ranging in length from one residue to polypeptides containing several hundred or more residues, as well as intersequence insertions of single or multiple amino acid residues. An example of a terminal insertion is an antibody with an N-terminal methionyl residue. Other insertional variants of antibody molecules include the fusion to the antibody N- or C-terminus to an enzyme (e.g., an enzyme for ADEPT) or a polypeptide that extends the serum half-life of the antibody.

[0231] In certain embodiments, the CD3-binding domains of the invention and antibodies comprising same can be modified to increase or decrease the extent to which the antibody is glycosylated. Addition or deletion of glycosylation sites to an anti-CD3 antibody of the invention can be readily achieved by altering the amino acid sequence such that one or more glycosylation sites are created or removed.

[0232] If the antibody contains an Fc region, the carbohydrate attached thereto can be altered. Antibodies produced by mammalian cells often contain branched, biantennary oligosaccharides, which are typically N-linked to Asn297 in the CH2 domain of the Fc region. See, e.g., Wright et al. TIBTECH 15:26-32 (1997). The oligosaccharides can contain various carbohydrates, such as mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, and can also contain fucose attached to GlcNAc in the "stem" of the biantennary ol...

Claims

1. An anti-cluster of differentiation 3 (CD3) antibody or an antigen-binding antibody fragment thereof, (A) a heavy chain variable region (VH) comprising CDRH1, CDRH2 and CDRH3; (B) a light chain variable region (VL) comprising CDRL1, CDRL2 and CDRL3; where: (A) the amino acid sequences of the CDRH1, CDRH2, and CDRH3 are, respectively, the amino acid sequences of CDRH1, CDRH2, and CDRH3 contained in SEQ ID NO: 1090; and (B) the amino acid sequences of the CDRL1, CDRL2 and CDRL3 are the amino acid sequences of CDRL1, CDRL2 and CDRL3 contained in SEQ ID NO: 1098, respectively; An anti-CD3 antibody or an antigen-binding antibody fragment thereof.

2. The anti-CD3 antibody or antigen-binding antibody fragment thereof of claim 1, wherein the CDRH1, the CDRH2, the CDRH3, the CDRL1, the CDRL2 and the CDRL3 comprise the amino acid sequences of SEQ ID NOs: 1091, 1093, 1095, 1099, 1101, and 1103, respectively.

3. (A) the VH comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 1090; and / or (B) the VL comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 1098; 3. An anti-CD3 antibody or an antigen-binding antibody fragment thereof according to claim 1 or 2.

4. (A) the VH comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 1090, and / or (B) the VL comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 1098; 3. An anti-CD3 antibody or an antigen-binding antibody fragment thereof according to claim 1 or 2.

5. (A) the VH comprises the amino acid sequence of SEQ ID NO: 1090; and / or (B) the VL comprises the amino acid sequence of SEQ ID NO: 1098; 3. An anti-CD3 antibody or an antigen-binding antibody fragment thereof according to claim 1 or 2.

6. An anti-CD3 antibody or an antigen-binding antibody fragment thereof described in claim 1 or 2, wherein the amino acid sequences of the VH and VL consist of SEQ ID NOs: 1090 and 1098, respectively.

7. scFv fragment, diabody, minibody, scFv-Fc, Fab fragment, Fab' fragment, Fab'-SH fragment, F(ab') ２ The anti-CD3 antibody or antigen-binding antibody fragment thereof according to any one of claims 1 to 6, comprising a CD4 fragment, a CD5 fragment, and / or an Fv fragment.

8. (A) a heavy chain comprising a VH and a heavy chain constant region (CH); (B) a light chain comprising a VL and a light chain constant region (CL); The anti-CD3 antibody or antigen-binding antibody fragment thereof according to any one of claims 1 to 7,

9. Demonstrating an enhanced development potential profile; The anti-CD3 antibody or antigen-binding antibody fragment thereof according to any one of claims 1 to 8.

10. eliciting T cell activation or T cell killing while exhibiting a reduced risk of inducing cytokine release syndrome (CRS) or a reduced tendency to elicit cytokine production; The anti-CD3 antibody or antigen-binding antibody fragment thereof according to any one of claims 1 to 9.

11. (i) a multispecific antibody or antibody fragment; (ii) a bispecific antibody or antibody fragment; (iii) at least a second antigen-binding domain that specifically binds to an oncological target, an immuno-oncological target, a neurodegenerative disease target, an autoimmune disorder target, an infectious disease target, a metabolic disease target, a cognitive disorder target, a blood-brain barrier target, or a hematological disorder target; (iv) 17-IA, 4-1BB, 4Dc, 6-keto-PGF1a, 8-iso-PGF2a, 8-oxo-dG, Al adenosine receptor, A33, ACE, ACE-2, activin, activin A, activin AB, activin B, activin C, activin RIA, activin RIA ALK-2, activin RIB ALK-4, activin RIIA, activin RUB, ADAM, ADAM10, ADAM12, ADAM15, ADAM17 / T ACE, ADAM8, ADAM9, ADAMTS, ADAMTS4, ADAMTS5, addressin, aFGF, ALCAM, ALK, ALK-1, ALK-7, alpha-l-antitrypsin, alpha-V / beta-1 antagonist, ANG, Ang, APAF-1, APE, APJ, APP, APRIL, AR, ARC, ART, Artemin, anti-Id, ASPARTIC, atrial Natriuretic factor, av / b3 integrin, Axl, b2M, B7-1, B7-2, B7-H, B-lymphocyte stimulatory factor (BlyS), BACE, BACE-1, Bad, BAFF, BAFF-R, Bag-1, BAK, Bax, BCA-1, BCAM, Bel, BCMA, BDNF, b-ECGF, bFGF, BID, Bik, BIM, BLC, BL-CAM, BLK, BMP, BMP-2 BMP-2a, BMP-3 osteogenin, BMP-4 BMP-2b, BMP-5, BMP-6Vgr-1, BMP-7 (OP-1), BMP-8 (BMP-8a, OP-2), BMPR, BMPR-IA (ALK-3), BMPR-IB (ALK-6), BRK-2, RPK-1, BMPR-II (BRK-3), BMPs, b-NGF, BOK, bombesin, bone-derived neurotrophic factor, BPDE, BPDE-DNA, BTC, complement factor 3 (C3), C3a, C4, C5, C5a, CIO, CA125, CAD-8, calcitonin, cAMP, carcinoembryonic antigen (CEA), tumor-associated antigen, cathepsin A, cathepsin B Cathepsin B, cathepsin C / DPPI, cathepsin D, cathepsin E, cathepsin H, cathepsin L, cathepsin O, cathepsin S, cathepsin V, cathepsin X / Z / P, CBL, CCI, CCK2, CCL, CCL1, CCL11, CCL12, CCL13, CCL14, CCL15, CCL16, CCL17, CCL18, CCL19, CCL2, CCL20, CCL21, CCL22, CCL23, CCL24, CCL25, CCL26, CCL27, CCL28, CCL3, CCL4, CCL5, CCL6, CCL7, CCL8, CCL9 / 10, CCR, CCR1, CCR10, CCR10, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CD1, CD2, CD4, CD5, CD6, CD7, CD8, C D10, CDlla, CDllb, CDllc, CD13, CD14, CD15, CD16, CD18, CD19, CD20, CD21, CD22, CD23, CD25, CD27L, CD28, CD29, CD30, CD30L, CD32, CD33 ( p67 protein), CD34, CD38, CD40, CD40L, CD44, CD45, CD46, CD49a, CD52, CD54, CD55, CD56, CD61, CD64, CD66e, CD74, CD80 (B7-1), CD89, CD95, CD123, CD137, CD138, CD140a, CD146, CD147, CD148, CD152, CD164, CEACAM5, CFTR, cGMP, CINC, botulinum toxin, Clostridium perfringens toxin, CKb8-1, CLC, CMV, CMVUL, CNTF, CNTN-1, COX, C-Ret, CRG-2, CT-1, CTACK, CTGF, CTLA-4, CX3CL1, CX3CR1, CXCL, CXCL1, CXCL2, CXCL3, CXCL 4, CXCL5, CXCL6, CXCL7, CXCL8, CXCL9, CXCL10, CXCL11, CXCL12, CXCL13, CXCL14, CXCL15, CXCL16, CXCR, CXCR1, CXCR 2, CXCR3, CXCR4, CXCR5, CXCR6, cytokeratin tumor-associated antigen, DAN, DCC, DcR3, DC-SIGN, decay-accelerating factor, des(1-3)-IGF-I (brain IGF-1), Dhh, digoxin, DNAM-1, Dnase, Dpp, DPPIV / CD26, Dtk, ECAD, EDA, EDA-A1, EDA-A2, EDAR, EGF, EGFR (ErbB-1), EMA, EMMPRIN, EN A, endothelin receptor, enkephalinase, eNOS, Eot, eotaxinl, EpCAM, ephrinB2 / EphB4, EPO, ERCC, E-selectin, ET-1, Factor Ila, Factor VII, Factor VIIIc, Factor IX, fibroblast activation protein (FAP), Fas, FcRl, FEN-1, ferritin, FGF, FGF-19, FGF-2, FGF3, FGF-8, FGFR, FGFR-3, Fibrin, FL, FLIP, Flt-3, Flt-4, follicle-stimulating hormone, fractalkine, FZD1, FZD2, FZD3, FZD4, FZD5, FZD6, FZD7, FZD8, FZD9, FZD10, G250, Gas 6, GCP-2, GCSF, GD2, GD3, GDF, GDF-1, GDF-3 (Vgr-2), GDF-5 (BMP-14, CDMP-1), GDF-6 (BMP-13, CDMP-2), GDF-7 (BMP-12, CDMP-3), GDF-8 (myostatin), GDF-9, GDF-15 (MIC-1), GDNF, GFAP, GFRa-1, GFR-alpha1, GFR-alpha2, GFR-alpha3, GITR, glucagon, Glut 4, glycoprotein Ilb / IIIa (GP Ilb / IIIa), GM-CSF, gpl30, gp72, GRO, growth hormone releasing factor, hapten (NP-cap or NIP-cap), HB-EGF, HCC, HCMV gBenvelope glycoprotein, HCMV gH envelope glycoprotein, HCMV UL, hematopoietic growth factor (HGF), Hep B gpl20, heparanase, Her2, Her2 / neu (ErbB-2), Her3 (ErbB-3), Her4 (ErbB-4), herpes simplex virus (HSV) gB glycoprotein, HSV gD glycoprotein, HGFA, high molecular weight melanoma-associated antigen (HMW-MAA), HIV gpl20, HIV IIIB gp 120 V3 loop, HLA, HLA-DR, HM1.24, HMFG PEM, HRG, Hrk, human cardiac myosin, human cytomegalovirus (HCMV), human growth hormone (HGH), HVEM, 1-309, IAP, ICAM, ICAM-1, ICAM-3, ICE, ICOS, IFNg, Ig, IgA receptor, IgE, IGF, IGF Binding protein, IGF-1R, IGFBP, IGF-I, IGF-II, IL, IL-1, IL-1R, IL-2, IL-2R, IL-4, IL-4R, IL-5, IL-5R, IL-6, IL-6R, IL-8, IL-9, IL-10, IL-12, IL-13, IL-15, IL-18, IL-18R, IL-23, interferon (INF)-alpha, INF-beta, INF-gamma, inhibin, iNOS, insulin A-chain, insulin B-chain, insulin-like growth factor 1, integrin alpha 2, integrin alpha 3, integrin alpha 4, integrin alpha 4 / beta 1, integrin, alpha 4 / beta 7, integrin alpha 5 (alpha V), integrin alpha 5 / beta 1, integrin alpha 5 / beta 3, integrin alpha 6, integrin beta 1, integrin beta 2, interferon gamma, IP-10, 1-TAC, JE, kallikrein 2, kallikrein 5, kallikrein 6, kallikrein 11, kallikrein 12, kallikrein 14, kallikrein 15, kallikrein LI, kallikrein L2, kallikrein L3, kallikrein L4, KC, KDR, keratinocyte growth factor (KGF), laminin 5, LAMP, LAP, LAP (TGF-1), latent TGF-1, latent TGF-1bpl, LBP, LDGF, LECT2, Lefty, Lewis-Y antigen, Lewis-Y related antigen, LFA-1, LFA-3, Lfo, LIF, LIGHT, lipoprotein, LIX, LKN, Lptn, L-selectin, LT-a, LT-b, LTB4, LTBP-1, pulmonary surfactant, progesterone, lymphotoxin beta receptor, Mac-1, MAdCAM, MAG, MAP2, MARC, MCAM, MCAM, MCK-2, MCP, M-CSF, MDC, Mer, metalloproteinase, MGDF receptor, MGMT, MHC (HLA-DR), MIF, MIG, MIP, MIP-1-alpha, MK, MMAC1, MMP, MMP-1, MMP-10, MMP-11, MMP-12, MMP-13, MMP-14, MMP-15, MMP-2, MMP-24, MMP-3, MMP-7, MMP-8, MMP-9, MPIF, Mpo, MSK, MSP, mucin (Mucl), MUC18, Müllerian inhibitory factor, Mug, MuSK, NAIP, NAP, NCAD, N-cadherin, NCA 90, NCAM, neprilysin, neurotrophin-3, -4, or -6, neurturin, nerve growth factor (NGF), NGFR, NGF-beta, nNOS, NO, NOS, Npn, NRG-3, NT, NTN, OB, OGG1, OPG, OPN, OSM, OX40L, OX40R, pl50, p95, PADPr, Parathyroid hormone, PARC, PARP, PBR, PBSF, PCAD, P-cadherin, PCNA, PDGF, PDGF, PDK-1, PECAM, PEM, PF4, PGE, PGF, PGI2, PGJ2, PIN, PLA2, placental alkaline phosphatase (PLAP), PIGF, PLP, PP14, proinsulin, prorelaxin, protein C, PS, PSA, PSCA, prostate specific membrane antigen (PSMA), PTEN, PTHrp, Ptk, PTN, R51, RANK, RANKL, RANTES, relaxin A-chain, relaxin B-chain, renin, respiratory syncytial virus (RSV) F, RSVFgp, Ret, rheumatoid factor, RLIP76, RPA2, RSK, S100, SCF / KL, SDF-1, SERINE, serum albumin, sFRP-3, Shh, SIGIRR, SK-1, SLAM, SLPI, SMAC, SMDF, SMOH, SOD, SPARC, Stat, STEAP, STEAP-II, TACE, TACI, TAG-72 (tumor-associated glycoprotein-72), TARC, TCA-3, T cell receptor (e.g., T cell receptor alpha / beta), TdT, TECK, TEM1, TEM5, TEM7, TEM8, TERT, testicular PLAP-like alkaline phosphatase, TfR, TGF, TGF-alpha, TGF-beta, TGF-beta pan-specific (Pan Specific), TGF-beta RI (ALK-5), TGF-beta RII, TGF-beta Rllb, TGF-beta RIII, TGF-beta 1, TGF-beta 2, TGF-beta 3, TGF-beta 4, TGF-beta 5, Thrombin, Thymic Ck-1, Thyroid Stimulating Hormone, Tie, TIMP, TIQ, TMEFF2, Tmpo, TMPRSS2, TNF, TNF-alpha, TNF-alpha beta, TNF-beta 2, TNFc, TNF-RI, TNF-RII, TNFRSF10A (TRAIL Rl Apo-2, DR4), TNFRSFIOB (TRAIL R2 DR5, KILLER, TRICK-2A, TRICK-B), TNFRSF10C (TRAIL R3 DcRl, LIT, TRID), TNFRSF10D (TRAIL R4 DcR2, TRUNDD), TNFRSF11A (RANK ODF R, TRANCE R), TNFRSFllB (OPG OCIF, TR1), TNFRSF12 (TWEAK R FN14), TNFRSF13B (TACI), TNFRSF13C (BAFF R), T DASH14(DHASH) DASH、DYS S、SY2)、SYSYS16(SYS) 07SYS、SYSYS17(SYSYS)、SYSYS 18(SYS) SHYSY、SYSYS10(SYS3 WHY、WHYWH、WHYWHYWHYWHYWHYWHYWHYWHYWHY 1000000000000000000000000000000000000000000000000000000 H1200、F75806、SHOSH2 6(SYS3)、SYSYS3(SYS THIS SUCH、SHIS S6、SHOSCHS4(SYS)0 SIS33、SHIS1 S) 、WHYS 5(SH40 50)、SWHYS6 DA11、HAR1、DAY5)、DYSCHYS(DYS3 H68、S6)、S6S67(S027)、 FASHION(SHR30)、SHASHES(400) HR137、HARSH、SHRYSY21 DASH) 、SHASH22(DASH) 2 SHY2)、SYSYS23(SYSYS) SHYS1)、SHYSYS25(GY3 FASH3、SHASH、SHASH、SHASH 、SHAKE1)、SHASE100(SHARE). DAY2リガンド、LOY2)、SHAS11(SHAS SHASE RHリガンドDAY 、DAYリガンド)、DAYS12(SIR). DA3リガンド、DAY3リガンド)、DAYDAY13(SYS) LOVE2)、SIGNIFICANCE130 LOVE SY、 LOVE1 LOVE LOVE LOVE LOVE LOVE LOVE LIKEリガンド、THE)、SHASE15(30). SUCH SUCH 18(SUSHリガンドSUBJECTリガンド、CHASE) FASHION、DYS、SYSYS)、SYSYSYS LIKE、SYSYS1)、SYSYS(LOVE THIS、D33) THIS 4(S40リガンドW34、SH1)、SHOSIS5(CHA40リガンドH154、N39、NH11、13 、WATCHES、DAYSJS(THEリガンドDA11リガンド 、DA1リガンド) DASH 7(KS27リガンドCHR706、SHQS8(CHR33リガンドDA153)、DJSJN(401リガンドCD137 ligand), TP-1, t-PA, Tpo, TRAIL, TRAIL R, TRAIL-R1, TRAIL-R2, TRANCE, transfer receptor, TRF, Trk, TROP-2, TSG, TSLP, tumor-associated antigen CA 125, tumor-associated antigen expressing Lewis Y-related carbohydrate, TWEAK, TXB2, Ung, uPAR, uPAR-1, urokinase, VCAM, VCAM-1, VECAD, VE-cadherin, VE-cadherin-2, VEFGR-1 (flt-1), VEGF, VEGFR, VEGFR-3 (flt-4), VEGI, VIM, viral antigen, VLA, VLA-1, VLA-4, VNR integrin, von Willebrand Revland factor, WIF-1, WNT1, WNT2, WNT2B / 13, WNT3, WNT3A, WNT4, WNT5A, WNT5B, WNT6, WNT7A, WNT7B, WNT8A, WNT8B, WN T9A, WNT9A, WNT9B, WNT10A, WNT10B, WNT11, WNT16, XCL1, XCL2, XCR1, XCR1, XEDAR, XIAP, XPD, CTLA4 (cytotoxic at least a second antigen-binding domain that specifically binds to an antigen selected from the group consisting of T lymphocyte antigen-4, PD1 (programmed cell death protein 1), PD-L1 (programmed cell death ligand 1), LAG-3 (lymphocyte activation gene-3), TIM-3 (T cell immunoglobulin and mucin protein-3), hormone receptors, and growth factors; (v) BCMA, CTLA4 (cytotoxic lymphocyte antigen-4), PD1 (programmed cell death protein 1), PD-L1 (programmed cell death ligand 1), LAG-3 (lymphocyte activation at least a second antigen binding domain that specifically binds to an antigen selected from the group consisting of: FcγRIIIa (CD16), FcγRIIa (CD32a), FcγRIIb (CD32b), FcγRI (CD64), Toll-like receptors (TLRs), TLR4, TLR9, cytokines, IL-2, IL-5, IL-13, IL-6, IL-17, IL-12, IL-23, TNFa, TGFb, cytokine receptors, IL-2R, chemokines, chemokine receptors, growth factors, VEGF, and HGF; and / or (vi) at least a second antigen-binding domain that specifically binds to an antigen, wherein the antibody contains a multispecific format selected from the group consisting of Fab-Fc-scFv, "bottle-opener", Mab-scFv, Mab-Fv, dual scFv, central Fv, central scFv, one-arm central scFv, Fab-Fab, Fab-Fv, mAb-Fv, mAb-Fab, DART, BiTE, common light chain-IgG, TandAb, Cross-Mab, SEED, BEAT, TrioMab, and DuetMab. The anti-CD3 antibody or antigen-binding antibody fragment thereof according to any one of claims 1 to 10,

12. An isolated nucleic acid encoding the anti-CD3 antibody or antigen-binding antibody fragment thereof according to any one of claims 1 to 11.

13. 13. An expression vector comprising the isolated nucleic acid of claim 12.

14. A host cell transfected, transformed or transduced with the nucleic acid of claim 12 or the expression vector of claim 13.

15. (A) one or more anti-CD3 antibodies or antigen-binding antibody fragments thereof according to any one of claims 1 to 11, one or more nucleic acids according to claim 12, or one or more expression vectors according to claim 13; (B) a pharma- ceutically acceptable carrier and / or excipient; 13. A pharmaceutical composition comprising:

16. A composition for treating or delaying the progression of a disorder or preventing or reducing the risk of onset of a disorder in a mammal in need thereof, the composition comprising an anti-CD3 antibody or an antigen-binding antibody fragment thereof described in any one of claims 1 to 11.

17. Use of the anti-CD3 antibody or antigen-binding antibody fragment thereof according to any one of claims 1 to 11 in the manufacture of a medicament.

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