anti-HER2 antibodies

Abstract

To provide chimeric antigen receptors comprising anti-HER2 monoclonal antibodies and anti-HER2 antigen-binding domains, capable of inducing apoptosis in HER2-expressing cells.SOLUTION: The present invention provides chimeric antigen receptors comprising HER2-binding monoclonal antibodies and anti-HER2 antigen-binding domains, comprising light chain CDRs and heavy chain CDRs with specific amino acid sequences, or comprising light chain variable regions with at least 85% sequence identity to specific amino acid sequences and heavy chain variable regions with at least 85% sequence identity to specific amino acid sequences.SELECTED DRAWING: Figure 15

Description

[Technical Field]

[0001] This application claims priority from SG10201801219V, filed February 13, 2018, the contents and elements of which are incorporated herein by reference for all purposes. FIELD OF THE INVENTION The present invention relates to an antibody that binds to the HER2 protein. [Background technology]

[0002] Breast cancer is the most common cause of cancer death in women, with over 400,000 deaths per year worldwide. In Singapore, breast cancer is the most frequent cancer in women, affecting over 8,000 women from 2007 to 2011, accounting for approximately 30% of cancer cases in Singaporean women and the leading cause of cancer death in Singaporean women (Singapore Cancer Registry, Interim Annual Registry Report, Trends in Cancer Incidence in Singapore).

[0003] The introduction of trastuzumab (Herceptin®), a monoclonal antibody that targets epidermal growth factor receptor (HER2 / HER-2)-positive cancers, the most aggressive form of breast cancer, accounting for approximately 20% to 30% of cases, marked a turning point in breast cancer treatment. HER2 is a member of the epidermal growth factor receptor family, which includes epidermal growth factor receptor (EGFR / HER1), HER3, and HER4. Either homo- or heterodimerization of these receptor tyrosine kinases leads to phosphorylation of intracellular domain residues, triggering downstream signaling cascades including the RAS / Raf and Akt pathways. HER2 dimerization is known to result in increased cell survival, proliferation, and resistance to apoptosis.

[0004] Trastuzumab is a humanized antibody approved by the FDA in 1998 for the treatment of HER2-positive breast cancer. Trastuzumab binds to domain IV of HER2, and while the antibody's mechanism of action is still poorly understood, several effects have been observed, including cell cycle arrest, immune-mediated cytolysis, and inhibition of angiogenesis. However, response rates to trastuzumab treatment remain low, ranging from 15% to 30%. While the use of trastuzumab in combination with chemotherapy prolongs the duration of response, the majority of responders develop resistance within one year.

[0005] The mechanisms of resistance have been intensively studied but have not been clinically confirmed. Possible causes of resistance include reduced trastuzumab binding, e.g., shielding of the trastuzumab binding site by the MUC4 glycoprotein upon association with HER2, signaling via alternative HER22-independent pathways, e.g., insulin-like growth factor-I receptor (IGF-IR), and constitutive HER2 signaling as a result of mutations in downstream signaling molecules such as PTEN and PI3K.

[0006] Furthermore, there is evidence for an important role of HER3 / HER2 heterodimers in breast cancer development, and the inability of trastuzumab to inhibit such heterodimerization may be a factor in resistance to treatment.

[0007] To address this, a second-generation HER2 humanized monoclonal antibody, pertuzumab (Perjeta®), was developed. Pertuzumab (Perjeta®) binds to an epitope in HER2 domain II, which inhibits receptor dimerization. A phase III clinical study comparing the use of pertuzumab in combination with trastuzumab with the antimitotic drug docetaxel (Taxotere®) in HER2-positive patients showed a significant improvement in overall survival with the new regimen, prompting the FDA in 2012 to approve the combination for the treatment of patients with HER2-positive metastatic breast cancer.

[0008] The prevalence of HER2-associated cancers and the high risk of resistance to current therapies highlight the importance of developing new therapeutics to treat these cancers. Summary of the Invention [Means for solving the problem]

[0009] The present invention relates to antibodies or antigen-binding fragments that bind to HER2. Heavy and light chain polypeptides are also disclosed. The antibodies, antigen-binding fragments, and polypeptides may be provided in isolated and / or purified form and may be formulated into compositions suitable for use in research, therapy, and diagnosis.

[0010] In one aspect of the present invention, an antibody or antigen-binding fragment is provided, the amino acid sequence of which is selected from the group consisting of amino acid sequences i) to iii), or amino acid sequences iv) to vi), or preferably amino acid sequences i) to vi): i) LC-CDR1: GLSSGSVSTX1X2YX3S (SEQ ID NO: 9); ii) LC-CDR2: X4TNX5RSS (SEQ ID NO: 10); iii) LC-CDR3: X6LYX7GX8GIX9V (SEQ ID NO: 11); iv) HC-CDR1:X 10 X 11 X 12 X 13 X 14 X 15 X 16 (SEQ ID NO: 12); v) HC-CDR2:X 17 X 18 X 19 X 20 X 21 X 22 GX 23 TX 24 YX 25 X 26 X 27 X 28 X 29 X 30 (SEQ ID NO: 13); vi) HC-CDR3:X 31 X 32 X 33 X 34 X 35 SX 36 X 37 YX 38 X 39 X 40 X 41 X 42 X 43 (SEQ ID NO: 14); Alternatively, variants thereof, in which one, two or three amino acids in one or more of sequences (i) to (vi) are substituted with another amino acid (wherein X1 = G or S, X2 = H or Y, X3 = A or P, X4 = N, S or T, X5 = T, S or I, X6 = V or M, X7 = V or M, X8 = D or S, X9 = W or S, and X 10 = S or absent, X 11 = S or absent, X 12 = S or G, X 13 = Y or N, and X 14 = Y or W, and X 15 = I or W, X 16 = H, G or S, and X 17 = I or absent, X 18 = I, S or E, and X 19 = N or I, and X 20 = P, Y or N, and X 21 = G, Y or H, and X 22 = N or S, and X 23 = D or S, X 24 = N or Y, and X 25 = A or N, X 26 = Q or P, and X 27 = R or S, and X 28 = F or L, and X 29 = Q or K, and X 30 = G or S, and X 31 =E or absent, X 32 = I, Y or M, and X 33= A or G, and X 34 = S, P, I or A, and X 35 = Y, D or N, and X 36 = G or S, and X 37 = G or S, and X 38 = L or absent, X 39 = V or Y, and X 40 = A, D or G, and X 41 = F, M or Y, and X 42 =D or absent, X 43 = I, V, or absent) may include:

[0011] In some embodiments, LC-CDR1 is GLSSGSVTGHYAS (SEQ ID NO: 15), GLSSGSVTGYYPS (SEQ ID NO: 21), or GLSSGSVST SYYPS (SEQ ID NO: 27).

[0012] In some embodiments, LC-CDR2 is one of NTNTRSS (SEQ ID NO: 16), STNSRSS (SEQ ID NO: 22), TTNIRSS (SEQ ID NO: 28), or STNTRSS (SEQ ID NO: 33).

[0013] In some embodiments, the LC-CDR3 is one of VLYVGDGIWV (SEQ ID NO: 17), VLYMGSGISV (SEQ ID NO: 23), MLYMGSGIWV (SEQ ID NO: 29), or VLYMGSGIWV (SEQ ID NO: 34).

[0014] In some embodiments, HC-CDR1 is one of SYYIH (SEQ ID NO: 18), SSSYYWG (SEQ ID NO: 24), GYYWS (SEQ ID NO: 30), or SSNWWS (SEQ ID NO: 35).

[0015] In some embodiments, HC-CDR2 is one of IINPGNGDTNYAQRFQG (SEQ ID NO: 19), SIYYSGSTYYNPSLKS (SEQ ID NO: 25), EINHSGSTNYNPSLKS (SEQ ID NO: 31), or EIYHSGSTNYNPSLKS (SEQ ID NO: 36).

[0016] In some embodiments, the HC-CDR3 is one of EIASYSGSYYDY (SEQ ID NO: 20), YAPDSSGYLVAFDI (SEQ ID NO: 26), MGINSGGYLYGMDV (SEQ ID NO: 32), or MGANSGGYLYGMDV (SEQ ID NO: 37).

[0017] In some embodiments, the antibody, or antigen-binding fragment, has the following CDRs: LC-CDR1: GLSSGSVSTX1X2YX3S (SEQ ID NO: 9), LC-CDR2: X4TNX5RSS (SEQ ID NO: 10), LC-CDR3: X6LYX7GX8GIX9V (SEQ ID NO: 11) (wherein X1 = G or S, X2 = H or Y, X3 = A or P, X4 = N, S or T, X5 = T, S or I, X6 = V or M, X7 = V or M, X8 = D or S, and X9 = W or S). The antibody may comprise at least one light chain variable region incorporating:

[0018] In some embodiments, the antibody, or antigen-binding fragment, has the following CDRs: LC-CDR1: GLSSGSVSTGHYAS (SEQ ID NO: 15), LC-CDR2: NTNTRSS (SEQ ID NO: 16), LC-CDR3: VLYVGDGIWV (SEQ ID NO: 17) The antibody may comprise at least one light chain variable region incorporating:

[0019] In some embodiments, the antibody, or antigen-binding fragment, has the following CDRs: LC-CDR1: GLSSGSVSTGYYPS (SEQ ID NO: 21), LC-CDR2: STNSRSS (SEQ ID NO: 22), LC-CDR3: VLYMGSGISV (SEQ ID NO: 23) The antibody may comprise at least one light chain variable region incorporating:

[0020] In some embodiments, the antibody, or antigen-binding fragment, has the following CDRs: LC-CDR1: GLSSGSVSTSYYPS (SEQ ID NO: 27), LC-CDR2: TTNIRSS (SEQ ID NO: 28), LC-CDR3: MLYMGSGIWV (SEQ ID NO: 29) The antibody may comprise at least one light chain variable region incorporating:

[0021] In some embodiments, the antibody, or antigen-binding fragment, has the following CDRs: LC-CDR1: GLSSGSVSTSYYPS (SEQ ID NO: 27), LC-CDR2: STNTRSS (SEQ ID NO: 33), LC-CDR3: VLYMGSGIWV (SEQ ID NO: 34) The antibody may comprise at least one light chain variable region incorporating:

[0022] In some embodiments, the antibody, or antigen-binding fragment, has the following CDRs: HC-CDR1:X 10 X 11 X 12 X 13 X 14 X 15 X 16 (SEQ ID NO: 12), HC-CDR2:X 17 X 18 X 19 X 20 X 21 X 22 GX 23 TX 24 YX 25 X 26 X 27 X 28 X29 X 30 (SEQ ID NO: 13), HC-CDR3:X 31 X 32 X 33 X 34 X 35 SX 36 X 37 YX 38 X 39 X 40 X 41 X 42 X 43 (SEQ ID NO: 14) (In the formula, X 10 = S or absent, X 11 = S or absent, X 12 = S or G, X 13 = Y or N, and X 14 = Y or W, and X 15 = I or W, X 16 = H, G or S, and X 17 = I or absent, X 18 = I, S or E, and X 19 = N or I, and X 20 = P, Y or N, and X 21 = G, Y or H, and X 22 = N or S, and X 23 = D or S, X 24 = N or Y, and X 25 = A or N, X 26 = Q or P, and X 27 = R or S, and X 28 = F or L, and X 29 = Q or K, and X 30 = G or S, and X 31 =E or absent, X 32 = I, Y or M, and X 33 = A or G, and X 34 = S, P, I or A, and X 35 = Y, D or N, and X 36 = G or S, and X 37 = G or S, and X 38= L or absent, X 39 = V or Y, and X 40 = A, D or G, and X 41 = F, M or Y, and X 42 =D or absent, X 43 = I, V, or absent) The antibody may comprise at least one heavy chain variable region incorporating:

[0023] In some embodiments, the antibody, or antigen-binding fragment, has the following CDRs: HC-CDR1: SYYIH (SEQ ID NO: 18), HC-CDR2: IINPGNGDTNYAQRFQG (SEQ ID NO: 19), HC-CDR3: EIASYSGSYYDY (SEQ ID NO: 20) The heavy chain variable region may comprise at least one heavy chain variable region incorporating:

[0024] In some embodiments, the antibody, or antigen-binding fragment, has the following CDRs: HC-CDR1: SSSYYWG (SEQ ID NO: 24), HC-CDR2: SIYYSGSTYYNPSLKS (SEQ ID NO: 25), HC-CDR3: YAPDSSGYLVAFDI (SEQ ID NO: 26) The antibody may comprise at least one heavy chain variable region incorporating:

[0025] In some embodiments, the antibody, or antigen-binding fragment, has the following CDRs: HC-CDR1: GYYWS (SEQ ID NO: 30), HC-CDR2: EINHSGSTNYNPSLKS (SEQ ID NO: 31), HC-CDR3: MGINSGGYLYGMDV (SEQ ID NO: 32) The heavy chain variable region may comprise at least one heavy chain variable region incorporating:

[0026] In some embodiments, the antibody, or antigen-binding fragment, has the following CDRs: HC-CDR1: SSNWWS (SEQ ID NO: 35), HC-CDR2: EIYHSGSTNYNPSLKS (SEQ ID NO: 36), HC-CDR3: MGANSGGYLYGMDV (SEQ ID NO: 37) The heavy chain variable region may comprise at least one heavy chain variable region incorporating:

[0027] The antibody may comprise at least one light chain variable region (V) comprising an amino acid sequence of one of SEQ ID NOs: 1, 15, 16, 17; or 2, 21, 22, 23; or 3, 27, 28, 29; or 4, 33, 34, 35, or an amino acid sequence having at least 70%, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to one of SEQ ID NOs: 1, 15, 16, 17; or 2, 21, 22, 23; or 3, 27, 28, 29; or 4, 33, 34, 35. L ).

[0028] The antibody may comprise at least one heavy chain variable region (V) comprising an amino acid sequence of one of SEQ ID NOs: 5, 18, 19, 20; or 6, 24, 25, 26; or 7, 30, 31, 32; or 8, 36, 37, 38, or an amino acid sequence having at least 70%, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to one of SEQ ID NOs: 5, 18, 19, 20; or 6, 24, 25, 26; or 7, 30, 31, 32; or 8, 36, 37, 38. H ).

[0029] The antibody may comprise at least one light chain variable region (V) comprising an amino acid sequence of one of SEQ ID NOs: 1, 15, 16, 17; or 2, 21, 22, 23; or 3, 27, 28, 29; or 4, 33, 34, 35, or an amino acid sequence having at least 70%, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to one of SEQ ID NOs: 1, 15, 16, 17; or 2, 21, 22, 23; or 3, 27, 28, 29; or 4, 33, 34, 35. L ), and at least one heavy chain variable region (V) comprising the amino acid sequence of one of SEQ ID NOs: 5, 18, 19, 20; or 6, 24, 25, 26; or 7, 30, 31, 32; or 8, 36, 37, 38, or an amino acid sequence having at least 70%, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to one of SEQ ID NOs: 5, 18, 19, 20; or 6, 24, 25, 26; or 7, 30, 31, 32; or 8, 36, 37, 38. H ).

[0030] The antibodies, antigen-binding fragments, or polypeptides provided herein may optionally bind HER2, optionally human or murine HER2. The antibodies, antigen-binding fragments, or polypeptides may specifically bind to human, rhesus, and / or murine HER2. The antibodies, antigen-binding fragments, or polypeptides may optionally have amino acid sequence components as described above. The antibodies, antigen-binding fragments, or polypeptides may be IgG or may be derived from IgG. The antibodies, antigen-binding fragments, or polypeptides may have a molecular weight of approximately 140-160 kDa, preferably approximately 150 kDa. In one embodiment, an optionally isolated in vitro complex is provided comprising an antibody, antigen-binding fragment, or polypeptide described herein bound to HER2.

[0031] In one aspect of the present invention, an isolated light chain variable region polypeptide is provided, the light chain variable region polypeptide comprising the following CDRs: LC-CDR1: GLSSGSVSTX1X2YX3S (SEQ ID NO: 9), LC-CDR2: X4TNX5RSS (SEQ ID NO: 10), LC-CDR3: X6LYX7GX8GIX9V (SEQ ID NO: 11) wherein X1=G or S, X2=H or Y, X3=A or P, X4=N, S or T, X5=T, S or I, and X6=V or M; X7=V or M, X8=D or S, X9=W or S) Includes.

[0032] In some embodiments, the isolated light chain variable region polypeptide is provided in combination with a heavy chain variable region polypeptide described herein. In some embodiments, LC-CDR1 is one of GLSSGSVSTGHYAS (SEQ ID NO: 15), GLSSGSVSTGYYPS (SEQ ID NO: 21), or GLSSGSVSTSYYPS (SEQ ID NO: 27). In some embodiments, LC-CDR2 is one of NTNTRSS (SEQ ID NO: 16), STNSRSS (SEQ ID NO: 22), TTNIRSS (SEQ ID NO: 28), or STNTRSS (SEQ ID NO: 33). In some embodiments, LC-CDR3 is one of VLYVGDGIWV (SEQ ID NO: 17), VLYMGSGISV (SEQ ID NO: 23), MLYMGSGIWV (SEQ ID NO: 29), or VLYMGSGIWV (SEQ ID NO: 34). In some embodiments, the isolated light chain variable region polypeptide is capable of binding to HER2.

[0033] In one aspect of the present invention, a light chain variable region polypeptide is provided, comprising an amino acid sequence having at least 85% sequence identity to a light chain sequence: SEQ ID NO: 1, 2, 3, or 4 (FIGS. 1A-1D). In some embodiments, the isolated light chain variable region polypeptide is capable of binding to HER2.

[0034] In one aspect of the present invention, an isolated heavy chain variable region polypeptide is provided, the heavy chain variable region polypeptide comprising the following CDRs: HC-CDR1:X 10 X 11 X 12 X 13 X 14 X 15 X 16 (SEQ ID NO: 12), HC-CDR2:X 17 X 18 X 19 X 20 X 21 X 22 GX 23 TX 24 YX 25 X 26 X 27 X 28 X 29 X 30 (SEQ ID NO: 13), HC-CDR3:X 31 X 32 X 33 X 34 X 35 SX 36 X 37 YX 38 X 39 X 40 X 41 X 42 X 43 (SEQ ID NO: 14) (In the formula, X 10 = S or absent, X 11 = S or absent, X 12 = S or G, X 13 = Y or N, and X 14 = Y or W, and X 15 = I or W, X 16 = H, G or S, and X 17 = I or absent, X 18 = I, S or E, and X 19 = N or I, and X 20 = P, Y or N, and X 21 = G, Y or H, and X 22= N or S, and X 23 = D or S, X 24 = N or Y, and X 25 = A or N, X 26 = Q or P, and X 27 = R or S, and X 28 = F or L, and X 29 = Q or K, and X 30 = G or S, and X 31 =E or absent, X 32 = I, Y or M, and X 33 = A or G, and X 34 = S, P, I or A, and X 35 = Y, D or N, and X 36 = G or S, and X 37 = G or S, and X 38 = L or absent, X 39 = V or Y, and X 40 = A, D or G, and X 41 = F, M or Y, and X 42 =D or absent, X 43 = I, V, or absent) Includes.

[0035] In some embodiments, the isolated heavy chain variable region polypeptide is provided in combination with a light chain variable region polypeptide described herein. In some embodiments, HC-CDR1 is one of SYYIH (SEQ ID NO: 18), SSSYYWG (SEQ ID NO: 24), GYYWS (SEQ ID NO: 30), or SSNWWS (SEQ ID NO: 35). In some embodiments, HC-CDR2 is one of IINPGNGDTNYAQRFQG (SEQ ID NO: 19), SIYYSGSTYYNPSLKS (SEQ ID NO: 25), EINHSGSTNYNPSLKS (SEQ ID NO: 31), or EIYHSGSTNYNPSLKS (SEQ ID NO: 36). In some embodiments, HC-CDR3 is one of IINPGNGDTNYAQRFQG (SEQ ID NO: 19), SIYYSGSTYYNPSLKS (SEQ ID NO: 25), EINHSGSTNYNPSLKS (SEQ ID NO: 31), or EIYHSGSTNYNPSLKS (SEQ ID NO: 36). is one of EIASYSGSYYDY (SEQ ID NO: 20), YAPDSSGYLVAFDI (SEQ ID NO: 26), MGINSGGYLYGMDV (SEQ ID NO: 32), or MGANSGGYLYGMDV (SEQ ID NO: 37). In some embodiments, the isolated heavy chain variable region polypeptide is capable of binding to HER2.

[0036] In one aspect of the invention, an isolated heavy chain variable region polypeptide is provided, comprising an amino acid sequence having at least 85% sequence identity to the heavy chain sequence of SEQ ID NO: 5, 6, 7, or 8 (Figures 2A-2D). In some embodiments, the isolated heavy chain variable region polypeptide is capable of binding to HER2.

[0037] In one embodiment of the invention, an antibody, or antigen-binding fragment, is provided, the antibody, or antigen-binding fragment, comprising heavy and light chain variable region sequences, wherein: The light chains each have one of LC-CDR1:GLSSGSVSTX1X2YX3S (SEQ ID NO: 9), GLSSGSVSTGHYAS (SEQ ID NO: 15), GLSSGSVSTGYYPS (SEQ ID NO: 21), or GLSSGSVSTSYYPS (SEQ ID NO: 27), LC-CDR2:X4TNX5RSS (SEQ ID NO: 10), NTNTRSS (SEQ ID NO: 16), STNSRSS (SEQ ID NO: 22), TTNIRSS (SEQ ID NO: 28), or STNTRSS (SEQ ID NO: 33), and LC-CDR3:X6LYX7GX8GIX9V (SEQ ID NO: 11), VLYVGDGIWV (SEQ ID NO: 17), VLYMGSGISV (SEQ ID NO: 23), MLYMGSGIWV (SEQ ID NO: 29), or VLYMGSGIWV (SEQ ID NO: 34). (wherein X1 = G or S, X2 = H or Y, X3 = A or P, X4 = N, S or T, X5 = T, S or I, X6 = V or M, X7 = V or M, X8 = D or S, and X9 = W or S). LC-CDR1, LC-CDR2, and LC-CDR3 have at least 85% overall sequence identity to The heavy chains are each HC-CDR1:X10 X 11 X 12 X 13 X 14 X 15 X 16 (SEQ ID NO: 12), SYYIH (SEQ ID NO: 18), SSSYYWG (SEQ ID NO: 24), GYYWS (SEQ ID NO: 30), or SSNWWS (SEQ ID NO: 35), HC-CDR2:X 17 X 18 X 19 X 20 X 21 X 22 GX 23 TX 24 YX 25 X 26 X 27 X 28 X 29 X 30 (SEQ ID NO: 13), IINPGNGDTNYAQRFQG (SEQ ID NO: 19), SIYYSGSTYYNPSLKS (SEQ ID NO: 25), EINHSGSTNYNPSLKS (SEQ ID NO: 31), or EIYHSGSTNYNPSLKS (SEQ ID NO: 36), and one of HC-CDR3:X 31 X 32 X 33 X 34 X 35 SX 36 X 37 YX 38 X 39 X 40 X 41 X 42 X 43 (SEQ ID NO: 14), EIASYSGSYYDY (SEQ ID NO: 20), YAPDSSGYLVAFDI (SEQ ID NO: 26), MGINSGGYLYGMDV (SEQ ID NO: 32), or MGANSGGYLYGMDV (SEQ ID NO: 37) (In the formula, X 10 = S or absent, X 11 = S or absent, X 12 = S or G, X 13 = Y or N, and X 14 = Y or W, and X 15 = I or W, X 16 = H, G or S, and X 17= I or absent, X 18 = I, S or E, and X 19 = N or I, and X 20 = P, Y or N, and X 21 = G, Y or H, and X 22 = N or S, and X 23 = D or S, X 24 = N or Y, and X 25 = A or N, X 26 = Q or P, and X 27 = R or S, and X 28 = F or L, and X 29 = Q or K, and X 30 = G or S, and X 31 =E or absent, X 32 = I, Y or M, and X 33 = A or G, and X 34 = S, P, I or A, and X 35 = Y, D or N, and X 36 = G or S, and X 37 = G or S, and X 38 = L or absent, X 39 = V or Y, and X 40 = A, D or G, and X 41 = F, M or Y, and X 42 =D or absent, X 43 = I, V, or absent) and HC-CDR1, HC-CDR2, and HC-CDR3, which have at least 85% overall sequence identity to the CDR1, HC-CDR2, and HC-CDR3.

[0038] In some embodiments, the degree of sequence identity may be one of 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0039] In another aspect of the invention, an antibody or antigen-binding fragment capable of binding to HER2, optionally isolated, is provided, comprising heavy and light chain variable region sequences, wherein: the light chain sequence has at least 85% sequence identity to the light chain sequence of SEQ ID NO: 1, 2, 3, or 4 (Figures 1A-1D); The heavy chain sequence has at least 85% sequence identity to the heavy chain sequence of SEQ ID NO: 5, 6, 7, or 8 (Figures 2A-2D).

[0040] In some embodiments, the degree of sequence identity may be one of 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0041] In another aspect of the present invention, the amino acid sequences i) to iii), or the amino acid sequences iv) to vi), or preferably the amino acid sequences i) to vi): i) LC-CDR1: RSSQSLLHSNGFNYLD (SEQ ID NO: 74), RSSQSLLHSDGNKYLD (SEQ ID NO: 84), RSSQSLVYSDGNTYLN (SEQ ID NO: 93), RSSQSLLHSNGYNYLD (SEQ ID NO: 102), RSSQSLLHSNGNTYLD (SEQ ID NO: 110), RASQSVRNNLA (SEQ ID NO: 120), GSTTGAVTSGHYPS (SEQ ID NO: 138), RASQSVSSSYLA (SEQ ID NO: 161), RSSQSLQHSNGYQYLD (SEQ ID NO: 170), TGRSANIGGFDVQ (SEQ ID NO: 180) sequence number 186), ALTSGSVSTSYYPS (SEQ ID NO: 194), TSSQSLVYSDGNTYLN (SEQ ID NO: 211), RSSQSLLRSDGYNFVD (SEQ ID NO: 217), RASQGISSWLA (SEQ ID NO: 226), RASRSVGKYLA (SEQ ID NO: 247), GLSSGSVSTTYYPS (SEQ ID NO: 255), TLRSGINVGTYRIY (SEQ ID NO: 261), RASQSVSSYLA (SEQ ID NO: 270), GLTSGAVSSSYYPS (SEQ ID NO: 279), TGNNNNVGFAGAA (SEQ ID NO: 287); ii) LC-CDR2: one of LGSNRAS (SEQ ID NO: 75), KVSNRDS (SEQ ID NO: 94), SGSNRAS (SEQ ID NO: 111); YASTRAT (SEQ ID NO: 121), STSNKHS (SEQ ID NO: 139), LGSHRAS (SEQ ID NO: 146), LGSNRAP (SEQ ID NO: 155), GASSRAT (SEQ ID NO: 162), LGSFRAS (SEQ ID NO: 171), DNSNRPS (SEQ ID NO: 187), STNLRSS (SEQ ID NO: 195), KVSDRDS (SEQ ID NO: 204), KVSKRDS (SEQ ID NO: 212), LGSDRAS (SEQ ID NO: 218), AASSLQS (SEQ ID NO: 227), YKSDSDKQQGS (SEQ ID NO: 262), DASTRAS (SEQ ID NO: 248), STNTRSS (SEQ ID NO: 33), DASNRAT (SEQ ID NO: 271), NTDIRFS (SEQ ID NO: 280), RNNDRPS (SEQ ID NO: 288); iii) LC-CDR3: MQGLQTPYT (SEQ ID NO: 76), MLGTHWPPMYI (SEQ ID NO: 85), MQGTHWPLT (SEQ ID NO: 95), MQALQTPWT (SEQ ID NO: 103), MQGTHWPPT (SEQ ID NO: 112), QHYGSSRT (SEQ ID NO: 122), MAGLQTPRLT (SEQ ID NO: 130), LLYYGGARV (SEQ ID NO: 140), MQALQTPLT (SEQ ID NO: 147), QQYGSSPRT (SEQ ID NO: 163), MHALSTPPWT (SEQ ID NO: 172), MQGTHWPGT (SEQ ID NO: 180), GTWDSYLNIWV (SEQ ID NO: 188), ELYMGSGISV (SEQ ID NO: 196) ), MQGTHWPQT (SEQ ID NO: 205), MQALQTPRT (SEQ ID NO: 219), QQANSFPPT (SEQ ID NO: 228), QQYGSSSA (SEQ ID NO: 233), MIWHSSAWV (SEQ ID NO: 263), QHYGTSPPFI (SEQ ID NO: 249), VLYMGNGISV (SEQ ID NO: 256), QQRSNWPLT (SEQ ID NO: 272), VLYMGSGISV (SEQ ID NO: 23), SAWDSSLKVQV (SEQ ID NO: 289), iv) HC-CDR1: one of SYGMH (SEQ ID NO: 77), SAAAAWN (SEQ ID NO: 86), SYAMH (SEQ ID NO: 96), SFAMN (SEQ ID NO: 113), SYGIS (SEQ ID NO: 123), SYAMS (SEQ ID NO: 131), SYAIS (SEQ ID NO: 148), TYTMH (SEQ ID NO: 173), GYYWS (SEQ ID NO: 30), SYWIG (SEQ ID NO: 197), NYGMH (SEQ ID NO: 234), SYAIH (SEQ ID NO: 241), SSSYYWG (SEQ ID NO: 24), DYYIH (SEQ ID NO: 281); v) HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), RTYYRSKWYSEYAVSVKS (SEQ ID NO: 87), WINAGNGNTKYSQKFQG (SEQ ID NO: 104), TIGGSGDSTFYADPVKG (SEQ ID NO: 114), WISAYNGNTNYAQKLQG (SEQ ID NO: 124), AISGSGGSTYYADSVKG (SEQ ID NO: 132), GIIPIFGTANYAQKFQG (SEQ ID NO: 149), GINWNGGSTGYADSVKG (SEQ ID NO: 164), WITPGNGNTHYSQNFQG (SEQ ID NO: 165) 174), EINHSGSTNYNPSLKS (SEQ ID NO: 31), IIYPGDSDTRYSPSFQG (SEQ ID NO: 198), AISSNGGSTYYADSVKG (SEQ ID NO: 206), VISYDESNKYYADSVKG (SEQ ID NO: 220), FISYDGTNKYYADSVKG (SEQ ID NO: 235), SIYYSGSTYYNPSLKS (SEQ ID NO: 25), VIWYDGSNKYYADSVKG (SEQ ID NO: 264), SISSSSSSYIYYADSVKG (SEQ ID NO: 273), WVSAYNGDTNYAQKFQG (SEQ ID NO: 282); vi) HC-CDR3: DLFAVVGYYYYYGMDV (SEQ ID NO: 79), GSIFDV (SEQ ID NO: 88), SRGYYGMDV (SEQ ID NO: 97), GGYLVGY (SEQ ID NO: 105), AYGSGGHYFFAY (SEQ ID NO: 115), DWGSSWSDY (SEQ ID NO: 125), TYYDFWSGRVGAFDI (SEQ ID NO: 133), DRGYYGMDV (SEQ ID NO: 141), GRGSGYPDTWFWFDP ​​(SEQ ID NO: 150), SYGSGSYRSHAFDI (SEQ ID NO: 156), GLVPAASMDV (SEQ ID NO: 165), SRVGALDY (SEQ ID NO: 175) , SRGYSGYDN (SEQ ID NO: 181), GLPYYYFDY (SEQ ID NO: 189), LGYGVPLPEYFDL (SEQ ID NO: 199), EPSGSWSYLYYYYYGMDV (SEQ ID NO: 221), HYGDYYYYYGMDV (SEQ ID NO: 236), VYGYGLHYYGMDV (SEQ ID NO: 242), SYDSSGYYYFDY (SEQ ID NO: 250), YAPDSSGYLVAFDI (SEQ ID NO: 26), MTTEDY (SEQ ID NO: 265), DGSAWSRPY (SEQ ID NO: 274), EIASYSGSYYDY (SEQ ID NO: 20), GADWNSDY (SEQ ID NO: 290), Alternatively, there is provided an antibody, antigen-binding fragment or polypeptide capable of binding to HER2, optionally isolated, having a variant thereof in which one, two or three amino acids in one or more of sequences (i) to (vi) are substituted with other amino acids.

[0042] In another aspect of the invention, an antibody, antigen-binding fragment, or polypeptide capable of binding to HER2, optionally isolated, is provided, comprising heavy and light chain variable region sequences, wherein: The light chain contains LC-CDR1: RSSQSLLHSNGFNYLD (SEQ ID NO: 74), RSSQSLLHSDGNKYLD (SEQ ID NO: 84), RSSQSLVYSDGNTYLN (SEQ ID NO: 93), RSSQSLLHSNGYNYLD (SEQ ID NO: 102), RSSQSLLHSNGNTYLD (SEQ ID NO: 110), RASQSVRNNLA (SEQ ID NO: 120), GSTTGAVTSGHYPS (SEQ ID NO: 138), RASQSVSSSYLA (SEQ ID NO: 140). SEQ ID NO: 161), RSSQSLQHSNGYQYLD (SEQ ID NO: 170), TGRSANIGGFDVQ (SEQ ID NO: 186), ALTSGSVSTSYYPS (SEQ ID NO: 194), TSSQSLVYSDGNTYLN (SEQ ID NO: 211), RSSQSLLRSDGYNFVD (SEQ ID NO: 217), RASQGISSWLA (SEQ ID NO: 226), RASRSVGKYLA (SEQ ID NO: 247), GLSSGSVSTTYYPS (SEQ ID NO: 255), TLRSGINVGTYRIY (SEQ ID NO: 261), RASQSVSSYLA (SEQ ID NO: 270), GLTSGAVSSSYYPS (SEQ ID NO: 279), TGNNNNVGFAGAA (SEQ ID NO: 287), LC-CDR2: one of LGSNRAS (SEQ ID NO: 75), KVSNRDS (SEQ ID NO: 94), SGSNRAS (SEQ ID NO: 111); YASTRAT (SEQ ID NO: 121), STSNKHS (SEQ ID NO: 139), LGSHRAS (SEQ ID NO: 146), LGSNRAP (SEQ ID NO: 155), GASSRAT (SEQ ID NO: 162), LGSFRAS (SEQ ID NO: 171), DNSNRPS (SEQ ID NO: 187), STNLRSS (SEQ ID NO: 195), KVSDRDS (SEQ ID NO: 204), KVSKRDS (SEQ ID NO: 212), LGSDRAS (SEQ ID NO: 218), AASSLQS (SEQ ID NO: 227), YKSDSDKQQGS (SEQ ID NO: 262), DASTRAS (SEQ ID NO: 248), STNTRSS (SEQ ID NO: 33), DASNRAT (SEQ ID NO: 271), NTDIRFS (SEQ ID NO: 280), RNNDRPS (SEQ ID NO: 288), LC-CDR3: MQGLQTPYT (SEQ ID NO: 76), MLGTHWPPMYI (SEQ ID NO: 85), MQGTHWPLT (SEQ ID NO: 95), MQALQTPWT (SEQ ID NO: 103), MQGTHWPPT (SEQ ID NO: 112), QHYGSSRT (SEQ ID NO: 122), MAGLQTPRLT (SEQ ID NO: 130), LLYYGGARV (SEQ ID NO: 140), MQALQTPLT (SEQ ID NO: 147), QQYGSSPRT (SEQ ID NO: 163), MHALSTPPWT (SEQ ID NO: 172), MQGTHWPGT (SEQ ID NO: 180), one of GTWDSYLNIWV (SEQ ID NO: 188), ELYMGSGISV (SEQ ID NO: 196), MQGTHWPQT (SEQ ID NO: 205), MQALQTPRT (SEQ ID NO: 219), QQANSFPPT (SEQ ID NO: 228), QQYGSSSA (SEQ ID NO: 233), MIWHSSAWV (SEQ ID NO: 263), QHYGTSPPFI (SEQ ID NO: 249), VLYMGNGISV (SEQ ID NO: 256), QQRSNWPLT (SEQ ID NO: 272), VLYMGSGISV (SEQ ID NO: 23), SAWDSSLKVQV (SEQ ID NO: 289); LC-CDR1, LC-CDR2, and LC-CDR3 have at least 85% overall sequence identity to The heavy chain is one of HC-CDR1: SYGMH (SEQ ID NO: 77), SAAAAWN (SEQ ID NO: 86), SYAMH (SEQ ID NO: 96), SFAMN (SEQ ID NO: 113), SYGIS (SEQ ID NO: 123), SYAMS (SEQ ID NO: 131), SYAIS (SEQ ID NO: 148), TYTMH (SEQ ID NO: 173), GYYWS (SEQ ID NO: 30), SYWIG (SEQ ID NO: 197), NYGMH (SEQ ID NO: 234), SYAIH (SEQ ID NO: 241), SSSYYWG (SEQ ID NO: 24), DYYIH (SEQ ID NO: 281); HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), RTYYRSKWYSEYAVSVKS (SEQ ID NO: 87), WINAGNGNTKYSQKFQG (SEQ ID NO: 104), TIGGSGDSTFYADPVKG (SEQ ID NO: 114), WISAYNGNTNYAQKLQG (SEQ ID NO: 124), AISGSGGSTYYADSVKG (SEQ ID NO: 132), GIIPIFGTANYAQKFQG (SEQ ID NO: 149), GINWNGGSTGYADSVKG (SEQ ID NO: 164), WITPGNGNTHYSQNFQ G (SEQ ID NO: 174), EINHSGSTNYNPSLKS (SEQ ID NO: 31), IIYPGDSDTRYSPSFQG (SEQ ID NO: 198), AISSNGGSTYYADSVKG (SEQ ID NO: 206), VISYDESNKYYADSVKG (SEQ ID NO: 220), FISYDGTNKYYADSVKG (SEQ ID NO: 235), SIYYSGSTYYNPSLKS (SEQ ID NO: 25), VIWYDGSNKYYADSVKG (SEQ ID NO: 264), SISSSSSYIYYADSVKG (SEQ ID NO: 273), WVSAYNGDTNYAQKFQG (SEQ ID NO: 282), HC-CDR3: DLFAVVGYYYYYGMDV (SEQ ID NO: 79), GSIFDV (SEQ ID NO: 88), SRGYYGMDV (SEQ ID NO: 97), GGYLVGY (SEQ ID NO: 105), AYGSGGHYFFAY (SEQ ID NO: 115), DWGSSWSDY (SEQ ID NO: 125), TYYDFWSGRVGAFDI (SEQ ID NO: 133), DRGYYGMDV (SEQ ID NO: 141), GRGSGYPDTWFWFDP ​​(SEQ ID NO: 150), SYGSGSYRSHAFDI (SEQ ID NO: 156), GLVPAASMDV (SEQ ID NO: 165), SRVGALDY (SEQ ID NO: 175), S one of RGYSGYDN (SEQ ID NO: 181), GLPYYYFDY (SEQ ID NO: 189), LGYGVPLPEYFDL (SEQ ID NO: 199), EPSGSWSYLYYYYYGMDV (SEQ ID NO: 221), HYGDYYYYYGMDV (SEQ ID NO: 236), VYGYGLHYYGMDV (SEQ ID NO: 242), SYDSSGYYYFDY (SEQ ID NO: 250), YAPDSSGYLVAFDI (SEQ ID NO: 26), MTTEDY (SEQ ID NO: 265), DGSAWSRPY (SEQ ID NO: 274), EIASYSGSYYDY (SEQ ID NO: 20), GADWNSDY (SEQ ID NO: 290); and HC-CDR1, HC-CDR2, and HC-CDR3, which have at least 85% overall sequence identity to the CDR1, HC-CDR2, and HC-CDR3.

[0043] In some embodiments, the degree of sequence identity may be one of 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0044] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSNGFNYLD (SEQ ID NO: 74), LC-CDR2: LGSNRAS (SEQ ID NO: 75), LC-CDR3: MQGLQTPYT (SEQ ID NO: 76). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYGMH (SEQ ID NO: 77), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: DLFAVVGYYYYYGMDV (SEQ ID NO: 79). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSNGFNYLD (SEQ ID NO: 74), LC-CDR2: LGSNRAS (SEQ ID NO: 75), LC-CDR3: MQGLQTPYT (SEQ ID NO: 76), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYGMH (SEQ ID NO: 77), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: DLFAVVGYYYYYGMDV (SEQ ID NO: 79).

[0045] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSDGNKYLD (SEQ ID NO: 84), LC-CDR2: LGSNRAS (SEQ ID NO: 75), LC-CDR3: MLGTHWPPMYI (SEQ ID NO: 85). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SAAAAWN (SEQ ID NO: 86), HC-CDR2: RTYYRSKWYSEYAVSVKS (SEQ ID NO: 87), HC-CDR3: GSIFDV (SEQ ID NO: 88). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSDGNKYLD (SEQ ID NO: 84), LC-CDR2: LGSNRAS (SEQ ID NO: 75), LC-CDR3: MLGTHWPPMYI (SEQ ID NO: 85), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SAAAAWN (SEQ ID NO: 86), HC-CDR2: RTYYRSKWYSEYAVSVKS (SEQ ID NO: 87), HC-CDR3: GSIFDV (SEQ ID NO: 88).

[0046] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLVYSDGNTYLN (SEQ ID NO: 93), LC-CDR2: KVSNRDS (SEQ ID NO: 94), LC-CDR3: MQGTHWPLT (SEQ ID NO: 95). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SRGYYGMDV (SEQ ID NO: 97). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLVYSDGNTYLN (SEQ ID NO: 93), LC-CDR2: KVSNRDS (SEQ ID NO: 94), LC-CDR3: MQGTHWPLT (SEQ ID NO: 95), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SRGYYGMDV (SEQ ID NO: 97).

[0047] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSNRAS (SEQ ID NO: 75), LC-CDR3: MQALQTPWT (SEQ ID NO: 103). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: WINAGNGNTKYSQKFQG (SEQ ID NO: 104), HC-CDR3: GGYLVGY (SEQ ID NO: 105). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSNRAS (SEQ ID NO: 75), LC-CDR3: MQALQTPWT (SEQ ID NO: 103), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: WINAGNGNTKYSQKFQG (SEQ ID NO: 104), HC-CDR3: GGYLVGY (SEQ ID NO: 105).

[0048] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSNGNTYLD (SEQ ID NO: 110), LC-CDR2: SGSNRAS (SEQ ID NO: 111), LC-CDR3: MQGTHWPPT (SEQ ID NO: 112). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SFAMN (SEQ ID NO: 113), HC-CDR2: TIGGSGDSTFYADPVKG (SEQ ID NO: 114), HC-CDR3: AYGSGGHYFFAY (SEQ ID NO: 115). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSNGNTYLD (SEQ ID NO: 110), LC-CDR2: SGSNRAS (SEQ ID NO: 111), LC-CDR3: MQGTHWPPT (SEQ ID NO: 112), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SFAMN (SEQ ID NO: 113), HC-CDR2: TIGGSGDSTFYADPVKG (SEQ ID NO: 114), HC-CDR3: AYGSGGHYFFAY (SEQ ID NO: 115).

[0049] In some embodiments, the antibody, antigen-binding fragment, or polypeptide comprises the following CDRs: LC-CDR1: RASQSVRNNLA (SEQ ID NO: 120), LC-CDR2: YASTRAT (SEQ ID NO: 121), LC-CDR3: QHYGSSRT (SEQ ID NO: 122). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYGIS (SEQ ID NO: 123), HC-CDR2: WISAYNGNTNYAQKLQG (SEQ ID NO: 124), HC-CDR3: DWGSSWSDY (SEQ ID NO: 125). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RASQSVRNNLA (SEQ ID NO: 120), LC-CDR2: YASTRAT (SEQ ID NO: 121), LC-CDR3: QHYGSSRT (SEQ ID NO: 122), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYGIS (SEQ ID NO: 123), HC-CDR2: WISAYNGNTNYAQKLQG (SEQ ID NO: 124), HC-CDR3: DWGSSWSDY (SEQ ID NO: 125).

[0050] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSNRAS (SEQ ID NO: 75), LC-CDR3: MAGLQTPRLT (SEQ ID NO: 130). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMS (SEQ ID NO: 131), HC-CDR2: AISGSGGSTYYADSVKG (SEQ ID NO: 132), HC-CDR3: TYYDFWSGRVGAFDI (SEQ ID NO: 133). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSNRAS (SEQ ID NO: 75), LC-CDR3: MAGLQTPRLT (SEQ ID NO: 130), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMS (SEQ ID NO: 131), HC-CDR2: AISGSGGSTYYADSVKG (SEQ ID NO: 132), HC-CDR3: TYYDFWSGRVGAFDI (SEQ ID NO: 133).

[0051] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: GSTTGAVTSGHYPS (SEQ ID NO: 138), LC-CDR2: STSNKHS (SEQ ID NO: 139), LC-CDR3: LLYYGGARV (SEQ ID NO: 140). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYGMH (SEQ ID NO: 77), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: DRGYYGMDV (SEQ ID NO: 141). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: GSTTGAVTSGHYPS (SEQ ID NO: 138), LC-CDR2: STSNKHS (SEQ ID NO: 139), LC-CDR3: LLYYGGARV (SEQ ID NO: 140), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYGMH (SEQ ID NO: 77), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: DRGYYGMDV (SEQ ID NO: 141).

[0052] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSHRAS (SEQ ID NO: 146), LC-CDR3: MQALQTPLT (SEQ ID NO: 147). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: HC-CDR1: SYAIS (SEQ ID NO: 148), HC-CDR2: GIIPIFGTANYAQKFQG (SEQ ID NO: 149), HC-CDR3: GRGSGYPDTWFWFDP ​​(SEQ ID NO: 150). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSHRAS (SEQ ID NO: 146), LC-CDR3: MQALQTPLT (SEQ ID NO: 147), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAIS (SEQ ID NO: 148), HC-CDR2: GIIPIFGTANYAQKFQG (SEQ ID NO: 149), HC-CDR3: GRGSGYPDTWFWFDP ​​(SEQ ID NO: 150).

[0053] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSNRAP (SEQ ID NO: 155), LC-CDR3: MQGTHWPLT (SEQ ID NO: 95). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SYGSGSYRSHAFDI (SEQ ID NO: 156). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSNRAP (SEQ ID NO: 155), LC-CDR3: MQGTHWPLT (SEQ ID NO: 95), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SYGSGSYRSHAFDI (SEQ ID NO: 156).

[0054] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RASQSVSSSYLA (SEQ ID NO: 161), LC-CDR2: GASSRAT (SEQ ID NO: 162), LC-CDR3: QQYGSSPRT (SEQ ID NO: 163). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMS (SEQ ID NO: 131), HC-CDR2: GINWNGGSTGYADSVKG (SEQ ID NO: 164), HC-CDR3: GLVPAASMDV (SEQ ID NO: 165). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RASQSVSSSYLA (SEQ ID NO: 161), LC-CDR2: GASSRAT (SEQ ID NO: 162), LC-CDR3: QQYGSSPRT (SEQ ID NO: 163), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMS (SEQ ID NO: 131), HC-CDR2: GINWNGGSTGYADSVKG (SEQ ID NO: 164), HC-CDR3: GLVPAASMDV (SEQ ID NO: 165).

[0055] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLQHSNGYQYLD (SEQ ID NO: 170), LC-CDR2: LGSFRAS (SEQ ID NO: 171), LC-CDR3: MHALSTPPWT (SEQ ID NO: 172). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: TYTMH (SEQ ID NO: 173), HC-CDR2: WITPGNGNTHYSQNFQG (SEQ ID NO: 174), HC-CDR3: SRVGALDY (SEQ ID NO: 175). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLQHSNGYQYLD (SEQ ID NO: 170), LC-CDR2: LGSFRAS (SEQ ID NO: 171), LC-CDR3: MHALSTPPWT (SEQ ID NO: 172). It has at least one light chain variable region incorporating DR3:MHALSTPPWT (SEQ ID NO: 172) and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1:TYTMH (SEQ ID NO: 173), HC-CDR2:WITPGNGNTHYSQNFQG (SEQ ID NO: 174), HC-CDR3:SRVGALDY (SEQ ID NO: 175).

[0056] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLVYSDGNTYLN (SEQ ID NO: 93), LC-CDR2: KVSNRDS (SEQ ID NO: 94), LC-CDR3: MQGTHWPGT (SEQ ID NO: 180). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYGMH (SEQ ID NO: 77), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SRGYSGYDN (SEQ ID NO: 181). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLVYSDGNTYLN (SEQ ID NO: 93), LC-CDR2: KVSNRDS (SEQ ID NO: 94), LC-CDR3: MQGTHWPGT (SEQ ID NO: 180), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYGMH (SEQ ID NO: 77), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SRGYSGYDN (SEQ ID NO: 181).

[0057] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: TGRSANIGGFDVQ (SEQ ID NO: 186), LC-CDR2: DNSNRPS (SEQ ID NO: 187), LC-CDR3: GTWDSYLNIWV (SEQ ID NO: 188). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: GYYWS (SEQ ID NO: 30), HC-CDR2: EINHSGSTNYNPSLKS (SEQ ID NO: 31), HC-CDR3: GLPYYYFDY (SEQ ID NO: 189). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: TGRSANIGGFDVQ (SEQ ID NO: 186), LC-CDR2: DNSNRPS (SEQ ID NO: 187), LC-CDR3: GTWDSYLNIWV (SEQ ID NO: 188), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: GYYWS (SEQ ID NO: 30), HC-CDR2: EINHSGSTNYNPSLKS (SEQ ID NO: 31), HC-CDR3: GLPYYYFDY (SEQ ID NO: 189).

[0058] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: ALTSGSVSTSYYPS (SEQ ID NO: 194), LC-CDR2: STNLRSS (SEQ ID NO: 195), LC-CDR3: ELYMGSGISV (SEQ ID NO: 196). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYWIG (SEQ ID NO: 197), HC-CDR2: IIYPGDSDTRYSPSFQG (SEQ ID NO: 198), HC-CDR3: LGYGVPLPEYFDL (SEQ ID NO: 199). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: ALTSGSVSTSYYPS (SEQ ID NO: 194), LC-CDR2: STNLRSS (SEQ ID NO: 195), LC-CDR3: ELYMGSGISV (SEQ ID NO: 196), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYWIG (SEQ ID NO: 197), HC-CDR2: IIYPGDSDTRYSPSFQG (SEQ ID NO: 198), HC-CDR3: LGYGVPLPEYFDL (SEQ ID NO: 199).

[0059] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLVYSDGNTYLN (SEQ ID NO: 93), LC-CDR2: KVSDRDS (SEQ ID NO: 204), LC-CDR3: MQGTHWPQT (SEQ ID NO: 205). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: AISSNGGSTYYADSVKG (SEQ ID NO: 206), HC-CDR3: SRGYYGMDV (SEQ ID NO: 97). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLVYSDGNTYLN (SEQ ID NO: 93), LC-CDR2: KVSDRDS (SEQ ID NO: 204), LC-CDR3: MQGTHWPQT (SEQ ID NO: 205), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: AISSNGGSTYYADSVKG (SEQ ID NO: 206), HC-CDR3: SRGYYGMDV (SEQ ID NO: 97).

[0060] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: TSSQSLVYSDGNTYLN (SEQ ID NO: 211), LC-CDR2: KVSKRDS (SEQ ID NO: 212), LC-CDR3: MQGTHWPLT (SEQ ID NO: 95). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SRGYYGMDV (SEQ ID NO: 97). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: TSSQSLVYSDGNTYLN (SEQ ID NO: 211), LC-CDR2: KVSKRDS (SEQ ID NO: 212), LC-CDR3: MQGTHWPLT (SEQ ID NO: 95), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SRGYYGMDV (SEQ ID NO: 97).

[0061] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLRSDGYNFVD (SEQ ID NO: 217), LC-CDR2: LGSDRAS (SEQ ID NO: 218), LC-CDR3: MQALQTPRT (SEQ ID NO: 219). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDESNKYYADSVKG (SEQ ID NO: 220), HC-CDR3: EPSGSWSYLYYYYYGMDV (SEQ ID NO: 221). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLRSDGYNFVD (SEQ ID NO: 217), LC-CDR2: LGSDRAS (SEQ ID NO: 218), LC-CDR3: MQALQTPRT (SEQ ID NO: 219), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDESNKYYADSVKG (SEQ ID NO: 220), HC-CDR3: EPSGSWSYLYYYYYGMDV (SEQ ID NO: 221).

[0062] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has the following CDRs: LC-CDR1: RASQGISSWLA (SEQ ID NO: 226), LC-CDR2: AASSLQS (SEQ ID NO: 227), LC-CDR3: QQANSFPPT (SEQ ID NO: 228) In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SRGYYGMDV (SEQ ID NO: 97). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RASQGISSWLA (SEQ ID NO: 226), LC-CDR2: AASSLQS (SEQ ID NO: 227), LC-CDR3: QQANSFPPT (SEQ ID NO: 228), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SRGYYGMDV (SEQ ID NO: 97).

[0063] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RASQSVSSSYLA (SEQ ID NO: 161), LC-CDR2: GASSRAT (SEQ ID NO: 162), LC-CDR3: QQYGSSSA (SEQ ID NO: 233). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: NYGMH (SEQ ID NO: 234), HC-CDR2: FISYDGTNKYYADSVKG (SEQ ID NO: 235), HC-CDR3: HYGDYYYYYGMDV (SEQ ID NO: 236). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RASQSVSSSYLA (SEQ ID NO: 161), LC-CDR2: GASSRAT (SEQ ID NO: 162), LC-CDR3: QQYGSSSA (SEQ ID NO: 233), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: NYGMH (SEQ ID NO: 234), HC-CDR2: FISYDGTNKYYADSVKG (SEQ ID NO: 235), HC-CDR3: HYGDYYYYYGMDV (SEQ ID NO: 236).

[0064] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSNRAS (SEQ ID NO: 75), LC-CDR3: MQGTHWPGT (SEQ ID NO: 180). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAIH (SEQ ID NO: 241), HC-CDR2: AISSNGGSTYYADSVKG (SEQ ID NO: 206), HC-CDR3: VYGYGLHYYGMDV (SEQ ID NO: 242). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSNRAS (SEQ ID NO: 75), LC-CDR3: MQGTHWPGT (SEQ ID NO: 180), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAIH (SEQ ID NO: 241), HC-CDR2: AISSNGGSTYYADSVKG (SEQ ID NO: 206), HC-CDR3: VYGYGLHYYGMDV (SEQ ID NO: 242).

[0065] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RASRSVGKYLA (SEQ ID NO: 247), LC-CDR2: DASTRAS (SEQ ID NO: 248), LC-CDR3: QHYGTSPPFI (SEQ ID NO: 249). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: HC-CDR1: SYAIS (SEQ ID NO: 148), HC-CDR2: GIIPIFGTANYAQKFQG (SEQ ID NO: 149), HC-CDR3: SYDSSGYYYFDY (SEQ ID NO: 250). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RASRSVGKYLA (SEQ ID NO: 247), LC-CDR2: DASTRAS (SEQ ID NO: 248), LC-CDR3: QHYGTSPPFI (SEQ ID NO: 249), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAIS (SEQ ID NO: 148), HC-CDR2: GIIPIFGTANYAQKFQG (SEQ ID NO: 149), HC-CDR3: SYDSSGYYYFDY (SEQ ID NO: 250).

[0066] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: GLSSGSVSTTYYPS (SEQ ID NO: 255), LC-CDR2: STNTRSS (SEQ ID NO: 33), LC-CDR3: VLYMGNGISV (SEQ ID NO: 256). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SSSYYWG (SEQ ID NO: 24), HC-CDR2: SIYYSGSTYYNPSLKS (SEQ ID NO: 25), HC-CDR3: YAPDSSGYLVAFDI (SEQ ID NO: 26). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: GLSSGSVSTTYYPS (SEQ ID NO: 255), LC-CDR2: STNTRSS (SEQ ID NO: 33), LC-CDR3: VLYMGNGISV (SEQ ID NO: 256), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SSSYYWG (SEQ ID NO: 24), HC-CDR2: SIYYSGSTYYNPSLKS (SEQ ID NO: 25), HC-CDR3: YAPDSSGYLVAFDI (SEQ ID NO: 26).

[0067] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: TLRSGINVGTYRIY (SEQ ID NO: 261), LC-CDR2: YKSDSDKQQGS (SEQ ID NO: 262), LC-CDR3: MIWHSSAWV (SEQ ID NO: 263). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYGMH (SEQ ID NO: 77), HC-CDR2: VIWYDGSNKYYADSVKG (SEQ ID NO: 264), HC-CDR3: MTTEDY (SEQ ID NO: 265). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: TLRSGINVGTYRIY (SEQ ID NO: 261), LC-CDR2: YKSDSDKQQGS (SEQ ID NO: 262), LC-CDR3: MIWHSSAWV (SEQ ID NO: 263), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYGMH (SEQ ID NO: 77), HC-CDR2: VIWYDGSNKYYADSVKG (SEQ ID NO: 264), HC-CDR3: MTTEDY (SEQ ID NO: 265).

[0068] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RASQSVSSYLA (SEQ ID NO: 270), LC-CDR2: DASNRAT (SEQ ID NO: 271), LC-CDR3: QQRSNWPLT (SEQ ID NO: 272). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMS (SEQ ID NO: 131), HC-CDR2: SISSSSSYIYYADSVKG (SEQ ID NO: 273), HC-CDR3: DGSAWSRPY (SEQ ID NO: 274). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RASQSVSSYLA (SEQ ID NO: 270), LC-CDR2: DASNRAT (SEQ ID NO: 271), LC-CDR3: QQRSNWPLT (SEQ ID NO: 272), R: Has at least one heavy chain variable region incorporating HC-CDR1: SYAMS (SEQ ID NO: 131), HC-CDR2: SISSSSSYIYYADSVKG (SEQ ID NO: 273), HC-CDR3: DGSAWSRPY (SEQ ID NO: 274).

[0069] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: GLTSGAVSSSYYPS (SEQ ID NO: 279), LC-CDR2: NTDIRFS (SEQ ID NO: 280), LC-CDR3: VLYMGSGISV (SEQ ID NO: 23). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: DYYIH (SEQ ID NO: 281), HC-CDR2: WVSAYNGDTNYAQKFQG (SEQ ID NO: 282), HC-CDR3: EIASYSGSYYDY (SEQ ID NO: 20). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: GLTSGAVSSSYYPS (SEQ ID NO: 279), LC-CDR2: NTDIRFS (SEQ ID NO: 280), LC-CDR3: VLYMGSGISV (SEQ ID NO: 23), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: DYYIH (SEQ ID NO: 281), HC-CDR2: WVSAYNGDTNYAQKFQG (SEQ ID NO: 282), HC-CDR3: EIASYSGSYYDY (SEQ ID NO: 20).

[0070] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: TGNNNNVGFAGAA (SEQ ID NO: 287), LC-CDR2: RNNDRPS (SEQ ID NO: 288), LC-CDR3: SAWDSSLKVQV (SEQ ID NO: 289). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAIS (SEQ ID NO: 148), HC-CDR2: GIIPIFGTANYAQKFQG (SEQ ID NO: 149), HC-CDR3: GADWNSDY (SEQ ID NO: 290). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: TGNNNNVGFAGAA (SEQ ID NO: 287), LC-CDR2: RNNDRPS (SEQ ID NO: 288), LC-CDR3: SAWDSSLKVQV (SEQ ID NO: 289), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAIS (SEQ ID NO: 148), HC-CDR2: GIIPIFGTANYAQKFQG (SEQ ID NO: 149), HC-CDR3: GADWNSDY (SEQ ID NO: 290).

[0071] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSNRAP (SEQ ID NO: 155), LC-CDR3: MQGTHWPLT (SEQ ID NO: 95). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SYGSGSYRSHAFDI (SEQ ID NO: 156). In some embodiments, the antibody, antigen-binding fragment, or polypeptide has at least one light chain variable region incorporating the following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSNRAP (SEQ ID NO: 155), LC-CDR3: MQGTHWPLT (SEQ ID NO: 95), and at least one heavy chain variable region incorporating the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SYGSGSYRSHAFDI (SEQ ID NO: 156).

[0072] In another aspect of the invention, there is provided an antibody, antigen-binding fragment, or polypeptide capable of binding to HER2, optionally isolated, comprising heavy and light chain variable region sequences, wherein: the light chain sequence has at least 85% sequence identity to the light chain sequence of SEQ ID NO: 72, 82, 91, 100, 108, 118, 128, 136, 144, 153, 159, 168, 178, 184, 192, 202, 209, 215, 224, 231, 239, 245, 253, 259, 268, 277, 285, or 293; The heavy chain sequence has at least 85% sequence identity to the heavy chain sequence of SEQ ID NO: 73, 83, 92, 101, 109, 119, 129, 137, 145, 154, 160, 169, 179, 185, 193, 203, 210, 216, 225, 232, 240, 246, 254, 260, 269, 278, 286, or 294.

[0073] In some embodiments, the degree of sequence identity may be one of 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0074] In some embodiments, the following CDRs: LC-CDR1: RSSQSLLHSNGFNYLD (SEQ ID NO: 74), RSSQSLLHSDGNKYLD (SEQ ID NO: 84), RSSQSLVYSDGNTYLN (SEQ ID NO: 93), RSSQSLLHSNGYNYLD (SEQ ID NO: 102), RSSQSLLHSNGNTYLD (SEQ ID NO: 110), RASQSVRNNLA (SEQ ID NO: 120), GSTTGAVTSGHYPS (SEQ ID NO: 138), RASQSVSSSYLA (SEQ ID NO: 161), RSSQSLQHSNGYQYLD (SEQ ID NO: 170), TGRSANIGGFDVQ (SEQ ID NO: 186), ALTSGSVSTSYYPS (SEQ ID NO: 194), TSSQSLVYSDGNTYLN (SEQ ID NO: 211), RSSQSLLRSDGYNFVD (SEQ ID NO: 217), RASQGISSWLA (SEQ ID NO: 226), RASRSVGKYLA (SEQ ID NO: 247), GLSSGSVSTTYYPS (SEQ ID NO: 255), TLRSGINVGTYRIY (SEQ ID NO: 261), RASQSVSSYLA (SEQ ID NO: 270), GLTSGAVSSSYYPS (SEQ ID NO: 279), or TGNNNNVGFAGAA (SEQ ID NO: 287); LC-CDR2: one of LGSNRAS (SEQ ID NO:75), KVSNRDS (SEQ ID NO:94), SGSNRAS (SEQ ID NO:111); YASTRAT (SEQ ID NO:121), STSNKHS (SEQ ID NO:139), LGSHRAS (SEQ ID NO:146), LGSNRAP (SEQ ID NO:155), GASSRAT (SEQ ID NO:162), LGSFRAS (SEQ ID NO:171), DNSNRPS (SEQ ID NO:187), STNLRSS (SEQ ID NO:195), KVSDRDS (SEQ ID NO:204), KVSKRDS (SEQ ID NO:212), LGSDRAS (SEQ ID NO:218), AASSLQS (SEQ ID NO:227), YKSDSDKQQGS (SEQ ID NO:262), DASTRAS (SEQ ID NO:248), STNTRSS (SEQ ID NO:33), DASNRAT (SEQ ID NO:271), NTDIRFS (SEQ ID NO:280), or RNNDRPS (SEQ ID NO:288), LC-CDR3: MQGLQTPYT (SEQ ID NO: 76), MLGTHWPPMYI (SEQ ID NO: 85), MQGTHWPLT (SEQ ID NO: 95), MQALQTPWT (SEQ ID NO: 103), MQGTHWPPT (SEQ ID NO: 112), QHYGSSRT (SEQ ID NO: 122), MAGLQTPRLT (SEQ ID NO: 130), LLYYGGARV (SEQ ID NO: 140), MQALQTPLT (SEQ ID NO: 147), QQYGSSPRT (SEQ ID NO: 163), MH ALSTPPWT (SEQ ID NO: 172), MQGTHWPGT (SEQ ID NO: 180), GTWDSYLNIWV (SEQ ID NO: 188), ELYMGSGISV (SEQ ID NO: 196), MQGTHWPQT (SEQ ID NO: 205), MQALQTPRT (SEQ ID NO: 219), QQANSFPPT (SEQ ID NO: 228), QQYGSSSA (SEQ ID NO: 233), MIWHSSAWV (SEQ ID NO: 263), QHYGTSPPFI (SEQ ID NO: 249), VLYMGNGI SV (SEQ ID NO: 256), QQRSNWPLT (SEQ ID NO: 272), VLYMGSGISV (SEQ ID NO: 23), or SAWDSSLKVQV (SEQ ID NO: 289)

[0013] An isolated light chain variable region polypeptide is provided, comprising:

[0075] In some embodiments, the following CDRs: HC-CDR1: one of SYGMH (SEQ ID NO: 77), SAAAAWN (SEQ ID NO: 86), SYAMH (SEQ ID NO: 96), SFAMN (SEQ ID NO: 113), SYGIS (SEQ ID NO: 123), SYAMS (SEQ ID NO: 131), SYAIS (SEQ ID NO: 148), TYTMH (SEQ ID NO: 173), GYYWS (SEQ ID NO: 30), SYWIG (SEQ ID NO: 197), NYGMH (SEQ ID NO: 234), SYAIH (SEQ ID NO: 241), SSSYYWG (SEQ ID NO: 24), or DYYIH (SEQ ID NO: 281); VISYDGSNKYYADSVKG (SEQ ID NO: 78), RTYYRSKWYSEYAVSVKS (SEQ ID NO: 87), WINAGNGNTKYSQKFQG (SEQ ID NO: 104), TIGGSGDSTFYADPVKG (SEQ ID NO: 114), WISAYNGNTNYAQKLQG (SEQ ID NO: 124), AISGSGGSTYYADSVKG (SEQ ID NO: 132), GIIPIFGTANYAQKFQG (SEQ ID NO: 149), GINWNGGSTGYADSVKG (SEQ ID NO: 164), WITPGNGNTHYSQNFQG (SEQ ID NO: 174), EI one of NHSGSTNYNPSLKS (SEQ ID NO: 31), IIYPGDSDTRYSPSFQG (SEQ ID NO: 198), AISSNGGSTYYADSVKG (SEQ ID NO: 206), VISYDESNKYYADSVKG (SEQ ID NO: 220), FISYDGTNKYYADSVKG (SEQ ID NO: 235), SIYYSGSTYYNPSLKS (SEQ ID NO: 25), VIWYDGSNKYYADSVKG (SEQ ID NO: 264), SISSSSSSYIYYADSVKG (SEQ ID NO: 273), or WVSAYNGDTNYAQKFQG (SEQ ID NO: 282); HC-CDR3: DLFAVVGYYYYYGMDV (SEQ ID NO: 79), GSIFDV (SEQ ID NO: 88), SRGYYGMDV (SEQ ID NO: 97), GGYLVGY (SEQ ID NO: 105), AYGSGGHYFFAY (SEQ ID NO: 115), DWGSSWSDY (SEQ ID NO: 125), TYYDFWSGRVGAFDI (SEQ ID NO: 133), DRGYYGMDV (SEQ ID NO: 141), GRGSGYPDTWFWFDP ​​(SEQ ID NO: 150), SYGSGSYRSHAFDI (SEQ ID NO: 156), GLVPAASMDV (SEQ ID NO: 165), SRVGALDY (SEQ ID NO: 175), SR one of GYSGYDN (SEQ ID NO: 181), GLPYYYFDY (SEQ ID NO: 189), LGYGVPLPEYFDL (SEQ ID NO: 199), EPSGSWSYLYYYYYGMDV (SEQ ID NO: 221), HYGDYYYYYGMDV (SEQ ID NO: 236), VYGYGLHYYGMDV (SEQ ID NO: 242), SYDSSGYYYFDY (SEQ ID NO: 250), YAPDSSGYLVAFDI (SEQ ID NO: 26), MTTEDY (SEQ ID NO: 265), DGSAWSRPY (SEQ ID NO: 274), EIASYSGSYYDY (SEQ ID NO: 20), or GADWNSDY (SEQ ID NO: 290);

[0013] An isolated heavy chain variable region polypeptide is provided, comprising:

[0076] In some embodiments, an antibody, antigen-binding fragment, or polypeptide of the invention comprises any six of the CDR sequences described herein. In some embodiments, an antibody, antigen-binding fragment, or polypeptide comprises any three of the light chain ("LC") CDR sequences described herein and / or any three of the heavy chain ("HC") CDR sequences described herein.

[0077] In some embodiments, the antibody, antigen-binding fragment, or polypeptide further comprises a variable region light chain framework sequence between the CDRs according to the formula: LCFR1-CDR1-LCFR2-CDR2-LCFR3-CDR3-LCFR4. The framework sequence may be derived from a human consensus framework sequence.

[0078] In some embodiments, the antibody, antigen-binding fragment, or polypeptide has the formula: HCFR Further comprising a variable region heavy chain framework sequence between the CDRs according to the following: 1-CDR1-HCFR2-CDR2-HCFR3-CDR3-HCFR4. The framework sequence may be derived from a human consensus framework sequence.

[0079] In some embodiments, the antibody, antigen-binding fragment, or polypeptide may further comprise a human constant region selected from, for example, one of IgG1, IgG2, IgG3, and IgG4.

[0080] In some embodiments, the antibody, antigen-binding fragment, or polypeptide may further comprise a murine constant region selected from, for example, one of IgG1, IgG2A, IgG2B, and IgG3.

[0081] In another aspect of the invention, there is provided an antibody, antigen-binding fragment, or polypeptide conjugated to a drug moiety or a detectable moiety. The drug moiety may be an anti-cancer drug moiety.

[0082] In another aspect of the present invention, a composition is provided. The composition may be, for example, a pharmaceutical composition or a medicament. The composition may comprise an antibody, antigen-binding fragment, or polypeptide described herein. The composition may comprise an antibody, antigen-binding fragment, or polypeptide described herein and at least one pharmaceutically acceptable carrier, excipient, adjuvant, or diluent. The composition may comprise an antibody, antigen-binding fragment, or polypeptide described herein and at least one anti-cancer agent, optionally together with at least one pharmaceutically acceptable carrier, excipient, adjuvant, or diluent.

[0083] In another aspect of the present invention, there is provided an in vitro complex comprising an antibody, or antigen-binding fragment, or polypeptide described herein that binds to HER2. The in vitro complex may be isolated.

[0084] In any aspect of the invention, the antibodies, antigen-binding fragments, or polypeptides described herein can induce cell death by cell membrane disruption. In any aspect of the invention, the antibodies, antigen-binding fragments, or polypeptides described herein can induce cell death by apoptosis. The apoptotic process can proceed through the caspase 3 / 7 pathway and optionally involve PARP cleavage. The antibodies, antigen-binding fragments, or polypeptides described herein can induce activation of caspase 3 / 7 to induce PARP cleavage. Cell death caused by cell membrane disruption or apoptosis can be induced by the antibodies, antigen-binding fragments, or polypeptides described herein that bind to HER2.

[0085] In another aspect of the invention, there is provided an optionally isolated nucleic acid encoding an antibody, antigen-binding fragment, or polypeptide described herein. The nucleic acid may comprise one or more of SEQ ID NOs: 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, or 69, or a coding sequence that is degenerate as a result of the genetic code, or may have a nucleotide sequence having at least 70% identity thereto, and optionally 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity thereto. The nucleic acids are selected from the group consisting of SEQ ID NOs: 70, 71, 80, 81, 89, 90, 98, 99, 106, 107, 116, 117, 126, 127, 134, 135, 142, 143, 151, 152, 157, 158, 166, 167, 176, 177, 182, 183, 190, 191, 200, 201, 207, 208, 213, 214, 222, 223, 229, 230, 237, 238, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 300, 301, 302, 303, 304, 305, 306, 30 3, 244, 251, 252, 257, 258, 266, 267, 275, 276, 283, 284, 291, or 292, or coding sequences that are degenerate as a result of the genetic code, or may have a nucleotide sequence having at least 70% identity thereto, and optionally 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity thereto. A coding sequence that is degenerate as a result of the genetic code refers to a coding sequence that encodes an equivalent polypeptide sequence due to the redundancy of the genetic code.

[0086] In one aspect of the present invention, a vector is provided that contains the nucleic acid described herein. In another aspect of the present invention, a cell is provided that contains the vector. For example, the cell can be eukaryotic or mammalian, such as Chinese hamster ovary (CHO), human or non-human, or prokaryotic, such as Escherichia coli (E. coli).

[0087] In one aspect of the invention, there is provided a method of making an antibody, or antigen-binding fragment, or polypeptide described herein, the method comprising culturing a cell described herein under conditions suitable for expression of the antibody, or antigen-binding fragment, or polypeptide from a nucleic acid or vector described herein.

[0088] In another aspect of the invention, there is provided an antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, cell, or composition described herein for use in therapy or in a method of medical treatment. The antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, cell, or composition described herein may be used in a method of medical treatment or prophylaxis.

[0089] In another aspect of the present invention, an antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, cell, or composition described herein is provided for use in the treatment of cancer, e.g., an antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, cell, or composition described herein is provided for use in a method of treating or preventing cancer. In some embodiments, the cancer is a HER2-positive (HER2+) cancer. In some embodiments, the cancer comprises HER2-positive tumor cells. In some embodiments, the cancer comprises a HER2-positive tumor. In some embodiments, the treatment comprises administering the antibody, antigen-binding fragment, polypeptide, or composition in combination with a therapeutic agent. In some embodiments, the therapeutic agent is an anti-cancer agent. In some embodiments, the therapeutic agent is an anti-HER2 antibody. In some embodiments, the therapeutic agent may be trastuzumab or pertuzumab.

[0090] In another aspect of the present invention, there is provided a use of an antibody, antigen-binding fragment, polypeptide, or composition described herein in the manufacture of a medicament for use in treating cancer. In some embodiments, the cancer is a HER2-positive (HER2+) cancer. In some embodiments, the cancer comprises HER2-positive tumor cells. In some embodiments, the cancer comprises a HER2-positive tumor.

[0091] In another aspect of the present invention, methods of treating cancer are provided, the methods comprising administering to a patient suffering from cancer an antibody, antigen-binding fragment, polypeptide, or composition described herein. In some embodiments, the cancer is a HER2-positive (HER2+) cancer. In some embodiments, the cancer comprises HER2-positive tumor cells. In some embodiments, the cancer comprises a HER2-positive tumor. In some embodiments, the method comprises administering the antibody, antigen-binding fragment, polypeptide, or composition in combination with a therapeutic agent. In some embodiments, the therapeutic agent is an anti-cancer agent. In some embodiments, the therapeutic agent targets HER2. In some embodiments, the therapeutic agent is an anti-HER2 antibody. In some embodiments, the therapeutic agent may be trastuzumab or pertuzumab.

[0092] In another aspect of the present invention, there is provided a method for inhibiting the growth of tumor cells, the method comprising administering to the cells an antibody, antigen-binding fragment, polypeptide, or composition described herein. The method can be performed in vitro or in vivo. In some embodiments, there is provided a method for inhibiting the growth of tumor cells in a subject, the method comprising administering to the subject a therapeutically effective amount of an antibody, antigen-binding fragment, polypeptide, or composition described herein. Also provided is an antibody, antigen-binding fragment, polypeptide, or composition described herein for use in a method for inhibiting the growth of tumor cells in a subject. In some embodiments, the method for inhibiting the growth of tumor cells comprises administering the antibody, antigen-binding fragment, polypeptide, or composition in combination with a therapeutic agent described herein.

[0093] Also provided are methods of killing tumor cells, comprising administering to the cells an antibody, antigen-binding fragment, polypeptide, or composition described herein. The methods can be performed in vitro or in vivo. In some embodiments, methods of killing tumor cells in a subject are provided, comprising administering to the subject a therapeutically effective amount of an antibody, antigen-binding fragment, polypeptide, or composition described herein. In some embodiments, the method of killing tumor cells comprises administering the antibody, antigen-binding fragment, polypeptide, or composition in combination with a therapeutic agent described herein. Tumor cell death can be seen, for example, as a result of membrane disruption, cell lysis, induction of apoptosis, antibody-dependent cell-mediated cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), or through the action of a drug conjugated to the antibody, antigen-binding fragment, or polypeptide. Also provided are antibodies, antigen-binding fragments, polypeptides, or compositions described herein for use in the methods described herein, e.g., of killing tumor cells in a subject.

[0094] In another aspect of the invention, a method is provided that includes contacting a sample containing or suspected of containing HER2 with an antibody, antigen-binding fragment, or polypeptide described herein, and detecting the formation of a complex of the antibody, antigen-binding fragment, or polypeptide and HER2.

[0095] In another aspect of the invention, a method of diagnosing a disease or condition in a subject is provided, the method comprising contacting in vitro a sample from the subject with an antibody, antigen-binding fragment, or polypeptide described herein, and detecting the formation of a complex of the antibody, antigen-binding fragment, or polypeptide and HER2.

[0096] In another aspect of the invention, a method is provided for selecting or stratifying a subject for treatment with a HER2-targeted agent, the method comprising contacting in vitro a sample from the subject with an antibody, antigen-binding fragment, or polypeptide described herein, and detecting formation of a complex of the antibody, or antigen-binding fragment, and HER2.

[0097] In a further aspect of the present invention, there is provided the use of an antibody, antigen-binding fragment, or polypeptide described herein for the detection of HER2 in vitro. In another aspect of the present invention, there is provided the use of an antibody, antigen-binding fragment, or polypeptide described herein as an in vitro diagnostic agent.

[0098] In the methods of the invention, the antibody, antigen-binding fragment or polypeptide is prepared using the compositions described herein. It may also be provided as a composition. The present invention also provides a chimeric antigen receptor (CAR) comprising an antibody, antigen-binding fragment or polypeptide according to the invention.

[0099] The present invention also provides an in vitro complex, optionally isolated, comprising a CAR according to the present invention bound to HER2. The present invention also provides an optionally isolated nucleic acid encoding a CAR according to the present invention.

[0100] The present invention also provides an expression vector comprising a nucleic acid according to the present invention. The present invention also provides a cell comprising a CAR, a nucleic acid, or an expression vector according to the present invention.

[0101] The present invention also provides a composition comprising a CAR, a nucleic acid, an expression vector, or a cell according to the present invention. The present invention also provides a CAR, a nucleic acid, an expression vector, a cell, or a composition according to the present invention for use in a method of medical treatment or prevention.The present invention also provides a CAR, a nucleic acid, an expression vector, a cell, or a composition according to the present invention for use in a method of treating or preventing cancer.

[0102] In any aspect or embodiment, the antibody may be clone P1A3, P1C5, P1E4, or P1F1 described herein. In any aspect or embodiment, the antibody may be clone PFA4, PFB4, PFB5, PFC3, PFD4, PFE1, PFF5, or PFG3 described herein. In any aspect or embodiment, the antibody may be clone PFA1, PFA2, PFA5, PFB1, PFB2, PFB3, PFC2, PFC4, PFD1, PFD2, PFD3, PFE2, PFE5, PFF2, PFF3, PFF4, PFG1, PFG2, PFG4, or PFG5 described herein.

[0103] Embodiments and experiments illustrating the principles of the present invention will now be discussed with reference to the accompanying figures. [Brief explanation of the drawings]

[0104] [Figure 1] Light chain variable domain amino acid sequences for anti-HER2 antibody clones (A) P1A3, (B) P1C5, (C) P1E4, and (D) P1F1. CDRs are underlined. [Figure 2] Heavy chain variable domain amino acid sequences for anti-HER2 antibody clones (A) P1A3, (B) P1C5, (C) P1E4, and (D) P1F1. CDRs are underlined. [Figure 3] Binding to immobilized HER2 protein by bacterially produced crude Fab selected from an antibody phage display library. [Figure 4]Binding of P1A3, P1C5, P1E4, and P1F1 to HER-2, EGFR, and HER-3 immobilized on a microchip in an SPR assay. Trastuzumab was used as a HER2-positive binder control. [Figure 5] Binding of P1A3, P1C5, P1E4, and P1F1 to HER-2 overexpressing (A), low expressing (B), or non-expressing (C) cells. Trastuzumab was used as a positive binder control, and secondary antibody alone as a negative control. [Figure 6] Dose effect of P1A3, P1C5, P1E4, and P1F1 binding to HER-2 overexpressing cells SK-BR-3 or low-expressing cells BT-20 and MCF-7. Mean fluorescence intensity (MFI) is shown after incubation of cells in the presence of various concentrations of antibody. Binding was revealed by the use of a fluorochrome-labeled secondary antibody; trastuzumab was used as a positive binding control. [Figure 7] Binding of trastuzumab, P1A3, P1C5, P1E4, and P1F1 to HER-2 bound to immobilized antibodies. P1A3, P1C5, P1E4, and P1F1 bind to HER-2 bound to trastuzumab (first panel). Trastuzumab binds to HER-2 bound to one of SIgN's antibodies (all other panels). [Figure 8] Propidium iodide staining of BT-474 cells after incubation in the presence of lysis buffer (lysis positive control), trastuzumab, isotype control, or P1A3, P1C5, P1E4, or P1F1. Propidium iodide staining is evidence of cell lysis. [Figure 9] Apoptosis induced by P1E4. Propidium iodide and annexin-V staining of BT-474 cells (A) and SK-BR-3 cells (B) after incubation in the presence of P1E4 or trastuzumab. [Figure 10]Induction of apoptosis by P1E4 via caspase 3 / 7 and cleaved PARP. (A) Propidium iodide uptake after incubation of BT-474 cells in the presence of trastuzumab, pertuzumab, or P1E4. (B) Activation of caspase 3 / 7 after incubation of BT-474 cells in the presence of trastuzumab, pertuzumab, or P1E4. (C) Levels of cleaved PARP in BT-474 cells after incubation with trastuzumab, pertuzumab, or P1E4. [Figure 11] Kinetics of cell lysis after incubation of BT-474 cells in the presence of P1E4 or trastuzumab. Mean percentage of propidium iodide-positive cells ± SD is shown. [Figure 12] Figure 1 shows the cytostatic effect of P1E4, trastuzumab, and pertuzumab on B-474 tumor cell growth after incubation with 1 μg / mL, 5 μg / mL, or 25 μg / mL of antibody. MDA-MB-468 cell growth was used as a negative control. Shown is the mean cell growth ± SD compared to the growth of BT-474 cells in the absence of any treatment. [Figure 13] Cytostatic effect of the combination of P1E4, trastuzumab, and pertuzumab compared to single antibodies on B-474 tumor cell growth after incubation with 1 μg / mL, 5 μg / mL, or 25 μg / mL of antibody (A). MDA-MB-468 (HER-2 negative cells) cell growth was used as a negative control (B). Shown is the mean cell growth ± SD compared to the growth of BT-474 cells in the absence of any treatment. [Figure 14] ADCC induced by P1A3, P1C5, P1E4, and P1F1 in BT-474 cells compared to trastuzumab. Cells were incubated at an effector-to-target ratio of 10:1 in the presence of various antibody concentrations. Mean cell death ± SD is shown. [Figure 15]Tumor control and regression by P1E4 and trastuzumab in nude mice injected with BT-474 cells and treated with various doses of antibody. Mean tumor volume ± SEM of five mice per group is shown. The black line represents tumor size at the start of antibody treatment. [Figure 16] Long-term effective tumor control using anti-HER2 antibody combinations. Nude mice injected with HER-2+BT-474 cells were treated with 10 mg / kg antibody in combination with P1E4, trastuzumab, or pertuzumab antibodies. Mean tumor volume ± SEM for five mice per group is shown. The dotted black line represents tumor size at the start of antibody treatment. [Figure 17] Schematic of the construction of the anti-HER2 CAR showing the signal peptide (sp), anti-HER2 scFv, IgH hinge, CD28 transmembrane domain, and 4-1BB and CD3ζ domain signaling tails. [Figure 18] Graphs showing the characteristics of anti-HER2 CAR T cells, including clones A4, B4, B5, D4, E1, F5, and G3. (18A) Percent cytolysis of BT474 human breast cancer cells by CAR T cells. (18B) Cytokine (IFNγ and IL-2) production by CAR T cells. [Figure 19] Schematic diagram of anti-HER2 CAR mRNA. [Figure 20] Graphs showing characteristics of anti-HER2 CAR T cells containing clone C3. (20A) Percent cytolysis of BT474 human breast cancer cells by CAR T cells. (20B) Cytokine (IFNγ and IL-2) production by CAR T cells. DETAILED DESCRIPTION OF THE INVENTION

[0105] Antibodies and antigen-binding molecules The present invention provides antibodies and antigen-binding molecules capable of binding to HER2. The antibody of the present invention may optionally have a K in the range of 4 to 23 nM. D and preferably binds to HER2 (antigen), preferably human or murine HER2.

[0106] The antibodies according to the present invention may be provided in isolated form. The antibodies may be provided in substantially purified form. The antibody according to the invention may exhibit at least one of the following properties: a) a K of 1 μM or less, preferably ≦100 nM, ≦75 nM, ≦50 nM, ≦40 nM, ≦30 nM, ≦20 nM, ≦15 nM, ≦12.5 nM, ≦10 nM, ≦9 nM, ≦8 nM, ≦7 nM, ≦6 nM, ≦5 nM, ≦4 nM, ≦3 nM, ≦2 nM, ≦1 nM, ≦500 pM D binds to human HER2 (e.g., as determined by SPR); b) binds to HER2-overexpressing or HER2-expressing cells, and optionally does not bind to HER2-negative cells; c) binds to an epitope on HER2, optionally human HER2, that is different from the epitope on HER2 bound by trastuzumab (Herceptin®); d) inducing cell death of HER2-expressing cells, optionally by disruption of the cell membrane; e) inducing cell death of HER2-expressing cells, optionally by activation of the caspase 3 / 7 pathway and apoptosis through activation of cleaved PARP; f) induces cell death in HER2-expressing cells within 40 minutes of application; g) optionally inhibiting tumor growth in vivo; h) optionally in vivo, exhibiting synergistic effects on the inhibition of tumor growth when optionally combined with trastuzumab or pertuzumab.

[0107] By "antibody" we include fragments or derivatives thereof, or synthetic antibodies or synthetic antibody fragments. The fragments and derivatives may be referred to as "antigen-binding molecules" or "antigen-binding fragments."

[0108] "Antibody" or "antigen-binding molecule" refers to a molecule capable of binding to a target antigen, and includes monoclonal antibodies, polyclonal antibodies, monospecific and multispecific antibodies (e.g., bispecific antibodies), as well as antibody fragments (e.g., Fv, scFv, Fab, scFab, F(ab')2, Fab2, diabodies, triabodies, scFv-Fc, minibodies, single-domain antibodies (e.g., VhH), etc.) so long as they bind to the appropriate target molecule(s). The antibodies / antigen-binding molecules of the present invention comprise a portion capable of binding to a target antigen(s). In some embodiments, the portion capable of binding to a target antigen comprises an antibody heavy chain variable region (VH) and an antibody light chain variable region (VL) of an antibody capable of specifically binding to the target antigen. In some embodiments, the moiety capable of binding to a target antigen comprises or consists of an aptamer, e.g., a nucleic acid aptamer, capable of binding to the target antigen (reviewed, e.g., in Zhou and Rossi Nat Rev Drug Discov. 2017 16(3):181-202). In some embodiments, the moiety capable of binding to a target antigen comprises an antigen-binding peptide / polypeptide, e.g., a peptide aptamer, a thioredoxin, a monobody, anticalin, a Kunitz domain, an avimer, a knottin, a fynomer, an atrimer, a DARPin, an affibody, a nanobody (i.e., a single-domain antibody (dsAb)), an affilin, an armadillo repeat protein (ArmRP), an OBody, or the like. or fibronectin, as reviewed, e.g., in Reverdatto et al., Curr Top Med Chem. 2015; 15(12): 1082-1101, which are incorporated herein by reference in their entireties (see also, e.g., Boersma et al., J Biol Chem (2011) 286:41273-85 and Emanuel et al., Mabs (2011) 3:38-48).

[0109] Given current techniques related to monoclonal antibody technology, antibodies against almost any antigen can be prepared. The antigen-binding portion may be a portion of an antibody (e.g., an Fab fragment) or a synthetic antibody fragment (e.g., a single-chain Fv fragment [ScFv]). Suitable monoclonal antibodies against a selected antigen can be prepared by known techniques, such as those disclosed in "Monoclonal Antibodies: A manual of techniques," by H. Zola (CRC Press, 1988) and "Monoclonal Hybridoma Antibodies: Techniques and Applications," by J.G.R. Hurrell (CRC Press, 1982). Chimeric antibodies are discussed by Neuberger et al. (1988, 8th International Biotechnology Symposium Part 2, 792-799).

[0110] Monoclonal antibodies (mAbs) are useful in the methods of the invention and are homogenous populations of antibodies that specifically target a single epitope on an antigen. Polyclonal antibodies are useful in the methods of the invention. Monospecific polyclonal antibodies are preferred. Suitable polyclonal antibodies can be prepared using methods well known in the art.

[0111] Antigen-binding fragments of antibodies, such as Fab and Fab2 fragments, may be used and / or provided, as well as genetically engineered antibodies and antibody fragments. H ) and variable light chain (V L) domains are involved in antigen recognition, a fact first recognized by early protease digestion experiments. Further confirmation was found by the "humanization" of rodent antibodies: variable domains of rodent origin can be fused to constant domains of human origin such that the resulting antibody retains the antigen specificity of the rodent parent (Morrison et al. (1984) Proc. Natl. Acad. Sd. USA 81, 6851-6855).

[0112] As is known from experiments involving bacterial expression of antigen fragments, all of which contain one or more variable domains, the antigen specificity is conferred by the variable domains and is independent of the constant domains. These molecules include Fab-like molecules (Better et al. (1988) Science 240, 1041); Fv molecules (Skerra et al. (1988) Science 240, 1038); V H and V L These include single-chain Fv (ScFv) molecules, in which partner domains are linked via a flexible oligopeptide (Bird et al. (1988) Science 242, 423; Huston et al. (1988) Proc. Natl. Acad. Sd. USA 85, 5879), and single-domain antibodies (dAbs), which comprise isolated V domains (Ward et al. (1989) Nature 341, 544). For a review of the techniques involved in the synthesis of antibody fragments that retain their specific binding sites, see Winter & Milstein (1991) Nature 349, 293-299.

[0113] By "ScFv" we mean a molecule that is H and VL partner domains are covalently linked, for example by a flexible oligopeptide. Fab, Fv, ScFv and dAb fragments may all be expressed in and secreted from E. coli, thus allowing the facile production of large amounts of the said fragments. Synthetic antibodies that bind to HER2 may also be generated using phage display technology, as is well known in the art.

[0114] Antibodies generally contain six complementarity determining regions (CDRs): three in the heavy chain variable (VH) region: HC-CDR1, HC-CDR2, and HC-CDR3, and three in the light chain variable (VL) region: LC-CDR1, LC-CDR2, and LC-CDR3. Together, the six CDRs define the paratope of the antibody, the part of the antibody that binds to the target antigen.

[0115] The VH and VL regions comprise framework regions (FRs) on either side of each CDR, which provide a scaffold for the CDRs. From N- to C-terminus, the VH region comprises the following structure: N-terminus-[HC-FR1]-[HC-CDR1]-[HC-FR2]-[HC-CDR2]-[HC-FR3]-[HC-CDR3]-[HC-FR4]-C-terminus, and the VL region comprises the following structure: N-terminus-[LC-FR1]-[LC-CDR1]-[LC-FR2]-[LC-CDR2]-[LC-FR3]-[LC-CDR3]-[LC-FR4]-C-terminus.

[0116] those described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991); Chothia et al., J. Mol. Biol. 196:901-917 (1987); VBASE2 as described in Retter et al., Nucl. Acids Res. (2005) 33 (suppl 1): D671-D674; and IMGT as described in Lefranc et al., Dev. Comp. Immunol. (2003) 27:55-77. There are several different conventions for defining antibody CDRs and FRs, such as the international IMGT (ImMunoGeneTics) information system (LeFranc et al., Nucleic Acids Res. (2015) 43 (DatABASE issue):D413-22), which uses the V-DOMAIN numbering convention.

[0117] In some embodiments, the antibody / antigen-binding molecule comprises the CDRs of an antigen-binding molecule capable of binding to HER2. In some embodiments, the antibody / antigen-binding molecule comprises the FRs of an antigen-binding molecule capable of binding to HER2. In some embodiments, the antibody-binding molecule comprises the CDRs and FRs of an antigen-binding molecule capable of binding to HER2. That is, in some embodiments, the antigen-binding molecule comprises the VH region and VL region of an antigen-binding molecule capable of binding to HER2.

[0118] Antibodies may be produced by affinity maturation, which produces modified antibodies with improved affinity for the antigen compared to the unmodified parent antibody. Affinity-matured antibodies may be produced by procedures known in the art, such as Mark et al., Rio / Technology 10:779-783 (1992); Barbas et al., Proc Nat. Acad. Sci. USA 91:3809-3813 (1994); Schier et al., Gene 169:147-155 (1995); Yelton et al., J. Immunol. 155:1994-2004 (1995); Jackson et al., J. Immunol. 154(7):331 0-15 9 (1995); and Hawkins et al., J. Mol. Biol. 226:889-896 (1992). An antibody, antigen-binding fragment or polypeptide according to the invention may be subjected to affinity maturation.

[0119] Preferably, the antibody, antigen-binding fragment, or polypeptide according to the present invention exhibits specific binding to HER2. Preferably, the peptide binds the target with higher affinity and / or with longer duration than it binds to other targets.

[0120] Binding of an antibody, antigen-binding fragment, or polypeptide according to the present invention to a predetermined molecule can be determined by various methods, such as ELISA, surface plasmon resonance (SPR, see, e.g., Hearty et al., Methods Mol Biol (2012) 907:411-442), Bio-Layer Interferometry (see, e.g., Lad et al., (2015) J Biomol Screen 20(4): 498-507), flow cytometry, or by techniques well known to those skilled in the art, including radioimmunoassay (RIA) enzyme-linked immunosorbent assay. Through such analyses, binding to a predetermined target can be measured and quantified. In some embodiments, binding may be a response detected in a predetermined assay. The binding affinity of an antibody for its target is often determined by its dissociation constant (K D ) is described.

[0121] The degree of binding of an antibody to an unrelated target can be less than about 10% of the binding of the antibody to the target, as measured, for example, by ELISA, SPR, Bio-Layer interferometry, or by RIA. Alternatively, binding specificity can be determined by the K of an anti-HER2 antibody of the invention directed against another target molecule. D at least 0.1 orders of magnitude (i.e., 0.1 × 10 n , where n is an integer corresponding to the digit) D This may be reflected in terms of the binding affinity for binding to HER2 at 100 kJ / s, which may optionally be at least one of 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, or 2.0.

[0122] The antibodies, antigen-binding fragments, or polypeptides according to the invention have one dissociation constant (K) of ≦25 nM, ≦20 nM, ≦15 nM, ≦10 nM, ≦5 nM, ≦3 nM, ≦2 nM, ≦1.5 nM, ≦1.4 nM, ≦1.3 nM, ≦1.25 nM, ≦1.24 nM, ≦1.23 nM, ≦1.22 nM, ≦1.21 nM, ≦1.2 nM, ≦1.15 nM, ≦1.1 nM, ≦1.05 nM, ≦1 nM, ≦900 pM, ≦800 pM, ≦700 pM, ≦600 pM, or ≦500 pM, as determined by analysis according to SPR, Bio-Layer interferometry, or by RIA. D ) is preferred. D can range from about 0.1 nM to about 3 nM. D can range from about 0.1 nM to about 10 nM. D can range from about 0.1 nM to about 15 nM. D can range from about 0.1 nM to about 25 nM. D can range from about 1 nM to about 100 nM. D can be in the range of about 50 to about 500 nM.

[0123] The antibody, antigen-binding fragment, or polypeptide according to the present invention may exhibit high binding affinity to HER2. In some embodiments, the antibody, antigen-binding fragment, or polypeptide according to the present invention has a dissociation constant (K D ) The antibody, antigen-binding fragment, or polypeptide according to the present invention may exhibit moderate or low binding affinity to HER2. In some embodiments, the antibody, antigen-binding fragment, or polypeptide according to the present invention has a dissociation constant (K D In some embodiments, the antibody, antigen-binding fragment, or polypeptide according to the invention has a dissociation constant (K) of 14 to 310 nM. D In some embodiments, the antibody, antigen-binding fragment, or polypeptide according to the invention has a dissociation constant (K) of 25 to 310 nM. D In some embodiments, the antibody, antigen-binding fragment, or polypeptide according to the invention has a dissociation constant (K) of 150 to 310 nM. D)

[0124] The antibodies, antigen-binding fragments, or polypeptides of the present invention preferably bind to HER2-overexpressing or HER2-expressing cells. The antibodies, antigen-binding fragments, or polypeptides are preferably capable of binding to HER2-overexpressing cells. Preferably, the antibodies, antigen-binding fragments, or polypeptides described herein The binding fragment, or polypeptide, does not bind to HER2-negative cells, ie, cells that do not express HER2.

[0125] Preferably, the antibody, antigen-binding fragment, or polypeptide of the present invention binds to a HER2 epitope different from that bound by trastuzumab (Herceptin®). Trastuzumab has been reported to bind to the C-terminal part of subdomain IV of the HER2 extracellular domain (Cho et al., (2003) Nature 421: 756-760). Epitope binding assays can be performed by methods known in the art, for example, by SPR as described herein (see, for example, Hearty et al., Methods Mol Biol (2012) 907:411-442). Briefly, an antibody of interest is immobilized on an SPR microchip, recombinant HER-2 is applied to bind to the immobilized antibody, and then other antibodies of interest are applied and their binding is measured. Tracing the dendrogram can identify whether the antibody of interest shares a common epitope with trastuzumab (similar binding profile) or binds to a different epitope as described herein (different binding profile). In some embodiments, the antibody, antigen-binding fragment, or polypeptide of the invention preferably binds to a different HER2 epitope than that bound by pertuzumab (Perjeta).

[0126] Preferably, the antibody, antigen-binding fragment, or polypeptide of the present invention induces cell death of HER2-expressing cells. In some embodiments, the antibody, antigen-binding fragment, or polypeptide of the present invention induces cell death by disrupting the cell membrane. Cell death can be measured, for example, by staining with propidium iodide using methods well known in the art and described herein. Insertion of propidium iodide into cells indicates cell lysis.

[0127] In some embodiments, the antibodies, antigen-binding fragments, or polypeptides of the present invention induce cell death of HER2-expressing cells by apoptosis. Apoptosis can be measured using methods well known in the art and described herein, for example, by staining with Annexin-V and propidium iodide. Cells stained with Annexin-V are apoptotic.

[0128] The caspase 3 / 7 pathway is involved in the apoptosis process. After activation, caspase 3 / 7 activates cleaved PARP in the nucleus, which can subsequently induce the release of proapoptotic factors by mitochondria and block the process of DNA repair. In some embodiments, the antibody, antigen-binding fragment, or polypeptide of the present invention induces cell death of HER2-expressing cells by apoptosis via activation of the caspase 3 / 7 pathway and activation of cleaved PARP. Activation of the caspase 3 / 7 pathway and activation of cleaved PARP can be measured using methods well known in the art.

[0129] In some embodiments, the antibodies, antigen-binding fragments, or polypeptides of the invention induce cell death of HER2-expressing cells within 40 minutes of application. Induced cell death can be measured by propidium iodide uptake.

[0130] Preferably, the antibodies, antigen-binding fragments, or polypeptides of the invention inhibit tumor growth. In some embodiments, the inhibition of tumor growth is achieved in vivo. Tumor growth can be measured by methods well known in the art and described herein, for example, using a cell proliferation assay. In some embodiments, the antibodies, antigen-binding fragments, or polypeptides of the invention exhibit a synergistic effect on tumor growth inhibition. In some embodiments, the antibodies, antigen-binding fragments, or polypeptides of the invention exhibit a synergistic effect on tumor growth inhibition when used in combination with trastuzumab and / or pertuzumab. In some embodiments, the antibodies, antigen-binding fragments, or polypeptides of the invention exhibit a synergistic effect on tumor growth inhibition when used in combination with trastuzumab and / or pertuzumab. In some embodiments, the antibodies, antigen-binding fragments, or polypeptides of the invention are capable of inducing death of HER2-expressing cells with efficiency similar to that induced by trastuzumab or pertuzumab. In some embodiments, the antibodies, antigen-binding fragments, or polypeptides of the invention are capable of inducing death of HER2-expressing cells with increased efficiency relative to that induced by trastuzumab and / or pertuzumab. In some embodiments, the antibodies, antigen-binding fragments, or polypeptides of the invention are capable of inducing antibody-dependent cell-mediated cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC) as measured by, for example, LDH release using a Promega Cytotox 96 non-radioactive cytotoxicity assay kit.

[0131] In some embodiments, an antibody, antigen-binding fragment, or polypeptide of the invention is capable of reducing tumor size, e.g., compared to the size of a tumor not treated with the antibody, antigen-binding fragment, or polypeptide of the invention.

[0132] In some embodiments, the antibodies, antigen-binding fragments, or polypeptides of the invention are capable of conferring long-term protection against cancer, cancer cells, tumors, and / or tumor cells after administration of the antibody, antigen-binding fragment, or polypeptide has ceased. The protection conferred may be longer-lasting than other HER2 therapies known in the art, such as trastuzumab or pertuzumab. The present invention provides antigen-binding polypeptides. In aspects of the invention, the antigen-binding polypeptides are capable of binding to HER2. The polypeptides may be provided in isolated or substantially purified form.

[0133] "Antigen-binding polypeptide" refers to a polypeptide capable of binding to a target molecule, and includes monoclonal antibodies, polyclonal antibodies, monospecific and multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, so long as they exhibit binding to the appropriate target molecule(s). Antigen-binding polypeptide, as used herein, also refers to a noncovalent or covalent complex of more than one peptide (e.g., two, three, four, six, or eight polypeptides), such as a bispecific antigen-binding polypeptide comprising two heavy chain polypeptides and two light chain polypeptides.

[0134] The antigen-binding polypeptides described herein preferably exhibit specific binding to an appropriate target (e.g., HER2). As used herein, "specific binding" refers to binding that is selective for the antigen and that is distinguishable from non-specific binding to non-target antigens. Antigen-binding polypeptides that specifically bind to a target molecule preferably bind the target with higher affinity and / or with longer duration than they bind to other non-target molecules.

[0135] In some embodiments, the antigen-binding polypeptide binds to the same or overlapping epitope of a target molecule (i.e., HER2) as a reference antigen-binding polypeptide capable of binding to the target molecule. In some embodiments, the antigen-binding polypeptide exhibits competitive binding with the reference antigen-binding polypeptide capable of binding to the target molecule. Whether a given antigen-binding polypeptide exhibits such competitive binding may be determined by a variety of methods known to those skilled in the art, including competitive ELISA.

[0136] In some embodiments, the antigen-binding polypeptide comprises a complementarity-determining region (CDR) of the antigen-binding polypeptide capable of binding to a target molecule (i.e., HER2). Antibodies generally comprise six CDRs: three in the light chain variable region (VL): LC-CDR1, LC-CDR2, LC-CDR, and three in the heavy chain variable region (VH): HC-CDR1, HC-CDR2, and HC-CDR3. The six CDRs together define the antibody paratope, which is the part of the antibody that binds to the target molecule. Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991), those described in Chothia et al., J. Mol. Biol. 196:901-917 (1987), and VBASE2 as described in Retter et al., Nucl. Acids Res. (2005) 33 (suppl 1): D671-D674. Unless otherwise specified, the CDRs of the antigen-binding polypeptides described herein are defined according to Kabat et al., Sequences of Proteins of Immunological Interest, 5th Edition Defined according to Public Health Service, National Institutes of Health, Bethesda, MD (1991).

[0137] Where a polypeptide comprises more than one domain or region, it will be understood that it is preferred that the multiple domains / regions are present in the same polypeptide chain, i.e., the polypeptide comprises more than one domain or region and is a fusion polypeptide comprising the domains / regions.

[0138] The sequence of a monoclonal antibody (mAb) capable of binding to HER2 may be used to design and prepare antigen-binding polypeptides. Antigen-binding regions of antibodies, such as single-chain variable fragments (scFv), Fab and Fab2 fragments, may also be used / provided. An "antigen-binding region" is any fragment of an antibody capable of binding to a target for which a given antibody is specific.

[0139] In some embodiments, the antigen-binding polypeptide of the present invention is a HER2-binding polypeptide. In some embodiments, the antigen-binding polypeptide comprises or consists of a HER2-binding polypeptide. In some embodiments, the antigen-binding polypeptide comprises a heavy chain variable (VH) region comprising HC-CDR1, HC-CDR2, and HC-CDR3 of a HER2-binding antibody clone described herein, or a variant thereof in which one, two, or three amino acids in one or more of HC-CDR1, HC-CDR2, and HC-CDR3 are substituted with another amino acid. In some embodiments, the antigen-binding polypeptide comprises a light chain variable (VL) region comprising LC-CDR1, LC-CDR2, and LC-CDR3 of a HER2-binding antibody clone described herein, or a variant thereof in which one, two, or three amino acids in one or more of LC-CDR1, LC-CDR2, and LC-CDR3 are substituted with another amino acid. In some embodiments, the antigen-binding polypeptide comprises a VH region comprising HC-CDR1, HC-CDR2 and HC-CDR3 and a VL region comprising LC-CDR1, LC-CDR2 and LC-CDR3 of a HER2-binding antibody clone described herein, or a variant thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, HC-CDR3, LC-CDR1, LC-CDR2 or LC-CDR3 are substituted with another amino acid.

[0140] In some embodiments, the antigen-binding polypeptide comprises a VH region comprising or consisting of an amino acid sequence having at least 70%, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to a VH region of a HER2-binding antibody clone described herein. In some embodiments, the antigen-binding polypeptide comprises or consists of an amino acid sequence having at least 70%, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to a VL region of a HER2-binding antibody clone described herein. In some embodiments, the antigen-binding polypeptide comprises a VH region comprising or consisting of an amino acid sequence having at least 70%, and more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to a VH region of a HER2-binding antibody clone described herein, and a VL region comprising or consisting of an amino acid sequence having at least 70%, and more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to a VL region of a HER2-binding antibody clone described herein.

[0141] In some embodiments, the antigen-binding polypeptide may comprise a variant of a reference VL / VH region comprising, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 substitutions with respect to the amino acid sequence of the reference VL / VH region(s). In some embodiments, the substitution(s) are not in the CDRs. In some embodiments, the substitution(s) are in the framework region(s), i.e., in the amino acid sequence of the VL / VH region(s) other than the CDRs.

[0142] In some embodiments, substitutions are conservative substitutions, for example according to the following table: In some embodiments, amino acids in the same block in the middle column are substituted. In some embodiments, amino acids in the same line in the rightmost column are substituted.

[0143] [Table 1]

[0144] In some embodiments, HER2-binding antigen-binding polypeptides according to the invention comprise or consist of an amino acid sequence having at least 70%, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one of SEQ ID NOs: 1 to 4. In some embodiments, HER2-binding antigen-binding polypeptides according to the invention comprise or consist of an amino acid sequence having at least 70%, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one of SEQ ID NOs: 5 to 8.

[0145] In some embodiments, the HER2-binding antigen-binding polypeptide according to the invention is selected from the group consisting of SEQ ID NOs: 72, 82, 91, 100, 108, 118, 128, 136, 144, 153, 15 9, 168, 178, 184, 192, 202, 209, 215, 224, 231, 239, 245, 253, 259, 268, 277, 285, or 293, having at least 70%, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to one of In some embodiments, a HER2-binding antigen-binding polypeptide according to the invention comprises or consists of an amino acid sequence having at least 70%, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to one of SEQ ID NOs: 73, 83, 92, 101, 109, 119, 129, 137, 145, 154, 160, 169, 179, 185, 193, 203, 210, 216, 225, 232, 240, 246, 254, 260, 269, 278, 286, or 294.

[0146] In some embodiments, HER2-binding antigen-binding polypeptides according to the invention lack the HC-CDR1, HC-CDR2, HC-CDR3, LC-CDR1, LC-CDR2 and LC-CDR3 of one or more of the following clones: P1A3, P1C5, P1E4 and P1F1. In some embodiments, HER2-binding antigen-binding polypeptides according to the invention lack the VL domain sequence and / or VH domain sequence of one or more of said clones.

[0147] The VL and VH regions of the antigen-binding region of an antibody together comprise the Fv region. In some embodiments, the antigen-binding polypeptide according to the invention comprises or consists of an Fv region that binds to HER2.

[0148] In some embodiments, the polypeptide further comprises one or more antibody heavy chain constant regions (CH). In some embodiments, the polypeptide further comprises one or more antibody light chain constant regions (CL). In some embodiments, the polypeptide comprises an immunoglobulin (Ig) CH1, CH2, and / or CH3 region.

[0149] The VL and light chain constant (CL) region, and the VH region and heavy chain constant 1 (VH1) region of the antigen-binding region of an antibody together comprise a Fab region. In some embodiments, the antigen-binding polypeptide of the antigen-binding polypeptides described herein comprises or consists of a Fab region that binds to HER2.

[0150] In some embodiments, the polypeptide comprises one or more regions of an immunoglobulin heavy chain constant sequence. In some embodiments, the polypeptide comprises a CH1 region described herein. In some embodiments, the polypeptide comprises a CH1-CH2 hinge region described herein. In some embodiments, the polypeptide comprises a CH2 region described herein. In some embodiments, the polypeptide comprises a CH3 region described herein.

[0151] In some embodiments, the polypeptide lacks one or more regions of an immunoglobulin heavy chain constant sequence. In some embodiments, the polypeptide lacks a CH2 region. In some embodiments, the polypeptide lacks a CH3 region. In some embodiments, the polypeptide lacks a CH2 region and also lacks a CH3 region.

[0152] In some embodiments, the polypeptide according to the present invention is one of the following: (i) VH (ii) VL (iii) VH-CH1 (iv) VL-CL (v) VL-CH1 (vi) VH-CL (vii) VH-CH1-CH2-CH3 (viii) VL-CL-CH2-CH3 (ix) VL-CH1-CH2-CH3 (x) VH-CL-CH2-CH3 The amino acid sequence includes a structure from N-terminus to C-terminus according to one of the following:

[0153] The present invention also provides antibodies and antibody-binding molecules composed of the polypeptides of the present invention. In some embodiments, the antibody / antigen-binding molecules of the present invention comprise one of the following combinations of polypeptides: (A) VH+VL (B) VH-CH1+VL-CL (C) VL-CH1+VH-CL (D) VH-CH1-CH2-CH3+VL-CL (E) VH-CL-CH2-CH3+VL-CH1 (F) VL-CH1-CH2-CH3+VH-CL (G) VL-CL-CH2-CH3+VH-CH1 (H) VH-CH1-CH2-CH3+VL-CL-CH2-CH3 (I) VH-CL-CH2-CH3+VL-CH1-CH2-CH3 In some embodiments, the antigen-binding molecule comprises more than one of the polypeptide combinations shown in (A) to (I) above. For example, see (D) above. In some embodiments, the antigen-binding molecule comprises two polypeptides comprising the structure VH-CH1-CH2-CH3 and two polypeptides comprising the structure VL-CL.

[0154] In some embodiments, the antigen-binding polypeptides described herein comprise or consist of a whole antibody that binds to HER2. As used herein, "whole antibody" refers to an antibody having a structure substantially similar to that of an immunoglobulin (Ig). The various immunoglobulins and their structures are described, for example, in Schroeder and Cavacini J Allergy Clin Immunol. (2010) 125(202): S41-S52, which is incorporated herein by reference in its entirety.

[0155] Type G immunoglobulins (i.e., IgG) are approximately 150 kDa glycoproteins comprising two heavy chains and two light chains. From the N-terminus to the C-terminus, the heavy chain comprises a VH followed by a heavy chain constant region comprising three constant domains (CH1, CH2, and CH3); similarly, the light chain comprises a VL followed by a CL. Depending on the heavy chain, immunoglobulins can be classified as IgG (e.g., IgG1, IgG2, IgG3, IgG4), IgA (e.g., IgA1, IgA2), IgD, IgE, or IgM. Light chains may be kappa (κ) or lambda (λ). In some embodiments, the antigen-binding polypeptides described herein comprise or consist of IgG (e.g., IgG1, IgG2, IgG3, IgG4), IgA (e.g., IgA1, IgA2), IgD, IgE, or IgM that binds to HER2.

[0156] Antigen-binding polypeptides according to the invention may be provided in any suitable manner. In some aspects, the antibody is clone P1A3, or a variant of P1A3. P1A3 comprises the following CDR sequences: Light chain: LC-CDR1: GLSSGSVSTGHYAS (SEQ ID NO: 15) LC-CDR2: NTNTRSS (SEQ ID NO: 16) LC-CDR3: VLYVGDGIWV (SEQ ID NO: 17) Heavy chain: HC-CDR1:SYYIH (SEQ ID NO: 18) HC-CDR2: IINPGNGDTNYAQRFQG (SEQ ID NO: 19) HC-CDR3: EIASYSGSYYDY (SEQ ID NO: 20) Unless otherwise specified, the CDRs of the antigen-binding polypeptides described herein are defined according to Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991).

[0157] In some aspects, the antibody is clone P1C5, or a variant of P1C5. P1C5 comprises the following CDR sequences: Light chain: LC-CDR1: GLSSGSVSTGYYPS (SEQ ID NO: 21) LC-CDR2: STNSRSS (SEQ ID NO: 22) LC-CDR3: VLYMGSGISV (SEQ ID NO: 23) Heavy chain: HC-CDR1:SSSYYWG (SEQ ID NO: 24) HC-CDR2:SIYYSGSTYYNPSLKS (SEQ ID NO: 25) HC-CDR3: YAPDSSGYLVAFDI (SEQ ID NO: 26) In some aspects, the antibody is clone P1E4, or a variant of P1E4. P1E4 comprises the following CDR sequences: Light chain: LC-CDR1: GLSSGSVSTSYYPS (SEQ ID NO: 27) LC-CDR2: TTNIRSS (SEQ ID NO: 28) LC-CDR3: MLYMGSGIWV (SEQ ID NO: 29) Heavy chain: HC-CDR1: GYYWS (SEQ ID NO: 30) HC-CDR2: EINHSGSTNYNPSLKS (SEQ ID NO: 31) HC-CDR3:MGINSGGYLYGMDV (SEQ ID NO: 32) In some aspects, the antibody is clone P1F1, or a variant of P1F1. P1F1 comprises the following CDR sequences: Light chain: LC-CDR1: GLSSGSVSTSYYPS (SEQ ID NO: 27) LC-CDR2: STNTRSS (SEQ ID NO: 33) LC-CDR3: VLYMGSGIWV (SEQ ID NO: 34) Heavy chain: HC-CDR1:SSNWWS (SEQ ID NO: 35) HC-CDR2: EIYHSGSTNYNPSLKS (SEQ ID NO: 36) HC-CDR3:MGANSGGYLYGMDV (SEQ ID NO: 37) Antibodies according to the invention may comprise the CDRs of P1A3, P1C5, P1E4 or P1F1 or one of SEQ ID NOs: 1 and 5; SEQ ID NOs: 2 and 6; SEQ ID NOs: 3 and 7; or SEQ ID NOs: 4 and 8. In antibodies according to the invention, one or two or three or four of the six CDR sequences may be diverse. Variants may have one or two amino acid substitutions in one or two of the six CDR sequences.

[0158] P1A3 V L Chain and V H Chain (SEQ ID NOs: 1 and 5), V of P1C5 L Chain and V H Chain (SEQ ID NOs: 2 and 6), V of P1E4 L Chain and V H Chains (SEQ ID NOs: 3 and 7) and and P1F1 V L Chain and V H The amino acid sequences of the chains (SEQ ID NOS: 4 and 8) are shown in Figures 1A to 1D (V L ) and Figures 2A to 2D (V H The coding nucleotide sequences are shown in SEQ ID NOs: 39 and 43 (P1A3), SEQ ID NOs: 40 and 44 (P1C5), SEQ ID NOs: 41 and 45 (P1E4), SEQ ID NOs: 42 and 46 (P1F1), and in Figures 16 and 17.

[0159] In some embodiments, antibodies according to the invention can comprise one or more of the amino acid sequences described herein. Antibodies according to the invention can be encoded by one or more of the nucleotide sequences described herein. In some aspects, the antibody is clone A1, A2, A4, A5, B1, B2, B3, B4, B5, C2, C3, C4, D1, D2, D3, D4, E1, E2, E5, F2, F3, F4, F5, G1, G2, G3, G4, or G5 described herein, or a variant of said clones.

[0160] In some aspects and embodiments, the antibody, antigen-binding fragment, or polypeptide comprises at least one VL region according to one of the following (1) to (31): (1) (P1A3) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 15 LC-CDR2 having the amino acid sequence of SEQ ID NO: 16 LC-CDR3 having the amino acid sequence of SEQ ID NO: 17 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (2) (P1C5) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 21 LC-CDR2 having the amino acid sequence of SEQ ID NO: 22 LC-CDR3 having the amino acid sequence of SEQ ID NO: 23 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (3) (P1E4) A VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 27 LC-CDR2 having the amino acid sequence of SEQ ID NO: 28 LC-CDR3 having the amino acid sequence of SEQ ID NO: 29 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (4) (P1F1) A VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 27 LC-CDR2 having the amino acid sequence of SEQ ID NO: 33 LC-CDR3 having the amino acid sequence of SEQ ID NO: 34 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (5) (PFA1) A VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 74 LC-CDR2 having the amino acid sequence of SEQ ID NO: 75 LC-CDR3 having the amino acid sequence of SEQ ID NO: 76 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (6) (PFA2) A VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 84 LC-CDR2 having the amino acid sequence of SEQ ID NO: 75 LC-CDR3 having the amino acid sequence of SEQ ID NO: 85 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (7) (PFA4) A VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 93 LC-CDR2 having the amino acid sequence of SEQ ID NO: 94 LC-CDR3 having the amino acid sequence of SEQ ID NO: 95 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (8) (PFA5) A VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 102 LC-CDR2 having the amino acid sequence of SEQ ID NO: 75 LC-CDR3 having the amino acid sequence of SEQ ID NO: 103 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (9) (PFB1) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 110 LC-CDR2 having the amino acid sequence of SEQ ID NO: 111 LC-CDR3 having the amino acid sequence of SEQ ID NO: 112 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (10) (PFB2) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 120 LC-CDR2 having the amino acid sequence of SEQ ID NO: 121 LC-CDR3 having the amino acid sequence of SEQ ID NO: 122 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (11) (PFB3) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 102 LC-CDR2 having the amino acid sequence of SEQ ID NO: 75 LC-CDR3 having the amino acid sequence of SEQ ID NO: 130 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (12) (PFB4) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 138 LC-CDR2 having the amino acid sequence of SEQ ID NO: 139 LC-CDR3 having the amino acid sequence of SEQ ID NO: 140 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (13) (PFB5) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 102 LC-CDR2 having the amino acid sequence of SEQ ID NO: 146 LC-CDR3 having the amino acid sequence of SEQ ID NO: 147 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (14) (PFC2; PFG5) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 102 LC-CDR2 having the amino acid sequence of SEQ ID NO: 155 LC-CDR3 having the amino acid sequence of SEQ ID NO: 95 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (15)(PFC3) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 161 LC-CDR2 having the amino acid sequence of SEQ ID NO: 162 LC-CDR3 having the amino acid sequence of SEQ ID NO: 163 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (16)(PFC4) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 170 LC-CDR2 having the amino acid sequence of SEQ ID NO: 171 LC-CDR3 having the amino acid sequence of SEQ ID NO: 172 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (17) (PFD1) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 93 LC-CDR2 having the amino acid sequence of SEQ ID NO: 94 LC-CDR3 having the amino acid sequence of SEQ ID NO: 180 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (18) (PFD2) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 186 LC-CDR2 having the amino acid sequence of SEQ ID NO: 187 LC-CDR3 having the amino acid sequence of SEQ ID NO: 188 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (19) (PFD3) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 194 LC-CDR2 having the amino acid sequence of SEQ ID NO: 195 LC-CDR3 having the amino acid sequence of SEQ ID NO: 196 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (20) (PFD4) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 93 LC-CDR2 having the amino acid sequence of SEQ ID NO: 204 LC-CDR3 having the amino acid sequence of SEQ ID NO: 205 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (21) (PFE1) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 211 LC-CDR2 having the amino acid sequence of SEQ ID NO: 212 LC-CDR3 having the amino acid sequence of SEQ ID NO: 95 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (22) (PFE2) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 217 LC-CDR2 having the amino acid sequence of SEQ ID NO: 218 LC-CDR3 having the amino acid sequence of SEQ ID NO: 219 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (23) (PFE5) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 226 LC-CDR2 having the amino acid sequence of SEQ ID NO: 227 LC-CDR3 having the amino acid sequence of SEQ ID NO: 228 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (24) (PFF2) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 161 LC-CDR2 having the amino acid sequence of SEQ ID NO: 162 LC-CDR3 having the amino acid sequence of SEQ ID NO: 233 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (25) (PFF3) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 102 LC-CDR2 having the amino acid sequence of SEQ ID NO: 75 LC-CDR3 having the amino acid sequence of SEQ ID NO: 180 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (26) (PFF4) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 247 LC-CDR2 having the amino acid sequence of SEQ ID NO: 248 LC-CDR3 having the amino acid sequence of SEQ ID NO: 249 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (27) (PFF5) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 255 LC-CDR2 having the amino acid sequence of SEQ ID NO: 33 LC-CDR3 having the amino acid sequence of SEQ ID NO: 256 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (28) (PFG1) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 261 LC-CDR2 having the amino acid sequence of SEQ ID NO: 262 LC-CDR3 having the amino acid sequence of SEQ ID NO: 263 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (29) (PFG2) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 270 LC-CDR2 having the amino acid sequence of SEQ ID NO: 271 LC-CDR3 having the amino acid sequence of SEQ ID NO: 272 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (30) (PFG3) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 279 LC-CDR2 having the amino acid sequence of SEQ ID NO: 280 LC-CDR3 having the amino acid sequence of SEQ ID NO: 23 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid. (31) (PFG4) VL region incorporating the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 287 LC-CDR2 having the amino acid sequence of SEQ ID NO: 288 LC-CDR3 having the amino acid sequence of SEQ ID NO: 289 Alternatively, variants thereof in which one or two or three amino acids in one or more of LC-CDR1, LC-CDR2, or LC-CDR3 are substituted with another amino acid.

[0161] In some aspects and embodiments, the antibody, antigen-binding fragment, or polypeptide comprises at least one VL region according to one of the following (32) to (63): (32) (P1A3) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 1. (33) (P1C5) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 2. (34) (P1E4) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 3. (35) (P1F1) At least 70% sequence identity to the amino acid sequence of SEQ ID NO: 4, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or or a VL region comprising an amino acid sequence having 100% sequence identity to one of the VL regions. (36) (PFA1) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 72. (37) (PFA2) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 82. (38) (PFA4) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 91. (39) (PFA5) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 100. (40) (PFB1) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 108. (41) (PFB2) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 118. (42) (PFB3) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 128. (43) (PFB4) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 136. (44) (PFB5) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 144. (45) (PFC2) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 153. (46) (PFC3) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 159. (47) (PFC4) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 168. (48) (PFD1) at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 178; More preferably, a VL region comprising an amino acid sequence having at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to one of the VL regions. (49) (PFD2) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 184. (50) (PFD3) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 192. (51) (PFD4) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 202. (52) (PFE1) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 209. (53) (PFE2) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 215. (54) (PFE5) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 224. (55) (PFF2) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 231. (56) (PFF3) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 239. (57) (PFF4) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 245. (58) (PFF5) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 253. (59) (PFG1) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 259. (60) (PFG2) At least 70% sequence identity to the amino acid sequence of SEQ ID NO: 268, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% , or a VL region comprising an amino acid sequence with 100% sequence identity to one of the sequences. (61) (PFG3) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 277. (62) (PFG4) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 285. (63) (PFG5) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 293.

[0162] In some aspects and embodiments, the antibody, antigen-binding fragment, or polypeptide comprises at least one VH region according to one of the following (64) to (92): (64) (P1A3) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 18 HC-CDR2 having the amino acid sequence of SEQ ID NO: 19 HC-CDR3 having the amino acid sequence of SEQ ID NO: 20 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (65) (P1C5; PFF5) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 24 HC-CDR2 having the amino acid sequence of SEQ ID NO: 25 HC-CDR3 having the amino acid sequence of SEQ ID NO: 26 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (66) (P1E4) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 30 HC-CDR2 having the amino acid sequence of SEQ ID NO: 31 HC-CDR3 having the amino acid sequence of SEQ ID NO: 32 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (67) (P1F1) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 35 HC-CDR2 having the amino acid sequence of SEQ ID NO: 36 HC-CDR3 having the amino acid sequence of SEQ ID NO: 37 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (68) (PFA1) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 77 HC-CDR2 having the amino acid sequence of SEQ ID NO: 78 HC-CDR3 having the amino acid sequence of SEQ ID NO: 79 Alternatively, one, two, or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are replaced with another amino acid. , and their variants. (69) (PFA2) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 86 HC-CDR2 having the amino acid sequence of SEQ ID NO: 87 HC-CDR3 having the amino acid sequence of SEQ ID NO: 88 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (70) (PFA4) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 96 HC-CDR2 having the amino acid sequence of SEQ ID NO: 78 HC-CDR3 having the amino acid sequence of SEQ ID NO: 97 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (71) (PFA5) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 96 HC-CDR2 having the amino acid sequence of SEQ ID NO: 104 HC-CDR3 having the amino acid sequence of SEQ ID NO: 105 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (72) (PFB1) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 113 HC-CDR2 having the amino acid sequence of SEQ ID NO: 114 HC-CDR3 having the amino acid sequence of SEQ ID NO: 115 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (73) (PFB2) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 123 HC-CDR2 having the amino acid sequence of SEQ ID NO: 124 HC-CDR3 having the amino acid sequence of SEQ ID NO: 125 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (74) (PFB3) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 131 HC-CDR2 having the amino acid sequence of SEQ ID NO: 132 HC-CDR3 having the amino acid sequence of SEQ ID NO: 133 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (75) (PFB4) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 77 HC-CDR2 having the amino acid sequence of SEQ ID NO: 78 HC-CDR3 having the amino acid sequence of SEQ ID NO: 141 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (76) (PFB5) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 148 HC-CDR2 having the amino acid sequence of SEQ ID NO: 149 HC-CDR3 having the amino acid sequence of SEQ ID NO: 150 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (77) (PFC2; PFG5) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 96 HC-CDR2 having the amino acid sequence of SEQ ID NO: 78 HC-CDR3 having the amino acid sequence of SEQ ID NO: 156 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (78)(PFC3) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 131 HC-CDR2 having the amino acid sequence of SEQ ID NO: 164 HC-CDR3 having the amino acid sequence of SEQ ID NO: 165 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (79)(PFC4) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 173 HC-CDR2 having the amino acid sequence of SEQ ID NO: 174 HC-CDR3 having the amino acid sequence of SEQ ID NO: 175 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (80) (PFD1) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 77 HC-CDR2 having the amino acid sequence of SEQ ID NO: 78 HC-CDR3 having the amino acid sequence of SEQ ID NO: 181 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (81) (PFD2) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 30 HC-CDR2 having the amino acid sequence of SEQ ID NO: 31 HC-CDR3 having the amino acid sequence of SEQ ID NO: 189 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (82) (PFD3) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 197 HC-CDR2 having the amino acid sequence of SEQ ID NO: 198 HC-CDR3 having the amino acid sequence of SEQ ID NO: 199 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (83) (PFD4) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 96 HC-CDR2 having the amino acid sequence of SEQ ID NO: 206 HC-CDR3 having the amino acid sequence of SEQ ID NO: 97 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (84) (PFE1; PFE5) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 96 HC-CDR2 having the amino acid sequence of SEQ ID NO: 78 HC-CDR3 having the amino acid sequence of SEQ ID NO: 97 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (85) (PFE2) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 96 HC-CDR2 having the amino acid sequence of SEQ ID NO: 220 HC-CDR3 having the amino acid sequence of SEQ ID NO: 221 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (86) (PFF2) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 234 HC-CDR2 having the amino acid sequence of SEQ ID NO: 235 HC-CDR3 having the amino acid sequence of SEQ ID NO: 236 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (87) (PFF3) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 241 HC-CDR2 having the amino acid sequence of SEQ ID NO: 206 HC-CDR3 having the amino acid sequence of SEQ ID NO: 242 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (88) (PFF4) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 148 HC-CDR2 having the amino acid sequence of SEQ ID NO: 149 HC-CDR3 having the amino acid sequence of SEQ ID NO: 250 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (89) (PFG1) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 77 HC-CDR2 having the amino acid sequence of SEQ ID NO: 264 HC-CDR3 having the amino acid sequence of SEQ ID NO: 265 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (90) (PFG2) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 131 HC-CDR2 having the amino acid sequence of SEQ ID NO: 273 HC-CDR3 having the amino acid sequence of SEQ ID NO: 274 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (91) (PFG3) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 281 HC-CDR2 having the amino acid sequence of SEQ ID NO: 282 HC-CDR3 having the amino acid sequence of SEQ ID NO: 20 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid. (92) (PFG4) VH region incorporating the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 148 HC-CDR2 having the amino acid sequence of SEQ ID NO: 149 HC-CDR3 having the amino acid sequence of SEQ ID NO: 290 Alternatively, variants thereof in which one or two or three amino acids in one or more of HC-CDR1, HC-CDR2, or HC-CDR3 are substituted with another amino acid.

[0163] In some aspects and embodiments, the antibody, antigen-binding fragment, or polypeptide comprises at least one VH region according to one of the following (93) to (124): (93) (P1A3) A VH region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:5. (94) (P1C5) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 6. (95) (P1E4) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 7. (96) (P1F1) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 8. (97) (PFA1) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 73. (98) (PFA2) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 83. (99) (PFA4) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 92. (100) (PFA5) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 101. (101) (PFB1) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 109. (102) (PFB2) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 119. (103) (PFB3) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 129. (104) (PFB4) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 137. (105) (PFB5) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 145. (106) (PFC2) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 154. (107) (PFC3) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 160. (108) (PFC4) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 169. (109) (PFD1) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 179. (110) (PFD2) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 185. (111) (PFD3) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 193. (112) (PFD4) At least 70% sequence identity to the amino acid sequence of SEQ ID NO: 203, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89% %, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to one of the VL regions. (113) (PFE1) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 210. (114) (PFE2) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 216. (115) (PFE5) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 225. (116) (PFF2) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 232. (117) (PFF3) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 240. (118) (PFF4) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 246. (119) (PFF5) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 254. (120) (PFG1) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 260. (121) (PFG2) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 269. (122) (PFG3) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 278. (123) (PFG4) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 286. (124) (PFG5) A VL region comprising an amino acid sequence having at least 70% sequence identity, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 294.

[0164] In some embodiments, the antigen-binding molecule comprises at least one VL region according to any one of (1) to (63) above, and at least one VH region according to any one of (64) to (124) above: Additionally, light and heavy chain CDRs may be particularly useful in conjunction with a variety of framework regions. Thus, light and / or heavy chains having LC-CDR1 to LC-CDR3 or HC-CDR1 to HC-CDR3 may possess alternative framework regions. Suitable framework regions are well known in the art and are described, for example, in M. Lefranc & G. Lefranc (2001) "The Immunoglobulin Facts Book," Academic Press, incorporated herein by reference.

[0165] As used herein, antibodies may be any of the V of SEQ ID NOs: 1 and 5; SEQ ID NOs: 2 and 6; SEQ ID NOs: 3 and 7; or SEQ ID NOs: 4 and 8. L and / or V H V comprising an amino acid sequence having a high percent sequence identity to one or more of the amino acid sequences shown in Figures 1 and 2. L and / or V H It may have a chain.

[0166] For example, an antibody according to the present invention may bind HER2 and have one of the V sequences of SEQ ID NOs: 1 to 8. L Or V H and V comprising an amino acid sequence having at least 70%, more preferably at least 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to one of the amino acid sequences shown in Figures 1 and 2. L or V H Includes antibodies with chains.

[0167] In some embodiments, the antibodies, antigen-binding molecules, or polypeptides provided herein comprise the CDRs, V and V sequences of PFA4, PFB4, PFB5, PFC3, PFD4, PFE1, PFF5, or PFG3. L and / or V HIn some embodiments, the antibodies, antigen-binding molecules, or polypeptides provided herein comprise the CDRs, V and V chains of PFA4, PFB4, PFC3, PFD4, PFE1, PFF5, or PFG3. L and / or V H In some embodiments, the antibodies, antigen-binding molecules, or polypeptides provided herein comprise the CDRs, V chains, or CDRs of PFB4, PFC3, PFD4, PFE1, PFF5, or PFG3. L and / or V H In some embodiments, the antibodies, antigen-binding molecules, or polypeptides provided herein comprise the CDRs, V chains, or CDRs of PFB4, PFC3, PFE1, PFF5, or PFG3. L and / or V H In some embodiments, the antibodies, antigen-binding molecules, or polypeptides provided herein comprise the CDRs, V chains, or CDRs of PFB4, PFC3, PFE1, PFF5, or PFG3. L and / or V H Includes chains.

[0168] In some embodiments, the antibodies, antigen-binding molecules, or polypeptides provided herein comprise any of the CDRs, VDRs, or VDRs of P1A3, P1C5, P1E4, P1F1, PFA4, PFB4, PFC3, PFD4, or PFE1. L and / or V H Includes chains.

[0169] Multivalent antibodies Whole antibodies and F(ab')2 fragments are "bivalent." By "bivalent," we mean that the antibodies and F(ab')2 fragments have two antigen-binding sites. In contrast, Fab, Fv, ScFv, and dAb fragments are monovalent and have only one antigen-binding site.

[0170] The present application also provides antibodies or antigen-binding fragments that are capable of binding to HER2 and that are bispecific antibodies or bispecific antigen-binding fragments. Thus, bispecific antibodies or bispecific antigen-binding fragments may be isolated. The term "bispecific" means that an antigen-binding molecule is capable of specifically binding to at least two distinct antigenic determinants.

[0171] In some embodiments, the bispecific antibodies and bispecific antigen-binding fragments comprise an antigen-binding fragment or polypeptide according to the invention. In some embodiments, the bispecific antibodies and bispecific antigen-binding fragments comprise an antigen-binding fragment capable of binding to HER2, wherein the antigen-binding fragment capable of binding to HER2 comprises or consists of an antigen-binding fragment or polypeptide according to the invention.

[0172] In some embodiments, the bispecific antibody and bispecific antigen-binding fragment comprise an antigen-binding fragment capable of binding to HER2 and an antigen-binding fragment capable of binding to another target protein. In some embodiments, the bispecific antibody / antigen-binding fragment comprises an antigen-binding molecule capable of binding to HER2 and an antigen-binding molecule capable of binding to an antigen other than HER2. In some embodiments, the antigen other than HER2 is an immune cell surface molecule. In some embodiments, the antigen other than HER2 is a cancer cell antigen. In some embodiments, the antigen other than HER2 is a receptor molecule, for example, a cell surface receptor.

[0173] A cancer cell antigen is an antigen expressed or overexpressed by cancer cells. A cancer cell antigen may be any peptide / polypeptide, glycoprotein, lipoprotein, glycan, glycolipid, lipid, or fragment thereof. Expression of a cancer cell antigen may be associated with cancer. A cancer cell antigen may be aberrantly expressed by cancer cells (e.g., the cancer cell antigen may be expressed with abnormal localization) or may be expressed in an abnormal structure by cancer cells. A cancer cell antigen may be capable of eliciting an immune response. In some embodiments, the antigen is expressed on the cell surface of cancer cells (i.e., the cancer cell antigen is a cancer cell surface antigen). In some embodiments, the portion of the antigen bound by the antigen-binding molecule described herein is displayed on the external surface of the cancer cell (i.e., is extracellular). A cancer cell antigen may be a cancer-associated antigen. In some embodiments, a cancer cell antigen is an antigen whose expression is associated with the onset, progression, or severity of symptoms of cancer. A cancer-associated antigen may be associated with the cause or pathology of cancer, or may be aberrantly expressed as a result of cancer. In some embodiments, a cancer cell antigen is an antigen whose expression is upregulated (e.g., at the RNA and / or protein level) by cancer cells, e.g., when compared to the level of expression by comparable non-cancerous cells (e.g., non-cancerous cells derived from the same tissue / cell type). In some embodiments, a cancer-associated antigen may be preferentially expressed by cancerous cells and not expressed by comparable non-cancerous cells (e.g., non-cancerous cells derived from the same tissue / cell type). In some embodiments, a cancer-associated antigen may be the product of a mutated oncogene or a mutated suppressor gene. In some embodiments, a cancer-associated antigen may be the product of an overexpressed cellular protein, a cancer antigen produced by an oncogenic virus, an oncofetal antigen, or a cell surface glycolipid or glycoprotein.

[0174] The immune cell surface molecule may be any peptide / polypeptide, glycoprotein, lipoprotein, glycan, glycolipid, lipid, or fragment thereof that is expressed on the cell surface of an immune cell or on the cell surface of an immune cell. In some embodiments, the portion of the immune cell surface molecule that is bound by the antigen-binding molecule of the present invention is present on the external surface of the immune cell (i.e., is extracellular). The immune cell surface molecule may be expressed on the cell surface of any immune cell. In some embodiments, the immune cell may be a cell of hematopoietic origin, such as a neutrophil, eosinophil, basophil, dendritic cell, lymphocyte, or monocyte. The lymphocyte may be, for example, a T cell, a B cell, a natural killer (NK) cell, a NKT cell, or an innate lymphoid cell (ILC), or a precursor thereof (e.g., a thymocyte or a pre-B cell). In some embodiments, In some embodiments, the immune cell surface molecule may be a costimulatory molecule (e.g., CD28, OX40, 4-1BB, ICOS, or CD27) or its ligand. In some embodiments, the immune cell surface molecule may be a checkpoint inhibitor (e.g., PD-1, CTLA-4, LAG-3, TIM-3, TIGIT, or BTLA) or its ligand.

[0175] In some embodiments, the bispecific antibody / antigen-binding fragment comprises an antigen-binding fragment / molecule as provided herein and an antigen-binding fragment / molecule capable of binding to HER2. In some embodiments, the bispecific antibody / antigen-binding fragment comprises an antigen-binding fragment / molecule capable of binding HER2 at a first epitope and an antigen-binding fragment / molecule capable of binding HER2 at a second epitope. For example, the bispecific antibody / antigen-binding fragment comprises an antigen-binding fragment / molecule as provided herein and an antigen-binding fragment / molecule derived from trastuzumab and / or pertuzumab.

[0176] An antigen-binding fragment capable of binding to another target protein may be capable of binding to another protein other than HER2. The antibodies and antigen-binding polypeptides described herein may be multispecific. Multispecific antibodies and antigen-binding polypeptides may be provided in any suitable format, such as, for example, those described in Kontermann MAbs 2012, 4(2): 182-197, which is incorporated herein by reference in its entirety. For example, the antigen-binding polypeptide can be a bispecific antibody conjugate (e.g., IgG2, F(ab')2 or CovX-Body), bispecific IgG or IgG-like molecule (e.g., IgG, scFv4-Ig, IgG-scFv, scFv-IgG, DVD-Ig, IgG-sVD, sVD-IgG, 2in1-IgG, mAb2, or Tandemab common LC), asymmetric bispecific IgG or IgG-like molecule (e.g., kih IgG, kih IgG common LC, CrossMab, kih IgG-scFab, mAb-Fv, charge pair or SEED-body), small molecule bispecific antibody molecule (e.g., Diabody (Db), dsDb, DART, scDb, tandAbs, tandem scFv (taFv), tandem dAb / VHH, triple body, triple head, Fab-scFv, or F(ab')2-scFv2), bispecific Fc and C H The fusion protein may be a trispecific fusion protein (e.g., taFv-Fc, Di-diabody, scDb-CH3, scFv-Fc-scFv, HCAb-VHH, scFv-kih-Fc, or scFv-kih-CH3), or a bispecific fusion protein (e.g., scFv2-albumin, scDb-albumin, taFv-toxin, DNL-Fab3, DNL-Fab4-IgG, DNL-Fab4-IgG-cytokine 2). In particular, Kontermann MAbs 2012, 4(2): See Figure 2 at 182-19. "Multispecific" means that an antigen-binding molecule exhibits specific binding to more than one target.

[0177] Those skilled in the art can design and prepare bispecific antibodies. Methods for producing bispecific antibodies include linking antibodies or antibody fragments, for example, via a reducible disulfide or non-reducible thioether bond, as described, for example, in Segal and Bast, 2001. Production of Bispecific Antibodies. Current Protocols in Immunology., which is incorporated herein in its entirety. 14:IV:2.13:2.13.1-2.13.16. For example, Fab fragments may be chemically crosslinked through their hinge region SH-groups using, for example, N-succinimidyl-3-(-2-pyridyldithio)-propionate (SPDP) to generate disulfide-linked bispecific F(ab)2 heterodimers. Other methods for producing bispecific antibodies include fusing antibody-producing hybridomas, for example with polyethylene glycol, to produce quadroma cells capable of secreting bispecific antibodies, for example, as described in DM and Bast, BJ 2001. Production of Bispecific Antibodies. Current Protocols in Immunology. 14:IV:2.13:2.13.1-2.13.16.

[0178] Bispecific antibodies can also be recombinantly produced by expression from nucleic acid constructs encoding polypeptides for antigen-binding molecules, for example, as described in Antibody Engineering: Methods and Protocols, 2nd Edition (Humana Press, 2012), Chapter 40: Production of Bispecific Antibodies.: Diabodies and Tandem scFv (Hornig and Farber-Schwarz), or French, How to make bispecific antibodies, Methods Mol. Med. 2000; 40:333-339, both of which are incorporated herein in their entirety. For example, a DNA construct encoding the light and heavy chain variable domains for two antigen-binding fragments (i.e., light and heavy chain variable domains for an antigen-binding fragment capable of binding HER2, and light and heavy chain variable domains for an antigen-binding fragment capable of binding to another target protein / another epitope on HER2), and containing a sequence encoding an appropriate linker or dimerization domain between the antigen-binding fragments, can be prepared by molecular cloning techniques. The recombinant bispecific antibody can then be produced by expression (e.g., in vitro) of the construct in a suitable host cell (e.g., a mammalian host cell), and the expressed recombinant bispecific antibody can then be purified, if desired.

[0179] antibody conjugates The present invention also provides an antibody conjugate comprising an antibody, antigen-binding fragment or polypeptide according to the invention conjugated to a chemical moiety.

[0180] In some embodiments, the chemical moiety may be a moiety for providing a therapeutic effect. In some embodiments, the chemical moiety may be a drug moiety (e.g., a cytotoxic agent or a cytostatic agent). A cytotoxic agent is an agent that kills cells. A cytostatic agent is an agent that inhibits cell growth.

[0181] The use of antibody-drug conjugates can target the delivery of and promote intracellular accumulation of the drug moiety to cancer cells that express HER2. In some embodiments, the chemical moiety may be at least one anti-cancer agent. "At least one anti-cancer agent" refers to one, or two, or three, or four, or more agents effective in treating a malignant or cancerous disease, such as a cancer, as described herein. An anti-cancer agent may also be described herein as a therapeutic agent, an additional agent, or an additional therapeutic agent.

[0182] In some embodiments, the drug moiety may be a chemotherapeutic agent described herein below. In some embodiments, the drug moiety may be a hormone capable of controlling tumor growth, such as ethinyl estradiol, medroxyprogesterone, norethisterone, megestrol, diethylstilbestrol, or fosfestrol. In some embodiments, the drug moiety may be a hormone antagonist, such as tamoxifen, toremifene, aminoglutethimide, anastrozole, letrozole, exemestane, trilostane, goserelin, buserelin, goserelin, leuprorelin, cyproterone acetate, flutamide, or bicalutamide.

[0183] In some embodiments, the chemical moiety may be a detectable moiety. Suitable moieties and means for their detection are well known to those of skill in the art and include radioisotopes or Includes non-isotopic objects.

[0184] The antibody conjugate may include a linker that can be cleaved to release the drug at the target location. Chimeric Antigen Receptor The antibody, antigen-binding fragment, or polypeptide according to the present invention can be incorporated into a chimeric antigen receptor (CAR) capable of binding to HER2. Thus, the present invention also provides a CAR comprising the antibody, antigen-binding molecule, or polypeptide of the present invention.

[0185] Chimeric antigen receptor (CAR) is a recombinant receptor that provides both antigen binding function and T cell activation function. CAR structure and operation are outlined in, for example, Dotti et al., Immunol Rev (2014) 257(1), which is incorporated herein in its entirety. CAR comprises an antigen binding region linked to a cell membrane anchor region and a signal transduction region. An optional hinge region may provide separation between the antigen binding region and the cell membrane anchor region, and can act as a flexible linker.

[0186] The antibodies, antigen-binding fragments, or polypeptides according to the present invention may be used as the antigen-binding domain of a chimeric antigen receptor (CAR). In some embodiments, the CAR comprises a V-type antibody, antigen-binding fragment, or polypeptide according to any embodiment of the antibody, antigen-binding fragment, or polypeptide described herein. L Domains and V H Thus, the antigen to which the CAR of the present invention binds is HER2.

[0187] Also provided is an in vitro complex, optionally isolated, comprising a CAR bound to HER2. CARs may be combined with costimulatory ligands, chimeric costimulatory receptors, or cytokines to further enhance T cell potency, specificity, and safety (Sadelain et al., The basic principles of chimeric antigen receptor (CAR) design. Cancer Discov. 2013 April; 3(4): 388-398. doi:10.1158 / 2159-8290.CD-12-0548, specifically incorporated herein by reference).

[0188] The present invention provides an optionally isolated nucleic acid encoding the CAR described herein. Also provided is an expression vector comprising the nucleic acid. The present invention also provides a cell comprising the CAR, nucleic acid, or expression vector described herein. Also provided is a composition comprising the CAR, nucleic acid, expression vector, or cell described herein. CARs can be used to generate T cells targeted to tumors, for example, T cells targeted to tumor cells that express or overexpress HER2, for which the CAR is specific.

[0189] The cell may be an immune cell, for example, a T cell, an antigen-specific T cell (e.g., a virus-specific T cell), an antigen-specific CD4 T cell, an antigen-specific CD8 T cell, an effector memory CD4 T cell, an effector memory CD8 T cell, a central memory CD4 T cell, a central memory CD8 T cell, a cytotoxic CD8+ T cell (i.e., a CTL), an NK cell, or an antigen-specific NK cell.

[0190] Engineering a CAR into cells may be performed during in vitro culture for transduction and expansion, such as occurs during T cell expansion for adoptive T cell therapy. Transduction can utilize a variety of methods, but stable gene transfer is required to allow for persistent CAR expression in clonally expanded and remaining T cells.

[0191] In addition to the HER2 specificity determinants described herein, CD28 and tumor necrosis factor receptor CAR molecules can be further engineered to express costimulatory endodomes, such as those derived from tumor suppressor superfamily member 9 (TNFRSF9; 4-1BB), to promote T cell proliferation and persistence upon encountering tumor cells (Nishio and Dotti., OncoImmunology 4:2, e988098; February 2015).

[0192] CARs typically combine an antigen-binding domain with the intracellular domain of the CD3-zeta chain or FcyRI protein in a single chimeric protein. The structural features of CARs are described by Sjouke et al. (The pharmacology of second-generation chimeric antigen receptors. Nature Reviews Drug Discovery, 14, 499 509 (2015) doi:10.1038 / nrd4597). CARs typically have an extracellular antigen-binding domain linked to a transmembrane domain and an endodomain. An optional hinge or spacer domain may provide separation between the binding moiety and the transmembrane domain and act as a flexible linker.

[0193] According to the present invention, the antigen recognition domain of the CAR can be or can be derived from an antibody, antigen-binding fragment, or polypeptide capable of binding to HER2 as described herein. The antigen-binding region of the CAR of the present invention can be provided in any suitable format, e.g., scFv, scFab, etc.

[0194] In some embodiments, the antigen recognition domain of the CAR can be or be derived from clones P1A3, P1C5, P1E4, or P1F1 described herein. In some embodiments, a CAR can comprise an antibody, antigen-binding fragment, or polypeptide described herein that exhibits moderate or lower binding to HER2 as described herein. Moderate and / or lower affinity antigen-binding regions may be able to distinguish cancer cells that overexpress a target protein from normal cells that express physiological target levels, as described, for example, in Liu et al., Cancer Res. 2015; (75) (17) 3596-3607. This approach may specifically target cancer cells while sparing non-cancer cells, reducing off-target toxicity. In some embodiments, the antigen recognition domain of the CAR can be or be derived from clones PFA4, PFB4, PFB5, PFC3, PFD4, PFE1, PFF5, or PFG3 described herein. In some embodiments, the antigen recognition domain of the CAR can be or be derived from clones PFA4, PFB4, PFC3, PFD4, PFE1, PFF5, or PFG3 described herein.

[0195] The hinge or spacer region may be a flexible domain that allows the binding moiety to be oriented in various directions. The hinge or spacer region may be derived from the CH2CH3 region of IgG1, IgH or an immunoglobulin.

[0196] The transmembrane domain may be a hydrophobic alpha helix that spans the cell membrane. A transmembrane domain accompanied by an endodomain is commonly used. The transmembrane region may be referred to as a cell membrane anchor region. In some embodiments, the CAR comprises a cell membrane anchor region that comprises, consists of, or is derived from the transmembrane region amino acid sequence of one of CD3-zeta, CD4, CD8, or CD28. As used herein, a region "derived from" a reference amino acid sequence comprises an amino acid sequence that has at least 60%, for example, at least 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the reference sequence.

[0197] The endodomain is responsible for receptor clustering / dimerization after antigen binding and for initiating signal transduction to the cell. One commonly used transmembrane domain is the CD3-zeta transmembrane and endodomain. Optionally, intracellular domains from one or more costimulatory protein receptors, such as CD28 4-1BB, OX40, and ICOS, can be incorporated into the cytoplasmic end of the CAR to provide additional costimulatory signal transduction, which may be beneficial in terms of antitumor activity.

[0198] The CAR described herein may comprise an extracellular domain having an antigen recognition domain, a transmembrane domain, and a cytoplasmic domain. The transmembrane domain naturally associated with one of the domains in the CAR may be used, or the transmembrane domain may be selected or modified by amino acid substitution to prevent binding of such domain to the transmembrane domain of the same or a different surface membrane protein, thereby minimizing interaction with other membranes of the receptor complex. The cytoplasmic domain may be designed to include a CD28 and / or 4-1BB signaling domain by itself or may be combined with any other desired cytoplasmic domain(s). The cytoplasmic domain may also be designed to include a CD3-zeta signaling domain. For example, the cytoplasmic domain of a CAR may include, but is not limited to, CD3-zeta, 4-1BB, and CD28 signaling modules, and combinations thereof.

[0199] Also provided herein are CAR T cells comprising an antigen-binding fragment capable of binding to HER2 as a CAR, as described herein. The CAR T cells described herein can be generated by introducing a lentiviral vector containing a desired CAR, such as a CAR containing an anti-HER2, CD8a hinge and transmembrane domain, and a human 4-1BB and CD3 zeta signaling domain, into cells in vitro. The CAR T cells described herein can also be generated by electroporation of mRNA encoding the desired CAR. The CAR T cells of the present invention can replicate in vivo, resulting in long-term persistence that can lead to sustained tumor control.

[0200] In some embodiments, the CAR T according to the present invention promotes cytolysis of cells, e.g., cancer cells or tumor cells. The present invention provides a CAR, nucleic acid, expression vector, cell, or composition described herein for use in a method of medical treatment or prevention. Also provided herein is the administration of genetically modified T cells expressing a CAR, nucleic acid, or expression vector described herein for use in a method of medical treatment or prevention. In some embodiments, the CAR or T cells described herein are used to treat patients with cancer or at risk of having cancer. In some embodiments, the cancer to be treated is a cancer described herein. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer to be treated is ovarian, uterine, lung, for example, lung adenocarcinoma, or gastrointestinal, for example, stomach, salivary duct cancer.

[0201] Administration may be by lymphocyte infusion. Preferably, autologous lymphocyte infusion is used in the treatment, i.e., the lymphocytes are derived from the patient into whom they are introduced. Autologous PBMCs (peripheral blood mononuclear cells) are collected from the patient in need of treatment, and T cells are activated and expanded using methods known in the art, and then infused back into the patient. Cell generation, activation, and expansion may be performed in vitro or ex vivo. In some cases, cells may be obtained from an individual other than the patient and introduced into the patient, i.e., allogeneic cell therapy.

[0202] Adoptive T cell transfer is described, for example, in Chia WK et al., which is incorporated herein by reference in its entirety. , Molecular Therapy (2014), 22(1): 132-139, Kalos and June 2013, Immunity 39(1): 49-60, and Cobbold et al., (2005) J. Exp. Med. 202:379-386. Thus, those skilled in the art can determine appropriate reagents and procedures for adoptive transfer of immune cells as described herein.

[0203] Detection Method The antibodies, antigen-binding fragments, polypeptides, conjugates, CARs or cells described herein can be detectably labeled or at least detectable.For example, antibodies can be labeled with radioactive atoms, colored molecules, fluorescent molecules, or any other easily detectable molecules.Suitable detectable molecules include fluorescent proteins, luciferase, enzyme substrates, and radiolabels.Binding moieties can be directly labeled with detectable labels, or binding moieties can be indirectly labeled.For example, binding moieties can be unlabeled antibodies that can be detected by another antibody that is itself labeled.Alternatively, the secondary antibody can be bound to biotin, and the first antibody is indirectly labeled by using the binding of labeled streptavidin to biotin.

[0204] The antibodies, antigen-binding fragments, polypeptides, conjugates, CARs, or cells described herein may be used in methods involving the binding of antibodies or antigen-binding fragments to HER2. Such methods may include detecting a binding complex of an antibody, antigen-binding fragment, polypeptide, conjugate, CAR, or cell, and HER2. Thus, in one embodiment, a method is provided that includes contacting a sample containing HER2 or suspected of containing HER2 with an antibody, antigen-binding fragment, polypeptide, conjugate, CAR, or cell described herein, and detecting the formation of a complex of an antibody, antigen-binding fragment, polypeptide, conjugate, CAR, or cell, and HER2.

[0205] Suitable method formats, including sandwich assays, immunoassays such as ELISA, are well known in the art.The above-mentioned method can include labeling antibody, antigen-binding fragment, polypeptide, conjugate, CAR or cell, or HER2, or both, with detectable labels, such as fluorescent labels, luminescent labels, or radiolabels.HER2 expression can be measured in tissue samples obtained, for example, by biopsy, by immunohistochemistry (IHC) or fluorescence in situ hybridization (FISH).

[0206] This type of method can provide the basis for a diagnostic method for a disease or condition that requires the detection and / or quantification of HER2. Such a method can be performed in vitro on a patient sample or after processing of the patient sample. Once the sample is collected, the patient does not need to be present for the diagnostic in vitro method to be performed, and therefore the method is not performed on a human or animal body.

[0207] The method can include determining the amount of HER2 present in patient samples.The method can further include comparing the determined amount with a standard or reference value as part of the process of reaching a diagnosis.Other diagnostic tests can be used in conjunction with those described herein to improve the accuracy of diagnosis or prognosis, or to confirm the results obtained by using the tests described herein.

[0208] The level of HER2 present in a patient sample may indicate that the patient may respond to treatment with an anti-HER2 antibody, antigen-binding fragment, polypeptide, conjugate, CAR, or cell. The presence of a high level of HER2 in a sample may be used to select a patient for treatment with an anti-HER2 antibody, antigen-binding fragment, polypeptide, conjugate, CAR, or cell. Thus, the antibodies, antigen-binding fragments, polypeptides, conjugates, CARs, or cells of the present invention may be used to select a patient for treatment with an anti-HER2 antibody, antigen-binding fragment, polypeptide, conjugate, CAR, or cell. The tumor, CAR, or cells can be used to select patients for treatment with anti-HER2 therapy.

[0209] Detection of HER2 in a sample may be used for purposes of diagnosing a cancerous condition in a patient, diagnosing a predisposition to a cancerous condition, or to provide a prognosis (prognosis) of a cancerous condition. The diagnosis or prognosis may relate to an existing (previously diagnosed) cancerous condition, which may be benign or malignant, may relate to a suspected cancerous condition, or may relate to screening for a cancerous condition in a patient (who may be previously undiagnosed).

[0210] The sample may be taken from any tissue or body fluid. The sample may include a large volume of blood; a large volume of serum from an individual's blood, which may include the liquid portion of blood obtained after removing fibrin clots and blood cells; a tissue sample or biopsy; or cells isolated from said individual, or may be derived from these tissues.

[0211] The method according to the invention may preferably be carried out in vivo. The term "in vitro" is intended to include experiments using cells in culture, while the term "in vivo" is intended to include experiments using intact multicellular organisms.

[0212] The antibodies, antigen-binding fragments, polypeptides, conjugates, CARs, or cells described herein may be used, for example, in methods of detecting, localizing, or imaging cancer (e.g., tumors) in vivo.

[0213] The antibody, antigen-binding fragment, polypeptide, conjugate, CAR, or cell may be appropriately labeled, directly or indirectly, with a detectable label (e.g., a signal-generating label), such as a radioisotope or a non-isotopic entity, for detection purposes. Radioactive isotopes include iodine-123, iodine-126, iodine-131, iodine-133, bromine-77, technetium-99m, indium-111, indium-113m, gallium-67, gallium-68, ruthenium-95, ruthenium-97, ruthenium-103, ruthenium-105, mercury-207, mercury-203, rhenium-99m, rhenium-101, rhenium-105, scandium-47, tellurium-121m, tellurium-122m, tellurium-125m, thulium-165, thulium-167, thulium-168, copper-67, fluorine-18, yttrium-90, palladium-100, bismuth-217, and antimony-211. The non-isotopic entity may be selected from enzymes (e.g., peroxidase, alkaline phosphatase, glucose oxidase, beta-glucosidase, luciferase), dyes, haptens, radioluminescent agents, chemiluminescent agents (e.g., acridinium esters, luminol, isoluminol), bioluminescent agents, luminescent agents such as fluorescent (e.g., fluorescein, rhodamine, eosin, and NDB, green fluorescent protein (GFP) chelates of rare earths such as europium (Eu), terbium (Tb), and samarium (Sm), tetramethylrhodamine, Texas Red, 4-methylumbelliferone, 7-amino-4-methylcoumarin, Cy3, Cy5) or phosphorescent agents, antibodies, receptors, and ligands such as biotin, avidin, streptavidin, or digoxigenin.

[0214] Detection techniques are well known to those skilled in the art and can be selected to be consistent with the labeling agent. Suitable techniques include PCR amplification of the oligonucleotide tag, mass spectrometry, fluorescence or color detection upon enzymatic conversion of a substrate, for example, by a reporter protein, or detection of radioactivity.

[0215] In some embodiments, the methods involve administering the antibody, antigen fragment, polypeptide, conjugate, CAR, or cell to a subject, e.g., a subject diagnosed with or suffering from cancer. and detecting the antibody, antigen-binding fragment, polypeptide, conjugate, CAR, or cell. In some embodiments, the method includes detecting a signal from the antibody, antigen-binding fragment, polypeptide, conjugate, CAR, or cell. In some embodiments, the method includes converting the signal into an image.

[0216] The present invention also provides a method for selecting / stratifying a subject for treatment with an agent targeted to HER2 using an antibody, antigen-binding fragment, polypeptide, conjugate, CAR, or cell according to the present invention. In some embodiments, the method comprises contacting a sample from the subject with the antibody, antigen-binding fragment, polypeptide, conjugate, CAR, or cell. In some embodiments, the sample is contacted in vitro. In some embodiments, the subject is selected for treatment according to the present invention or identified as a subject who will benefit from such treatment based on, for example, detecting the presence of HER2 or a nucleic acid encoding HER2 in a sample obtained from the individual.

[0217] therapeutic use The antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, CAR and cell according to the present invention, and the composition comprising such agent can be provided for use in medical treatment method.The treatment can be provided to a subject with a disease or condition that requires treatment.The disease or condition can be cancer.The disease or condition can be cancer in which HER2 is overexpressed.

[0218] The present invention provides an antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, CAR, or cell according to the present invention for use in a method of medical treatment or prevention. The present invention also provides the use of an antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, CAR, or cell according to the present invention in the manufacture of a medicament for treating or preventing a disease or condition. The present invention also provides a method of treating or preventing a disease or condition, comprising the step of administering to a subject a therapeutically or prophylactically effective amount of an antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, CAR, or cell according to the present invention.

[0219] HER2 is a member of the epidermal growth factor receptor family and functions as a preferential heterodimerization signaling partner with other members of the EGFR family (EGFR / HER1, HER3, and HER4), playing an important role in all stages of cell development, including cell proliferation and differentiation (Karunagaran et al., (1996) EMBO J. 15: 254-264; Klapper et al., (1999) PNAS 96: As a key gene for cell survival, HER2 gene amplification and protein overexpression lead to malignant transformation (Neve et al., (2001) Ann Oncol. 12 Suppl 1():S9-13).

[0220] HER2 is expressed in numerous cancers, including breast, ovarian, gastric, endometrial, bladder, lung, colon, prostate, and head and neck cancers. HER2 overexpression is directly associated with poor clinical outcomes. HER2 overexpression occurs in approximately 15-30% of breast cancers and 10-30% of gastric cancers (Iqbal & Iqbal (2014) Molecular Biology International, vol. 2014, Article ID 852748).

[0221] In some embodiments, the cancer may comprise cells that overexpress HER2. In some embodiments, the cancer may comprise cells that express HER2 to which the antibody, antigen-binding fragment, or polypeptide is specific.

[0222] The antibodies, antigen-binding fragments, polypeptides, conjugates, nucleic acids, vectors, compositions, CARs, or cells according to the present invention can be provided for use in treating cancers that express or overexpress HER2. For example, tumor cells that overexpress HER2 can be killed directly by treatment with the antibodies, antigen-binding fragments, polypeptides, conjugates, CARs, or cells according to the present invention, for example, by cell lysis or apoptosis, by antibody-dependent cell-mediated cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), or using anti-HER2 antibody-drug conjugates.

[0223] "Treatment" can be, for example, reducing the onset or progression of a disease / condition, alleviating the symptoms of a disease / condition, or reducing the pathology of a disease / condition. Treating or alleviating a disease / condition can be effective in preventing the progression of the disease / condition, for example, preventing the condition from worsening or slowing the rate of onset. In some embodiments, treating or alleviating can lead to an improvement of the disease / condition, for example, a reduction in the symptoms of the disease / condition, or a reduction in some other correlate of the severity / activity of the disease / condition. Prevention / prophylaxis of a disease / condition can refer to preventing the condition from worsening, or preventing the onset of the disease / condition, for example, preventing an early stage disease / condition from developing into a later, chronic stage.

[0224] Treatment can be aimed at preventing the onset or progression of cancer.Therefore, antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, composition, CAR or cell can be used to formulate pharmaceutical composition or medicine, and subject can be treated prophylactically against the onset of disease state.This can be done before the onset of symptoms of disease state, and / or can be administered to subjects who are considered to be at a higher risk of onset or progression of cancer.

[0225] Treatment may aim to reduce the number of HER2-expressing cells, induce cell death of HER2-expressing cells, reduce tumor size, or inhibit / prevent tumor growth. In some embodiments, the cancer, cells, tumor, or subject to be treated has been determined, optionally in vitro, to be HER2-positive (HER2+). In some embodiments, the methods of treatment or prevention described herein include determining, optionally in vitro, whether HER2 is expressed by the cancer, cells, or tumor of the subject to be treated. The determination can be performed on a sample obtained from the subject.

[0226] In some embodiments, the cancer, cell, tumor or subject to be treated is unable to accept or does not respond to other therapies for HER2+ cancer.In some embodiments, the subject to be treated is unable to accept trastuzumab and / or pertuzumab, and / or it has been determined that the subject is unable to accept / cannot be treated with trastuzumab and / or pertuzumab.For example, trastuzumab is associated with cardiotoxic effects (Murray, CMAJ. 2006; 174(1): 36-37).

[0227] In some embodiments, the cancer, cell, tumor, or subject to be treated is already being treated with another therapy for HER2+ cancer. In some embodiments, the subject to be treated has already been administered trastuzumab and / or pertuzumab and / or has already been treated with trastuzumab and / or pertuzumab.

[0228] cancer Cancer is any unwanted cell proliferation (or any disease manifested by unwanted cell proliferation). Cancer can be a cancer of the liver (a disease of the liver), a neoplasia or tumor, or an unwanted cell proliferation, an increased risk of or predisposition to a neoplasia or tumor. Cancer can be benign or malignant, primary or secondary (metastatic). A neoplasia or tumor can be any abnormal growth or proliferation of cells and can be located in any tissue. Exemplary tissues include adrenal gland, adrenal medulla, anus, appendix, bladder, blood, bone, bone marrow, brain, breast, cecum, central nervous system (including or excluding the brain), cerebellum, cervix, colon, duodenum, endometrium, epithelial cells (e.g., renal epithelium), gallbladder, esophagus, glial cells, heart, ileum, jejunum, kidney, lacrimal gland, pharynx, liver, lung, lymph, lymph node, lymphoblast, maxilla, mediastinum, mesentery, uterine muscle, nasopharynx, omentum, oral cavity, ovary, pancreas, parotid gland, peripheral nervous system, peritoneum, pleura, prostate, salivary gland, sigmoid colon, skin, small intestine, soft tissue, spleen, stomach, testis, thymus, thyroid, tongue, tonsils, trachea, uterus, vulva, and leukocytes.

[0229] In some embodiments, the cancer to be treated is a breast cancer, hi some embodiments, the cancer to be treated is an ovarian, uterine, lung, e.g., adenocarcinoma of the lung, or gastrointestinal, e.g., stomach, salivary duct cancer.

[0230] The tumor to be treated may be a nervous system or non-nervous system tumor. Nervous system tumors may originate from either the central or peripheral nervous system, and may be, for example, glioma, medulloblastoma, meningioma, neurofibroma, ependymoma, schwannoma, neurofibrosarcoma, astrocytoma, and oligodendroglioma. Non-nervous system cancers / tumors may originate from any other non-nervous tissue, and examples include melanoma, mesothelioma, lymphoma, myeloma, leukemia, non-Hodgkin's lymphoma (NHL), Hodgkin's lymphoma, chronic myeloid leukemia (CML), acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), cutaneous T-cell lymphoma (CTCL), chronic lymphocytic leukemia (CLL), hepatocellular carcinoma, epidermoid carcinoma, prostate cancer, breast cancer, lung cancer, colon cancer, ovarian cancer, pancreatic cancer, thymic cancer, NSCLC, hematological cancer, and sarcoma.

[0231] In some embodiments, the cancer is a cancer that encodes or expresses HER2. In some embodiments, the cancer is a cancer that overexpresses HER2. As used herein, a "HER2-positive cancer" or "HER2+ cancer" is a cancer that expresses or overexpresses the HER2 gene and / or protein.

[0232] As used herein, a "cancer that encodes or expresses HER2" or a "cancer that overexpresses HER2" includes any cell that encodes or expresses HER2. In some embodiments, the cell may be a cell of a tumor. Such a cell may be identified as a HER2-positive cell or a HER2-positive tumor cell. As used herein, a "tumor cell" includes one or more tumor cells.

[0233] Cancers or cells that express or overexpress HER2 can be identified by methods including immunohistochemistry (IHC), fluorescent in situ hybridization (FISH), chromogenic in situ hybridization (CISH), and silver-enhanced in situ hybridization (SISH), as well as other methods known in the art (Wolff et al., 2011). Arch Pathol Lab Med. 2007;131(1):18-43, incorporated herein by reference in its entirety. Other methods include the HERmark® test, an assay that uses the VeraTag system to measure total HER2 and HER2 homodimers and heterodimers to predict the efficacy of anti-HER2 therapy (Shi et al. (2009) Diagn Mol Pathol. Mar;18(1):11-21, incorporated herein by reference in its entirety).

[0234] HER2 laboratory results by IHC are classified into three categories: positive, equivocal, and negative. Positive status is assigned a score of 3+, indicating presence of more than 30% of invasive tumor cells. The tumor cells show uniform and intense membrane staining. An equivocal status is assigned a score of 2+, indicating uneven or weakly intense complete membrane staining in at least 10% of the cells, or intense complete membrane staining in 30% or less of the tumor cells. A negative status is assigned a score of 1+ or 0, indicating weak or incomplete membrane staining of any proportion of the tumor cells, or no staining, respectively (Wolff et al., supra). In the context of the present invention, a score of 3+ or 2+ is considered to be "HER2 positive."

[0235] Fluorescent (FISH) or chromogenic (CISH) in situ hybridization for HER2 gene amplification has become an essential part of the diagnostic workup for patients with breast cancer. The principle of in situ hybridization is simple: a DNA probe complementary to the genomic sequence of interest is generated, labeled, and then hybridized to target tissue. FISH / CISH techniques can be applied to a wide variety of samples: cell lines, frozen tissues, paraffin-embedded tissues, and microtissue arrays. FISH utilizes fluorescence microscopy, whereas CISH uses light microscopy (Gutierrez and Schiff (2011) Arch Pathol Lab Med. 135(1): 55-62). CISH has several advantages over FISH in detecting gene amplification: 1) permanent staining, allowing samples to be preserved; 2) use of bright-field microscopy; 3) easy identification of target cells; and 4) tumor heterogeneity is easily assessed; however, CISH does not allow determination of actual gene copy number (Lambros et al., Hum Pathol. 2007 Aug; 38(8):1105-22).

[0236] HER2 FISH positivity is defined as an average of more than six HER2 gene copies per nucleus for a test system without an internal control probe or a HER2 / CEP17 ratio greater than 2.2, where CEP17 is a centromeric probe for chromosome 17, where the HER2 gene is located. The ambiguous range for a HER2 FISH assay is defined as a HER2 / CEP ratio of 1.8 to 2.2 or an average gene copy number of 4.0 to 6.0 for a system without an internal control. Negative HER2 FISH amplification is defined as a HER2 / CEP17 ratio less than 1.8 or an average of less than four HER2 gene copies per nucleus for a system without an internal control probe (Wolff et al., supra). In the context of the present invention, a positive or ambiguous FISH status is considered "HER2 positive."

[0237] Pharmaceutically useful compositions and pharmaceutical formulations The antibodies, antigen-binding fragments, polypeptides, conjugates, nucleic acids, vectors, CARs and cells according to the invention may be formulated as pharmaceutical compositions for clinical use and may include a pharmaceutically acceptable carrier, diluent, excipient or adjuvant.

[0238] The present invention also provides methods for producing pharmaceutically useful compositions, which may comprise one or more steps selected from isolating an antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, CAR, or cell described herein, and / or mixing the isolated antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, CAR, or cell described herein with a pharmaceutically acceptable carrier, adjuvant, excipient, or diluent.

[0239] For example, a further aspect of the invention relates to a method of formulating or producing a medicament or pharmaceutical composition for use in the treatment of cancer, comprising the step of mixing an antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, CAR, or cell described herein with a pharmaceutically acceptable carrier, adjuvant, excipient, or diluent, thereby formulating a pharmaceutical composition or medicament.

[0240] Antibodies, antigen-binding fragments, polypeptides, conjugates, nucleic acids, vectors, CARs, cells, medicaments, and pharmaceutical compositions according to aspects of the present invention can be formulated for administration by a number of routes, including, but not limited to, parenteral, intravenous, intraarterial, intramuscular, intratumoral, oral, and nasal. The medicaments and compositions may be formulated for injection or in solid form. Antibodies, antigen-binding fragments, polypeptides, conjugates, nucleic acids, vectors, CARs, cells, and therapeutic agents may be formulated in liquid or solid form. Liquid forms may be formulated for administration by injection into a selected region of the human or animal body.

[0241] The administration of antibodies, antigen-binding fragments, polypeptides, conjugates, nucleic acids, vectors, compositions, CARs, or cells is preferably in a "therapeutically effective amount," which is sufficient to show benefit to an individual. The actual amount administered, as well as the rate and time course of administration, will depend on the nature and severity of the disease being treated. Prescribing treatment, such as determining dosage, is within the responsibility of medical practitioners and other physicians and typically takes into account the disorder to be treated, the condition of the individual patient, the delivery site, the method of administration, and other factors known to physicians. Examples of the techniques and protocols mentioned above can be found in Remington's Pharmaceutical Sciences, 20th Edition, 2000, published by Lippincott, Williams & Wilkin, which is incorporated herein by reference.

[0242] The antibodies, antigen-binding fragments, polypeptides, conjugates, nucleic acids, vectors, CARs, cells, medicaments, and pharmaceutical compositions can be administered alone or in combination with other treatments, either simultaneously or sequentially, depending on the condition to be treated. The other treatments can be one or more therapeutic agents.

[0243] As used herein, the antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, CAR, cell or composition of the invention and the therapeutic agent can be administered simultaneously or sequentially. The therapeutic agent can be a chemotherapeutic agent.

[0244] In some embodiments, treatment with an antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, composition, CAR, or cell of the invention may be accompanied by chemotherapy.

[0245] In some embodiments, treatment with an antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, composition, CAR, or cell of the invention may be accompanied by at least one anti-cancer agent, where "at least one anti-cancer agent" refers to one, or two, or three, or four, or more agents that are effective in treating a malignant or cancerous disease, such as a cancer, as described herein.

[0246] Co-administration refers to the administration of an antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, composition, CAR, or cell together with a therapeutic or anti-cancer agent, e.g., as a pharmaceutical composition containing both agents (combined preparation) or shortly after each other, and optionally through the same route of administration, e.g., into the same artery, vein, or other blood vessel.

[0247] Sequential administration refers to the administration of one of antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, composition, CAR or cell, or therapeutic agent / anticancer agent, followed by the separate administration of another agent after a predetermined time interval.The two agents do not need to be administered by the same route, although this is the case in some embodiments.The time interval can be any time interval.

[0248] The therapeutic or anti-cancer agent described herein can be an anti-HER2 antibody or an agent that targets HER2. In some embodiments, the therapeutic or anti-cancer agent described herein is trastuzumab (Herceptin®), trastuzumab emtansine (Kadsi®), or trastuzumab serotonin (TMS). In some embodiments, the antibody, antigen-binding molecule, or polypeptide described herein is administered in combination, e.g., simultaneously or sequentially, with one or more of the above agents, such as trastuzumab (Herceptin®) and / or pertuzumab (Perjeta®).

[0249] The therapeutic or anti-cancer agents described herein may be formulated so as to be suitable for injection or infusion into a tumor or into the blood. Chemotherapy / radiation therapy Chemotherapy and radiotherapy refer to the treatment of cancer with drugs or ionizing radiation (eg, radiotherapy using X-rays or gamma rays), respectively.

[0250] A drug may be a chemical entity, e.g., a small molecule pharmaceutical, an antibiotic, a DNA intercalator, a protein inhibitor (e.g., a kinase inhibitor), or a biological agent, e.g., an antibody, an antibody fragment, a nucleic acid or peptide aptamer, a nucleic acid (e.g., DNA, RNA), a peptide, a polypeptide, or a protein. A drug may be formulated as a pharmaceutical composition or medicament. A formulation may include one or more drugs (e.g., one or more active agents) together with one or more pharmaceutically acceptable diluents, excipients, or carriers.

[0251] Treatment may involve the administration of more than one drug.Depending on the condition to be treated, drug may be administered alone or in combination with other treatment, simultaneously or sequentially.For example, chemotherapy may be a combination therapy that includes the administration of two drugs, one or more of which may be intended to treat cancer.

[0252] Chemotherapy may be administered by one or more routes of administration, such as parenteral, intravenous injection, oral, subcutaneous, intradermal, or intratumoral routes. Chemotherapy may be administered according to a treatment regimen, which may be a predetermined timetable, plan, scheme, or schedule of chemotherapy administration that may be prepared by a doctor or physician and may be tailored to the patient in need of treatment.

[0253] A treatment regimen may indicate one or more of: the type of chemotherapy to administer to the patient; the dose of each drug or radiation; the time interval between doses; the length of each treatment; and the number and nature of any treatment holidays, if any. For combination therapy, a single treatment regimen may be provided that indicates how each drug is to be administered.

[0254] Chemotherapeutic agents and biologics may be selected from: alkylating agents such as cisplatin, carboplatin, mechlorethamine, cyclophosphamide, chlorambucil, ifosfamide; purine or pyrimidine antimetabolites such as azathioprine or mercaptopurine; alkaloids and terpenoids such as vinca alkaloids (e.g., vincristine, vinblastine, vinorelbine, vindesine), podophyllotoxin, etoposide, teniposide, taxanes such as paclitaxel (Taxol), TM ), docetaxel; topoisomerase inhibitors, such as type I topoisomerase inhibitors, camptothecin, irinotecan and topotecan, or type II topoisomerase inhibitors, amsacrine, etoposide, etoposide phosphate, teniposide; antitumor antibiotics (e.g., anthracycline antibiotics), such as dactinomycin, doxorubicin (adriamycin), TM ), epirubicin, bleomycin, rapamycin; antibody-based agents, such as anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-TIM-3 antibodies, anti-CTLA-4, anti-4-1BB, anti-GITR, anti-CD27, anti-BLTA, anti-OX43, anti-VEGF, anti-TNFα, anti-IL-2, anti-GpIIb / I IIa, anti-CD-52, anti-CD20, anti-RSV, anti-HER2 / neu (erbB2), anti-TNF receptor, anti-EGFR antibodies, monoclonal antibodies or antibody fragments, examples of which include: cetuximab, panitumumab, infliximab, basiliximab, bevacizumab (Avastin®), abciximab, daclizumab, gemtuzumab, alemtuzumab, rituximab (MabThera®), palivizumab, trastuzumab, etanercept, adalimumab, nimotuzumab; EGFR inhibitors such as erlotinib, cetuximab and gefitinib; anti-angiogenic agents such as bevacizumab (Avastin®); cancer vaccines such as sipuleucel-T (Provenge®).

[0255] Additional chemotherapeutic agents include: 13-cis-retinoic acid, 2-chlorodeoxyadenosine, 5-azacytidine, 5-fluorouracil, 6-mercaptopurine, 6-thioguanine, Abraxane, Accutane®, Actinomycin-D, Adriamycin®, Adrsil®, Afinitor®, Agrylin®, Ala-Cort®, aldesleukin, alemtuzumab, ALIMTA, alitretinoin Alkaban-AQ (registered trademark), Alkeran (registered trademark), all-trans retinoic acid, alpha interferon, altretamine, amethopterin, amifostine, aminoglutethimide, anagrelide, anandrone (registered trademark), anastrozole, arabinosylcytosine, Aranesp (registered trademark), Alesia (registered trademark), Arimidex (registered trademark), Aromasin (registered trademark), Alanon (registered trademark), arsenic trioxide, asparaginase, ATRA, Avastin (registered trademark), azacitidine, BCG, BCNU, bendamustine, bevacizumab, bexarotene, BEXXAR (registered trademark), bicalutamide, BiCNU, Blenoxan (registered trademark), bleomycin, bortezomib, busulfan, Busulfex (registered trademark), calcium leucovorin, Campath (registered trademark), Camptosar (registered trademark), camptothecin-11, capecitabine, carac TM , carboplatin, carmustine, Casodex®, CC-5013, CCI-779, CCNU, CDDP, CeeNU, Cervizin®, cetuximab, chlorambucil, cisplatin, citrovorum factor, cladribine, cortisone, Cosmegen®, CPT-11, cyclophosphamide, Citadren®, cytarabine, Citosar-U®, Cytoxan®, Dacogen, dactinomycin, darbepoetin alfa, dasatinib, daunomycin, daunorubicin, daunorubicin hydrochloride, daunorubicin liposomal, Daunoxome®, Decadron, decitabine, Delta-Cortef®, Deltazone®, denileukin, diftitox, Depocyto TM, dexamethasone, dexamethasone acetate, dexamethasone sodium phosphate, dexasone, dexrazoxane, DHAD, DIC, Geodex, docetaxel, Doxil®, doxorubicin, doxorubicin liposomal, droxia TM , DTIC, DTIC-Dome (registered trademark), Duralon (registered trademark), Eligard TM , Elence TM , Eloxatin TM , Elspar (registered trademark), Encit (registered trademark), epirubicin, epoetin alfa, Erbitux, erlotinib, Erwinia L-asparaginase, estramustine, ethyol-etopofos (registered trademark), etoposide, etoposide phosphate, Eurexin (registered trademark), everolimus, Evista (registered trademark), exemestane, Faslodex (registered trademark), Femara (registered trademark), filgrastim, floxuridine, Fludara (registered trademark), fludarabine, Fluoroplex (registered trademark), fluorouracil, floxymesterone, flutamide, folinic acid, FUDR (registered trademark), fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, Gleevec TM , Gliadel® wafer, goserelin, granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, Herceptin®, Hexadrol, Hexalen®, hexamethylmelamine, HMM, Hicamtine®, Hydrea®, Hydrocort® acetate, hydrocortisone, hydrocortisone sodium phosphate, hydrocortisone sodium succinate, hydrocortone phosphate, hydroxyurea, Ibritz Momab, ibritumomab tiuxetan, idamycin®, idarubicin, Ifex®, IFN-alpha, ifosfamide, IL-11, IL-2, imatinib mesylate, imidazole carboxamide, interferon alpha, interferon alpha-2b (PEG conjugate), interleukin-2, interleukin-11, Intron A® (interferon alpha-2b), Iressa®, irinotecan, isotretinoin, ixabepilone, ixempra TM , Kidrolase, Lanacort®, Lapatinib, L-asparaginase, LCR, Lenalidomide, Letrozole, Leucovorin, Leukeran, Leukine TM , leuprolide, leurocristin, leustatin TM , Liposomal Ara-C, Liquid Pred®, Lomustine, L-PAM, L-Sarcolysin, Lupron®, Lupron Depot®, Matulan®, Maxidex, Mechlorethamine, Mechlorethamine Hydrochloride, Medralon®, Medrol®, Megase®, Megestrol, Megestrol Acetate, Melphalan, Mercaptopurine, Mesna, Mesunex TM , methotrexate, methotrexate sodium, methylprednisolone, methycorten (registered trademark), mitomycin, mitomycin-C, mitoxantrone, M-prednisol (registered trademark), MTC, MTX, Mustagen (registered trademark), Mustine, Mutamycin (registered trademark), Myleran (registered trademark), Mirocel TM , Mirotarg (registered trademark), Navelbine (registered trademark), Nelarabine, Neosar (registered trademark), Neulasta TM , Neumega (registered trademark), Neupogen (registered trademark), Nexavar (registered trademark), Nilandrone (registered trademark), Nilutamide, Nipent (registered trademark), Nitrogen Mustard, Novardex (registered trademark), Novantrone (registered trademark), Octreotide, Octreotide Acetate, Oncospar (registered trademark), Oncovin (registered trademark), Ontac (registered trademark), Onxal TM, Oplevelkin, Orapred®, Orazon®, oxaliplatin, paclitaxel, paclitaxel protein-bound, pamidronate, panitumumab, Panretin®, Paraplatin®, Pediapred®, PEG-interferon, pegaspargase, pegfilgrastim, PEG-Intron TM , PEG-L-asparaginase, PEMETREXED, pentostatin, phenylalanine mustard, Platinol®, Platinol-AQ®, prednisolone, prednisone, Prelon®, procarbazine, PROCRIT®, Proleukin®, carmustine implant, Prolife Prospan 20 including Prinetol®, raloxifene, Revlimid®, Rheumatrex®, Rituxan®, rituximab, Roferon-A® (interferon alpha-2a), Lubex®, rubidomycin hydrochloride, Sandostatin®, Sandostatin LAR®, Sargramostim, Solu-Cortef®, Solu-Medrol®, sorafenib, Sprycel® TM , STI-571, streptozocin, SU11248, sunitinib, Sutent (registered trademark), tamoxifen, Tarceva (registered trademark), Targretin (registered trademark), Taxol (registered trademark), Taxotere (registered trademark), Temodar (registered trademark), temozolomide, temsirolimus, teniposide, TESPA, thalidomide, Talomide (registered trademark), Terasis (registered trademark), thioguanine, thioguanine tabloid (registered trademark), thiophosphoamide, Thioplex (registered trademark), thiotepa, TICE (registered trademark), Toposar (registered trademark), topotecan, toremifene, Torisel (registered trademark), tositumomab, trastuzumab, Treanda (registered trademark), tretinoin, Trexar TM , Trisenox (registered trademark), TSPA, TYKERB (registered trademark), VCR, Vectivix TM , Velban (registered trademark), Velcade (registered trademark), Bepesid (registered trademark), Vesanoid (registered trademark), ViadurTM , Vidaza (registered trademark), vinblastine, vinblastine sulfate, Vincasar Pfs (registered trademark), vincristine, vinorelbine, vinorelbine tartrate, VLB, VM-26, vorinostat, VP-16, Bumon (registered trademark), Xeloda (registered trademark), Zanosar (registered trademark), Zevalin TM , Ginecard (registered trademark) ), Zoladex®, zoledronic acid, Zolinza, Zometa®.

[0256] In some embodiments, the antibody / antigen-binding fragment according to the invention is administered in combination with one or more of doxorubicin, epirubicin, paclitaxel, docetaxel, fluorouracil, cyclophosphamide, methotrexate, and capecitabine.

[0257] Medication regimen Multiple doses of the antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, CAR, or cell may be provided, and one or more or each of the doses may be accompanied by the simultaneous or sequential administration of another therapeutic agent.

[0258] The multiple doses may be separated by predetermined time intervals, which may be selected to be one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 days, or 1, 2, 3, 4, 5, or 6 months. For example, doses may be administered once every 7, 14, 21, or 28 days (plus or minus 3, 2, or 1 day).

[0259] kit In some aspects of the present invention, a kit of parts is provided. In some embodiments, the kit may include at least one container containing a predetermined amount of an antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, CAR, or cell described herein. The kit may provide the antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, CAR, or cell in the form of a medicament or pharmaceutical composition, and may be provided with instructions for administration to a patient to treat a specified disease or condition. The antibody, antigen-binding fragment, or polypeptide may be formulated to be suitable for injection or infusion into a tumor or blood.

[0260] In some embodiments, the kit may further include at least one container with a predetermined amount of another therapeutic agent (e.g., a chemotherapeutic or anti-cancer agent). In such embodiments, the kit may also include a second medicament or pharmaceutical composition so that the two medicaments or pharmaceutical compositions can be administered simultaneously or separately to provide a combined treatment for a particular disease or condition. The therapeutic agent or anti-cancer agent may also be formulated to be suitable for injection or infusion into a tumor or blood.

[0261] Subject The subject to be treated with the antibody, antigen-binding fragment, polypeptide, conjugate, nucleic acid, vector, CAR or cell according to the present invention may be any animal or human. The subject is preferably a mammal, more preferably a human. The subject may be a non-human mammal, but more preferably a human. The subject may be male or female. The subject may be a patient. The subject may be diagnosed with a disease or condition that requires treatment, or may be suspected of having such a disease / condition.

[0262] Sequence identity Percent (%) sequence identity is defined as the percent of amino acid residues in a candidate sequence that are identical to the residues in a given recited sequence (referenced by a SEQ ID NO) after aligning the sequences and introducing gaps, if necessary, to achieve maximum sequence identity, and does not take into account any conservative substitutions as part of the sequence identity. Sequence identity is preferably calculated based on each is calculated over the entire length of the sequence.

[0263] If the aligned sequences are of different lengths, the sequence identity of the shorter comparison sequence may be determined over the entire length of the longer given sequence, or if the comparison sequence is longer than the given sequence, the sequence identity of the comparison sequence may be determined over the entire length of the shorter given sequence.

[0264] For the purpose of determining percent amino acid sequence identity, alignment can be achieved by various methods known to those skilled in the art, for example, by using publicly available computer software, such as ClustalW 1.82, ClustalO, T-coffee or Megalign (DNASTAR) software.When using such software, preferably use default parameters, for example, gap penalty and extension penalty.The default parameters of ClustalW 1.82 are: Protein Gap Open Penalty=10.0, Protein Gap Extension Penalty=0.2, Protein matrix= Gonnet, Protein / DNA ENDGAP = -1, Protein / DNA GAPDIST = 4. The default parameters for ClustalO are described in Soding, J. (2005) 'Protein homology detection by HMM-HMM comparison'. Bioinformatics 21, 951-960.

[0265] Recombinant production The immunogens, antibodies, fragments, polypeptides, conjugates, and CARs according to the present invention may be produced by recombinant expression. Molecular biology techniques suitable for recombinant production are well known in the art, such as those set forth in Green and Sambrook, Molecular Cloning: A Laboratory Manual (4th Edition), Cold Spring Harbor Press, 2012, which is incorporated herein by reference in its entirety.

[0266] In some embodiments, the nucleic acids described herein are purified or isolated, e.g., from other nucleic acids or naturally occurring biological materials. In some embodiments, the nucleic acid(s) comprise or consist of DNA and / or RNA.

[0267] Expression may be from a nucleotide sequence. The nucleotide sequence may be contained in a vector. As used herein, a "vector" is an oligonucleotide molecule (DNA or RNA) used as a vehicle to transfer exogenous genetic material into a cell. The vector may be an expression vector for expressing exogenous genetic material in a cell. Such a vector may include a promoter sequence operably linked to a nucleotide sequence encoding the sequence to be expressed. The vector may also include a termination codon and an expression enhancer. Any suitable vector, promoter, enhancer, and termination codon known in the art may be used to express a peptide or polypeptide from a vector according to the present invention. In some embodiments, the vector may be a plasmid, MAC, virus, etc. In some embodiments, the vector may be a eukaryotic expression vector, e.g., a vector containing elements necessary for protein expression from the vector in a eukaryotic cell. In some embodiments, the vector may be a mammalian expression vector, e.g., including a cytomegalovirus (CMV) or SV40 promoter to drive protein expression.

[0268] The term "operably linked" includes the ability of a selected nucleotide sequence and a regulatory nucleotide sequence (e.g., a promoter and / or enhancer) to function in a controlled manner. This includes situations in which a nucleotide sequence is covalently linked (thereby forming an expression cassette) in such a manner as to achieve expression of the nucleotide sequence under the influence or control of the sequence. Thus, a regulatory sequence is operably linked to a selected nucleotide sequence if the regulatory sequence is capable of effecting transcription of the nucleotide sequence. The resulting transcript may then be translated into a desired peptide or polypeptide.

[0269] For recombinant production according to the present invention, any cell suitable for expression of a polypeptide may be used. The cell may be a prokaryotic or eukaryotic organism. In some embodiments, the cell is a prokaryotic cell, such as an archaeal or bacterial cell. In some embodiments, the bacterium may be a gram-negative bacterium, such as a bacterium of the Enterobacteriaceae family, such as Escherichia coli.

[0270] In some embodiments, the cell is a eukaryotic cell, such as a yeast cell, a plant cell, an insect cell, or a mammalian cell, such as a CHO, HEK, HeLa, or COS cell. In some embodiments, the host cell is a non-human cell, such as a non-human mammalian cell.

[0271] In some cases, the cells are not prokaryotic because prokaryotic cells do not allow the same folding or post-translational modifications as eukaryotic cells. Furthermore, much higher expression levels are possible in eukaryotes, and with the appropriate tags, it may be easier to purify proteins from eukaryotes. Specific plasmids that enhance secretion of proteins into the medium may also be available.

[0272] Production may involve the culture or fermentation of eukaryotic cells modified to express the peptide or polypeptide. Culture or fermentation may be carried out in a bioreactor provided with an appropriate supply of nutrients, air / oxygen, and / or growth factors. The secreted protein can be collected from the cells by partitioning the medium / fermentation broth, extracting the protein content, and separating the individual proteins to isolate the secreted peptide or polypeptide. Culture, fermentation, and separation techniques are well known to those of skill in the art and are described, for example, in Green and Sambrook, Molecular Cloning: A Laboratory Manual (4th ed.; incorporated herein by reference).

[0273] A bioreactor contains one or more vessels in which cells can be cultured. Cultivation in a bioreactor can occur continuously, with continuous flow of reactants into the reactor and continuous flow of cultured cells out of the reactor. Alternatively, cultivation can occur in batches. Bioreactors monitor and regulate environmental conditions, such as pH, oxygen, flow rates into and out of the vessel, and agitation within the vessel, to provide optimal conditions for the cells being cultured.

[0274] After culturing cells expressing an immunogen, antibody, fragment, polypeptide, conjugate, or CAR, the peptide / polypeptide of interest is preferably isolated. Any suitable method known in the art for isolating proteins from cell culture may be used. To isolate the peptide / polypeptide from the culture, it may be necessary to first separate the cultured cells from the medium containing the peptide / polypeptide of interest. If the peptide / polypeptide of interest is secreted from the cells, the cells may be separated from the medium containing the secreted peptide / polypeptide of interest by centrifugation. If the peptide / polypeptide of interest accumulates within the cells, it may be necessary to disrupt the cells before centrifugation, for example, using ultrasound, rapid freeze-thawing, or osmotic lysis. Centrifugation will produce a pellet containing the cultured cells, or a supernatant containing cell debris from the cultured cells, medium, and the peptide / polypeptide of interest.

[0275] It may then be desirable to isolate the peptide / polypeptide of interest from the supernatant or medium, which may also contain other proteins and non-protein components. A common approach to separating protein components from the supernatant or medium is by precipitation. Proteins of different solubilities are precipitated with different concentrations of a precipitant, such as ammonium sulfate. For example, low concentrations of the precipitant extract water-soluble proteins. Thus, by adding different, increasing concentrations of the precipitant, proteins of different solubilities can be distinguished. Dialysis can then be used to remove the ammonium sulfate from the separated proteins.

[0276] Other methods for distinguishing between different proteins are known in the art, such as ion exchange chromatography and size chromatography, and may be used as an alternative to precipitation or may follow precipitation.

[0277] Once the peptide / polypeptide of interest has been isolated from the culture, it may be desirable or necessary to concentrate the peptide or polypeptide. Several methods for concentrating proteins are known in the art, such as ultrafiltration or lyophilization.

[0278] array

[0279] [Table 2-1]

[0280] [Table 2-2]

[0281] [Table 2-3]

[0282] [Table 2-4]

[0283] [Table 2-5]

[0284] [Table 2-6]

[0285] [Table 2-7]

[0286] [Table 2-8]

[0287] [Table 2-9]

[0288] [Table 2-10]

[0289] [Table 2-11]

[0290] [Table 2-12]

[0291] [Table 2-13]

[0292] [Table 2-14]

[0293] [Table 2-15]

[0294] [Table 2-16]

[0295] The present invention includes combinations of the described aspects and preferred features except where such combinations are clearly impermissible or clearly avoided. The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0296] Aspects and embodiments of the present invention will now be illustrated, by way of example, with reference to the accompanying figures. Further aspects and embodiments will be apparent to those skilled in the art. All documents mentioned in this text are incorporated herein by reference.

[0297] Throughout this specification, including the claims that follow, unless the context requires otherwise, the word "comprises" and variations such as "comprises" and "comprising" mean the inclusion of a stated integer or step or integers or steps, but not any other integer. It will be understood that no single step, or integer or group of steps, is excluded.

[0298] It should be noted that, in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Ranges may also be expressed herein as from "about" one particular value and / or to "about" another particular value. When such a range is expressed, another embodiment includes from the one particular value and / or to the other particular value. Similarly, when values ​​are expressed as approximations, it will be understood that the particular value forms another embodiment by use of the antecedent "about." [Example]

[0299] Example 1: Discovery Anti-human HER-2 binding antibodies were isolated from a human Fab library by phage display in three rounds of biopanning. Binding of isolated Fabs from the human Fab library to immobilized HER-2 protein was measured by ELISA (Figure 3). The dissociation constant (K) of each clone from HER-2 was measured. d ) was measured by surface plasmon resonance (SPR). The results are shown in Table 1.

[0300] [Table 3]

[0301] Four clones were initially selected for further characterization: P1A3, P1C5, P1E4, and P1F1. Clones A4, B4, B5, C3, D4, E1, F5, and G3 were also selected for characterization and investigation of their utility in CARs.

[0302] Example 2: Specific binding to HER-2 Binding to HER-2 The interaction between four antibody clones (P1A3, P1C5, P1E4, and P1F1) and HER-2 was examined by SPR using soluble HER-2 bound to a microchip and flowing antibodies. To assess the specificity of binding, molecules closely related to HER-2, namely EGFR and HER-3, were also used as binding antigens. The humanized anti-HER-2 antibody trastuzumab was used as a positive control.

[0303] Four antibodies showed specific binding to human HER-2 and no apparent interaction with HER-3 or EGF-G (Fig. 4). Binding to HER2-expressing cells To further characterize the binding of the antibody to HER-2, the antibody was incubated with cells that constitutively overexpress HER-2, BT-474, and SK-BR-3, cells that express but do not overexpress HER-2, BT-20, and MCF-7, or the triple-negative cancer cell line MDA-MB-468, which does not express the antigen. Both BT-474 and SK-BR-3 cell lines are classified as 3+ for HER-2 expression, i.e., overexpressing. The BT-20 cell line is classified as a basal HER-2 cell line with an expression score of 0 / 1+, while MCF-7 has the same score and is classified as a luminal tumor cell line. MDA-MB-468 has a HER-2 expression score of 0 (Subik K et al. 2010, Breast Cancer (Auckl), 4:35-41).

[0304] After incubation, a secondary antibody was added and its binding to the cells was detected by flow cytometry. Trastuzumab was used as a positive control in all assays. The four anti-HER-2 antibodies bound to HER-2-overexpressing (Fig. 5A) and HER-2-expressing (Fig. 5B) cells in a manner similar to trastuzumab, whereas no binding was observed in HER-2-negative cells (Fig. 5C).

[0305] The antibodies were then incubated with HER-2 expressing cells at various concentrations to study the dose-effect binding profile. In these assays, E4 exhibited a binding profile most similar to that of trastuzumab (Figure 6).

[0306] Example 3: Binding to an epitope distinct from trastuzumab To assess whether the four anti-HER-2 antibodies P1A3, P1C5, P1E4, and P1F1 bind the same epitope as trastuzumab, we performed a binding assay using SPR. Briefly, the antibody of interest was immobilized on an SPR microchip, and then recombinant HER-2 was applied to bind to the immobilized antibody. Afterwards, all other antibodies were applied, and their binding was measured.

[0307] All four antibodies showed similar dendrograms, suggesting that they share a common epitope or bind to closely related epitopes. However, trastuzumab showed a different binding profile, demonstrating that the four antibodies do not share the epitope of trastuzumab (Figure 7).

[0308] The binding epitopes of clones A4, B4, B5, C3, D4, E1, F5 and G3 are evaluated by SPR as described above.Comparing the binding profiles with those of trastuzumab, clones A4, B4, B5, C3, D4, E1, F5 and G3 are found to bind to epitopes different from those of trastuzumab.

[0309] Example 4: Affinity for human HER-2 Affinity was measured by SPR. E4 showed the highest affinity for HER-2 among the four clones (Table 2).

[0310] [Table 4]

[0311] The binding properties of clones A4, B4, B5, C3, D4, E1, F5, and G3 were also determined by SPR. The association rate constant, dissociation rate constant (k a , k d ) and the equilibrium dissociation constant (K D ) was determined for anti-HER2 Fab molecules. Human HER2 protein (50 μg / mL) was immobilized on an SPR sensor chip and exposed to five different concentrations of each Fab at 2-fold or 3-fold dilutions across a flow cell. A 1:1 Langmuir model was used to derive the constants.

[0312] The results are shown in Table 3.

[0313] [Table 5]

[0314] Example 5: Mechanism of Action Membrane disruption HER2-overexpressing BT-474 cells were incubated with P1A3, P1C5, P1E4, or P1F1 and propidium iodide. Unlike trastuzumab, the four antibodies induced membrane disruption, allowing propidium iodide insertion and demonstrating cell death (Figure 8).

[0315] Induction of apoptosis HER2-overexpressing cell lines BT-474 and SK-BR-3 were incubated in the presence of P1E4 or trastuzumab and stained with annexin-V and propidium iodide. Apoptosis and death were measured by staining.

[0316] Cells incubated with P1E4 and showing propidium iodide staining, i.e., dying / dead cells, also showed staining with Annexin-V, indicating that P1E4 induces cell death by apoptosis (Figure 9). To further confirm the induction of apoptosis, BT-474 cells were incubated in the presence of P1E4, trastuzumab, or pertuzumab (another anti-HER-2 antibody) for subsequent measurement of propidium uptake, activation of the caspase 3 / 7 pathway, and activity of cleaved PARP. The caspase 3 / 7 pathway is involved in the apoptotic process. After activation, caspase 3 / 7 activates cleaved PARP in the nucleus, which in turn can induce the release of proapoptotic factors by mitochondria and inhibit the process of DNA repair.

[0317] As described above, incubation in the presence of P1E4 resulted in the uptake of propidium iodide (Figure 10A). P1E4 induced activation of caspase 3 / 7 (Figure 10B) and cleaved PARP (Figure 10C). Neither trastuzumab nor pertuzumab showed induction of this pathway in the assay, suggesting different mechanisms of action.

[0318] The kinetics of P1E4-induced cell death was analyzed. Briefly, BT-474 and SK-BR-3 cells were incubated with 10 μg / mL E4 or trastuzumab at 37°C. 10, 20, 30, 40, 50, and 60 minutes after antibody addition, cells were harvested and analyzed for induction of cell death by propidium iodide uptake.

[0319] Immediately after addition of P1E4, 10-20 min, cells began to die, whereas cells incubated with Trastuzumab did not show any signs of lysis (Fig. 11). Clones A3, C5, F1, A4, B4, B5, C3, D4, E1, F5 and G3 were evaluated for their mechanism of action and found to induce cell death, for example, by apoptosis.

[0320] Example 6: Efficacy in vitro antitumor activity To evaluate the activity of the antibody on tumor growth, BT-474 cells were incubated for 72 hours in the presence of various concentrations of P1E4. Trastuzumab and pertuzumab were used as positive controls capable of inhibiting tumor growth. Some cells were grown without antibody and served as a growth reference. The MDA-MB-468 cell line, a cell line that does not express HER-2, was used as a negative control. After 72 hours, cell numbers were measured using Promega's Celltiter 96 Aqueous One Solution cell proliferation assay (MTS).

[0321] P1E4 exhibited a cytostatic effect on B-474 tumor growth that was comparable to that of trastuzumab or pertuzumab (FIG. 12). Similar assays were performed to evaluate the potential synergistic effect of P1E4 and trastuzumab. The combination of P1E4 and trastuzumab exhibited greater cytostatic efficacy than either antibody alone, suggesting a synergistic effect (Figure 13).

[0322] Antibody-dependent cellular cytotoxicity of P1A3, P1C5, P1E4, or P1F1 was also measured and compared to that of trastuzumab. BT-474 cells were plated with various concentrations of antibody and PBMCs at an effector-to-target (E:T) ratio of 10:1. After 16 hours of incubation at 37°C, supernatants were collected and analyzed by Promega's C LDH release was assessed by ytotox 96 non-radioactive cytotoxicity assay kit.

[0323] All four antibodies were able to induce ADCC, with P1E4 showing the most potent ADCC effect. At high concentrations, P1E4 was able to induce cytotoxicity at rates similar to those of trastuzumab, but was less effective at lower doses (Figure 14).

[0324] Clones A4, B4, B5, C3, D4, E1, F5 and G3 were evaluated for their effect on tumor growth and were found to have a cytotoxic effect. In vivo antitumor efficacy P1E4 was tested for its ability to control tumor growth in vivo. Briefly, BT-474 cells were injected in nude mice. Tumors grew to approximately 125 mm 3 Once tumors reached a mass index (MI) of 10 mg / kg, i.e., were palpable, the antibodies were injected intravenously every 4 days from days 1 to 17 at 10 mg / kg for trastuzumab and 3.3, 10, 20, or 30 mg / kg for P1E4. Only one control group received vehicle at 10 mL / kg / injection. Tumor growth was then monitored every 3 to 4 days until day 31.

[0325] While doses up to 20 mg / kg appeared to control tumor growth during the treatment period, doses up to 20 mg / kg did not confer long-term protection, and tumors began to grow at the end of treatment. At the highest dose, P1E4 not only allowed a noticeable regression of tumor size, but also conferred long-term protection; regressed tumors did not grow back after treatment ended (Figure 15).

[0326] Combination treatment with P1E4, trastuzumab, or pertuzumab was tested for synergistic effects on tumor growth control in vivo. Similar to the above, nude mice bearing BT-474 xenografts were treated with 10 mg / kg of trastuzumab, pertuzumab, or P1E4 in combination with either of the two antibodies every 4 days from days 1 to 17, and tumor growth was monitored through day 39.

[0327] P1E4 in combination with either trastuzumab or pertuzumab is as effective as the combination of trastuzumab and pertuzumab in controlling tumor growth (Figure 16). Furthermore, combining 10 mg / kg P1E4 with either trastuzumab or pertuzumab (both at 10 mg / kg) was found to be as effective as 30 mg / kg P1E4 alone (Figure 15) and much more effective than trastuzumab alone (Figure 15) in conferring long-term protection (Figure 16).

[0328] Example 7: Generation and activity of chimeric antigen receptor (CAR) T cells Figure 17 shows a schematic diagram of the construction of an anti-HER2 CAR. The CAR consists of a signal peptide, an anti-HER2 scFv, an IgH hinge, a transmembrane domain of CD28, followed by a second-generation CAR signaling tail composed of 4-1BB and CD3ζ signaling domains.

[0329] Anti-HER2 CARs, including clones A4, B4, B5, C3, D4, E1, F5, and G3, were constructed and evaluated for antitumor activity. Lentiviral-based constructs T cells from healthy donors were transfected with TransAct TM TexMACS activated with (CD3 / CD28 agonist) and supplemented with 50 ng / mL IL TM 1–2 × 10 cells in culture medium for 48 h. 6 Upon stimulation, cells were cultured at a density of 1000 cells / mL. Upon stimulation, cells were cultured with 1000 cells / mL of HER2-encoding anti-HER2 CAR, including one of the clones A4, B4, B5, D4, E1, F5, and G3. The cells were transduced with a lentivirus.

[0330] 72 hours after transduction, antitumor activity (% cell lysis) was analyzed in real time using an electrical impedance-based tumor cell culture system (xCELLigence). BT474 cells (human breast cancer cells) were seeded in triplicate in electrode-coated 96-well plates (e-plates) at a density of 10,000 cells per well. 24 hours later, CAR T cells (effector cells) were added at a ratio of 3 effector cells to each target cell (BT474).

[0331] Cytokines (IFN-γ and IL-2) secreted into the medium after 48 h of co-culture were measured by ELISA. Data from triplicates are presented as mean ± SEM. The results are shown in Figure 18A and Figure 18B. CAR T cells containing one of each of the seven anti-HER2 CARs demonstrated tumor cell cytolysis, i.e., anti-tumor activity (18A), and increased IFN-γ and IL-2 secretion, i.e., T cell activation (18B).

[0332] Electroporation construct Figure 19 shows a schematic diagram of anti-HER2 CAR mRNA. CAR T cells were generated by mRNA electroporation. T cells from healthy donors were isolated, activated, and cultured for 48 hours as described above. Activated T cells were transfected with CAR mRNA containing clone C3 (1 × 10 6 T cells were electroporated with 2 μg of IFN-γ per T cell and assessed for their cytotoxic activity (%) 20 h after electroporation by the xCELLigence system. Cytokines (IFN-γ and IL-2) secreted into the medium after 48 h of co-culture were measured by ELISA. Data from triplicates are presented as mean ± SEM.

[0333] The results are shown in Figure 20A and Figure 20B. CAR T cells containing the anti-HER2 CAR (C3) demonstrated tumor cell cytolysis, i.e., anti-tumor activity (20A), and increased IFN-γ and IL-2 secretion, i.e., T cell activation (20B).

[0334] Selection for tumor cells Anti-HER2 CAR T cells according to the invention are evaluated for their ability to distinguish between cells that overexpress HER2 and cells with lower or physiological levels of HER2. CAR T cells with lower or intermediate affinity for HER2 may be able to distinguish between cells that express HER2 at high levels and cells that express HER2 at low levels (e.g., as described in Liu et al. Cancer Res. 2015; (75) (17) 3596-3607).

[0335] The anti-HER2 CAR T cells of the present invention are stimulated with HER2-overexpressing tumor cell lines, such as SK-BR3 (breast cancer), SK-OV3 (ovarian cancer), and / or BT-474 (breast cancer), or tumor cell lines expressing HER2 at low or undetectable levels, such as MCF7 (breast cancer), 293T (embryonic kidney), A549 (lung cancer), 624Mel (melanoma), PC3 (prostate cancer), MDA231 (breast cancer), and / or MDA468 (breast cancer). CAR T cell activation is assessed 24 or 48 hours later by upregulation of CD137 (4-1BB), secretion of IFNγ and IL-2, and / or induction of surface CD107a expression. T cells expressing a CAR containing the antigen-binding domain of trastuzumab (high affinity CAR) and T cells expressing a non-HER2-specific CAR are used as controls. CD137 and CD107a expression is measured by flow cytometry (eg, CD3+ gated) and cytokine secretion in the culture supernatants is measured by ELISA.

[0336] CAR T cells comprising the antigen-binding molecules of the present invention have been shown to be highly reactive to tumor cells with high HER2 expression, but have low or no reactivity to tumor cell lines with low or no HER2 expression. Anti-HER2 CAR T cells have also been shown to secrete higher levels of cytokines when exposed to cells expressing high levels of HER2, demonstrating higher T cell activity compared to that seen with cells expressing low levels or no HER2.

[0337] To evaluate whether anti-HER2 CAR T cells can distinguish between tumor cells and non-tumor cells, T cells are contacted with tumor cells that overexpress HER2 (e.g., BT-474) and cell lines that express HER2 at physiological levels, such as primary organ cell lines.T cells that express non-HER2 specific CAR and cell lines that express HER2 at high or low levels are used as controls.CAR T cells are stimulated with HER2-expressing cells for 4 hours.CAR T cell recognition and activation are evaluated by monitoring CD107a upregulation and analyzed by flow cytometry by gating on CD3+ cells.

[0338] CAR-T cells containing the antigen-binding molecules of the present invention are found to be selective for HER2-overexpressing cells. These cells have been shown to exhibit potent activity against HER2-overexpressing cells, but when contacted with organ cell lines, they exhibit no or weak activity. The present invention includes, but is not limited to, the following aspects. [Aspect 1] Amino acid sequences i) to iii), or amino acid sequences iv) to vi), or preferably amino acid sequences i) to vi): i) LC-CDR1: GLSSGSVSTX1X2YX3S (SEQ ID NO: 9); ii) LC-CDR2: X4TNX5RSS (SEQ ID NO: 10); iii) LC-CDR3: X6LYX7GX8GIX9V (SEQ ID NO: 11); iv) HC-CDR1:X 10 X 11 X 12 X 13 X 14 X 15 X 16 (SEQ ID NO: 12); v) HC-CDR2:X 17 X 18 X 19 X 20 X 21 X 22 GX 23 TX 24 YX 25 X 26 X 27 X 28 X 29 X 30 (SEQ ID NO: 13); vi) HC-CDR3:X 31 X 32 X 33 X 34 X 35 SX 36 X 37 YX 38 X 39 X 40 X 41 X 42 X 43 (SEQ ID NO: 14); Alternatively, variants thereof, in which one, two or three amino acids in one or more of sequences (i) to (vi) are substituted with another amino acid (wherein X1 = G or S, X2 = H or Y, X3 = A or P, X4 = N, S or T, X5 = T, S or I, X6 = V or M, X7 = V or M, X8 = D or S, X9 = W or S, and X 10 = S or absent, X 11 = S or absent, X 12 = S or G, X 13 = Y or N, and X 14 = Y or W, and X 15 = I or W, X 16 = H, G or S, and X17 = I or absent, X 18 = I, S or E, and X 19 = N or I, and X 20 = P, Y or N, and X 21 = G, Y or H, and X 22 = N or S, and X 23 = D or S, X 24 = N or Y, and X 25 = A or N, X 26 = Q or P, and X 27 = R or S, and X 28 = F or L, and X 29 = Q or K, and X 30 = G or S, and X 31 =E or absent, X 32 = I, Y or M, and X 33 = A or G, and X 34 = S, P, I or A, and X 35 = Y, D or N, and X 36 = G or S, and X 37 = G or S, and X 38 = L or absent, X 39 = V or Y, and X 40 = A, D or G, and X 41 = F, M or Y, and X 42 =D or absent, X 43 = I, V, or absent) An optionally isolated antibody or antigen-binding fragment capable of binding to HER2, having the following structure: [Aspect 2] 2. The antibody or antigen-binding fragment of embodiment 1, wherein LC-CDR1 is one of GLSSGSVSTGHYAS (SEQ ID NO: 15), GLSSGSVSTGYYPS (SEQ ID NO: 21), or GLSSGSVSTSYYPS (SEQ ID NO: 27). [Aspect 3] 3. The antibody or antigen-binding fragment of embodiment 1 or 2, wherein LC-CDR2 is one of NTNTRSS (SEQ ID NO: 16), STNSRSS (SEQ ID NO: 22), TTNIRSS (SEQ ID NO: 28), or STNTRSS (SEQ ID NO: 33). [Aspect 4] The antibody or antigen-binding fragment of any one of aspects 1 to 3, wherein LC-CDR3 is one of VLYVGDGIWV (SEQ ID NO: 17), VLYMGSGISV (SEQ ID NO: 23), MLYMGSGIWV (SEQ ID NO: 29), or VLYMGSGIWV (SEQ ID NO: 34). [Aspect 5] The antibody or antigen-binding fragment of any one of aspects 1 to 4, wherein HC-CDR1 is one of SYYIH (SEQ ID NO: 18), SSSYYWG (SEQ ID NO: 24), GYYWS (SEQ ID NO: 30), or SSNWWS (SEQ ID NO: 35). [Aspect 6] 6. The antibody or antigen-binding fragment of any one of aspects 1 to 5, wherein HC-CDR2 is one of IINPGNGDTNYAQRFQG (SEQ ID NO: 19), SIYYSGSTYYNPSLKS (SEQ ID NO: 25), EINHSGSTNYNPSLKS (SEQ ID NO: 31), or EIYHSGSTNYNPSLKS (SEQ ID NO: 36). [Aspect 7] The antibody or antigen-binding fragment of any one of embodiments 1 to 6, wherein HC-CDR3 is one of EIASYSGSYYDY (SEQ ID NO: 20), YAPDSSGYLVAFDI (SEQ ID NO: 26), MGINSGGYLYGMDV (SEQ ID NO: 32), or MGANSGGYLYGMDV (SEQ ID NO: 37). [Aspect 8] The following CDRs: LC-CDR1: GLSSGSVSTX1X2YX3S (SEQ ID NO: 9), LC-CDR2: X4TNX5RSS (SEQ ID NO: 10), LC-CDR3: X6LYX7GX8GIX9V (SEQ ID NO: 11) (wherein X1 = G or S, X2 = H or Y, X3 = A or P, X4 = N, S or T, X5 = T, S or I, X6 = V or M, X7 = V or M, X8 = D or S, and X9 = W or S). The antibody or antigen-binding fragment of any one of embodiments 1 to 7, having at least one light chain variable region incorporating: [Aspect 9] The following CDRs: LC-CDR1: GLSSGSVSTGHYAS (SEQ ID NO: 15), LC-CDR2: NTNTRSS (SEQ ID NO: 16), LC-CDR3: VLYVGDGIWV (SEQ ID NO: 17) The antibody or antigen-binding fragment of any one of embodiments 1 to 8, having at least one light chain variable region incorporating: [Aspect 10] The following CDRs: LC-CDR1: GLSSGSVSTGYYPS (SEQ ID NO: 21), LC-CDR2: STNSRSS (SEQ ID NO: 22), LC-CDR3: VLYMGSGISV (SEQ ID NO: 23) The antibody or antigen-binding fragment of any one of embodiments 1 to 8, having at least one light chain variable region incorporating: [Aspect 11] The following CDRs: LC-CDR1: GLSSGSVSTSYYPS (SEQ ID NO: 27), LC-CDR2: TTNIRSS (SEQ ID NO: 28), LC-CDR3: MLYMGSGIWV (SEQ ID NO: 29) The antibody or antigen-binding fragment of any one of embodiments 1 to 8, having at least one light chain variable region incorporating: [Aspect 12] The following CDRs: LC-CDR1: GLSSGSVSTSYYPS (SEQ ID NO: 27), LC-CDR2: STNTRSS (SEQ ID NO: 33), LC-CDR3: VLYMGSGIWV (SEQ ID NO: 34) The antibody or antigen-binding fragment of any one of embodiments 1 to 8, having at least one light chain variable region incorporating: [Aspect 13] The following CDRs: HC-CDR1:X 10 X 11 X 12 X 13 X 14 X 15 X 16 (SEQ ID NO: 12), HC-CDR2:X 17 X 18 X 19 X 20 X 21 X 22 GX 23 TX 24 YX 25 X 26 X 27 X 28 X 29 X 30 (SEQ ID NO: 13), HC-CDR3:X 31 X 32 X 33 X 34 X 35 SX 36 X 37 YX 38 X 39 X 40 X 41 X 42 X 43 (SEQ ID NO: 14) (In the formula, X 10 = S or absent, X 11 = S or absent, X 12 = S or G, X 13 = Y or N, and X 14 = Y or W, and X 15 = I or W, X 16 = H, G or S, and X 17 = I or absent, X 18 = I, S or E, and X 19 = N or I, and X 20= P, Y or N, and X 21 = G, Y or H, and X 22 = N or S, and X 23 = D or S, X 24 = N or Y, and X 25 = A or N, X 26 = Q or P, and X 27 = R or S, and X 28 = F or L, and X 29 = Q or K, and X 30 = G or S, and X 31 =E or absent, X 32 = I, Y or M, and X 33 = A or G, and X 34 = S, P, I or A, and X 35 = Y, D or N, and X 36 = G or S, and X 37 = G or S, and X 38 = L or absent, X 39 = V or Y, and X 40 = A, D or G, and X 41 = F, M or Y, and X 42 =D or absent, X 43 = I, V, or absent) 13. The antibody or antigen-binding fragment of any one of embodiments 1 to 12, having at least one heavy chain variable region incorporating: [Aspect 14] The following CDRs: HC-CDR1: SYYIH (SEQ ID NO: 18), HC-CDR2: IINPGNGDTNYAQRFQG (SEQ ID NO: 19), HC-CDR3: EIASYSGSYYDY (SEQ ID NO: 20) 14. The antibody or antigen-binding fragment of any one of embodiments 1 to 13, having at least one heavy chain variable region incorporating: [Aspect 15] The following CDRs: HC-CDR1: SSSYYWG (SEQ ID NO: 24), HC-CDR2: SIYYSGSTYYNPSLKS (SEQ ID NO: 25), HC-CDR3: YAPDSSGYLVAFDI (SEQ ID NO: 26) 14. The antibody or antigen-binding fragment of any one of embodiments 1 to 13, having at least one heavy chain variable region incorporating: [Aspect 16] The following CDRs: HC-CDR1: GYYWS (SEQ ID NO: 30), HC-CDR2: EINHSGSTNYNPSLKS (SEQ ID NO: 31), HC-CDR3: MGINSGGYLYGMDV (SEQ ID NO: 32) 14. The antibody or antigen-binding fragment of any one of embodiments 1 to 13, having at least one heavy chain variable region incorporating: [Aspect 17] The following CDRs: HC-CDR1: SSNWWS (SEQ ID NO: 35), HC-CDR2: EIYHSGSTNYNPSLKS (SEQ ID NO: 36), HC-CDR3: MGANSGGYLYGMDV (SEQ ID NO: 37) 14. The antibody or antigen-binding fragment of any one of embodiments 1 to 13, having at least one heavy chain variable region incorporating: [Aspect 18] 18. The antibody or antigen-binding fragment of any one of embodiments 1 to 17, which specifically binds to human, rhesus monkey, or mouse HER2. [Aspect 19] 19. The antibody or antigen-binding fragment of any one of embodiments 1 to 18, which induces cell death by apoptosis. [Aspect 20] CDR below: LC-CDR1: GLSSGSVSTX1X2YX3S (SEQ ID NO: 9), LC-CDR2: X4TNX5RSS (SEQ ID NO: 10), LC-CDR3: X6LYX7GX8GIX9V (SEQ ID NO: 11) (wherein X1 = G or S, X2 = H or Y, X3 = A or P, X4 = N, S or T, X5 = T, S or I, X6 = V or M, X7 = V or M, X8 = D or S, and X9 = W or S). 1. An isolated light chain variable region polypeptide comprising: [Aspect 21] 21. The isolated light chain variable region polypeptide of embodiment 20, wherein LC-CDR1 is one of GLSSGSVSTGHYAS (SEQ ID NO: 15), GLSSGSVSTGYYPS (SEQ ID NO: 21), or GLSSGSVSTSYYPS (SEQ ID NO: 27). [Aspect 22] 22. The isolated light chain variable region polypeptide of embodiment 20 or 21, wherein LC-CDR2 is one of NTNTRSS (SEQ ID NO: 16), STNSRSS (SEQ ID NO: 22), TTNIRSS (SEQ ID NO: 28), or STNTRSS (SEQ ID NO: 33). [Aspect 23] 23. The isolated light chain variable region polypeptide of any one of aspects 20 to 22, wherein LC-CDR3 is one of VLYVGDGIWV (SEQ ID NO: 17), VLYMGSGISV (SEQ ID NO: 23), MLYMGSGIWV (SEQ ID NO: 29), or VLYMGSGIWV (SEQ ID NO: 34). [Aspect 24] Light chain sequence: An isolated light chain variable region polypeptide comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 1, 2, 3, or 4 (Figures 1A-1D). [Aspect 25] The following CDRs: HC-CDR1:X 10 X 11 X 12 X 13 X 14 X 15 X 16 (SEQ ID NO: 12), HC-CDR2:X 17 X 18 X 19 X 20 X 21 X 22 GX 23 TX24 YX 25 X 26 X 27 X 28 X 29 X 30 (SEQ ID NO: 13), HC-CDR3:X 31 X 32 X 33 X 34 X 35 SX 36 X 37 YX 38 X 39 X 40 X 41 X 42 X 43 (SEQ ID NO: 14) (In the formula, X 10 = S or absent, X 11 = S or absent, X 12 = S or G, X 13 = Y or N, and X 14 = Y or W, and X 15 = I or W, X 16 = H, G or S, and X 17 = I or absent, X 18 = I, S or E, and X 19 = N or I, and X 20 = P, Y or N, and X 21 = G, Y or H, and X 22 = N or S, and X 23 = D or S, X 24 = N or Y, and X 25 = A or N, X 26 = Q or P, and X 27 = R or S, and X 28 = F or L, and X 29 = Q or K, and X 30 = G or S, and X 31 =E or absent, X 32 = I, Y or M, and X 33 = A or G, and X 34 = S, P, I or A, and X 35 = Y, D or N, and X36 = G or S, and X 37 = G or S, and X 38 = L or absent, X 39 = V or Y, and X 40 = A, D or G, and X 41 = F, M or Y, and X 42 =D or absent, X 43 = I, V, or absent) 1. An isolated heavy chain variable region polypeptide comprising: [Aspect 26] 26. The isolated heavy chain variable region polypeptide of embodiment 25, wherein HC-CDR1 is one of SYYIH (SEQ ID NO: 18), SSSYYWG (SEQ ID NO: 24), GYYWS (SEQ ID NO: 30), or SSNWWS (SEQ ID NO: 35). [Aspect 27] 27. The isolated heavy chain variable region polypeptide of embodiment 25 or 26, wherein HC-CDR2 is one of IINPGNGDTNYAQRFQG (SEQ ID NO: 19), SIYYSGSTYYNPSLKS (SEQ ID NO: 25), EINHSGSTNYNPSLKS (SEQ ID NO: 31), or EIYHSGSTNYNPSLKS (SEQ ID NO: 36). [Aspect 28] 28. The isolated heavy chain variable region polypeptide of any one of embodiments 25 to 27, wherein HC-CDR3 is one of EIASYSGSYYDY (SEQ ID NO: 20), YAPDSSGYLVAFDI (SEQ ID NO: 26), MGINSGGYLYGMDV (SEQ ID NO: 32), or MGANSGGYLYGMDV (SEQ ID NO: 37). [Aspect 29] Heavy chain sequence: An isolated heavy chain variable region polypeptide comprising an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 5, 6, 7, or 8 (Figures 2A-2D). [Aspect 30] 1. An antibody or antigen-binding fragment capable of binding to HER2, comprising heavy and light chain variable region sequences: the light chains each having LC-CDR1: one of GLSSGSVSTX1X2YX3S (SEQ ID NO: 9), GLSSGSVSTGHYAS (SEQ ID NO: 15), GLSSGSVSTGYYPS (SEQ ID NO: 21), or GLSSGSVSTSYYPS (SEQ ID NO: 27); LC-CDR2: one of X4TNX5RSS (SEQ ID NO: 10), NTNTRSS (SEQ ID NO: 16), STNSRSS (SEQ ID NO: 22), TTNIRSS (SEQ ID NO: 28), or STNTRSS (SEQ ID NO: 33); and LC-CDR3: one of X6LYX7GX8GIX9V (SEQ ID NO: 11), VLYVGDGIWV (SEQ ID NO: 17), VLYMGSGISV (SEQ ID NO: 23), MLYMGSGIWV (SEQ ID NO: 29), or VLYMGSGIWV (SEQ ID NO: 34). (wherein X1 = G or S, X2 = H or Y, X3 = A or P, X4 = N, S or T, X5 = T, S or I, X6 = V or M, X7 = V or M, X8 = D or S, and X9 = W or S). LC-CDR1, LC-CDR2, and LC-CDR3 have at least 85% overall sequence identity to The heavy chains are each HC-CDR1:X 10 X 11 X 12 X 13 X 14 X 15 X 16 (SEQ ID NO: 12), SYYIH (SEQ ID NO: 18), SSSYYWG (SEQ ID NO: 24), GYYWS (SEQ ID NO: 30), or SSNWWS (SEQ ID NO: 35), HC-CDR2:X 17 X 18 X 19 X 20 X 21 X 22 GX 23 TX 24 YX 25 X 26 X 27 X 28 X 29 X 30(SEQ ID NO: 13), IINPGNGDTNYAQRFQG (SEQ ID NO: 19), SIYYSGSTYYNPSLKS (SEQ ID NO: 25), EINHSGSTNYNPSLKS (SEQ ID NO: 31), or EIYHSGSTNYNPSLKS (SEQ ID NO: 36), and one of HC-CDR3:X 31 X 32 X 33 X 34 X 35 SX 36 X 37 YX 38 X 39 X 40 X 41 X 42 X 43 (SEQ ID NO: 14), EIASYSGSYYDY (SEQ ID NO: 20), YAPDSSGYLVAFDI (SEQ ID NO: 26), MGINSGGYLYGMDV (SEQ ID NO: 32), or MGANSGGYLYGMDV (SEQ ID NO: 37) (In the formula, X 10 = S or absent, X 11 = S or absent, X 12 = S or G, X 13 = Y or N, and X 14 = Y or W, and X 15 = I or W, X 16 = H, G or S, and X 17 = I or absent, X 18 = I, S or E, and X 19 = N or I, and X 20 = P, Y or N, and X 21 = G, Y or H, and X 22 = N or S, and X 23 = D or S, X 24 = N or Y, and X 25 = A or N, X 26 = Q or P, and X 27 = R or S, and X 28 = F or L, and X 29 = Q or K, and X 30 = G or S, and X 31 =E or absent, X 32= I, Y or M, and X 33 = A or G, and X 34 = S, P, I or A, and X 35 = Y, D or N, and X 36 = G or S, and X 37 = G or S, and X 38 = L or absent, X 39 = V or Y, and X 40 = A, D or G, and X 41 = F, M or Y, and X 42 =D or absent, X 43 = I, V, or absent) an antibody or antigen-binding fragment comprising HC-CDR1, HC-CDR2, and HC-CDR3 that have at least 85% overall sequence identity to the [Aspect 31] An antibody or antigen-binding fragment capable of binding to HER2, optionally isolated, comprising heavy and light chain variable region sequences, the light chain sequence has at least 85% sequence identity to the light chain sequence of SEQ ID NO: 1, 2, 3, or 4 (Figures 1A-1D); An antibody or antigen-binding fragment, wherein the heavy chain sequence has at least 85% sequence identity to the heavy chain sequence of SEQ ID NO: 5, 6, 7, or 8 (Figures 2A-2D). [Aspect 32] Amino acid sequences i) to iii), or amino acid sequences iv) to vi), or preferably amino acid sequences i) to vi): i) LC-CDR1: RSSQSLLHSNGFNYLD (SEQ ID NO: 74), RSSQSLLHSDGNKYLD (SEQ ID NO: 84), RSSQSLVYSDGNTYLN (SEQ ID NO: 93), RSSQSLLHSNGYNYLD (SEQ ID NO: 102), RSSQSLLHSNGNTYLD (SEQ ID NO: 110), RASQSVRNNLA (SEQ ID NO: 120), GSTTGAVTSGHYPS (SEQ ID NO: 138), RASQSVSSSYLA (SEQ ID NO: 161), RSSQSLQHSNGYQYLD (SEQ ID NO: 170), TGRSANIGGFDVQ (SEQ ID NO: 180) sequence number 186), ALTSGSVSTSYYPS (SEQ ID NO: 194), TSSQSLVYSDGNTYLN (SEQ ID NO: 211), RSSQSLLRSDGYNFVD (SEQ ID NO: 217), RASQGISSWLA (SEQ ID NO: 226), RASRSVGKYLA (SEQ ID NO: 247), GLSSGSVSTTYYPS (SEQ ID NO: 255), TLRSGINVGTYRIY (SEQ ID NO: 261), RASQSVSSYLA (SEQ ID NO: 270), GLTSGAVSSSYYPS (SEQ ID NO: 279), TGNNNNVGFAGAA (SEQ ID NO: 287); ii) LC-CDR2: one of LGSNRAS (SEQ ID NO: 75), KVSNRDS (SEQ ID NO: 94), SGSNRAS (SEQ ID NO: 111); YASTRAT (SEQ ID NO: 121), STSNKHS (SEQ ID NO: 139), LGSHRAS (SEQ ID NO: 146), LGSNRAP (SEQ ID NO: 155), GASSRAT (SEQ ID NO: 162), LGSFRAS (SEQ ID NO: 171), DNSNRPS (SEQ ID NO: 187), STNLRSS (SEQ ID NO: 195), KVSDRDS (SEQ ID NO: 204), KVSKRDS (SEQ ID NO: 212), LGSDRAS (SEQ ID NO: 218), AASSLQS (SEQ ID NO: 227), YKSDSDKQQGS (SEQ ID NO: 262), DASTRAS (SEQ ID NO: 248), STNTRSS (SEQ ID NO: 33), DASNRAT (SEQ ID NO: 271), NTDIRFS (SEQ ID NO: 280), RNNDRPS (SEQ ID NO: 288); iii) LC-CDR3: MQGLQTPYT (SEQ ID NO: 76), MLGTHWPPMYI (SEQ ID NO: 85), MQGTHWPLT (SEQ ID NO: 95), MQALQTPWT (SEQ ID NO: 103), MQGTHWPPT (SEQ ID NO: 112), QHYGSSRT (SEQ ID NO: 122), MAGLQTPRLT (SEQ ID NO: 130), LLYYGGARV (SEQ ID NO: 140), MQALQTPLT (SEQ ID NO: 147), QQYGSSPRT (SEQ ID NO: 163), MHALSTPPWT (SEQ ID NO: 172), MQGTHWPGT (SEQ ID NO: 180) ), GTWDSYLNIWV (SEQ ID NO: 188), ELYMGSGISV (SEQ ID NO: 196), MQGTHWPQT (SEQ ID NO: 205), MQALQTPRT (SEQ ID NO: 219), QQANSFPPT (SEQ ID NO: 228), QQYGSSSA (SEQ ID NO: 233), MIWHSSAWV (SEQ ID NO: 263), QHYGTSPPFI (SEQ ID NO: 249), VLYMGNGISV (SEQ ID NO: 256), QQRSNWPLT (SEQ ID NO: 272), VLYMGSGISV (SEQ ID NO: 23), SAWDSSLKVQV (SEQ ID NO: 289), iv) HC-CDR1: one of SYGMH (SEQ ID NO: 77), SAAAAWN (SEQ ID NO: 86), SYAMH (SEQ ID NO: 96), SFAMN (SEQ ID NO: 113), SYGIS (SEQ ID NO: 123), SYAMS (SEQ ID NO: 131), SYAIS (SEQ ID NO: 148), TYTMH (SEQ ID NO: 173), GYYWS (SEQ ID NO: 30), SYWIG (SEQ ID NO: 197), NYGMH (SEQ ID NO: 234), SYAIH (SEQ ID NO: 241), SSSYYWG (SEQ ID NO: 24), DYYIH (SEQ ID NO: 281); v) HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), RTYYRSKWYSEYAVSVKS (SEQ ID NO: 87), WINAGNGNTKYSQKFQG (SEQ ID NO: 104), TIGGSGDSTFYADPVKG (SEQ ID NO: 114), WISAYNGNTNYAQKLQG (SEQ ID NO: 124), AISGSGGSTYYADSVKG (SEQ ID NO: 132), GIIPIFGTANYAQKFQG (SEQ ID NO: 149), GINWNGGSTGYADSVKG (SEQ ID NO: 164), WITPGNGNTHYSQNFQG (SEQ ID NO: 165) 174), EINHSGSTNYNPSLKS (SEQ ID NO: 31), IIYPGDSDTRYSPSFQG (SEQ ID NO: 198), AISSNGGSTYYADSVKG (SEQ ID NO: 206), VISYDESNKYYADSVKG (SEQ ID NO: 220), FISYDGTNKYYADSVKG (SEQ ID NO: 235), SIYYSGSTYYNPSLKS (SEQ ID NO: 25), VIWYDGSNKYYADSVKG (SEQ ID NO: 264), SISSSSSSYIYYADSVKG (SEQ ID NO: 273), WVSAYNGDTNYAQKFQG (SEQ ID NO: 282); vi) HC-CDR3: DLFAVVGYYYYYGMDV (SEQ ID NO: 79), GSIFDV (SEQ ID NO: 88), SRGYYGMDV (SEQ ID NO: 97), GGYLVGY (SEQ ID NO: 105), AYGSGGHYFFAY (SEQ ID NO: 115), DWGSSWSDY (SEQ ID NO: 125), TYYDFWSGRVGAFDI (SEQ ID NO: 133), DRGYYGMDV (SEQ ID NO: 141), GRGSGYPDTWFWFDP ​​(SEQ ID NO: 150), SYGSGSYRSHAFDI (SEQ ID NO: 156), GLVPAASMDV (SEQ ID NO: 165), SRVGALDY (SEQ ID NO: 175) , SRGYSGYDN (SEQ ID NO: 181), GLPYYYFDY (SEQ ID NO: 189), LGYGVPLPEYFDL (SEQ ID NO: 199), EPSGSWSYLYYYYYGMDV (SEQ ID NO: 221), HYGDYYYYYGMDV (SEQ ID NO: 236), VYGYGLHYYGMDV (SEQ ID NO: 242), SYDSSGYYYFDY (SEQ ID NO: 250), YAPDSSGYLVAFDI (SEQ ID NO: 26), MTTEDY (SEQ ID NO: 265), DGSAWSRPY (SEQ ID NO: 274), EIASYSGSYYDY (SEQ ID NO: 20), GADWNSDY (SEQ ID NO: 290), or a variant thereof in which one, two or three amino acids in one or more of sequences (i) to (vi) are replaced with another amino acid. 10. An antibody, antigen-binding fragment or polypeptide capable of binding to HER2, optionally isolated, having the following structure: [Aspect 33] An antibody, antigen-binding fragment or polypeptide capable of binding to HER2, optionally isolated, comprising heavy and light chain variable region sequences: The light chain is LC-CDR1: RSSQSLLHSNGFNYLD (SEQ ID NO: 74), RSSQSLLHSDGNKYLD (SEQ ID NO: 84), RSSQSLVYSDGNTYLN (SEQ ID NO: 93), RSSQSLLHSNGYNYLD (SEQ ID NO: 102), RSSQSLLHSNGNTYLD (SEQ ID NO: 110), RASQSVRNNLA (SEQ ID NO: 120), GSTTGAVTSGHYPS (SEQ ID NO: 138), RASQSVSSSYLA (SEQ ID NO: 161), RSSQSLQHSNGYQYLD (SEQ ID NO: 170), TGRSANIGGFDVQ ( SEQ ID NO: 186), ALTSGSVSTSYYPS (SEQ ID NO: 194), TSSQSLVYSDGNTYLN (SEQ ID NO: 211), RSSQSLLRSDGYNFVD (SEQ ID NO: 217), RASQGISSWLA (SEQ ID NO: 226), RASRSVGKYLA (SEQ ID NO: 247), GLSSGSVSTTYYPS (SEQ ID NO: 255), TLRSGINVGTYRIY (SEQ ID NO: 261), RASQSVSSYLA (SEQ ID NO: 270), GLTSGAVSSSYYPS (SEQ ID NO: 279), TGNNNNVGFAGAA (SEQ ID NO: 287), LC-CDR2: one of LGSNRAS (SEQ ID NO: 75), KVSNRDS (SEQ ID NO: 94), SGSNRAS (SEQ ID NO: 111); YASTRAT (SEQ ID NO: 121), STSNKHS (SEQ ID NO: 139), LGSHRAS (SEQ ID NO: 146), LGSNRAP (SEQ ID NO: 155), GASSRAT (SEQ ID NO: 162), LGSFRAS (SEQ ID NO: 171), DNSNRPS (SEQ ID NO: 187), STNLRSS (SEQ ID NO: 195), KVSDRDS (SEQ ID NO: 204), KVSKRDS (SEQ ID NO: 212), LGSDRAS (SEQ ID NO: 218), AASSLQS (SEQ ID NO: 227), YKSDSDKQQGS (SEQ ID NO: 262), DASTRAS (SEQ ID NO: 248), STNTRSS (SEQ ID NO: 33), DASNRAT (SEQ ID NO: 271), NTDIRFS (SEQ ID NO: 280), RNNDRPS (SEQ ID NO: 288), LC-CDR3: MQGLQTPYT (SEQ ID NO: 76), MLGTHWPPMYI (SEQ ID NO: 85), MQGTHWPLT (SEQ ID NO: 95), MQALQTPWT (SEQ ID NO: 103), MQGTHWPPT (SEQ ID NO: 112), QHYGSSRT (SEQ ID NO: 122), MAGLQTPRLT (SEQ ID NO: 130), LLYYGGARV (SEQ ID NO: 140), MQALQTPLT (SEQ ID NO: 147), QQYGSSPRT (SEQ ID NO: 163), MHALSTPPWT (SEQ ID NO: 172), MQGTHWPGT (SEQ ID NO: 180), one of GTWDSYLNIWV (SEQ ID NO: 188), ELYMGSGISV (SEQ ID NO: 196), MQGTHWPQT (SEQ ID NO: 205), MQALQTPRT (SEQ ID NO: 219), QQANSFPPT (SEQ ID NO: 228), QQYGSSSA (SEQ ID NO: 233), MIWHSSAWV (SEQ ID NO: 263), QHYGTSPPFI (SEQ ID NO: 249), VLYMGNGISV (SEQ ID NO: 256), QQRSNWPLT (SEQ ID NO: 272), VLYMGSGISV (SEQ ID NO: 23), SAWDSSLKVQV (SEQ ID NO: 289); LC-CDR1, LC-CDR2, and LC-CDR3 have at least 85% overall sequence identity to the heavy chain is one of HC-CDR1: SYGMH (SEQ ID NO: 77), SAAAAWN (SEQ ID NO: 86), SYAMH (SEQ ID NO: 96), SFAMN (SEQ ID NO: 113), SYGIS (SEQ ID NO: 123), SYAMS (SEQ ID NO: 131), SYAIS (SEQ ID NO: 148), TYTMH (SEQ ID NO: 173), GYYWS (SEQ ID NO: 30), SYWIG (SEQ ID NO: 197), NYGMH (SEQ ID NO: 234), SYAIH (SEQ ID NO: 241), SSSYYWG (SEQ ID NO: 24), DYYIH (SEQ ID NO: 281); HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), RTYYRSKWYSEYAVSVKS (SEQ ID NO: 87), WINAGNGNTKYSQKFQG (SEQ ID NO: 104), TIGGSGDSTFYADPVKG (SEQ ID NO: 114), WISAYNGNTNYAQKLQG (SEQ ID NO: 124), AISGSGGSTYYADSVKG (SEQ ID NO: 132), GIIPIFGTANYAQKFQG (SEQ ID NO: 149), GINWNGGSTGYADSVKG (SEQ ID NO: 164), WITPGNGNTHYSQNFQG (SEQ ID NO: 1 74), EINHSGSTNYNPSLKS (SEQ ID NO: 31), IIYPGDSDTRYSPSFQG (SEQ ID NO: 198), AISSNGGSTYYADSVKG (SEQ ID NO: 206), VISYDESNKYYADSVKG (SEQ ID NO: 220), FISYDGTNKYYADSVKG (SEQ ID NO: 235), SIYYSGSTYYNPSLKS (SEQ ID NO: 25), VIWYDGSNKYYADSVKG (SEQ ID NO: 264), SISSSSSSYIYYADSVKG (SEQ ID NO: 273), WVSAYNGDTNYAQKFQG (SEQ ID NO: 282); HC-CDR3: DLFAVVGYYYYYGMDV (SEQ ID NO: 79), GSIFDV (SEQ ID NO: 88), SRGYYGMDV (SEQ ID NO: 97), GGYLVGY (SEQ ID NO: 105), AYGSGGHYFFAY (SEQ ID NO: 115), DWGSSWSDY (SEQ ID NO: 125), TYYDFWSGRVGAFDI (SEQ ID NO: 133), DRGYYGMDV (SEQ ID NO: 141), GRGSGYPDTWFWFDP ​​(SEQ ID NO: 150), SYGSGSYRSHAFDI (SEQ ID NO: 156), GLVPAASMDV (SEQ ID NO: 165), SRVGALDY (SEQ ID NO: 175), S one of RGYSGYDN (SEQ ID NO: 181), GLPYYYFDY (SEQ ID NO: 189), LGYGVPLPEYFDL (SEQ ID NO: 199), EPSGSWSYLYYYYYGMDV (SEQ ID NO: 221), HYGDYYYYYGMDV (SEQ ID NO: 236), VYGYGLHYYGMDV (SEQ ID NO: 242), SYDSSGYYYFDY (SEQ ID NO: 250), YAPDSSGYLVAFDI (SEQ ID NO: 26), MTTEDY (SEQ ID NO: 265), DGSAWSRPY (SEQ ID NO: 274), EIASYSGSYYDY (SEQ ID NO: 20), GADWNSDY (SEQ ID NO: 290); an antibody, antigen-binding fragment or polypeptide comprising HC-CDR1, HC-CDR2, and HC-CDR3 having at least 85% overall sequence identity to [Aspect 34] The following CDRs: LC-CDR1: RSSQSLLHSNGFNYLD (SEQ ID NO: 74), LC-CDR2: LGSNRAS (SEQ ID NO: 75), LC-CDR3: MQGLQTPYT (SEQ ID NO: 76) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYGMH (SEQ ID NO: 77), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: DLFAVVGYYYYYGMDV (SEQ ID NO: 79) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 35] The following CDRs: LC-CDR1: RSSQSLLHSDGNKYLD (SEQ ID NO: 84), LC-CDR2: LGSNRAS (SEQ ID NO: 75), LC-CDR3: MLGTHWPPMYI (SEQ ID NO: 85) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SAAAAWN (SEQ ID NO: 86), HC-CDR2: RTYYRSKWYSEYAVSVKS (SEQ ID NO: 87), HC-CDR3: GSIFDV (SEQ ID NO: 88) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 36] The following CDRs: LC-CDR1: RSSQSLVYSDGNTYLN (SEQ ID NO: 93), LC-CDR2: KVSNRDS (SEQ ID NO: 94), LC-CDR3: MQGTHWPLT (SEQ ID NO: 95) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SRGYYGMDV (SEQ ID NO: 97) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 37] The following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSNRAS (SEQ ID NO: 75), LC-CDR3: MQALQTPWT (SEQ ID NO: 103) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: WINAGNGNTKYSQKFQG (SEQ ID NO: 104), HC-CDR3: GGYLVGY (SEQ ID NO: 105) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 38] The following CDRs: LC-CDR1: RSSQSLLHSNGNTYLD (SEQ ID NO: 110), LC-CDR2: SGSNRAS (SEQ ID NO: 111), LC-CDR3: MQGTHWPPT (SEQ ID NO: 112) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SFAMN (SEQ ID NO: 113), HC-CDR2: TIGGSGDSTFYADPVKG (SEQ ID NO: 114), HC-CDR3: AYGSGGHYFFAY (SEQ ID NO: 115) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 39] The following CDRs: LC-CDR1: RASQSVRNNLA (SEQ ID NO: 120), LC-CDR2: YASTRAT (SEQ ID NO: 121), LC-CDR3: QHYGSSRT (SEQ ID NO: 122) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYGIS (SEQ ID NO: 123), HC-CDR2: WISAYNGNTNYAQKLQG (SEQ ID NO: 124), HC-CDR3: DWGSSWSDY (SEQ ID NO: 125) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 40] The following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSNRAS (SEQ ID NO: 75), LC-CDR3: MAGLQTPRLT (SEQ ID NO: 130) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYAMS (SEQ ID NO: 131), HC-CDR2: AISGSGGSTYYADSVKG (SEQ ID NO: 132), HC-CDR3: TYYDFWSGRVGAFDI (SEQ ID NO: 133) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 41] The following CDRs: LC-CDR1: GSTTGAVTSGHYPS (SEQ ID NO: 138), LC-CDR2: STSNKHS (SEQ ID NO: 139), LC-CDR3: LLYYGGARV (SEQ ID NO: 140) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYGMH (SEQ ID NO: 77), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: DRGYYGMDV (SEQ ID NO: 141) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 42] The following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSHRAS (SEQ ID NO: 146), LC-CDR3: MQALQTPLT (SEQ ID NO: 147) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYAIS (SEQ ID NO: 148), HC-CDR2: GIIPIFGTANYAQKFQG (SEQ ID NO: 149), HC-CDR3: GRGSGYPDTWFWFDP ​​(SEQ ID NO: 150) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 43] The following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSNRAP (SEQ ID NO: 155), LC-CDR3: MQGTHWPLT (SEQ ID NO: 95) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SYGSGSYRSHAFDI (SEQ ID NO: 156) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 44] The following CDRs: LC-CDR1: RASQSVSSSYLA (SEQ ID NO: 161), LC-CDR2: GASSRAT (SEQ ID NO: 162), LC-CDR3: QQYGSSPRT (SEQ ID NO: 163) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYAMS (SEQ ID NO: 131), HC-CDR2: GINWNGGSTGYADSVKG (SEQ ID NO: 164), HC-CDR3: GLVPAASMDV (SEQ ID NO: 165) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 45] The following CDRs: LC-CDR1: RSSQSLQHSNGYQYLD (SEQ ID NO: 170), LC-CDR2: LGSFRAS (SEQ ID NO: 171), LC-CDR3: MHALSTPPWT (SEQ ID NO: 172) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1:TYTMH (SEQ ID NO: 173), HC-CDR2: WITPGNGNTHYSQNFQG (SEQ ID NO: 174), HC-CDR3: SRVGALDY (SEQ ID NO: 175) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 46] The following CDRs: LC-CDR1: RSSQSLVYSDGNTYLN (SEQ ID NO: 93), LC-CDR2: KVSNRDS (SEQ ID NO: 94), LC-CDR3: MQGTHWPGT (SEQ ID NO: 180) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYGMH (SEQ ID NO: 77), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SRGYSGYDN (SEQ ID NO: 181) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 47] The following CDRs: LC-CDR1: TGRSANIGGFDVQ (SEQ ID NO: 186), LC-CDR2: DNSNRPS (SEQ ID NO: 187), LC-CDR3: GTWDSYLNIWV (SEQ ID NO: 188) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: GYYWS (SEQ ID NO: 30), HC-CDR2: EINHSGSTNYNPSLKS (SEQ ID NO: 31), HC-CDR3: GLPYYYFDY (SEQ ID NO: 189) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 48] The following CDRs: LC-CDR1: ALTSGSVSTSYYPS (SEQ ID NO: 194), LC-CDR2: STNLRSS (SEQ ID NO: 195), LC-CDR3: ELYMGSGISV (SEQ ID NO: 196) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYWIG (SEQ ID NO: 197), HC-CDR2: IIYPGDSDTRYSPSFQG (SEQ ID NO: 198), HC-CDR3: LGYGVPLPEYFDL (SEQ ID NO: 199) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 49] The following CDRs: LC-CDR1: RSSQSLVYSDGNTYLN (SEQ ID NO: 93), LC-CDR2: KVSDRDS (SEQ ID NO: 204), LC-CDR3: MQGTHWPQT (SEQ ID NO: 205) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: AISSNGGSTYYADSVKG (SEQ ID NO: 206), HC-CDR3: SRGYYGMDV (SEQ ID NO: 97) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 50] The following CDRs: LC-CDR1: TSSQSLVYSDGNTYLN (SEQ ID NO: 211), LC-CDR2: KVSKRDS (SEQ ID NO: 212), LC-CDR3: MQGTHWPLT (SEQ ID NO: 95) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SRGYYGMDV (SEQ ID NO: 97) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 51] The following CDRs: LC-CDR1: RSSQSLLRSDGYNFVD (SEQ ID NO: 217), LC-CDR2: LGSDRAS (SEQ ID NO: 218), LC-CDR3: MQALQTPRT (SEQ ID NO: 219) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDESNKYYADSVKG (SEQ ID NO: 220), HC-CDR3: EPSGSWSYLYYYYYGMDV (SEQ ID NO: 221) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 52] The following CDRs: LC-CDR1: RASQGISSWLA (SEQ ID NO: 226), LC-CDR2: AASSLQS (SEQ ID NO: 227), LC-CDR3: QQANSFPPT (SEQ ID NO: 228) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SRGYYGMDV (SEQ ID NO: 97) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 53] The following CDRs: LC-CDR1: RASQSVSSSYLA (SEQ ID NO: 161), LC-CDR2: GASSRAT (SEQ ID NO: 162), LC-CDR3: QQYGSSSA (SEQ ID NO: 233) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: NYGMH (SEQ ID NO: 234), HC-CDR2: FISYDGTNKYYADSVKG (SEQ ID NO: 235), HC-CDR3: HYGDYYYYYGMDV (SEQ ID NO: 236) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 54] The following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSNRAS (SEQ ID NO: 75), LC-CDR3: MQGTHWPGT (SEQ ID NO: 180) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYAIH (SEQ ID NO: 241), HC-CDR2: AISSNGGSTYYADSVKG (SEQ ID NO: 206), HC-CDR3: VYGYGLHYYGMDV (SEQ ID NO: 242) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 55] The following CDRs: LC-CDR1: RASRSVGKYLA (SEQ ID NO: 247), LC-CDR2: DASTRAS (SEQ ID NO: 248), LC-CDR3: QHYGTSPPFI (SEQ ID NO: 249) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYAIS (SEQ ID NO: 148), HC-CDR2: GIIPIFGTANYAQKFQG (SEQ ID NO: 149), HC-CDR3: SYDSSGYYYFDY (SEQ ID NO: 250) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 56] The following CDRs: LC-CDR1: GLSSGSVSTTYYPS (SEQ ID NO: 255), LC-CDR2: STNTRSS (SEQ ID NO: 33), LC-CDR3: VLYMGNGISV (SEQ ID NO: 256) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SSSYYWG (SEQ ID NO: 24), HC-CDR2: SIYYSGSTYYNPSLKS (SEQ ID NO: 25), HC-CDR3: YAPDSSGYLVAFDI (SEQ ID NO: 26) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 57] The following CDRs: LC-CDR1: TLRSGINVGTYRIY (SEQ ID NO: 261), LC-CDR2: YKSDSDKQQGS (SEQ ID NO: 262), LC-CDR3: MIWHSSAWV (SEQ ID NO: 263) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYGMH (SEQ ID NO: 77), HC-CDR2: VIWYDGSNKYYADSVKG (SEQ ID NO: 264), HC-CDR3: MTTEDY (SEQ ID NO: 265) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 58] The following CDRs: LC-CDR1: RASQSVSSYLA (SEQ ID NO: 270), LC-CDR2: DASNRAT (SEQ ID NO: 271), LC-CDR3: QQRSNWPLT (SEQ ID NO: 272) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYAMS (SEQ ID NO: 131), HC-CDR2:SISSSSSYIYYADSVKG (SEQ ID NO: 273), HC-CDR3: DGSAWSRPY (SEQ ID NO: 274) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 59] The following CDRs: LC-CDR1: GLTSGAVSSSYYPS (SEQ ID NO: 279), LC-CDR2: NTDIRFS (SEQ ID NO: 280), LC-CDR3: VLYMGSGISV (SEQ ID NO: 23) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1:DYYIH (SEQ ID NO: 281), HC-CDR2: WVSAYNGDTNYAQKFQG (SEQ ID NO: 282), HC-CDR3: EIASYSGSYYDY (SEQ ID NO: 20) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 60] The following CDRs: LC-CDR1: TGNNNNVGFAGAA (SEQ ID NO: 287), LC-CDR2: RNNDRPS (SEQ ID NO: 288), LC-CDR3: SAWDSSLKVQV (SEQ ID NO: 289) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYAIS (SEQ ID NO: 148), HC-CDR2: GIIPIFGTANYAQKFQG (SEQ ID NO: 149), HC-CDR3: GADWNSDY (SEQ ID NO: 290) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 61] The following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSNRAP (SEQ ID NO: 155), LC-CDR3: MQGTHWPLT (SEQ ID NO: 95) and having at least one light chain variable region incorporating and / or the following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SYGSGSYRSHAFDI (SEQ ID NO: 156) 34. The antibody, antigen-binding fragment, or polypeptide of embodiment 32 or 33, having at least one heavy chain variable region incorporating: [Aspect 62] An antibody, antigen-binding fragment or polypeptide capable of binding to HER2, optionally isolated, comprising heavy and light chain variable region sequences: the light chain sequence has at least 85% sequence identity to the light chain sequence of SEQ ID NO: 72, 82, 91, 100, 108, 118, 128, 136, 144, 153, 159, 168, 178, 184, 192, 202, 209, 215, 224, 231, 239, 245, 253, 259, 268, 277, 285, or 293; An antibody, antigen-binding fragment or polypeptide, wherein the heavy chain sequence has at least 85% sequence identity to the heavy chain sequence of SEQ ID NO: 73, 83, 92, 101, 109, 119, 129, 137, 145, 154, 160, 169, 179, 185, 193, 203, 210, 216, 225, 232, 240, 246, 254, 260, 269, 278, 286, or 294. [Aspect 63] 63. The antibody, antigen-binding fragment or polypeptide of any one of embodiments 1 to 62, conjugated to a drug moiety or a detectable moiety. [Aspect 64] 64. The antibody, antigen-binding fragment, or polypeptide of embodiment 63, wherein the drug moiety comprises an anticancer drug moiety. [Aspect 65] 65. A composition comprising the antibody, antigen-binding fragment or polypeptide of any one of embodiments 1 to 64, and at least one pharmaceutically acceptable carrier. [Aspect 66] A composition comprising the antibody, antigen-binding fragment, or polypeptide of any one of embodiments 1 to 64, and at least one anti-cancer agent. [Aspect 67] 67. The composition of embodiment 66, further comprising at least one pharmaceutically acceptable carrier. [Aspect 68] 65. An optionally isolated in vitro complex comprising the antibody, antigen-binding fragment or polypeptide of any one of embodiments 1 to 64, bound to HER2. [Aspect 69] 65. An optionally isolated nucleic acid encoding the antibody, antigen-binding fragment or polypeptide of any one of embodiments 1 to 64. [Aspect 70] 70. A vector comprising the nucleic acid of embodiment 69. [Aspect 71] A cell comprising the nucleic acid of embodiment 69 or the vector of embodiment 70. [Aspect 72] A method for producing the antibody, antigen-binding fragment, or polypeptide of any one of embodiments 1 to 64, the method comprising culturing a cell of embodiment 71 under conditions suitable for expression of the antibody, antigen-binding fragment, or polypeptide from the nucleic acid or vector. [Aspect 73] 68. The antibody, antigen-binding fragment, polypeptide or composition according to any one of embodiments 1 to 67 for use in therapy or in a method of medical treatment. [Aspect 74] 68. The antibody, antigen-binding fragment, polypeptide or composition according to any one of embodiments 1 to 67 for use in the treatment of cancer. [Aspect 75] 75. The antibody, antigen-binding fragment, polypeptide or composition for use according to embodiment 74, wherein the cancer is a HER2-positive cancer. [Aspect 76] 76. The antibody, antigen-binding fragment, polypeptide or composition for use according to embodiment 74 or 75, wherein the cancer comprises HER2-positive tumor cells. [Aspect 77] 77. The antibody, antigen-binding fragment, polypeptide or composition for use according to any one of embodiments 73 to 76, wherein treatment comprises administering the antibody, antigen-binding fragment, polypeptide or composition in combination with a therapeutic agent. [Aspect 78] 78. The antibody, antigen-binding fragment, polypeptide or composition for use according to embodiment 77, wherein the therapeutic agent is an anti-HER2 antibody. [Aspect 79] 68. Use of the antibody, antigen-binding fragment, polypeptide or composition according to any one of embodiments 1 to 67 in the manufacture of a medicament for use in the treatment of cancer. [Aspect 80] 80. The use of the antibody, antigen-binding fragment, polypeptide or composition according to embodiment 79, wherein the cancer is a HER2-positive cancer. [Aspect 81] 81. The use of the antibody, antigen-binding fragment, polypeptide or composition of embodiment 79 or 80, wherein the cancer comprises HER2-positive tumor cells. [Aspect 82] A method of treating cancer, comprising the step of administering to a patient suffering from cancer the antibody, antigen-binding fragment, polypeptide or composition according to any one of embodiments 1 to 67. [Aspect 83] 83. The method of treating cancer according to embodiment 82, wherein the cancer is a HER2-positive cancer. [Aspect 84] 84. The method of treating cancer of embodiment 82 or 83, wherein the cancer comprises HER2-positive tumor cells. [Aspect 85] 85. A method of treating cancer according to any one of embodiments 82 to 84, comprising administering the antibody, antigen-binding fragment, polypeptide or composition in combination with a therapeutic agent. [Aspect 86] 86. The method of treating cancer according to embodiment 85, wherein the therapeutic agent is an anti-HER2 antibody. [Aspect 87] A method of inhibiting tumor cell growth in a subject, comprising administering to the subject a therapeutically effective amount of the antibody, antigen-binding fragment, polypeptide, or composition of any one of embodiments 1 to 67. [Aspect 88] A method of killing tumor cells in a subject, comprising administering to the subject a therapeutically effective amount of the antibody, antigen-binding fragment, polypeptide, or composition of any one of embodiments 1 to 67. [Aspect 89] 65. A method comprising the steps of contacting a sample containing or suspected of containing HER2 with the antibody, antigen-binding fragment, or polypeptide of any one of embodiments 1 to 64, and detecting formation of a complex of the antibody, antigen-binding fragment, or polypeptide, and HER2. [Aspect 90] A method of diagnosing a disease or condition in a subject, the method comprising the steps of contacting in vitro a sample from the subject with the antibody, antigen-binding fragment, or polypeptide of any one of embodiments 1 to 64, and detecting formation of a complex of the antibody, antigen-binding fragment, or polypeptide, and HER2. [Aspect 91] 65. A method of selecting or stratifying a subject for treatment with a HER2-targeted agent, the method comprising the steps of contacting in vitro a sample from the subject with the antibody, antigen-binding fragment, or polypeptide of any one of embodiments 1 to 64, and detecting formation of a complex of the antibody or antigen-binding fragment and HER2. [Aspect 92] 65. Use of the antibody, antigen-binding fragment or polypeptide according to any one of embodiments 1 to 64 for the detection of HER2 in vitro. [Aspect 93] 65. Use of the antibody, antigen-binding fragment or polypeptide according to any one of embodiments 1 to 64 as an in vitro diagnostic agent. [Aspect 94] 65. A chimeric antigen receptor (CAR) comprising the antibody, antigen-binding fragment, or polypeptide of any one of embodiments 1 to 64. [Aspect 95] 95. An optionally isolated in vitro complex comprising a CAR according to embodiment 94 bound to HER2. [Aspect 96] 95. An optionally isolated nucleic acid encoding a CAR according to embodiment 94. [Aspect 97] 97. An expression vector comprising the nucleic acid of embodiment 96. [Aspect 98] 98. A cell comprising a CAR according to embodiment 94, a nucleic acid according to embodiment 96, or an expression vector according to embodiment 97. [Aspect 99] 99. A composition comprising a CAR according to embodiment 94, a nucleic acid according to embodiment 96, an expression vector according to embodiment 97, or a cell according to embodiment 98. [Aspect 100] 96. A CAR according to embodiment 94, a nucleic acid according to embodiment 96, an expression vector according to embodiment 97, a cell according to embodiment 98, or a composition according to embodiment 99 for use in a method of medical treatment or prevention. [Aspect 101] 97. A CAR according to embodiment 94, a nucleic acid according to embodiment 96, an expression vector according to embodiment 97, a cell according to embodiment 98, or a composition according to embodiment 99 for use in a method for treating cancer.

Claims

1. An antibody or antigen-binding fragment capable of binding to HER2, comprising: (i) the following CDRs: LC-CDR1: RSSQSLVYSDGNTYLN (SEQ ID NO: 93), LC-CDR2: KVSNRDS (SEQ ID NO: 94), LC-CDR3: MQGTHWPLT (SEQ ID NO: 95) a light chain variable region incorporating The following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SRGYYGMDV (SEQ ID NO: 97) a heavy chain variable region incorporating (ii) the following CDRs: LC-CDR1: GSTTGAVTSGHYPS (SEQ ID NO: 138), LC-CDR2: STSNKHS (SEQ ID NO: 139), LC-CDR3: LLYYGGARV (SEQ ID NO: 140) a light chain variable region incorporating The following CDRs: HC-CDR1: SYGMH (SEQ ID NO: 77), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: DRGYYGMDV (SEQ ID NO: 141) a heavy chain variable region incorporating (iii) the following CDRs: LC-CDR1: RSSQSLLHSNGYNYLD (SEQ ID NO: 102), LC-CDR2: LGSHRAS (SEQ ID NO: 146), LC-CDR3: MQALQTPLT (SEQ ID NO: 147) a light chain variable region incorporating The following CDRs: HC-CDR1: SYAIS (SEQ ID NO: 148), HC-CDR2: GIIPIFGTANYAQKFQG (SEQ ID NO: 149), HC-CDR3: GRGSGYPDTWFWFDP ​​(SEQ ID NO: 150) a heavy chain variable region incorporating (iv) the following CDRs: LC-CDR1: RASQSVSSSYLA (SEQ ID NO: 161), LC-CDR2: GASSRAT (SEQ ID NO: 162), LC-CDR3: QQYGSSPRT (SEQ ID NO: 163) a light chain variable region incorporating The following CDRs: HC-CDR1: SYAMS (SEQ ID NO: 131), HC-CDR2: GINWNGGSTGYADSVKG (SEQ ID NO: 164), HC-CDR3: GLVPAASMDV (SEQ ID NO: 165) a heavy chain variable region incorporating (v) the following CDRs: LC-CDR1: RSSQSLVYSDGNTYLN (SEQ ID NO: 93), LC-CDR2: KVSDRDS (SEQ ID NO: 204), LC-CDR3: MQGTHWPQT (SEQ ID NO: 205) a light chain variable region incorporating The following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: AISSNGGSTYYADSVKG (SEQ ID NO: 206), HC-CDR3: SRGYYGMDV (SEQ ID NO: 97) a heavy chain variable region incorporating (vi) the following CDRs: LC-CDR1: TSSQSLVYSDGNTYLN (SEQ ID NO: 211), LC-CDR2: KVSKRDS (SEQ ID NO: 212), LC-CDR3: MQGTHWPLT (SEQ ID NO: 95) a light chain variable region incorporating The following CDRs: HC-CDR1: SYAMH (SEQ ID NO: 96), HC-CDR2: VISYDGSNKYYADSVKG (SEQ ID NO: 78), HC-CDR3: SRGYYGMDV (SEQ ID NO: 97) a heavy chain variable region incorporating (vii) the following CDRs: LC-CDR1: GLTSGAVSSSYYPS (SEQ ID NO: 279), LC-CDR2: NTDIRFS (SEQ ID NO: 280), LC-CDR3: VLYMGSGISV (SEQ ID NO: 23) a light chain variable region incorporating The following CDRs: HC-CDR1: DYYIH (SEQ ID NO: 281), HC-CDR2: WVSAYNGDTNYAQKFQG (SEQ ID NO: 282), HC-CDR3: EIASYSGSYYDY (SEQ ID NO: 20) Heavy chain variable region incorporating The antibody or antigen-binding fragment thereof, comprising:

2. the antibody or antigen-binding fragment: (i) a light chain variable region having at least 90% sequence identity to the light chain variable region of SEQ ID NO: 91; and a heavy chain variable region having at least 90% sequence identity to the heavy chain variable region of SEQ ID NO: 92; or (ii) a light chain variable region having at least 90% sequence identity to the light chain variable region of SEQ ID NO: 136; and a heavy chain variable region having at least 90% sequence identity to the heavy chain variable region of SEQ ID NO: 137; or (iii) a light chain variable region having at least 90% sequence identity to the light chain variable region of SEQ ID NO: 144; and a heavy chain variable region having at least 90% sequence identity to the heavy chain variable region of SEQ ID NO: 145; or (iv) a light chain variable region having at least 90% sequence identity to the light chain variable region of SEQ ID NO: 159; and a heavy chain variable region having at least 90% sequence identity to the heavy chain variable region of SEQ ID NO: 160; or (v) a light chain variable region having at least 90% sequence identity to the light chain variable region of SEQ ID NO: 202; and a heavy chain variable region having at least 90% sequence identity to the heavy chain variable region of SEQ ID NO: 203; or (vi) a light chain variable region having at least 90% sequence identity to the light chain variable region of SEQ ID NO: 209; and a heavy chain variable region having at least 90% sequence identity to the heavy chain variable region of SEQ ID NO: 210; or (vii) a light chain variable region having at least 90% sequence identity to the light chain variable region of SEQ ID NO: 277; and A heavy chain variable region having at least 90% sequence identity to the heavy chain variable region of SEQ ID NO: 278 The antibody or antigen-binding fragment of claim 1 , comprising:

3. 3. The antibody or antigen-binding fragment of claim 1 or claim 2, which specifically binds to human, rhesus monkey, or mouse HER2.

4. The antibody or antigen-binding fragment of any one of claims 1 to 3 conjugated to a drug moiety or a detectable moiety.

5. The antibody or antigen-binding fragment of claim 4 , wherein the drug moiety comprises an anticancer drug moiety.

6. A chimeric antigen receptor (CAR) comprising the antibody or antigen-binding fragment of any one of claims 1 to 3.

7. An in vitro complex comprising the antibody or antigen-binding fragment of any one of claims 1 to 5, or the CAR of claim 6, bound to HER2.

8. A nucleic acid encoding the antibody or antigen-binding fragment according to any one of claims 1 to 5, or the CAR according to claim 6.

9. A vector comprising the nucleic acid of claim 8.

10. A cell comprising the nucleic acid of claim 8 or the vector of claim 9.

11. A method for producing the antibody or antigen-binding fragment of any one of claims 1 to 5, or the CAR of claim 6, comprising the step of culturing the cell of claim 10 under conditions suitable for expression of the antibody, antigen-binding fragment, or CAR from the nucleic acid or vector.

12. A composition comprising the antibody or antigen-binding fragment of any one of claims 1 to 5, or the cell of claim 10, and at least one pharmaceutically acceptable carrier.

13. An antibody or antigen-binding fragment according to any one of claims 1 to 5, a cell according to claim 10, or a composition according to claim 12 for use in therapy or in a method of medical treatment.

14. Use of the antibody or antigen-binding fragment of any one of claims 1 to 5, the cell of claim 10, or the composition of claim 12 in the manufacture of a medicament for use in the treatment of cancer.

15. 15. The use according to claim 14, wherein the cancer is a HER2-positive cancer.

16. 16. The use of claim 15, wherein the cancer comprises HER2-positive tumor cells.

17. A method comprising the steps of contacting in vitro a sample containing or suspected of containing HER2 with the antibody or antigen-binding fragment of any one of claims 1 to 5, and detecting the formation of a complex comprising the antibody or antigen-binding fragment and HER2.

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