Antibodies recognizing Tau

Abstract

The present invention provides antibodies that specifically bind to tau. The antibody inhibits or delays the pathology associated with tau and the associated symptom deterioration.

Description

[0001] Divisional Application Instructions

[0002] This application is a divisional application of the invention patent application filed on February 7, 2020, with application number 202080018281.7 and invention title "Antibody that Recognizes Tau".

[0003] Cross-reference to related applications

[0004] This application claims the benefit of U.S. Provisional Application No. 62 / 803,334, filed February 8, 2019, U.S. Provisional Application No. 62 / 813,124, filed March 3, 2019, and U.S. Provisional Application No. 62 / 855,434, filed May 31, 2019, pursuant to 35 USC 119(e), each of which is incorporated herein by reference in its entirety for all purposes.

[0005] References to sequence lists

[0006] The sequence list written in file 50887-0029CN2_SL_ST26.xml is 276 kilobytes long and was created on January 25, 2025 (the actual sequence content was created on February 7, 2020), and is incorporated by reference accordingly. Background Technology

[0007] Tau is a well-known human protein that can exist in phosphorylated forms (see, for example, Goedert, Proc. Natl. Acad. Sci. USA 85:4051-4055 (1988); Goedert, EMBO J. 8:393-399 (1989); Lee, Neuron 2:1615-1624 (1989); Goedert, Neuron 3:519-526 (1989); Andreadis, Biochemistry 31:10626-10633 (1992). Tau has been reported to play a role in stabilizing microtubules, particularly in the central nervous system. Total tau (t-tau, i.e., phosphorylated and non-phosphorylated forms) and phosphorylated tau (p-tau, i.e., phosphorylated tau) are released by the brain in response to neuronal damage and neurodegeneration, and its levels have been reported to be elevated in the CSF of Alzheimer's patients relative to the general population (Jack et al., Lancet). Neurol 9:119-28(2010)).

[0008] Tau is a major component of neurofibrillary tangles, which, along with plaques, are a hallmark feature of Alzheimer's disease. The tangles form abnormal fibrils with a diameter of 10 nm, occurring in pairs, spirally wound, and exhibiting a regular periodicity of 80 nm. Tau within the neurofibrillary tangles is aberrantly phosphorylated (hyperphosphorylated), with phosphate ester groups attached to specific sites on the molecule. Severe involvement of neurofibrillary tangles has been observed in layer II neurons of the entorhinal cortex, CA1 and subunculus regions of the hippocampus, the amygdala, and deep layers of the neocortex (layers III, V, and superficial VI) in Alzheimer's disease. Furthermore, hyperphosphorylated tau has been reported to interfere with microtubule assembly, which may accelerate the destruction of neuronal networks.

[0009] Tau inclusion bodies are a key part of the neuropathology of several neurodegenerative diseases, including Alzheimer's disease, frontotemporal degeneration, progressive supranuclear palsy, and Pick's disease. Summary of the Invention

[0010] In one aspect, the present invention provides an isolated monoclonal antibody that competes with antibody 9F5 for binding to human tau. In some such antibodies, the heavy chain CDR-H3 has an amino acid sequence comprising SEQ ID NO:10. In some such antibodies, the heavy chain CDR-H1 has an amino acid sequence comprising SEQ ID NO:8. In some such antibodies, the light chains CDR CDR-L1, CDR-L2, and CDR-L3 have amino acid sequences comprising SEQ ID NO:12, 13, and 14, respectively. In some such antibodies, the heavy chain CDR-H1 has an amino acid sequence comprising SEQ ID NO:8. Some of these antibodies share the same epitope as 9F5 binding to human tau.

[0011] Some of these antibodies comprise three light chain CDRs and three heavy chain CDRs of monoclonal antibody 9F5, wherein 9F5 is a mouse antibody characterized by having a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:7 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:11.

[0012] In some such antibodies, the three heavy chain CDRs CDR-H1, CDR-H2, and CDR-H3 are as defined by the Kabat / Chothia complex (SEQ ID NO: 8, 9, and 10, respectively), except that position H28 can be occupied by N or T, position H51 by I or V, position H54 by N or D, and position H56 by D or E; and the three light chain CDRs CDR-L1, CDR-L2, and CDR-L3 are as defined by the Kabat / Chothia complex (SEQ ID NO: 8, 9, and 10, respectively). NO:12, 13 and 14), the difference is that position L27b is occupied by L, D, T or Q, position L27c is occupied by L, D, G, S, E, T, N, A, P or I, position L30 can be occupied by I, Y, E, K, G or Q, position L31 can be occupied by T, N or G, position L33 is occupied by L, N, T, S, R or G, position L51 can be occupied by M, G, E, D, K or I, position L54 can be occupied by L, R, G or T, position L89 is occupied by A or G, position L92 is occupied by L, D, E, G, Q, T or I and position L93 is occupied by E or G.

[0013] In some antibodies, CDR-H1 has the amino acid sequence comprising SEQ ID NO:50. In some antibodies, CDR-H2 has the amino acid sequence comprising SEQ ID NO:51. In some antibodies, CDR-H2 has the amino acid sequence comprising SEQ ID NO:52. In some antibodies, CDR-L1 has the amino acid sequence comprising any one of SEQ ID NO:53, SEQ ID NO:54, and SEQ ID NO:172-193. In some antibodies, CDR-L2 has the amino acid sequence comprising any one of SEQ ID NO:55 and SEQ ID NO:194-205. In some antibodies, CDR-L3 has the amino acid sequence comprising any one of SEQ ID NO:206-213. In some antibodies, CDR-H1 has the amino acid sequence comprising SEQ ID NO:50, and CDR-H2 has the amino acid sequence comprising SEQ ID NO:51. In some antibodies, CDR-L1 has an amino acid sequence containing SEQ ID NO:53, and CDR-L2 has an amino acid sequence containing SEQ ID NO:55. In some antibodies, CDR-L1 has an amino acid sequence containing SEQ ID NO:54, and CDR-L2 has an amino acid sequence containing SEQ ID NO:55.

[0014] Some antibodies are 9F5 or its chimeric, decorated, or humanized forms. In some antibodies, the variable heavy chain has ≥85% identity with the human sequence. In some antibodies, the variable light chain has ≥85% identity with the human sequence. In some antibodies, each of the variable heavy and variable light chains has ≥85% identity with the human germline sequence. Some antibodies are humanized antibodies.

[0015] Some antibodies are humanized or chimeric 9F5 antibodies that specifically bind to human tau, wherein 9F5 is a mouse antibody characterized by the mature heavy chain variable region of SEQ ID NO:7 and the mature light chain variable region of SEQ ID NO:11. Some antibodies comprise humanized mature heavy chain variable regions containing the three heavy chain CDRs of 9F5 and humanized mature light chain variable regions containing the three light chain CDRs of 9F5. In some antibodies, the CDR is defined as selected from the group consisting of Kabat, Chothia, Kabat / Chothia complex, AbM, and Contact.

[0016] In some antibodies, the humanized mature heavy chain variable region comprises three Kabat / Chothia complex heavy chain CDRs of 9F5 (SEQ ID NO: 8-10), and the humanized mature light chain variable region comprises three Kabat / Chothia complex light chain CDRs of 9F5 (SEQ ID NO: 12-14). In some antibodies, the humanized mature heavy chain variable region comprises three Kabat heavy chain CDRs of 9F5 (SEQ ID NO: 40, SEQ ID NO: 9, and SEQ ID NO: 10), and the humanized mature light chain variable region comprises three Kabat light chain CDRs of 9F5 (SEQ ID NO: 12-14). In some antibodies, the humanized mature heavy chain variable region comprises three Chothia heavy chain CDRs of 9F5 (SEQ ID NO: 41, SEQ ID NO: 42, and SEQ ID NO: 10), and the humanized mature light chain variable region comprises three Chothia light chain CDRs of 9F5 (SEQ ID NO: 12-14). In some antibodies, the humanized mature heavy chain variable region contains three AbM heavy chain CDRs of 9F5 (SEQ ID NO:8, SEQ ID NO:43, and SEQ ID NO:10), and the humanized mature light chain variable region contains three AbM light chain CDRs of 9F5 (SEQ ID NO:12-14). In some antibodies, the humanized mature heavy chain variable region contains three Contact heavy chain CDRs of 9F5 (SEQ ID NO:44-46), and the humanized mature light chain variable region contains three Contact light chain CDRs of 9F5 (SEQ ID NO:47-49).

[0017] For example, antibodies can be humanized antibodies, decorative antibodies, or chimeric antibodies.

[0018] Some of these antibodies contain a humanized mature heavy chain variable region having an amino acid sequence that is at least 90% identical to any of SEQ ID NO:15-22 and SEQ ID NO:109-129, and a humanized mature light chain variable region having an amino acid sequence that is at least 90% identical to any of SEQ ID NO:23-29, SEQ ID NO:61-108, and SEQ ID NO:130-171.

[0019] In some antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H1 is occupied by E, H17 by T, H20 by I, H69 by M, H75 by T, H93 by T, H94 by T, and H109 by V. In some antibodies, positions H1, H17, H20, H69, H75, H94, and H109 are occupied by E, T, I, M, T, T, T, and V, respectively.

[0020] In some antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H66 is occupied by R, and H81 is occupied by E. In some antibodies, positions H66 and H81 are occupied by R and E, respectively.

[0021] In some antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H23 is occupied by I, and H83 is occupied by R. In some antibodies, positions H23 and H83 are occupied by K and R, respectively.

[0022] In some antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H43 is occupied by K, H51 by V, H76 by D, M80 by M, and H108 by L. In some antibodies, positions H43, H51, H76, H80, and H108 are occupied by K, V, D, M, and L, respectively.

[0023] In some antibodies, the H28 position in the VH region is occupied by T.

[0024] In some antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H54 is occupied by D, and H56 is occupied by E. In some antibodies, positions H54 and H56 are occupied by D and E, respectively.

[0025] In some antibodies, position H40 in the VH region is occupied by A.

[0026] In some antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H5 is occupied by V, H11 by V, H12 by K, H38 by R, and H42 by G. In some antibodies, positions H5, H11, H12, H38, and H42 are occupied by V, V, K, R, and G, respectively.

[0027] In some antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H1 occupied by Q or E, H5 occupied by Q or V, H11 occupied by L or V, H12 occupied by V or K, H17 occupied by S or T, H20 occupied by L or I, H23 occupied by T or K, H28 occupied by N or T, H38 occupied by K, R, or Q, H40 occupied by R or A, H42 occupied by E or G, H43 occupied by Q or K, H48 occupied by I or M, H51 occupied by I or V, H54 occupied by N or D, and H56 occupied by... D or E is occupied, H66 is occupied by K or R, H69 is occupied by I or M, H75 is occupied by S or T, H76 is occupied by N or D, H79 is occupied by Y, Q, D, N or G, H80 is occupied by L, M, P, D, G or E, H81 is occupied by Q or E, H82 is occupied by L, P, K, R, E or N, H82a is occupied by S or G, H82c is occupied by L, G, D or S, H83 is occupied by T or R, H93 is occupied by A or T, H94 is occupied by S or T, H108 is occupied by T or L, and H109 is occupied by L or V.

[0028] In some antibodies, the VH region positions H1, H17, H20, H69, H75, H93, H94, and H109 are occupied by E, T, I, M, T, T, T, and V, respectively. In other antibodies, the VH region positions H1, H17, H20, H66, H69, H75, H81, H93, H94, and H109 are occupied by E, T, I, R, M, T, E, T, T, and V, respectively. In still other antibodies, the VH region positions H1, H17, H20, H23, H28, H66, H69, H75, H81, H83, H93, H94, and H109 are occupied by E, T, I, K, T, R, M, T, E, R, T, T, and V, respectively. In some antibodies, the positions H1, H17, H20, H23, H28, H43, H51, H54, H56, H66, H69, H75, H76, H80, H81, H83, H93, H94, H108, and H109 in the VH region are occupied by E, T, I, K, T, K, V, D, E, R, M, T, D, M, E, R, T, T, L, and V, respectively. In some antibodies, the positions H1, H17, H20, H23, H28, H40, H43, H48, H51, H54, H56, H66, H69, H75, H76, H80, H81, H83, H93, H94, H108, and H109 in the VH region are occupied by E, T, I, K, T, A, K, M, V, D, E, R, M, T, D, M, E, R, T, T, L, and V, respectively. In some antibodies, the positions H1, H5, H11, H12, H17, H20, H23, H38, H40, H42, H43, H51, H54, H56, H66, H69, H75, H76, H80, H81, H83, H93, H94, H108, and H109 in the VH region are occupied by E, V, V, K, T, I, K, R, A, G, K, V, D, E, R, M, T, D, M, E, R, T, T, L, and V, respectively. In some antibodies, H1, H5, H11, H12, H17, H20, H23, H38, H40, H42, H43, H51, H66, H69, H75, H76, H80, H81, H83, H93, H94, H108, and H109 in the VH region are occupied by E, V, V, K, T, I, K, R, A, G, K, V, R, M, T, D, M, E, R, T, T, L, and V, respectively.

[0029] In some antibodies, positions H1, H5, H11, H12, H17, H20, H23, H38, H42, H43, H66, H69, H75, H80, H81, H83, H93, H94, H108, and H109 in the VH region are occupied by E, V, V, K, T, I, K, Q, G, K, R, M, T, M, E, R, T, T, L, and V, respectively. In some antibodies, the heavy chain variable region contains the amino acid sequence of SEQ ID NO:127.

[0030] In some antibodies, positions H1, H5, H11, H12, H17, H20, H23, H38, H42, H43, H66, H69, H75, H80, H81, H83, H93, H94, H108, and H109 in the VH region are occupied by E, V, V, K, T, I, K, K, E, K, R, M, T, M, E, R, T, T, L, and V, respectively. In some antibodies, the heavy chain variable region contains the amino acid sequence of SEQ ID NO:128.

[0031] In some antibodies, the positions H1, H5, H11, H12, H17, H20, H23, H38, H42, H43, H66, H69, H75, H80, H81, H82c, H83, H93, H94, H108, and H109 in the VH region are occupied by E, V, V, K, T, I, K, K, E, K, R, M, T, M, E, G, R, T, T, L, and V, respectively.

[0032] In some antibodies, position H80 in the VH region is occupied by P. In some antibodies, position H80 in the VH region is occupied by D. In some antibodies, position H82c in the VH region is occupied by G. In some antibodies, position H82c in the VH region is occupied by D. In some antibodies, position H82 in the VH region is occupied by P. In some antibodies, position H80 in the VH region is occupied by G. In some antibodies, position H82 in the VH region is occupied by K. In some antibodies, position H82 in the VH region is occupied by R. In some antibodies, position H82 in the VH region is occupied by E. In some antibodies, position H82 in the VH region is occupied by N.

[0033] In some antibodies, position H79 in the VH region is occupied by D. In some antibodies, position H79 in the VH region is occupied by N. In some antibodies, position H79 in the VH region is occupied by G. In some antibodies, position H80 in the VH region is occupied by E. In some antibodies, position H80 in the VH region is occupied by G. In some antibodies, position H82c in the VH region is occupied by S. In some antibodies, position H79 in the VH region is occupied by Q. In some antibodies, position H82a in the VH region is occupied by G.

[0034] In some antibodies, at least one of the following positions in the VL region is occupied by a specified amino acid: L7 is occupied by S, L8 by P, L15 by P, and L100 by Q. In some antibodies, positions L7, L8, L15, and L100 are occupied by S, P, P, and Q, respectively.

[0035] In some antibodies, position L66 in the VL region is occupied by G. In other antibodies, position L64 in the VL region is occupied by S.

[0036] In some antibodies, position L17 in the VL region is occupied by E. In some antibodies, at least one of the following positions in the VL region is occupied by a specified amino acid: L11 is occupied by L, L51 is occupied by G, and L54 is occupied by R.

[0037] In some antibodies, positions L11, L51, and L54 are occupied by L, G, and R, respectively. In some antibodies, position L30 in the VL region is occupied by Y.

[0038] In some antibodies, at least one of the following positions in the VL region is occupied by a specified amino acid: L3 occupied by V or Q, L7 occupied by A or S, L8 occupied by A or P, L9 occupied by F or L, L11 occupied by N or L, L15 occupied by L or P, L17 occupied by T or E, L18 occupied by S or P, L27b occupied by L, D, T, or Q, L27c occupied by L, D, G, S, E, T, N, A, P, or I, L30 occupied by I, Y, E, K, or G, L31 occupied by T, N, or G, L33 occupied by L, N, T, S, R, or G, L37 occupied by L, Q, G, or I, and L39 occupied by R or... K is occupied, L51 is occupied by M, G, E, D, K or I, L54 is occupied by R, G or T, L60 is occupied by N or D, L64 is occupied by G or S, L66 is occupied by E or G, L73 is occupied by L, P or G, L74 is occupied by R or K, L75 is occupied by I, D, P, Q or G, L76 is occupied by S, P or G, L77 is occupied by R or D, L78 is occupied by V, R, D, E, P, K, G or Q, L85 is occupied by V or G, L86 is occupied by Y or T, L89 is occupied by A or G, L92 is occupied by L, D, E, G, Q, T or I, L93 is occupied by E or G, L100 is occupied by G or Q.

[0039] In some antibodies, positions L64 and L66 in the VL region are occupied by S and G, respectively. In some antibodies, positions L7, L8, L15, L64, L66, and L100 in the VL region are occupied by S, P, P, S, G, and Q, respectively. In some antibodies, positions L7, L8, L15, L17, L66, and L100 in the VL region are occupied by S, P, P, E, G, and Q, respectively. In some antibodies, positions L7, L8, L11, L15, L17, L51, L54, L66, and L100 in the VL region are occupied by S, P, L, P, E, G, R, G, and Q, respectively.

[0040] In some antibodies, the light chain variable region contains the amino acid sequence of any one of SEQ ID NO:133, 135-137, 142-144, 149, 158, 159, and 168. In some antibodies, the light chain variable region contains the amino acid sequence of SEQ ID NO:133. In some antibodies, the light chain variable region contains the amino acid sequence of SEQ ID NO:137. In some antibodies, the light chain variable region contains the amino acid sequence of SEQ ID NO:149. In some antibodies, the light chain variable region contains the amino acid sequence of SEQ ID NO:159.

[0041] In some antibodies, positions L7, L8, L11, L15, L17, L30, L51, L54, L66, and L100 in the VL region are occupied by S, P, L, P, E, Y, G, R, G, and Q, respectively. In some antibodies, positions L7, L8, L11, L15, L17, L30, L51, L54, and L100 in the VL region are occupied by S, P, L, P, E, Y, G, R, and Q, respectively. In some antibodies, positions L7, L8, L9, L11, L15, L17, L18, L31, L39, L51, L54, L60, L66, L74, and L100 in the VL region are occupied by S, P, L, L, P, E, P, N, K, G, R, D, G, K, and Q, respectively.

[0042] In some antibodies, positions L7, L8, L11, L15, L17, L39, L60, L64, L66, L74, and L100 in the VL region are occupied by S, P, L, P, E, K, N, S, G, K, and Q, respectively. In some antibodies, position L3 in the VL region is occupied by Q. In some antibodies, position L27c in the VL region is occupied by D, G, I, L, or S; position L37 in the VL region is occupied by G, I, L, or Q; position L51 in the VL region is occupied by E, G, I, K, or M; position L54 in the VL region is occupied by G, L, R, or T; and position L92 in the VL region is occupied by G, I, or L. In some antibodies, position L27c in the VL region is occupied by D or S, position L37 in the VL region is occupied by G, L or Q, position L51 in the VL region is occupied by G or K, position L54 in the VL region is occupied by R, and position L92 in the VL region is occupied by I.

[0043] In some antibodies, position L27c in the VL region is occupied by D, position L37 in the VL region is occupied by G, and position L51 in the VL region is occupied by G. In some antibodies, the heavy chain variable region has an amino acid sequence containing SEQ ID NO:127, and the light chain variable region has an amino acid sequence containing SEQ ID NO:149.

[0044] In some antibodies, position L27c in the VL region is occupied by D, position L37 in the VL region is occupied by Q, and position L51 in the VL region is occupied by G. In some antibodies, the heavy chain variable region has an amino acid sequence containing SEQ ID NO:127, and the light chain variable region has an amino acid sequence containing SEQ ID NO:137.

[0045] In some antibodies, position L27c in the VL region is occupied by S, position L37 in the VL region is occupied by L, and position L51 in the VL region is occupied by G. In some antibodies, the heavy chain variable region has an amino acid sequence containing SEQ ID NO:127, and the light chain variable region has an amino acid sequence containing SEQ ID NO:159.

[0046] In some antibodies, position L27c in the VL region is occupied by D, position L37 in the VL region is occupied by Q, and position L51 in the VL region is occupied by K. In some antibodies, the heavy chain variable region has an amino acid sequence containing SEQ ID NO:127, and the light chain variable region has an amino acid sequence containing SEQ ID NO:138.

[0047] In some antibodies, position L27c in the VL region is occupied by S, position L37 in the VL region is occupied by Q, and position L51 in the VL region is occupied by G. In some antibodies, the heavy chain variable region has an amino acid sequence containing SEQ ID NO:127, and the light chain variable region has an amino acid sequence containing SEQ ID NO:133.

[0048] In some antibodies, positions L7, L8, L11, L15, L17, L39, L60, L64, L66, L74, and L100 in the VL region are occupied by S, P, L, P, E, K, D, S, G, K, and Q, respectively. In some antibodies, position L3 in the VL region is occupied by Q. In some antibodies, position L27c in the VL region is occupied by G or S, position L37 in the VL region is occupied by G, I, or Q, position L51 in the VL region is occupied by G, I, or K, position L54 in the VL region is occupied by G or R, and position L92 in the VL region is occupied by G, I, or L.

[0049] In some antibodies, position L27c in the VL region is occupied by G, position L37 in the VL region is occupied by G, position L51 in the VL region is occupied by G, and position L54 in the VL region is occupied by R. In some antibodies, position L92 in the VL region is occupied by I. In some antibodies, the heavy chain variable region has an amino acid sequence containing SEQ ID NO:129, and the light chain variable region has an amino acid sequence containing SEQ ID NO:168.

[0050] In some antibodies, position L51 in the VL region is occupied by E. In some antibodies, position L51 in the VL region is occupied by D. In some antibodies, position L27c in the VL region is occupied by D. In some antibodies, position L27c in the VL region is occupied by G. In some antibodies, position L27c in the VL region is occupied by S. In some antibodies, position L27c in the VL region is occupied by E. In some antibodies, position L30 in the VL region is occupied by E. In some antibodies, position L30 in the VL region is occupied by K. In some antibodies, position L27c in the VL region is occupied by T.

[0051] In some antibodies, position L27c in the VL region is occupied by N. In some antibodies, position L27b in the VL region is occupied by D. In some antibodies, position L30 in the VL region is occupied by G. In some antibodies, position L33 in the VL region is occupied by N. In some antibodies, position L27c in the VL region is occupied by A. In some antibodies, position L33 in the VL region is occupied by T. In some antibodies, position L33 in the VL region is occupied by S. In some antibodies, position L33 in the VL region is occupied by R. In some antibodies, position L30 in the VL region is occupied by Q. In some antibodies, position L27b in the VL region is occupied by T.

[0052] In some antibodies, position L31 in the VL region is occupied by G. In some antibodies, position L27b in the VL region is occupied by Q. In some antibodies, position L33 in the VL region is occupied by G. In some antibodies, position L27c in the VL region is occupied by P. In some antibodies, position L78 in the VL region is occupied by R. In some antibodies, position L75 in the VL region is occupied by D. In some antibodies, position L78 in the VL region is occupied by D. In some antibodies, position L78 in the VL region is occupied by E. In some antibodies, position L78 in the VL region is occupied by P. In some antibodies, position L78 in the VL region is occupied by K.

[0053] In some antibodies, position L77 in the VL region is occupied by D. In some antibodies, position L78 in the VL region is occupied by G. In some antibodies, position L76 in the VL region is occupied by P. In some antibodies, position L75 in the VL region is occupied by P. In some antibodies, position L75 in the VL region is occupied by Q. In some antibodies, position L75 in the VL region is occupied by G. In some antibodies, position L73 in the VL region is occupied by P. In some antibodies, position L73 in the VL region is occupied by G. In some antibodies, position L78 in the VL region is occupied by Q. In some antibodies, position L76 in the VL region is occupied by G.

[0054] In some antibodies, position L92 in the VL region is occupied by D. In some antibodies, position L86 in the VL region is occupied by T. In some antibodies, position L92 in the VL region is occupied by E. In some antibodies, position L92 in the VL region is occupied by G. In some antibodies, position L92 in the VL region is occupied by Q. In some antibodies, position L93 in the VL region is occupied by G. In some antibodies, position L85 in the VL region is occupied by G. In some antibodies, position L92 in the VL region is occupied by T. In some antibodies, position L89 in the VL region is occupied by G.

[0055] In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, G, G, and I, respectively. (This is repeated four times in the original text.)

[0056] In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, D, Q, G, R, and I, respectively. (This text is repeated five times in the original.)

[0057] In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, G, G, R, and I, respectively. (This text is repeated four times in the original.)

[0058] In some antibodies, positions L3, L27c, L37, L51, and L54 in the VL region are occupied by Q, G, G, G, and T, respectively. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, G, G, T, and I, respectively. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, D, G, G, R, and I, respectively. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, I, G, and I, respectively. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, I, G, and I, respectively.

[0059] In some antibodies, positions L3, L27c, L37, L51, and L54 in the VL region are occupied by Q, S, Q, I, and G, respectively. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, E, R, and I, respectively. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, I, E, R, and I, respectively. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, I, E, R, and G, respectively. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, I, E, R, and G, respectively.

[0060] In some antibodies, positions L3, L27c, L37, L51, and L54 in the VL region are occupied by Q, I, I, E, and R, respectively. In some antibodies, positions L3, L37, L51, L54, and L92 in the VL region are occupied by Q, Q, G, R, and I, respectively. In some antibodies, positions L3, L27c, L51, L54, and L92 in the VL region are occupied by Q, S, G, R, and I, respectively. In some antibodies, positions L3, L27c, L37, L54, and L92 in the VL region are occupied by Q, S, Q, G, and I, respectively. In some antibodies, positions L3, L27c, L37, L51, and L92 in the VL region are occupied by Q, S, Q, G, and I, respectively.

[0061] In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, G, G, and I. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, G, R, and I, respectively. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, G, R, and G, respectively. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, Q, G, R, and I, respectively. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, Q, K, R, and I, respectively.

[0062] In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, K, G, and I, respectively. (This text is repeated four times in the original.)

[0063] Some antibodies comprise a mature heavy chain variable region having an amino acid sequence that is at least 95% identical to any of SEQ ID NO:15-22 and SEQ ID NO:109-129, and a mature light chain variable region having an amino acid sequence that is at least 95% identical to any of SEQ ID NO:23-29, SEQ ID NO:61-108, and SEQ ID NO:130-171. Some antibodies comprise a mature heavy chain variable region having an amino acid sequence that is at least 98% identical to any of SEQ ID NO:15-22 and SEQ ID NO:109-129, and a mature light chain variable region having an amino acid sequence that is at least 98% identical to any of SEQ ID NO:23-29, SEQ ID NO:61-108, and SEQ ID NO:130-171. In some antibodies, the mature heavy chain variable region has the amino acid sequence of any one of SEQ ID NO:15-22 and SEQ ID NO:109-129, and the mature light chain variable region has the amino acid sequence of any one of SEQ ID NO:23-29, SEQ ID NO:61-108 and SEQ ID NO:130-171.

[0064] In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:15, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:23. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:15, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:24. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:15, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:25. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:15, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:26. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:15, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:27. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:15, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:28. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:15, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:29.

[0065] In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:16, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:23. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:16, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:24. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:16, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:25. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:16, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:26. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:16, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:27. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:16, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:28. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:16, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:29.

[0066] In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:17, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:23. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:17, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:24. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:17, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:25. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:17, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:26. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:17, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:27. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:17, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:28. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:17, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:29.

[0067] In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:18, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:23. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:18, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:24. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:18, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:25. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:18, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:26. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:18, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:27. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:18, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:28. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:18, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:29.

[0068] In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:19, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:23. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:19, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:24. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:19, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:25. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:19, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:26. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:19, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:27. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:19, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:28. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:19, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:29.

[0069] In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:20, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:23. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:20, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:24. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:20, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:25. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:20, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:26. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:20, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:27. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:20, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:28. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:20, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:29.

[0070] In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:21, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:23. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:21, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:24. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:21, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:25. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:21, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:26. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:21, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:27. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:21, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:28. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:21, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:29.

[0071] In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:22, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:23. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:22, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:24. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:22, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:25. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:22, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:26. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:22, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:27. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:22, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:28. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:22, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:29.

[0072] In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:149. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:142. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:159. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:148.

[0073] In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:137. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:145. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:136. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:138.

[0074] In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:158. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:143. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:144. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:133.

[0075] In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:160. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:161. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:139. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:128, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:168.

[0076] Some of these antibodies comprise three light chain CDRs and three heavy chain CDRs of monoclonal antibody 10C12, wherein 10C12 is a mouse antibody characterized by having a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:7 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:11.

[0077] In some of these antibodies, the three heavy chain CDRs CDR-H1, CDR-H2, and CDR-H3 are as defined by the Kabat / Chothia complex (SEQ ID NO: 8, 9, and 10, respectively), and the three light chain CDRs CDR-L1, CDR-L2, and CDR-L3 are as defined by the Kabat / Chothia complex (SEQ ID NO: 12, 13, and 14, respectively).

[0078] Some antibodies are 10C12 or their chimeric, decorated, or humanized forms. In some antibodies, the variable heavy chain has ≥85% identity with the human sequence. In some antibodies, the variable light chain has ≥85% identity with the human sequence. In some antibodies, each of the variable heavy chain and variable light chain has ≥85% identity with the human germline sequence. Some antibodies are humanized antibodies.

[0079] Some antibodies are humanized or chimeric 10C12 antibodies that specifically bind to human tau, wherein 10C12 is a mouse antibody characterized by the mature heavy chain variable region of SEQ ID NO:7 and the mature light chain variable region of SEQ ID NO:11. Some antibodies comprise humanized mature heavy chain variable regions containing three heavy chain CDRs of 10C12 and humanized mature light chain variable regions containing three light chain CDRs of 10C12. In some antibodies, the definition of CDR is selected from the group consisting of Kabat, Chothia, Kabat / Chothia complex, AbM, and Contact.

[0080] In some antibodies, the humanized mature heavy chain variable region comprises three Kabat / Chothia complex heavy chain CDRs of 10C12 (SEQ ID NO: 8-10), and the humanized mature light chain variable region comprises three Kabat / Chothia complex light chain CDRs of 10C12 (SEQ ID NO: 12-14). In some antibodies, the humanized mature heavy chain variable region comprises three Kabat heavy chain CDRs of 10C12 (SEQ ID NO: 40, SEQ ID NO: 9, and SEQ ID NO: 10), and the humanized mature light chain variable region comprises three Kabat light chain CDRs of 10C12 (SEQ ID NO: 12-14). In some antibodies, the humanized mature heavy chain variable region comprises three Chothia heavy chain CDRs of 10C12 (SEQ ID NO:41, SEQ ID NO:42, and SEQ ID NO:10), and the humanized mature light chain variable region comprises three Chothia light chain CDRs of 10C12 (SEQ ID NO:12-14). In some antibodies, the humanized mature heavy chain variable region comprises three AbM heavy chain CDRs of 10C12 (SEQ ID NO:8, SEQ ID NO:43, and SEQ ID NO:10), and the humanized mature light chain variable region comprises three AbM light chain CDRs of 10C12 (SEQ ID NO:12-14). In some antibodies, the humanized mature heavy chain variable region contains three Contact heavy chain CDRs of 10C12 (SEQ ID NO:44-46), and the humanized mature light chain variable region contains three Contact light chain CDRs of 10C12 (SEQ ID NO:47-49).

[0081] For example, antibodies can be humanized antibodies, decorative antibodies, or chimeric antibodies.

[0082] Some of these antibodies contain a humanized mature heavy chain variable region having an amino acid sequence that is at least 90% identical to any of SEQ ID NO:214-215 and a humanized mature light chain variable region having an amino acid sequence that is at least 90% identical to any of SEQ ID NO:216-217.

[0083] In some antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H24 is occupied by A, H48 by I, H67 by A, H69 by M, H93 by T, and H94 by T. In some antibodies, positions H24, H48, H67, H69, H93, and H94 are occupied by A, I, A, M, T, and T, respectively.

[0084] In some antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H1 is occupied by Q or E, H24 is occupied by A, H48 is occupied by I, H67 is occupied by A, H69 is occupied by M, H93 is occupied by T, and H94 is occupied by T.

[0085] In some antibodies, positions H24, H48, H67, H69, H93, and H94 are occupied by A, I, A, M, T, and T, respectively. In other antibodies, positions H1, H24, H48, H67, H69, H93, and H94 are occupied by E, A, I, A, M, T, and T, respectively.

[0086] In some antibodies, position L64 in the VL region is occupied by S.

[0087] In some antibodies, at least one of the following positions in the VL region is occupied by a specified amino acid: L64 is occupied by S, and L104 is occupied by V or L. In some antibodies, position L64 is occupied by S. In some antibodies, positions L64 and L104 in the VL region are occupied by S and L, respectively.

[0088] Some antibodies comprise a mature heavy chain variable region having an amino acid sequence that is at least 95% identical to any of SEQ ID NO:214-215 and a mature light chain variable region having an amino acid sequence that is at least 95% identical to any of SEQ ID NO:216-217. Some antibodies comprise a mature heavy chain variable region having an amino acid sequence that is at least 98% identical to any of SEQ ID NO:214-215 and a mature light chain variable region having an amino acid sequence that is at least 98% identical to any of SEQ ID NO:216-217. In some antibodies, the mature heavy chain variable region has an amino acid sequence of any of SEQ ID NO:214-215, and the mature light chain variable region has an amino acid sequence of any of SEQ ID NO:216-217.

[0089] In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:214, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:216. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:214, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:217. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:215, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:216. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:215, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:217.

[0090] Some of these antibodies comprise three light chain CDRs and three heavy chain CDRs of monoclonal antibody 12C4, wherein 12C4 is a mouse antibody characterized by having a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:219 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:11.

[0091] In some of these antibodies, the three heavy chain CDRs CDR-H1, CDR-H2, and CDR-H3 are as defined by the Kabat / Chothia complex (SEQ ID NO: 8, 220, and 10, respectively), and the three light chain CDRs CDR-L1, CDR-L2, and CDR-L3 are as defined by the Kabat / Chothia complex (SEQ ID NO: 12, 13, and 14, respectively).

[0092] Some antibodies are 12C4 or their chimeric, decorated, or humanized forms. In some antibodies, the variable heavy chain has ≥85% identity with the human sequence. In some antibodies, the variable light chain has ≥85% identity with the human sequence. In some antibodies, each of the variable heavy chain and variable light chain has ≥85% identity with the human germline sequence. Some antibodies are humanized antibodies.

[0093] Some antibodies are humanized or chimeric 12C4 antibodies that specifically bind to human tau, wherein 12C4 is a mouse antibody characterized by the mature heavy chain variable region of SEQ ID NO:219 and the mature light chain variable region of SEQ ID NO:11. Some antibodies comprise humanized mature heavy chain variable regions containing three heavy chain CDRs of 12C4 and humanized mature light chain variable regions containing three light chain CDRs of 12C4. In some antibodies, the definition of CDR is selected from the group consisting of Kabat, Chothia, Kabat / Chothia complex, AbM, and Contact.

[0094] In some antibodies, the humanized mature heavy chain variable region comprises three Kabat / Chothia complex heavy chain CDRs of 12C4 (SEQ ID NO: 8, 220, and 10), and the humanized mature light chain variable region comprises three Kabat / Chothia complex light chain CDRs of 12C4 (SEQ ID NO: 12-14). In some antibodies, the humanized mature heavy chain variable region comprises three Kabat heavy chain CDRs of 12C4 (SEQ ID NO: 40, SEQ ID NO: 220, and SEQ ID NO: 10), and the humanized mature light chain variable region comprises three Kabat light chain CDRs of 12C4 (SEQ ID NO: 12-14). In some antibodies, the humanized mature heavy chain variable region comprises three Chothia heavy chain CDRs of 12C4 (SEQ ID NO:41, SEQ ID NO:42, and SEQ ID NO:10), and the humanized mature light chain variable region comprises three Chothia light chain CDRs of 12C4 (SEQ ID NO:12-14). In some antibodies, the humanized mature heavy chain variable region comprises three AbM heavy chain CDRs of 12C4 (SEQ ID NO:8, SEQ ID NO:257, and SEQ ID NO:10), and the humanized mature light chain variable region comprises three AbM light chain CDRs of 12C4 (SEQ ID NO:12-14). In some antibodies, the humanized mature heavy chain variable region comprises three Contact heavy chain CDRs of 12C4 (SEQ ID NO:44, 258, and 46), and the humanized mature light chain variable region comprises three Contact light chain CDRs of 12C4 (SEQ ID NO:47-49).

[0095] For example, antibodies can be humanized antibodies, decorative antibodies, or chimeric antibodies.

[0096] Some of these antibodies contain a humanized mature heavy chain variable region having an amino acid sequence that is at least 90% identical to any of SEQ ID NO:221-222 and a humanized mature light chain variable region having an amino acid sequence that is at least 90% identical to any of SEQ ID NO:223-224.

[0097] In some antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H1 is occupied by Q or E, H48 by M or I, H93 by A or T, and H94 by R or T. In some antibodies, positions H1, H48, H93, and H94 in the VH region are occupied by E, I, T, and T, respectively.

[0098] In some antibodies, positions in the VL region are occupied by specific amino acids: L64 is occupied by G or S, and L104 is occupied by V or L. In other antibodies, positions L64 and L104 in the VL region are occupied by S and L, respectively.

[0099] Some antibodies comprise a mature heavy chain variable region having an amino acid sequence that is at least 95% identical to any of SEQ ID NO:221-222 and a mature light chain variable region having an amino acid sequence that is at least 95% identical to any of SEQ ID NO:223-224. Some antibodies comprise a mature heavy chain variable region having an amino acid sequence that is at least 98% identical to any of SEQ ID NO:221-222 and a mature light chain variable region having an amino acid sequence that is at least 98% identical to any of SEQ ID NO:223-224. In some antibodies, the mature heavy chain variable region has an amino acid sequence of any of SEQ ID NO:221-222, and the mature light chain variable region has an amino acid sequence of any of SEQ ID NO:223-224.

[0100] In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:221, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:223. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:221, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:224. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:222, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:223. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:222, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:224.

[0101] Some of these antibodies comprise three light chain CDRs and three heavy chain CDRs of the monoclonal antibody 17C12, wherein 17C12 is a mouse antibody characterized by having a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:225 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:228.

[0102] In some of these antibodies, the three heavy chain CDRs CDR-H1, CDR-H2, and CDR-H3 are as defined by the Kabat / Chothia complex (SEQ ID NO: 226, 227, and 10, respectively), and the three light chain CDRs CDR-L1, CDR-L2, and CDR-L3 are as defined by the Kabat / Chothia complex (SEQ ID NO: 229, 230, and 231, respectively).

[0103] Some antibodies are 17C12 or their chimeric, decorated, or humanized forms. In some antibodies, the variable heavy chain has ≥85% identity with the human sequence. In some antibodies, the variable light chain has ≥85% identity with the human sequence. In some antibodies, each of the variable heavy chain and variable light chain has ≥85% identity with the human germline sequence. Some antibodies are humanized antibodies.

[0104] Some antibodies are humanized or chimeric 17C12 antibodies that specifically bind to human tau, wherein 17C12 is a mouse antibody characterized by the mature heavy chain variable region of SEQ ID NO:225 and the mature light chain variable region of SEQ ID NO:228. Some antibodies comprise humanized mature heavy chain variable regions containing three heavy chain CDRs of 17C12 and humanized mature light chain variable regions containing three light chain CDRs of 17C12. In some antibodies, the definition of CDR is selected from the group consisting of Kabat, Chothia, Kabat / Chothia complex, AbM, and Contact.

[0105] In some antibodies, the humanized mature heavy chain variable region comprises three Kabat / Chothia complex heavy chain CDRs of 17C12 (SEQ ID NO: 226, 227, and 10), and the humanized mature light chain variable region comprises three Kabat / Chothia complex light chain CDRs of 17C12 (SEQ ID NO: 229-231). In some antibodies, the humanized mature heavy chain variable region comprises three Kabat heavy chain CDRs of 17C12 (SEQ ID NO: 40, SEQ ID NO: 227, and SEQ ID NO: 10), and the humanized mature light chain variable region comprises three Kabat light chain CDRs of 17C12 (SEQ ID NO: 229-231). In some antibodies, the humanized mature heavy chain variable region comprises three Chothia heavy chain CDRs of 17C12 (SEQ ID NO:259, SEQ ID NO:42, and SEQ ID NO:10), and the humanized mature light chain variable region comprises three Chothia light chain CDRs of 17C12 (SEQ ID NO:229-231). In some antibodies, the humanized mature heavy chain variable region comprises three AbM heavy chain CDRs of 17C12 (SEQ ID NO:226, SEQ ID NO:260, and SEQ ID NO:10), and the humanized mature light chain variable region comprises three AbM light chain CDRs of 17C12 (SEQ ID NO:229-231). In some antibodies, the humanized mature heavy chain variable region contains three Contact heavy chain CDRs of 17C12 (SEQ ID NO:44, SEQ ID NO:261 and SEQ ID NO:46), and the humanized mature light chain variable region contains three Contact light chain CDRs of 17C12 (SEQ ID NO:262-264).

[0106] For example, antibodies can be humanized antibodies, decorative antibodies, or chimeric antibodies.

[0107] Some of these antibodies contain a humanized mature heavy chain variable region having an amino acid sequence that is at least 90% identical to any of SEQ ID NO:232-233 and a humanized mature light chain variable region having an amino acid sequence that is at least 90% identical to any of SEQ ID NO:234-235.

[0108] In some antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H2 is occupied by I, H24 by A, H48 by I, H67 by A, H69 by M, H93 by T, and H94 by T. In some antibodies, positions H2, H24, H48, H67, H69, H93, and H94 are occupied by E, A, I, A, M, T, and T, respectively.

[0109] In some antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H1 occupied by Q or E, H2 occupied by I, H24 occupied by A, H48 occupied by I, H67 occupied by A, H69 occupied by M, H93 occupied by T, H94 occupied by T, H108 occupied by T or L, and H113 occupied by R or S. In some antibodies, positions H2, H24, H48, H67, H69, H93, and H94 in the VH region are occupied by E, A, I, A, M, T, and T, respectively. In some antibodies, positions H1, H2, H24, H48, H67, H69, H93, H94, H108, and H113 in the VH region are occupied by E, I, A, I, A, M, T, T, L, and S, respectively.

[0110] In some antibodies, at least one of the following positions in the VL region is occupied by a specified amino acid: L2 is occupied by V, and L36 is occupied by L. In some antibodies, positions L2 and L36 are occupied by V and L, respectively.

[0111] In some antibodies, at least one of the following positions in the VL region is occupied by a specified amino acid: L2 is occupied by V, L36 is occupied by L, and L43 is occupied by P or S. In some antibodies, positions L2 and L36 in the VL region are occupied by V and L, respectively. In some antibodies, positions L2, L36, and L43 in the VL region are occupied by V, L, and S, respectively.

[0112] Some antibodies comprise a mature heavy chain variable region having an amino acid sequence that is at least 95% identical to any of SEQ ID NO:232-233 and a mature light chain variable region having an amino acid sequence that is at least 95% identical to any of SEQ ID NO:234-235. Some antibodies comprise a mature heavy chain variable region having an amino acid sequence that is at least 98% identical to any of SEQ ID NO:232-233 and a mature light chain variable region having an amino acid sequence that is at least 98% identical to any of SEQ ID NO:234-235. In some antibodies, the mature heavy chain variable region has an amino acid sequence of any of SEQ ID NO:232-233, and the mature light chain variable region has an amino acid sequence of any of SEQ ID NO:234-235.

[0113] In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:232, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:234. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:232, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:235. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:233, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:234. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:233, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:235.

[0114] Some of these antibodies comprise three light chain CDRs and three heavy chain CDRs of monoclonal antibody 14H3, wherein 14H3 is a mouse antibody characterized by having a heavy chain variable region containing the amino acid sequence of SEQ ID NO:240 and a light chain variable region containing the amino acid sequence of SEQ ID NO:244.

[0115] In some such antibodies, the three heavy chain CDRs CDR-H1, CDR-H2, and CDR-H3 are as defined by the Kabat / Chothia complex (SEQ ID NO: 241, 242, and 243, respectively), except that position H35B can be occupied by G or S, and the three light chain CDRs CDR-L1, CDR-L2, and CDR-L3 are as defined by the Kabat / Chothia complex (SEQ ID NO: 245, 246, and 247, respectively). In some antibodies, CDR-H1 has an amino acid sequence including SEQ ID NO: 277.

[0116] Some antibodies are 14H3 or its chimeric, decorated, or humanized forms. In some antibodies, the variable heavy chain has ≥85% identity with the human sequence. In some antibodies, the variable light chain has ≥85% identity with the human sequence. In some antibodies, each of the variable heavy chain and variable light chain has ≥85% identity with the human germline sequence. Some antibodies are humanized antibodies.

[0117] Some antibodies are humanized or chimeric 14H3 antibodies that specifically bind to human tau, wherein 14H3 is a mouse antibody characterized by the mature heavy chain variable region of SEQ ID NO:240 and the mature light chain variable region of SEQ ID NO:244. Some antibodies comprise humanized mature heavy chain variable regions containing three heavy chain CDRs of 14H3 and humanized mature light chain variable regions containing three light chain CDRs of 14H3. In some antibodies, the definition of CDR is selected from the group consisting of Kabat, Chothia, Kabat / Chothia complex, AbM, and Contact.

[0118] In some antibodies, the humanized mature heavy chain variable region comprises three Kabat / Chothia complex heavy chain CDRs of 14H3 (SEQ ID NO:241-243), and the humanized mature light chain variable region comprises three Kabat / Chothia complex light chain CDRs of 14H3 (SEQ ID NO:245-247). In some antibodies, the humanized mature heavy chain variable region comprises three Kabat heavy chain CDRs of 14H3 (SEQ ID NO:265, SEQ ID NO:242, and SEQ ID NO:243), and the humanized mature light chain variable region comprises three Kabat light chain CDRs of 14H3 (SEQ ID NO:245-247). In some antibodies, the humanized mature heavy chain variable region contains three Chothia heavy chain CDRs of 14H3 (SEQ ID NO:266, SEQ ID NO:267, and SEQ ID NO:243), and the humanized mature light chain variable region contains three Chothia light chain CDRs of 14H3 (SEQ ID NO:245-247). In some antibodies, the humanized mature heavy chain variable region contains three AbM heavy chain CDRs of 14H3 (SEQ ID NO:241, SEQ ID NO:268, and SEQ ID NO:243), and the humanized mature light chain variable region contains three AbM light chain CDRs of 14H3 (SEQ ID NO:245-247). In some antibodies, the humanized mature heavy chain variable region contains three Contact heavy chain CDRs of 14H3 (SEQ ID NO: 269-271), and the humanized mature light chain variable region contains three Contact light chain CDRs of 14H3 (SEQ ID NO: 272-274).

[0119] For example, antibodies can be humanized antibodies, decorative antibodies, or chimeric antibodies.

[0120] Some of these antibodies contain a humanized mature heavy chain variable region having an amino acid sequence that is at least 90% identical to any of SEQ ID NO:248-249 and a humanized mature light chain variable region having an amino acid sequence that is at least 90% identical to any of SEQ ID NO:250-251.

[0121] In some antibodies, the H35B position in the VH region is occupied by S.

[0122] In some antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H35B is occupied by S, H108 is occupied by M or L, and H113 is occupied by L or S. In some antibodies, position H35B in the VH region is occupied by S. In some antibodies, positions H35B, H108, and H113 in the VH region are occupied by S, L, and S, respectively.

[0123] In some antibodies, at least one of the following positions in the VL region is occupied by a specified amino acid: L2 is occupied by V, and L87 is occupied by F. In some antibodies, positions L2 and L87 are occupied by V and F, respectively.

[0124] In some antibodies, at least one of the following positions in the VL region is occupied by a specified amino acid: L2 is occupied by V, L7 by T or S, L37 by L or Q, L87 by F, L100 by G or Q, and L104 by V or L. In some antibodies, positions L2 and L87 in the VL region are occupied by V and F, respectively. In some antibodies, positions L2, L7, L37, L87, L100, and L104 in the VL region are occupied by V, S, Q, F, Q, and L, respectively.

[0125] Some antibodies comprise a mature heavy chain variable region having an amino acid sequence that is at least 95% identical to any of SEQ ID NO:248-249 and a mature light chain variable region having an amino acid sequence that is at least 95% identical to any of SEQ ID NO:250-251. Some antibodies comprise a mature heavy chain variable region having an amino acid sequence that is at least 98% identical to any of SEQ ID NO:248-249 and a mature light chain variable region having an amino acid sequence that is at least 98% identical to any of SEQ ID NO:250-251. In some antibodies, the mature heavy chain variable region has an amino acid sequence of any of SEQ ID NO:248-249, and the mature light chain variable region has an amino acid sequence of any of SEQ ID NO:250-251.

[0126] In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:248, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:250. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:248, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:251. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:249, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:250. In some antibodies, the mature heavy chain variable region has the amino acid sequence of SEQ ID NO:249, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:251.

[0127] For example, an antibody can be a chimeric antibody. For example, an antibody can be a decorative antibody.

[0128] Antibodies can be intact antibodies. Antibodies can be binding fragments. In one embodiment, the binding fragment is a single-chain antibody, a Fab, or a Fab'2 fragment. Antibodies can be Fab fragments or single-chain Fv. Some antibodies have a human IgG1 isotype, while others may have a human IgG2 or IgG4 isotype. Some antibodies have a mature light chain variable region fused to a light chain constant region and a mature heavy chain variable region fused to a heavy chain constant region. The heavy chain constant region of some antibodies is a mutant form of the natural human heavy chain constant region, which binds less to the Fcγ receptor compared to the natural human heavy chain constant region. In some antibodies, the heavy chain constant region belongs to the IgG1 isotype.

[0129] Some antibodies may have at least one mutation in the constant region, such as mutations that reduce complement fixation or activation through the constant region, for example, mutations at one or more of EU numbers 241, 264, 265, 270, 296, 297, 318, 320, 322, 329, and 331. Some antibodies have an alanine residue at positions 318, 320, and 322. Some antibodies may be at least 95% w / w pure. Antibodies may be conjugated to therapeutic agents, cytotoxic agents, cell growth inhibitors, neurotrophic agents, or neuroprotective agents.

[0130] In another aspect, the present invention provides a pharmaceutical composition comprising any antibody and pharmaceutically acceptable carrier disclosed herein.

[0131] In another aspect, the present invention provides a nucleic acid encoding the heavy and / or light chains of any antibody disclosed herein, a recombinant expression vector comprising said nucleic acid, and a host cell transformed with said recombinant expression vector. Some nucleic acids have sequences comprising any of SEQ ID NO: 38-39.

[0132] In another aspect, the present invention provides a vector comprising a nucleic acid encoding a mature heavy chain variable region and a mature light chain variable region, said nucleic acid being operatively linked to one or more regulatory sequences to achieve expression of any antibody disclosed herein in mammalian cells; a recombinant expression vector comprising said nucleic acid; a host cell transformed with said recombinant expression vector; and a host cell transformed with said nucleic acid. Some nucleic acids further encode a heavy chain constant region fused to the mature heavy chain variable region and a light chain constant region fused to the mature light chain variable region. In some vectors, the antibody is scFv. In some vectors, the antibody is a Fab fragment. In some vectors, one or more regulatory sequences include one or more promoters, enhancers, ribosome binding sites, and transcription termination signals. In some vectors, the nucleic acid further encodes a signal peptide fused to the mature heavy chain and light chain variable regions. In some vectors, the nucleic acid is codon-optimized for expression in host cells. In some vectors, one or more regulatory sequences include a eukaryotic promoter. In some vectors, the nucleic acid further encodes a selection gene.

[0133] In another aspect, the present invention provides a method for expressing antibodies in mammalian cells, the method comprising incorporating the nucleic acids disclosed herein into the genome of a transgenic animal, thereby expressing the antibodies.

[0134] In another aspect, the present invention provides a first vector and a second vector comprising nucleic acids encoding mature heavy chain variable regions and mature light chain variable regions, respectively, each nucleic acid being operatively linked to one or more regulatory sequences to achieve expression of any antibody disclosed herein in mammalian cells; and a host cell comprising the nucleic acids. In some of the first and second vectors, the nucleic acids further encode a heavy chain constant region fused to the mature heavy chain variable region and a light chain constant region fused to the mature light chain variable region, respectively.

[0135] In another aspect, the present invention provides a method for expressing antibodies in mammalian cells, the method comprising incorporating any nucleic acid disclosed herein into the genome of a transgenic animal, thereby expressing the antibody.

[0136] In another aspect, the present invention provides a method for humanizing any non-human antibody described herein, such as mouse antibody 9F5, wherein 9F5 is characterized by the mature heavy chain variable region of SEQ ID NO:7 and the mature light chain variable region of SEQ ID NO:11; for example, mouse antibody 10C12, wherein 10C12 is characterized by the mature heavy chain variable region of SEQ ID NO:7 and the mature light chain variable region of SEQ ID NO:11; for example, mouse antibody 2D11, wherein 2D11 is characterized by the mature heavy chain variable region of SEQ ID NO:7 and the mature light chain variable region of SEQ ID NO:11; for example, mouse antibody 12C4, wherein 12C4 is characterized by the mature heavy chain variable region of SEQ ID NO:219 and the mature light chain variable region of SEQ ID NO:11; for example, mouse antibody 17C12, wherein 17C12 is characterized by the mature heavy chain variable region of SEQ ID NO:225 and the mature light chain variable region of SEQ ID NO:228; for example, mouse antibody 14H3, wherein 14H3 is characterized by the mature heavy chain variable region of SEQ ID NO:7 and the mature light chain variable region of SEQ ID NO:11. The mature heavy chain variable region of SEQ ID NO:240 and the mature light chain variable region of SEQ ID NO:244. Such methods may include selecting one or more receptor antibodies, identifying amino acid residues of a mouse antibody to be retained; synthesizing nucleic acids encoding a humanized heavy chain containing a mouse antibody heavy chain CDR and a humanized light chain encoding a mouse antibody light chain CDR, and expressing said nucleic acids in host cells to produce a humanized antibody.

[0137] Methods for producing antibodies, such as humanized, chimeric, or veneered antibodies, for example, humanized, chimeric, or veneered forms of 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3, are also provided. In such methods, cells transformed with nucleic acids encoding the heavy and light chains of the antibody are cultured to induce antibody secretion. The antibody can then be purified from the cell culture medium.

[0138] Cell lines that produce any of the antibodies disclosed herein can be generated by introducing a vector encoding the heavy and light chains of the antibody and a selection marker into the cells, allowing the cells to proliferate under conditions that select cells with a vector that increases the copy number, isolating single cells from the selected cells, and constructing a cell library from single-cell clones selected based on antibody yield.

[0139] Some cells can proliferate under selective conditions and be screened for naturally expressed and secreted at least 100 mg / L / 10 6Cell lines per 24 hours. Single cells can be isolated from selected cells. Cell libraries can then be constructed from single-cell clones. Single cells can be selected based on desired properties, such as antibody yield. Exemplary cell lines are those expressing 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3.

[0140] The present invention also provides a method for inhibiting or reducing tau accumulation in subjects suffering from or at risk of developing tau-mediated amyloidosis, the method comprising administering an effective regimen of the antibodies disclosed herein to the subject to inhibit or reduce tau accumulation in the subject. Exemplary antibodies include humanized forms of 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3.

[0141] Methods for treating or preventing tau-related diseases in subjects are also provided, the methods comprising administering an effective regimen of the antibody disclosed herein and thereby treating or preventing the disease. Examples of this disease include Alzheimer's disease, Down syndrome, mild cognitive impairment, primary age-related tauopathy, post-encephalitis Parkinson's disease, post-traumatic dementia or boxer's dementia, Pick's disease, type C Niemann-Pick disease, supranuclear palsy, frontotemporal dementia, frontotemporal degeneration, aurophilic granuloma, gliotropic tauopathy, amyotrophic lateral sclerosis / parkinsonism dementia complex of Guam, corticobasal degeneration (CBD), dementia with Lewy bodies, Lewy body variant of Alzheimer's disease (LBVAD), chronic traumatic encephalopathy (CTE), gliotropic tauopathy (GGT), Parkinson's disease, or progressive supranuclear palsy (PSP). In some approaches, the tau-related disease is Alzheimer's disease. In some methods, the patient is an ApoE4 carrier.

[0142] Methods for reducing the aberrant spread of tau are also provided, which include effective protocols for administering the antibodies disclosed herein and thereby reducing the spread of tau.

[0143] A method for inducing phagocytosis of tau is also provided, the method comprising an effective regimen of administering the antibody disclosed herein and thereby inducing phagocytosis of tau.

[0144] Methods for inhibiting tau aggregation or deposition are also provided, which include effective regimens of applying the antibodies disclosed herein to inhibit tau aggregation or deposition.

[0145] A method for inhibiting tau tangle formation is also provided, which includes an effective regimen for administering the antibody disclosed herein.

[0146] This invention also provides a method for detecting tau protein deposits in subjects suffering from or at risk of tau accumulation or deposition, the method comprising administering an antibody disclosed herein to the subject and detecting antibodies binding to tau in the subject. Examples of such diseases are Alzheimer's disease, Down syndrome, mild cognitive impairment, primary age-related tau proteinopathy, post-encephalitis Parkinson's disease, post-traumatic dementia or boxer's dementia, Pick's disease, Niemann-Pick disease type C, supranuclear palsy, frontotemporal dementia, frontotemporal degeneration, aerobatic granuloma, glioblastoma, amyotrophic lateral sclerosis / Parkinson's dementia relapse, corticobasal degeneration (CBD), Lewy body dementia, Lewy body variant of Alzheimer's disease (LBVAD), chronic traumatic encephalopathy (CTE), glioblastoma (GGT), Parkinson's disease, or progressive supranuclear palsy (PSP). In some embodiments, the antibody is administered to the subject via intravenous injection. In some embodiments, the antibody is administered directly to the subject's brain via intracranial injection or by drilling a hole through the subject's skull. In some embodiments, the antibody is labeled. In some embodiments, the antibody is labeled with fluorescent, paramagnetic, or radioactive labels. In some embodiments, the radioactive labeling is detected using positron emission tomography (PET) or single-photon emission computed tomography (SPECT).

[0147] The present invention also provides a method for measuring the efficacy of treatment in a subject being treated for a disease associated with tau aggregation or deposition, the method comprising: measuring a first level of tau protein deposits in the subject prior to treatment by administering an antibody disclosed herein, and detecting a first amount of tau-binding antibodies in the subject; administering treatment to the subject; measuring a second level of tau protein deposits in the subject after treatment by administering the antibody to the subject, and detecting tau-binding antibodies in the subject; wherein a decrease in tau protein deposit levels indicates a positive response to treatment.

[0148] The present invention also provides a method for measuring the efficacy of treatment in a subject being treated for a disease associated with tau aggregation or deposition, the method comprising: measuring a first level of tau protein deposits in the subject prior to treatment by administering an antibody disclosed herein, and detecting a first amount of tau-binding antibody in the subject; administering treatment to the subject; measuring a second level of tau protein deposits in the subject after treatment by administering the antibody to the subject, and detecting a second amount of tau-binding antibody in the subject; wherein no change in the level of tau protein deposits or a slight increase in tau protein deposits indicates a positive response to treatment.

[0149] This invention also provides an isolated monoclonal antibody that specifically binds to a peptide composed of residues (Q / E)IVYK(S / P) (SEQ ID NO: 56). This invention also provides an isolated monoclonal antibody that specifically binds to a peptide composed of residues QIVYKP (SEQ ID NO: 57). This invention also provides an isolated monoclonal antibody that specifically binds to a peptide composed of residues EIVYKSP (SEQ ID NO: 58). This invention also provides an isolated monoclonal antibody that specifically binds to a peptide composed of residues EIVYKS (SEQ ID NO: 277).

[0150] The present invention also provides an isolated monoclonal antibody that specifically binds to the polypeptide of SEQ ID NO:1 at an epitope including at least one residue from residues 307-312 of SEQ ID NO:1. Some of these antibodies bind to the epitope from residues 307-312 of SEQ ID NO:1. The present invention also provides an isolated monoclonal antibody that specifically binds to the polypeptide of SEQ ID NO:1 at an epitope including at least one residue from residues 391-397 of SEQ ID NO:1. Some of these antibodies bind to the epitope from residues 391-397 of SEQ ID NO:1. The present invention also provides an isolated monoclonal antibody that specifically binds to the polypeptide of SEQ ID NO:1 at an epitope including at least one residue from residues 391-396 of SEQ ID NO:1. Some of these antibodies bind to the epitope from residues 391-396 of SEQ ID NO:1. The present invention also provides an isolated monoclonal antibody that specifically binds to the polypeptide of SEQ ID NO:1 at an epitope comprising at least one residue from residues 307-312 of SEQ ID NO:1 and at least one residue from residues 391-397 of SEQ ID NO:1. The present invention also provides an isolated monoclonal antibody that specifically binds to the polypeptide of SEQ ID NO:1 at an epitope comprising at least one residue from residues 307-312 of SEQ ID NO:1 and at least one residue from residues 391-396 of SEQ ID NO:1.

[0151] The present invention also provides a method for treating or preventing tau-related diseases in a subject, the method comprising administering an immunogen comprising a tau peptide of up to 20 consecutive amino acids of SEQ ID NO:1, specifically bound to antibodies 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3, wherein the peptide induces the formation of antibodies specifically bound to tau in the subject. In some such methods, the tau peptide consists of 4-7 consecutive amino acids from residues 307-312 of SEQ ID NO:1, residues 391-397 of SEQ ID NO:1, or residues 391-396 of SEQ ID NO:1. In some such methods, the tau peptide consists of residue (Q / E)IVYK(S / P) (SEQ ID NO:56). In some such methods, the tau peptide consists of residue QIVYKP (SEQ ID NO:57). In some of these methods, the tau peptide consists of the residue EIVYKSP (SEQ ID NO: 58). In some of these methods, the tau peptide consists of the residue EIVYKS (SEQ ID NO: 277). In some of these methods, the tau peptide is attached to a heteroconjugate molecule.

[0152] The present invention also provides a method for generating an antibody that specifically binds to an epitope comprising (Q / E)IVYK(S / P) (SEQ ID NO:56), the method comprising immunizing an animal with tau or a fragment thereof, and screening for antibodies that specifically bind to the epitope. In some such methods, the animal is immunized with 383 amino acid human tau (4R0N). In some such methods, the human tau contains the P301S mutation. In some such methods, the human tau is recombinant with an N-terminal His tag.

[0153] In some such methods, screening is performed on a 15-amino acid peptide comprising QIVYKP (SEQ ID NO:57), EIVYKSP (SEQ ID NO:58), EIVYKS (SEQ ID NO:277), or any other common motif represented by (Q / E)IVYK(S / P) (SEQ ID NO:56). In some such methods, the peptide comprises QIVYKP (SEQ ID NO:57), EIVYKSP (SEQ ID NO:58), or EIVYKS (SEQ ID NO:277).

[0154] In some of these methods, animals are immunized with a tau fragment linked to a vector containing a peptide represented by (Q / E)IVYK(S / P) (SEQ ID NO:56). In some of these methods, the peptide is QIVYKP (SEQ ID NO:57), EIVYKSP (SEQ ID NO:58), or EIVYKS (SEQ ID NO:277). Attached Figure Description

[0155] Figure 1A and Figure 1B The heavy chain variable region (SEQ ID NO:7) of the mouse 9F5 antibody and the heavy chain variable regions (hu9F5VHv1, hu9F5VHv2, hu9F5VHv3, hu9F5VHv4, hu9F5VHv5, hu9F5VHv6, hu9F5VHv7 and hu9F5VHv8) of the humanized 9F5 antibody were compared with the human germline heavy chain variable region sequence IGHV1-69-2*01 (SEQ ID NO:33), the human receptor heavy chain variable region sequence AAN16432-VH_huFrwk (AAN16432_H; SEQ ID NO:31), and the human receptor heavy chain variable region sequence 2RCS-VH_huFrwk (2RCS_H; SEQ ID NO:32). hu9F5VHv1 is SEQ ID NO:15, hu9F5VHv2 is SEQ ID NO:16, hu9F5VHv3 is SEQ ID NO:17, hu9F5VHv4 is SEQ ID NO:18, hu9F5VHv5 is SEQ ID NO:19, hu9F5VHv6 is SEQ ID NO:20, hu9F5VHv7 is SEQ ID NO:21, and hu9F5VHv8 is SEQ ID NO:22. The CDR of mouse 9F5VH as defined by the Kabat / Chothia complex is shown in bold.

[0156] Figure 2A and Figure 2BThe light chain variable region (SEQ ID NO:11) of the mouse 9F5 antibody and the light chain variable regions (hu9F5VLv1, hu9F5VLv2, hu9F5VLv3, hu9F5VLv4, hu9F5VLv5, hu9F5VLv6 and hu9F5VLv7) of the humanized form of the 9F5 antibody were depicted and compared with the human germline light chain variable region sequences IGKV2-28*01&_IGKJ2*01 (IGKV2-28*01_IGKJ2*01; SEQ ID NO:37), human receptor CAB51297-VL_huFrwk (CAB51297_L; SEQ ID NO:35), and human receptor 1911357B-VL_huFRwk (1911357B_L; SEQ ID NO:36). hu9F5VLv1 is SEQ ID NO:23, hu9F5VLv2 is SEQ ID NO:24, hu9F5VLv3 is SEQ ID NO:25, hu9F5VLv4 is SEQ ID NO:26, hu9F5VLv5 is SEQ ID NO:27, hu9F5VLv6 is SEQ ID NO:28, and hu9F5VLv7 is SEQ ID NO:29. The CDR of mouse 9F5VL as defined by Kabat is shown in bold.

[0157] Figure 3 The results of an assay demonstrating the blocking of tau neuronal internalization by the mouse 9F5 antibody were described.

[0158] Figure 4A and Figure 4BThe heavy chain variable region (SEQ ID NO:7) of the mouse 9F5 antibody and the heavy chain variable regions (hu9F5VHv1, hu9F5VHv2, hu9F5VHv3, hu9F5VHv4, hu9F5VHv5, hu9F5VHv6, hu9F5VHv7, hu9F5VHv8, hu9F5VHv9 and hu9F5VHv10) of the human germline heavy chain variable region sequence IGHV1-69-2*01 (SEQ ID NO:33), the human receptor heavy chain variable region sequence AAN16432-VH_huFrwk (AAN16432_H; SEQ ID NO:31), and the human receptor heavy chain variable region sequence 2RCS-VH_huFrwk (2RCS_H; SEQ ID NO:32) were compared. hu9F5VHv1 is SEQ ID NO:15, hu9F5VHv2 is SEQ ID NO:16, hu9F5VHv3 is SEQ ID NO:17, hu9F5VHv4 is SEQ ID NO:18, hu9F5VHv5 is SEQ ID NO:19, hu9F5VHv6 is SEQ ID NO:20, hu9F5VHv7 is SEQ ID NO:21, hu9F5VHv8 is SEQ ID NO:22, hu9F5VHv9 is SEQ ID NO:127, and hu9F5VHv10 is SEQ ID NO:128. The CDR of mouse 9F5VH as defined by the Kabat / Chothia complex is shown in bold. Residues identical to those in mouse 9F5VH are indicated by a period (.).

[0159] Figure 5A and Figure 5BThe light chain variable region of the mouse 9F5 antibody (SEQ ID NO:11) and the light chain variable regions (hu9F5VLv1, hu9F5VLv2, hu9F5VLv3, hu9F5VLv4, hu9F5VLv5, hu9F5VLv6, hu9F5VLv7, hu9F5VLv8 and hu9F5VLv9) of the humanized form of the 9F5 antibody were depicted, along with the human germline light chain variable region sequences IGKV2-28*01&_IGKJ2*01 (IGKV2-28*01_IGKJ2*01; SEQ ID NO:37), human receptor CAB51297-VL_huFrwk (CAB51297_L; SEQ ID NO:35), and human receptor 1911357B-VL_huFRwk (1911357B_L; SEQ ID NO:37). The comparison is for SEQ ID NO:36. hu9F5VLv1 is SEQ ID NO:23, hu9F5VLv2 is SEQ ID NO:24, hu9F5VLv3 is SEQ ID NO:25, hu9F5VLv4 is SEQ ID NO:26, hu9F5VLv5 is SEQ ID NO:27, hu9F5VLv6 is SEQ ID NO:28, hu9F5VLv7 is SEQ ID NO:2, hu9F5VLv8 is SEQ ID NO:130, and hu9F5VLv9 is SEQ ID NO:131. The CDR of mouse 9F5VL as defined by Kabat is shown in bold. Residues identical to those in mouse 9F5VL are indicated by a ".".

[0160] Figure 6A , Figure 6B and Figure 6CDepicts the alignment of the light chain variable region of hu9F5VLv8 with the light chain variable region of the humanized form of the 9F5 antibody: hu9F5VLv8_DIM1 (SEQ ID NO:132), hu9F5VLv8_DIM2 (SEQ ID NO:133), hu9F5VLv8_DIM3 (SEQ ID NO:134), hu9F5VLv8_DIM4 (SEQ ID NO:135), hu9F5VLv8_DIM5 (SEQ ID NO:136), hu9F5VLv8_DIM6 (SEQ ID NO:137), hu9F5VLv8_DIM7 (SEQ ID NO:138), hu9F5VLv8_DIM8 (SEQ ID NO:139), hu9F5VLv8_DIM9 (SEQ ID NO:140), hu9F5VLv8_DIM10 (SEQ ID NO:141), hu9F5VLv8_DIM11 (SEQ ID NO:142), hu9F5VLv8_DIM12 (SEQ ID NO:143), hu9F5VLv8_DIM13 (SEQ ID NO:144), hu9F5VLv8_DIM14 (SEQ ID NO:145), hu9F5VLv8_DIM15 (SEQ ID NO:146), hu9F5VLv8_DIM16 (SEQ ID NO:147), hu9F5VLv8_DIM17 (SEQ ID NO:148), hu9F5VLv8_DIM18 (SEQ ID NO:149), hu9F5VLv8_DIM19 (SEQ ID NO:150), hu9F5VLv8_DIM20 (SEQ ID NO:151), hu9F5VLv8_DIM21 (SEQ ID NO:152), hu9F5VLv8_DIM22 (SEQ ID NO:153), hu9F5VLv8_DIM23 (SEQ ID NO:154), hu9F5VLv8_DIM24 (SEQ ID NO:155), hu9F5VLv8_DIM25 (SEQ ID NO:156), hu9F5VLv8_DIM26 (SEQ ID NO:157), hu9F5VLv9_DIM1 (SEQ ID NO:162), hu9F5VLv9_DIM2 (SEQ ID NO:163), hu9F5VLv9_DIM4 (SEQ ID NO:164), hu9F5VLv9_DIM5 (SEQ ID NO:165), hu9F5VLv9_DIM8 (SEQ ID NO:166), hu9F5VLv9_DIM 10(SEQThe following are listed: hu9F5VLv9_DIM11 (SEQ ID NO:168), hu9F5VLv9_DIM13 (SEQ ID NO:169), hu9F5VLv9_DIM19 (SEQ ID NO:170), hu9F5VLv9_DIM20 (SEQ ID NO:171), hu9F5VLv8_DIM27 (SEQ ID NO:158), hu9F5VLv8_DIM28 (SEQ ID NO:159), hu9F5VLv8_DIM29 (SEQ ID NO:160), and hu9F5VLv8_DIM30 (SEQ ID NO:161). The CDR for hu9F5VLv8 as defined by Kabat is shown in bold.

[0161] Figure 7 The heavy chain variable region of the mouse 10C12 antibody (SEQ ID NO:7) was depicted. Figure 7 The heavy chain variable regions (hu10C12VHv1 and hu10C12VHv2) of the humanized forms of the 10C12 antibody, labeled m10C12VH, were compared with the human germline heavy chain variable region sequence IGHV1-69-2*01 (SEQ ID NO:33) and the human receptor heavy chain variable region sequence CAC20421VH (SEQ ID NO:218). hu10C12VHv1 is SEQ ID NO:214 and hu10C12VHv2 is SEQ ID NO:215. The CDR of mouse 10C12VH as defined by the Kabat / Chothia complex is shown in bold.

[0162] Figure 8 Alignments were drawn between the mouse 10C12 light chain variable region (SEQ ID NO:11) and the humanized light chain variable regions (hu10C12VLv1 and hu10C12VLv2) of the 10C12 antibody and the human germline light chain variable region sequences IGKV2-28*01&_IGKJ2*01 (SEQ ID NO:37) and the human receptor CAB51297-VL_huFrwk (SEQ ID NO:35). hu10C12VLv1 is SEQ ID NO:216 and hu10C12VLv2 is SEQ ID NO:217. The CDR of mouse 10C12VL as defined by Kabat is shown in bold.

[0163] Figure 9The heavy chain variable region (SEQ ID NO:219) of the mouse 12C4 antibody and the heavy chain variable regions (hu12C4VHv1 and hu12C4VHv2) of the humanized form of the 12C4 antibody were depicted and compared with the human germline heavy chain variable region sequence IGHV1-69-2*01 (SEQ ID NO:33) and the human receptor heavy chain variable region sequence CAC20421VH (SEQ ID NO:218). hu12C4VHv1 is SEQ ID NO:221 and hu12C4VHv2 is SEQ ID NO:222. The CDR of mouse 12C4VH as defined by the Kabat / Chothia complex is shown in bold.

[0164] Figure 10 The light chain variable region (SEQ ID NO:11) of the mouse 12C4 antibody and the light chain variable regions (hu12C4VLv1 and huvVLv2) of the humanized form of the 12C4 antibody were depicted and compared with the human germline light chain variable region sequences IGKV2-28*01&_IGKJ2*01 (SEQ ID NO:37) and the human receptor CAB51297 (SEQ ID NO:35). hu12C4VLv1 is SEQ ID NO:223 and hu12C4VLv2 is SEQ ID NO:224. The CDR of mouse 12C4VL as defined by Kabat is shown in bold.

[0165] Figure 11 The heavy chain variable region (SEQ ID NO:225) of the mouse 17C12 antibody and the heavy chain variable regions (hu17C12VHv1 and hu17C12VHv2) of the humanized form of the 17C12 antibody were depicted and compared with the human germline heavy chain variable region sequence IGHV1-69-2*01 (SEQ ID NO:33) and the human receptor heavy chain variable region sequence CAC20421VH (SEQ ID NO:218). hu17C12VHv1 is SEQ ID NO:232 and hu17C12VHv2 is SEQ ID NO:233. The CDR of mouse 17C12VH as defined by the Kabat / Chothia complex is shown in bold.

[0166] Figure 12The light chain variable region (SEQ ID NO:228) of the mouse 17C12 antibody and the light chain variable regions (hu17C12VLv1 and hu17C12VLv2) of the humanized form of the 17C12 antibody were depicted and compared with the human germline light chain variable region sequences IGKV2-29*02&IGKJ4*01 (SEQ ID NO:239) and the human receptor QDO16713VL (SEQ ID NO:238). hu17C12VLv1 is SEQ ID NO:234 ​​and hu17C12VLv2 is SEQ ID NO:235. The CDR of mouse 17C12VL as defined by Kabat is shown in bold.

[0167] Figure 13 The heavy chain variable region (SEQ ID NO:240) of the mouse 14H3 antibody and the heavy chain variable regions (hu14H3VHv1 and hu14H3VHv2) of the humanized form of the 14H3 antibody were depicted and compared with the human germline heavy chain variable region sequences IGHV2-70*04 & IGHJ4*01 (SEQ ID NO:254) and the human receptor heavy chain variable region sequence QDJ57937VH hFrwk (SEQ ID NO:253). hu14H3VHv1 is SEQ ID NO:248 and hu14H3VHv2 is SEQ ID NO:249. The CDR of mouse 14H3VH as defined by the Kabat / Chothia complex is shown in bold.

[0168] Figure 14 The light chain variable region (SEQ ID NO:244) of the mouse 14H3 antibody and the light chain variable regions (hu14H3VLv1 and hu14H3VLv2) of the humanized form of the 14H3 antibody were depicted and compared with the human germline light chain variable region sequences IGKV2-28*01&_IGKJ2*01 (IGKV2-28*01_IGKJ2*01; SEQ ID NO:37) and the human receptor ABC66914VL_hFwrk (SEQ ID NO:256). hu14H3VLv1 is SEQ ID NO:250 and hu14H3VLv2 is SEQ ID NO:251. The CDR of mouse 14H3VL as defined by Kabat is shown in bold.

[0169] Figure 15 The results of assays showing that mouse antibodies 10C12, 12C4, 2D11, 17C12, 14H3 and 9F5 block neuronal internalization of tau were described.

[0170] Figure 16The results of assays (neuronal viability) showing that the mice 10C12, 12C4, 2D11 and 9F5 antibodies prevented tau toxicity in primary neurons were depicted.

[0171] Figure 17 The results depict the assay (LDH release) showing that the mouse 10C12, mouse 12C4, mouse 2D11 and mouse 9F5 antibodies prevent tau toxicity in primary neurons.

[0172] Figure 18 The results of Western blot assays showing the detection of tau protein in samples from the brains of Alzheimer's disease patients using mouse 10C12, mouse 12C4, mouse 2D11, mouse 17C12, mouse 14H3 and mouse 9F5 antibodies are depicted.

[0173] Figure 19 The results of immunoprecipitation assays using mouse 10C12, mouse 12C4, mouse 2D11, mouse 17C12, mouse 14H3 and mouse 9F5 antibodies and samples from the brains of Alzheimer's disease patients are depicted.

[0174] Figure 20 The results of a determination measuring the ability of the 9F5 humanized variant to withstand agglomeration induced by agitation stress are described.

[0175] Figure 21 The results of a determination measuring the ability of the humanized 9F5 variant to tolerate low pH exposure are described.

[0176] Figure 22 The results of the determination of the ability of the humanized variant of 9F5 to aggregate under simulated high concentration conditions are described.

[0177] Figures 23A to 23F The results of immunohistochemical assays using control, mouse 2D11, mouse 9F5, mouse 12C4, mouse 14H3 and mouse 17C12 are depicted.

[0178] Figure 24 The heavy chain variable regions of mouse 9F5 antibody (SEQ ID NO:7), mouse 10C12 antibody (SEQ ID NO:7), mouse 2D11 antibody (SEQ ID NO:7), mouse 12C4 antibody (SEQ ID NO:219), mouse 14H3 antibody (SEQ ID NO:240), and mouse 17C12 antibody (SEQ ID NO:225) are depicted. The CDR of mouse 9F5VH as defined by the Kabat / Chothia complex is shown in bold.

[0179] Figure 25The alignment of the light chain variable regions of mouse 9F5 antibody (SEQ ID NO:11), mouse 10C12 antibody (SEQ ID NO:11), mouse 2D11 antibody (SEQ ID NO:11), mouse 12C4 antibody (SEQ ID NO:11), mouse 14H3 antibody (SEQ ID NO:244), and mouse 17C12 antibody (SEQ ID NO:228) is depicted. The CDR of mouse 9F5VL as defined by Kabat is shown in bold.

[0180] Sequence Summary

[0181] SEQ ID NO:1 lists the amino acid sequence of the human tau isotype (Swiss-Prot P10636-8).

[0182] SEQ ID NO:2 lists the amino acid sequence of the human tau isotype (Swiss-Prot P10636-7).

[0183] SEQ ID NO:3 lists the amino acid sequence of the isotype of human tau (Swiss-Prot P10636-6) (4R0N human tau).

[0184] SEQ ID NO:4 lists the amino acid sequence of the human tau isotype (Swiss-Prot P10636-5).

[0185] SEQ ID NO:5 lists the amino acid sequence of the human tau isotype (Swiss-Prot P10636-4).

[0186] SEQ ID NO:6 lists the amino acid sequence of the human tau isotype (Swiss-Prot P10636-2).

[0187] SEQ ID NO:7 lists the amino acid sequence of the heavy chain variable region of the mouse 9F5 antibody.

[0188] SEQ ID NO:8 lists the amino acid sequence of the Kabat / Chothia complex CDR-H1 of the mouse 9F5 antibody.

[0189] SEQ ID NO:9 lists the amino acid sequence of the mouse 9F5 antibody Kabat CDR-H2.

[0190] SEQ ID NO:10 lists the amino acid sequence of the mouse 9F5 antibody Kabat CDR-H3.

[0191] SEQ ID NO:11 lists the amino acid sequence of the light chain variable region of the mouse 9F5 antibody.

[0192] SEQ ID NO:12 lists the amino acid sequence of the Kabat CDR-L1 antibody against mouse 9F5.

[0193] SEQ ID NO:13 lists the amino acid sequence of the mouse 9F5 antibody Kabat CDR-L2.

[0194] SEQ ID NO:14 lists the amino acid sequence of the Kabat CDR-L3 antibody against mouse 9F5.

[0195] SEQ ID NO:15 lists the amino acid sequence of the humanized heavy chain variable region hu9F5VHv1.

[0196] SEQ ID NO:16 lists the amino acid sequence of the humanized heavy chain variable region hu9F5VHv2.

[0197] SEQ ID NO:17 lists the amino acid sequence of the humanized heavy chain variable region hu9F5VHv3.

[0198] SEQ ID NO:18 lists the amino acid sequence of the humanized heavy chain variable region hu9F5VHv4.

[0199] SEQ ID NO:19 lists the amino acid sequence of the humanized heavy chain variable region hu9F5VHv5:

[0200] SEQ ID NO:20 lists the amino acid sequence of the humanized heavy chain variable region hu9F5VHv6.

[0201] SEQ ID NO:21 lists the amino acid sequence of the humanized heavy chain variable region hu9F5VHv7.

[0202] SEQ ID NO:22 lists the amino acid sequence of the humanized heavy chain variable region hu9F5VHv8.

[0203] SEQ ID NO:23 lists the amino acid sequence of the humanized light chain variable region hu9F5VLv1.

[0204] SEQ ID NO:24 lists the amino acid sequence of the humanized light chain variable region hu9F5VLv2.

[0205] SEQ ID NO:25 lists the amino acid sequence of the humanized light chain variable region hu9F5VLv3.

[0206] SEQ ID NO:26 lists the amino acid sequence of the humanized light chain variable region hu9F5VLv4.

[0207] SEQ ID NO:27 lists the amino acid sequence of the humanized light chain variable region of the humanized 9F5 antibody hu9F5VLv5.

[0208] SEQ ID NO:28 lists the amino acid sequence of the humanized light chain variable region hu9F5VLv6.

[0209] SEQ ID NO:29 lists the amino acid sequence of the humanized light chain variable region hu9F5VLv7.

[0210] SEQ ID NO:30 lists the amino acid sequence of the heavy chain variable region structural model PDB.#5OBF-VH_mSt.

[0211] SEQ ID NO:31 lists the amino acid sequence of the heavy chain variable region receptor, GenBank accession number AAN16432-VH_huFrwk.

[0212] SEQ ID NO:32 lists the amino acid sequence of the heavy chain variable region receptor PDB#2RCS-VH_huFrwk.

[0213] SEQ ID NO:33 lists the amino acid sequence of the heavy chain variable region germline sequence IMGT#IGHV1-69-2*01.

[0214] SEQ ID NO:34 lists the amino acid sequence of the light chain variable region structural model PDB#5OBF-VL_mSt.

[0215] SEQ ID NO:35 lists the amino acid sequence of the light chain variable region receptor, GenBank accession number CAB51297-VL_huFrwk.

[0216] SEQ ID NO:36 lists the amino acid sequence of the light chain variable region receptor GenBank accession number 1911357B-VL_huFrwk.

[0217] SEQ ID NO:37 lists the amino acid sequence of the light chain variable region germline sequence IMGT#IGKV2-28*01&IGKJ2*01.

[0218] SEQ ID NO:38 lists the nucleic acid sequence encoding the heavy chain variable region of the mouse 9F5 antibody.

[0219] SEQ ID NO:39 lists the nucleic acid sequence encoding the light chain variable region of the mouse 9F5 antibody.

[0220] SEQ ID NO:40 lists the amino acid sequence of the Kabat CDR-H1 of the mouse 9F5 antibody.

[0221] SEQ ID NO:41 lists the amino acid sequence of Chothia CDR-H1, a mouse 9F5 antibody.

[0222] SEQ ID NO:42 lists the amino acid sequence of the mouse 9F5 antibody Chothia CDR-H2.

[0223] SEQ ID NO:43 lists the amino acid sequence of the mouse 9F5 antibody AbM CDR-H2.

[0224] SEQ ID NO:44 lists the amino acid sequence of Contact CDR-H1 of the mouse 9F5 antibody.

[0225] SEQ ID NO:45 lists the amino acid sequence of the mouse 9F5 antibody Contact CDR-H2.

[0226] SEQ ID NO:46 lists the amino acid sequence of the mouse 9F5 antibody Contact CDR-H3.

[0227] SEQ ID NO:47 lists the amino acid sequence of the mouse 9F5 antibody Contact CDR-L1.

[0228] SEQ ID NO:48 lists the amino acid sequence of the mouse 9F5 antibody Contact CDR-L2.

[0229] SEQ ID NO:49 lists the amino acid sequence of the mouse 9F5 antibody Contact CDR-L3.

[0230] SEQ ID NO:50 lists the amino acid sequence of an alternative Kabat-Chothia complex CDR-H1 (present in hu9F5VHv4, hu9F5VHv5 and hu9F5VHv6) of the humanized 9F5 antibody.

[0231] SEQ ID NO:51 lists the amino acid sequence of an alternative Kabat CDR-H2 (present in hu9F5VHv5, hu9F5VHv6 and hu9F5VHv7) of the humanized 9F5 antibody.

[0232] SEQ ID NO:52 lists the amino acid sequence of an alternative Kabat CDR-H2 (present in hu9F5VHv8) of the humanized 9F5 antibody.

[0233] SEQ ID NO:53 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv5 and hu9F5VLv6) for the humanized 9F5 antibody.

[0234] SEQ ID NO:54 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv7) for the humanized 9F5 antibody.

[0235] SEQ ID NO:55 lists alternative Kabat CDR-L2 for humanized 9F5 antibodies (present in hu9F5VLv4, hu9F5VLv5, hu9F5VLv6, hu9F5VLv7, hu9F5VLv8_DIM2, hu9F5VLv8_DIM4, hu9F5VLv8_DIM5, hu9F5VLv8_DIM6, hu9F5VLv8_DIM11, hu9F5VLv8_DIM12, hu9 The amino acid sequences of F5VLv8_DIM13, hu9F5VLv8_DIM18, hu9F5VLv8_DIM27, hu9F5VLv8_DIM28, hu9F5VLv9_DIM2, hu9F5VLv9_DIM4, hu9F5VLv9_DIM5, hu9F5VLv9_DIM11 and hu9F5VLv9_DIM13.

[0236] SEQ ID NO:56 lists the amino acid sequence of the epitope of antibody 9F5.

[0237] SEQ ID NO:57 lists the amino acid sequence of the common motif of the peptide bound by antibody 9F5.

[0238] SEQ ID NO:58 lists the amino acid sequence of the common motif of the peptide bound by antibody 9F5.

[0239] SEQ ID NO:59 lists the amino acid sequence of the linker.

[0240] SEQ ID NO:60 lists the amino acid sequence of the HA control peptide.

[0241] SEQ ID NO:61 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_M51E).

[0242] SEQ ID NO:62 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_M51D).

[0243] SEQ ID NO:63 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27cD).

[0244] SEQ ID NO:64 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27cG).

[0245] SEQ ID NO:65 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27cS).

[0246] SEQ ID NO:66 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27cE).

[0247] SEQ ID NO:67 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_I30E).

[0248] SEQ ID NO:68 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_I30K).

[0249] SEQ ID NO:69 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27cT).

[0250] SEQ ID NO:70 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27cN).

[0251] SEQ ID NO:71 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27bD).

[0252] SEQ ID NO:72 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_I30G).

[0253] SEQ ID NO:73 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L33N).

[0254] SEQ ID NO:74 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27cA).

[0255] SEQ ID NO:75 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L33T).

[0256] SEQ ID NO:76 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L33S).

[0257] SEQ ID NO:77 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L33R).

[0258] SEQ ID NO:78 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_I30Q).

[0259] SEQ ID NO:79 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27bT).

[0260] SEQ ID NO:80 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_T31G).

[0261] SEQ ID NO:81 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27bQ).

[0262] SEQ ID NO:82 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L33G).

[0263] SEQ ID NO:83 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27cP).

[0264] SEQ ID NO:84 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_V78R).

[0265] SEQ ID NO:85 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_I75D).

[0266] SEQ ID NO:86 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_V78D).

[0267] SEQ ID NO:87 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_V78E).

[0268] SEQ ID NO:88 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_V78P).

[0269] SEQ ID NO:89 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_V78K).

[0270] SEQ ID NO:90 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_R77D).

[0271] SEQ ID NO:91 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_V78G).

[0272] SEQ ID NO:92 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_S76P).

[0273] SEQ ID NO:93 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_I75P).

[0274] SEQ ID NO:94 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_I75Q).

[0275] SEQ ID NO:95 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_I75G).

[0276] SEQ ID NO:96 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L73P).

[0277] SEQ ID NO:97 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L73G).

[0278] SEQ ID NO:98 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_V78Q).

[0279] SEQ ID NO:99 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_S76G).

[0280] SEQ ID NO:100 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L92D).

[0281] SEQ ID NO:101 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_Y86T).

[0282] SEQ ID NO:102 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L92E).

[0283] SEQ ID NO:103 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L92G).

[0284] SEQ ID NO:104 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L92Q).

[0285] SEQ ID NO:105 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L93G).

[0286] SEQ ID NO:106 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_V85G).

[0287] SEQ ID NO:107 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L92T).

[0288] SEQ ID NO:108 lists the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_A89G).

[0289] SEQ ID NO:109 lists the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L80P).

[0290] SEQ ID NO:110 lists the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L80D).

[0291] SEQ ID NO:111 lists the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L82cG).

[0292] SEQ ID NO:112 lists the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L82cD).

[0293] SEQ ID NO:113 lists the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L82P).

[0294] SEQ ID NO:114 lists the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L80G).

[0295] SEQ ID NO:115 lists the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L82K).

[0296] SEQ ID NO:116 lists the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L82R).

[0297] SEQ ID NO:117 lists the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L82E).

[0298] SEQ ID NO:118 lists the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L82N).

[0299] SEQ ID NO:119 lists the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_Y79D).

[0300] SEQ ID NO:120 lists the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_Y79N).

[0301] SEQ ID NO:121 lists the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_Y79G).

[0302] SEQ ID NO:122 lists the amino acid sequence of a variant of the hu9F5VHv5 heavy chain variable region (also known as hu9F5VHv5_M80E).

[0303] SEQ ID NO:123 lists the amino acid sequence of a variant of the hu9F5VHv5 heavy chain variable region (also known as hu9F5VHv5_M80G).

[0304] SEQ ID NO:124 lists the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L82cS).

[0305] SEQ ID NO:125 lists the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_Y79Q).

[0306] SEQ ID NO:126 lists the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_S82aG).

[0307] SEQ ID NO:127 lists the amino acid sequence of the heavy chain variable region hu9F5VHv9.

[0308] SEQ ID NO:128 lists the amino acid sequence of the heavy chain variable region hu9F5VHv10 (also known as hu9F5VHv9_Q38K_G42E).

[0309] SEQ ID NO:129 lists the amino acid sequence of the heavy chain variable region hu9F5VHv10_L82cG.

[0310] SEQ ID NO:130 lists the amino acid sequence of the light chain variable region hu9F5VLv8.

[0311] SEQ ID NO:131 lists the amino acid sequence of the light chain variable region hu9F5VLv9 (also known as hu9F5VLv8_N60D).

[0312] SEQ ID NO:132 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54G, L92I, also known as hu9F5VLv8_DIM1).

[0313] SEQ ID NO:133 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54R, L92I, also known as hu9F5VLv8_DIM2).

[0314] SEQ ID NO:134 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54T, L92I, also known as hu9F5VLv8_DIM3).

[0315] SEQ ID NO:135 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54R, L92G, also known as hu9F5VLv8_DIM4).

[0316] SEQ ID NO:136 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37Q, M51G, L54R, L92I, also known as hu9F5VLv8_DIM5).

[0317] SEQ ID NO:137 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cD, L37Q, M51G, L54R, L92I, also known as hu9F5VLv8_DIM6).

[0318] SEQ ID NO:138 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cD, L37Q, M51K, L54R, L92I, also known as hu9F5VLv8_DIM7).

[0319] SEQ ID NO:139 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37Q, M51K, L54R, L92I, also known as hu9F5VLv8_DIM8).

[0320] SEQ ID NO:140 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37Q, M51K, L54G, L92I, also known as hu9F5VLv8_DIM9).

[0321] SEQ ID NO:141 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, M51K, L54G, L92I, also known as hu9F5VLv8_DIM10).

[0322] SEQ ID NO:142 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54R, L92I, also known as hu9F5VLv8_DIM11).

[0323] SEQ ID NO:143 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54R, L92G, also known as hu9F5VLv8_DIM12).

[0324] SEQ ID NO:144 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54R, also known as hu9F5VLv8_DIM13).

[0325] SEQ ID NO:145 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54T, L92I, also known as hu9F5VLv8_DIM14).

[0326] SEQ ID NO:146 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54T, L92G, also known as hu9F5VLv8_DIM15).

[0327] SEQ ID NO:147 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54T, also known as hu9F5VLv8_DIM16).

[0328] SEQ ID NO:148 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37G, M51G, L54T, L92I, also known as hu9F5VLv8_DIM17).

[0329] SEQ ID NO:149 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cD, L37G, M51G, L54R, L92I, also known as hu9F5VLv8_DIM18).

[0330] SEQ ID NO:150 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37I, M51I, L54R, L92I, also known as hu9F5VLv8_DIM19).

[0331] SEQ ID NO:151 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, M51I, L54G, L92I, also known as hu9F5VLv8_DIM20).

[0332] SEQ ID NO:152 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, M51I, L54G, also known as hu9F5VLv8_DIM21).

[0333] SEQ ID NO:153 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, M51E, L54R, L92I, also known as hu9F5VLv8_DIM22).

[0334] Variants of the variable region of the light chain of SEQ ID NO:154hu9F5VLv8 (hu9F5VLv8_V3Q, L27cG, L37Q, M51E, L54G, L92I, also known as hu9F5VLv8_DIM23).

[0335] SEQ ID NO:155 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37I, M51E, L54R, L92I, also known as hu9F5VLv8_DIM24).

[0336] SEQ ID NO:156 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37I, M51E, L54R, L92G, also known as hu9F5VLv8_DIM25).

[0337] SEQ ID NO:157 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cI, L37I, M51E, L54R, also known as hu9F5VLv8_DIM26).

[0338] SEQ ID NO:158 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L37Q, M51G, L54R, L92I, also known as hu9F5VLv8_DIM27).

[0339] SEQ ID NO:159 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, M51G, L54R, L92I, also known as hu9F5VLv8_DIM28).

[0340] SEQ ID NO:160 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, L54R, L92I, also known as hu9F5VLv8_DIM29).

[0341] SEQ ID NO:161 lists the amino acid sequences of variants of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L92I, also known as hu9F5VLv8_DIM30).

[0342] SEQ ID NO:162 lists the amino acid sequences of variants of the hu9F5VLv9 light chain variable region (hu9F5VLv9_V3Q, L27cS, L37Q, M51G, L54G, L92I, also known as hu9F5VLv9_DIM1).

[0343] SEQ ID NO:163 lists the amino acid sequences of variants of the hu9F5VLv9 light chain variable region (hu9F5VLv9_V3Q, L27cS, L37Q, M51G, L54R, L92I, also known as hu9F5VLv9_DIM2).

[0344] SEQ ID NO:164 lists the amino acid sequences of variants of the hu9F5VLv9 light chain variable region (hu9F5VLv9_V3Q, L27cS, L37Q, M51G, L54R, L92G, also known as hu9F5VLv9_DIM4).

[0345] SEQ ID NO:165 lists the amino acid sequences of variants of the hu9F5VLv9 light chain variable region (hu9F5VLv9_V3Q, L27cG, L37Q, M51G, L54R, L92I, also known as hu9F5VLv9_DIM5).

[0346] SEQ ID NO:166 lists the amino acid sequences of variants of the hu9F5VLv9 light chain variable region (hu9F5VLv9_V3Q, L27cG, L37Q, M51K, L54R, L92I, also known as hu9F5VLv9_DIM8).

[0347] SEQ ID NO:167 lists the amino acid sequences of variants of the hu9F5VLv9 light chain variable region (hu9F5VLv9_V3Q, L27cS, L37Q, M51K, L54G, L92I, also known as hu9F5VLv9_DIM10).

[0348] SEQ ID NO:168 lists the amino acid sequences of variants of the hu9F5VLv9 light chain variable region (hu9F5VLv9_V3Q, L27cG, L37G, M51G, L54R, L92I, also known as hu9F5VLv9_DIM11).

[0349] SEQ ID NO:169 lists the amino acid sequences of variants of the hu9F5VLv9 light chain variable region (hu9F5VLv9_V3Q, L27cG, L37G, M51G, L54R, also known as hu9F5VLv9_DIM13).

[0350] SEQ ID NO:170 lists the amino acid sequences of variants of the hu9F5VLv9 light chain variable region (hu9F5VLv9_V3Q, L27cS, L37I, M51I, L54R, L92I, also known as hu9F5VLv9_DIM19).

[0351] SEQ ID NO:171 lists the amino acid sequences of variants of the hu9F5VLv9 light chain variable region (hu9F5VLv9_V3Q, L27cS, L37Q, M51I, L54G, L92I, also known as hu9F5VLv9_DIM20).

[0352] SEQ ID NO:172 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_L27bD) of the humanized 9F5 antibody.

[0353] SEQ ID NO:173 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_L27bT) of the humanized 9F5 antibody.

[0354] SEQ ID NO:174 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_L27bQ) of the humanized 9F5 antibody.

[0355] SEQ ID NO:175 lists the amino acid sequence of alternative Kabat CDR-L1 (present in hu9F5VLv2_L27cD, hu9F5VLv8_DIM6, hu9F5VLv8_DIM7 and hu9F5VLv8_DIM18) of the humanized 9F5 antibody.

[0356] SEQ ID NO:176 lists alternative Kabat antibodies for humanized 9F5. The amino acid sequence of CDR-L1 (present in hu9F5VLv2_L27cG, hu9F5VLv8_DIM5, hu9F5VLv8_DIM8, hu9F5VLv8_DIM9, hu9F5VLv8_DIM11, hu9F5VLv8_DIM12, hu9F5VLv8_DIM13, hu9F5VLv8_DIM14, hu9F5VLv8_DIM15, hu9F5VLv8_DIM16, hu9F5VLv8_DIM23, hu9F5VLv8_DIM24, hu9F5VLv8_DIM25, hu9F5VLv9_DIM5, hu9F5VLv9_DIM8, hu9F5VLv9_DIM11 and hu9F5VLv9_DIM13).

[0357] SEQ ID NO:177 lists alternative Kabat antibodies for humanized 9F5. The amino acid sequence of CDR-L1 (present in hu9F5VLv2_L27cS, hu9F5VLv8_DIM1, hu9F5VLv8_DIM2, hu9F5VLv8_DIM19, hu9F5VLv8_DIM20, hu9F5VLv8_DIM21, hu9F5VLv8_DIM22, hu9F5VLv8_DIM28, hu9F5VLv8_DIM29, hu9F5VLv8_DIM30, hu9F5VLv9_DIM1, hu9F5VLv9_DIM2, hu9F5VLv9_DIM4, hu9F5VLv9_DIM10, hu9F5VLv9_DIM19 and hu9F5VLv9_DIM20).

[0358] SEQ ID NO:178 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_L27cE) of the humanized 9F5 antibody.

[0359] SEQ ID NO:179 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_L27cT) of the humanized 9F5 antibody.

[0360] SEQ ID NO:180 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_L27cN) of the humanized 9F5 antibody.

[0361] SEQ ID NO:181 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_L27cA) of the humanized 9F5 antibody.

[0362] SEQ ID NO:182 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_L27cP) of the humanized 9F5 antibody.

[0363] SEQ ID NO:183 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv8_DIM26) of the humanized 9F5 antibody.

[0364] SEQ ID NO:184 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_I30E) for the humanized 9F5 antibody.

[0365] SEQ ID NO:185 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_I30K) for the humanized 9F5 antibody.

[0366] SEQ ID NO:186 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_I30G) for the humanized 9F5 antibody.

[0367] SEQ ID NO:187 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_I30Q) for the humanized 9F5 antibody.

[0368] SEQ ID NO:188 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_T31G) for the humanized 9F5 antibody.

[0369] SEQ ID NO:189 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_L33N) of the humanized 9F5 antibody.

[0370] SEQ ID NO:190 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_L33T) of the humanized 9F5 antibody.

[0371] SEQ ID NO:191 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_L33S) of the humanized 9F5 antibody.

[0372] SEQ ID NO:192 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_L33R) of the humanized 9F5 antibody.

[0373] SEQ ID NO:193 lists the amino acid sequence of an alternative Kabat CDR-L1 (present in hu9F5VLv2_L33G) of the humanized 9F5 antibody.

[0374] SEQ ID NO:194 lists the amino acid sequence of an alternative Kabat CDR-L2 (present in hu9F5VLv2_M51E) of the humanized 9F5 antibody.

[0375] SEQ ID NO:195 lists the amino acid sequence of an alternative Kabat CDR-L2 (present in hu9F5VLv2_M51D) of the humanized 9F5 antibody.

[0376] SEQ ID NO:196 lists the amino acid sequence of an alternative Kabat CDR-L2 (present in hu9F5VLv8_DIM30) of the humanized 9F5 antibody.

[0377] SEQ ID NO:197 lists the amino acid sequence of an alternative Kabat CDR-L2 (present in hu9F5VLv8_DIM29) of the humanized 9F5 antibody.

[0378] SEQ ID NO:198 lists the amino acid sequence of an alternative Kabat CDR-L2 (present in hu9F5VLv8_DIM1 and hu9F5VLv9_DIM1) of the humanized 9F5 antibody.

[0379] SEQ ID NO:199 lists the amino acid sequence of alternative Kabat CDR-L2 (present in hu9F5VLv8_DIM3, hu9F5VLv8_DIM14, hu9F5VLv8_DIM15, hu9F5VLv8_DIM16 and hu9F5VLv8_DIM17) of the humanized 9F5 antibody.

[0380] SEQ ID NO:200 lists the amino acid sequence of alternative Kabat CDR-L2 (present in hu9F5VLv8_DIM7, hu9F5VLv8_DIM8 and hu9F5VLv9_DIM8) of humanized 9F5 antibody.

[0381] SEQ ID NO:201 lists the amino acid sequence of alternative Kabat CDR-L2 (present in hu9F5VLv8_DIM9, hu9F5VLv8_DIM10 and hu9F5VLv9_DIM10) of the humanized 9F5 antibody.

[0382] SEQ ID NO:202 lists the amino acid sequence of an alternative Kabat CDR-L2 (present in hu9F5VLv8_DIM19 and hu9F5VLv9_DIM19) of the humanized 9F5 antibody.

[0383] SEQ ID NO:203 lists the amino acid sequence of alternative Kabat CDR-L2 (present in hu9F5VLv8_DIM20, hu9F5VLv8_DIM21 and hu9F5VLv9_DIM20) of humanized 9F5 antibody.

[0384] SEQ ID NO:204 lists the amino acid sequence of alternative Kabat CDR-L2 (present in hu9F5VLv8_DIM22, hu9F5VLv8_DIM24, hu9F5VLv8_DIM25 and hu9F5VLv8_DIM26) of the humanized 9F5 antibody.

[0385] SEQ ID NO:205 lists the amino acid sequence of an alternative Kabat CDR-L2 (present in hu9F5VLv8_DIM23) of the humanized 9F5 antibody.

[0386] SEQ ID NO:206 lists the amino acid sequence of an alternative Kabat CDR-L3 (present in hu9F5VLv2_A89G) of the humanized 9F5 antibody.

[0387] SEQ ID NO:207 lists the amino acid sequence of an alternative Kabat CDR-L3 (present in hu9F5VLv2_L92D) of the humanized 9F5 antibody.

[0388] SEQ ID NO:208 lists the amino acid sequence of an alternative Kabat CDR-L3 (present in hu9F5VLv2_L92E) of the humanized 9F5 antibody.

[0389] SEQ ID NO:209 lists the amino acid sequence of alternative Kabat CDR-L3 (present in hu9F5VLv8_DIM4, hu9F5VLv8_DIM12, hu9F5VLv8_DIM15, hu9F5VLv8_DIM25 and hu9F5VLv9_DIM4) of the humanized 9F5 antibody.

[0390] SEQ ID NO:210 lists the amino acid sequence of an alternative Kabat CDR-L3 (present in hu9F5VLv2_L92Q) of the humanized 9F5 antibody.

[0391] SEQ ID NO:211 lists the amino acid sequence of an alternative Kabat CDR-L3 (present in hu9F5VLv2_L92T) of the humanized 9F5 antibody.

[0392] SEQ ID NO:212 lists alternative Kabat CDR-L3 for humanized 9F5 antibodies (present in hu9F5VLv8_DIM1, hu9F5VLv8_DIM2, hu9F5VLv8_DIM3, hu9F5VLv8_DIM5, hu9F5VLv8_DIM6, hu9F5VLv8_DIM7, hu9F5VLv8_DIM8, hu9F5VLv8_DIM9, hu9F5VLv8_DIM10, hu9F5VLv8_DIM11, hu9F5VLv8_DIM14, hu9F5VLv8_DIM17, hu9F5VLv8_DIM18, hu9F5VLv8_DIM19, hu9F5VLv8_DIM20). The amino acid sequences of hu9F5VLv8_DIM22, hu9F5VLv8_DIM23, hu9F5VLv8_DIM24, hu9F5VLv8_DIM27, hu9F5VLv8_DIM28, hu9F5VLv8_DIM29, hu9F5VLv8_DIM30, hu9F5VLv9_DIM1, hu9F5VLv9_DIM2, hu9F5VLv9_DIM5, hu9F5VLv9_DIM8, hu9F5VLv9_DIM10, hu9F5VLv9_DIM11, hu9F5VLv9_DIM19 and u9F5VLv9_DIM20.

[0393] SEQ ID NO:213 lists the amino acid sequence of an alternative Kabat CDR-L3 (present in hu9F5VLv2_L93G) of the humanized 9F5 antibody.

[0394] SEQ ID NO:214 lists the amino acid sequence of the humanized heavy chain variable region hu10C12VHv1.

[0395] SEQ ID NO:215 lists the amino acid sequence of the humanized heavy chain variable region hu10C12VHv2.

[0396] SEQ ID NO:216 lists the amino acid sequence of the humanized light chain variable region hu10C12VLv1.

[0397] SEQ ID NO:217 lists the amino acid sequence of the humanized light chain variable region hu10C12VLv2.

[0398] SEQ ID NO:218 lists the amino acid sequence of the heavy chain variable region receptor CAC20421-VH_huFrwk.

[0399] SEQ ID NO:219 lists the amino acid sequence of the heavy chain variable region of the mouse 12C4 antibody.

[0400] SEQ ID NO:220 lists the amino acid sequence of the mouse 12C4 antibody Kabat CDR-H2.

[0401] SEQ ID NO:221 lists the amino acid sequence of the humanized heavy chain variable region hu12C4VHv1.

[0402] SEQ ID NO:222 lists the amino acid sequence of the humanized heavy chain variable region hu12C4VHv2.

[0403] SEQ ID NO:223 lists the amino acid sequence of the humanized light chain variable region hu12C4VLv1.

[0404] SEQ ID NO:224 lists the amino acid sequence of the humanized light chain variable region hu12C4VLv2.

[0405] SEQ ID NO:225 lists the amino acid sequence of the heavy chain variable region of the mouse 17C12 antibody.

[0406] SEQ ID NO:226 lists the amino acid sequence of the Kabat-Chothia complex CDR H1 of the mouse 17C12 antibody.

[0407] SEQ ID NO:227 lists the amino acid sequence of the Kabat CDR H2 antibody against mouse 17C12.

[0408] SEQ ID NO:228 lists the amino acid sequence of the light chain variable region of the mouse 17C12 antibody.

[0409] SEQ ID NO:229 lists the amino acid sequence of Kabat CDR-L1 of the mouse 17C12 antibody.

[0410] SEQ ID NO:230 lists the amino acid sequence of the mouse 17C12 antibody Kabat CDR-L2.

[0411] SEQ ID NO:231 lists the amino acid sequence of the mouse 17C12 antibody Kabat CDR-L3.

[0412] SEQ ID NO:232 lists the amino acid sequence of the humanized heavy chain variable region hu17C12VHv1.

[0413] SEQ ID NO:233 lists the amino acid sequence of the humanized heavy chain variable region hu17C12VHv2.

[0414] SEQ ID NO:234 ​​lists the amino acid sequence of the humanized light chain variable region hu17C12VLv1.

[0415] SEQ ID NO:235 lists the amino acid sequence of the humanized light chain variable region hu17C12VLv2.

[0416] SEQ ID NO:236 lists the amino acid sequence of the heavy chain variable region structural model 3PP3-VH_mSt.

[0417] SEQ ID NO:237 lists the amino acid sequence of the light chain variable region structural model 3PP3-VL_mSt.

[0418] SEQ ID NO:238 lists the amino acid sequence of the light chain variable region receptor QDO16713-VL_huFrwk.

[0419] SEQ ID NO:239 lists the amino acid sequence of the light chain variable region germline sequence IGKV2-29*02&IGKJ4*01.

[0420] SEQ ID NO:240 lists the amino acid sequence of the heavy chain variable region of the mouse 14H3 antibody.

[0421] SEQ ID NO:241 lists the amino acid sequence of the Kabat-Chothia complex CDR H1 of the mouse 14H3 antibody.

[0422] SEQ ID NO:242 lists the amino acid sequence of the Kabat CDR H2 of the mouse 14H3 antibody.

[0423] SEQ ID NO:243 lists the amino acid sequence of the Kabat CDR H3 antibody against mouse 14H3.

[0424] SEQ ID NO:244 lists the amino acid sequence of the light chain variable region of the mouse 14H3 antibody.

[0425] SEQ ID NO:245 lists the amino acid sequence of Kabat CDR L1 of the mouse 14H3 antibody.

[0426] SEQ ID NO:246 lists the amino acid sequence of the Kabat CDR L2 of the mouse 14H3 antibody.

[0427] SEQ ID NO:247 lists the amino acid sequence of Kabat CDR L3 of the mouse 14H3 antibody.

[0428] SEQ ID NO:248 lists the amino acid sequence of the humanized heavy chain variable region hu14H3VHv1.

[0429] SEQ ID NO:249 lists the amino acid sequence of the humanized heavy chain variable region hu14H3VHv2.

[0430] SEQ ID NO:250 lists the amino acid sequence of the humanized light chain variable region hu14H3VLv1.

[0431] SEQ ID NO:251 lists the amino acid sequence of the humanized light chain variable region hu14H3VLv2.

[0432] SEQ ID NO:252 lists the amino acid sequence of the heavy chain variable region structural model 2VQ1-VH_mSt.

[0433] SEQ ID NO:253 lists the amino acid sequence of the heavy chain variable region receptor QDJ57937-VH_huFrwk.

[0434] SEQ ID NO:254 lists the amino acid sequence of the heavy chain variable region germline sequence IGHV1-70*04&IGHJ4*01.

[0435] SEQ ID NO:255 lists the amino acid sequence of the light chain variable region structural model 2VQ1-VL_mSt.

[0436] SEQ ID NO:256 lists the amino acid sequence of the light chain variable region receptor ABC66914-VL_huFrwk.

[0437] SEQ ID NO:257 lists the amino acid sequence of the mouse 12C4 antibody AbM CDR-H2.

[0438] SEQ ID NO:258 lists the amino acid sequence of the mouse 12C4 antibody Contact CDR-H2.

[0439] SEQ ID NO:259 lists the amino acid sequence of Chothia CDR-H1, a mouse 17C12 antibody.

[0440] SEQ ID NO:260 lists the amino acid sequence of the mouse 17C12 antibody AbM CDR-H2.

[0441] SEQ ID NO:261 lists the amino acid sequence of the mouse 17C12 antibody Contact CDR-H2.

[0442] SEQ ID NO:262 lists the amino acid sequence of the mouse 17C12 antibody Contact CDR-L1.

[0443] SEQ ID NO:263 lists the amino acid sequence of the mouse 17C12 antibody Contact CDR-L2.

[0444] SEQ ID NO:264 lists the amino acid sequence of the mouse 17C12 antibody Contact CDR-L3.

[0445] SEQ ID NO:265 lists the amino acid sequence of the Kabat CDR-H1 antibody against mouse 14H3.

[0446] SEQ ID NO:266 lists the amino acid sequence of Chothia CDR-H1, a mouse 14H3 antibody.

[0447] SEQ ID NO:267 lists the amino acid sequence of the mouse 14H3 antibody, Chothia CDR-H2.

[0448] SEQ ID NO:268 lists the amino acid sequence of the mouse 14H3 antibody AbM CDR-H2.

[0449] SEQ ID NO:269 lists the amino acid sequence of the mouse 14H3 antibody Contact CDR-H1.

[0450] SEQ ID NO:270 lists the amino acid sequence of the mouse 14H3 antibody Contact CDR-H2.

[0451] SEQ ID NO:271 lists the amino acid sequence of the mouse 14H3 antibody Contact CDR-H3.

[0452] SEQ ID NO:272 lists the amino acid sequence of the mouse 14H3 antibody Contact CDR-L1.

[0453] SEQ ID NO:273 lists the amino acid sequence of the mouse 14H3 antibody Contact CDR-L2.

[0454] SEQ ID NO:274 lists the amino acid sequence of the mouse 14H3 antibody Contact CDR-L3.

[0455] SEQ ID NO:275 lists the amino acid sequence of an alternative Kabat-Chothia complex CDR H1 (present in hu14H3VHv1 and hu14H3VHv2) of the humanized 14H3 antibody.

[0456] SEQ ID NO:276 lists the amino acid sequence of the common motif of the peptide bound by antibodies 9F5, 10C12, 2D11, 12C4, 17C12 and 14H3.

[0457] SEQ ID NO:277 lists the amino acid sequence of the common motif of the peptide bound by antibody 2D11.

[0458] definition

[0459] Monoclonal antibodies or other biological entities are typically supplied in isolated form. This means that the antibody or other biological entity is generally at least 50% w / w free of interfering proteins and other contaminants resulting from its production or purification, but this does not preclude the possibility of combining the monoclonal antibody with an excess of one or more pharmaceutically acceptable carriers or other media intended to facilitate its use. Sometimes, monoclonal antibodies are at least 60%, 70%, 80%, 90%, 95%, or 99% w / w free of interfering proteins and contaminants resulting from their production or purification. Typically, the isolated monoclonal antibody or other biological entity is the major macromolecule remaining after its purification.

[0460] The specific binding of an antibody to its target antigen means at least 10 6 10 7 10 8 10 9 10 10 10 11 Or 10 12 M -1 The specific binding has a high affinity and / or binding strength. Specific binding is detectably higher and can be distinguished from nonspecific binding to at least one unrelated target. Specific binding can result from the formation of bonds between specific functional groups or specific spatial coordination (e.g., lock-and-key), while nonspecific binding is typically the result of van der Waals forces. However, specific binding does not necessarily mean that the antibody binds to only one target.

[0461] The basic antibody structural unit is a tetramer of subunits. Each tetramer consists of two pairs of identical polypeptide chains, each pair having a "light" chain (approximately 25 kDa) and a "heavy" chain (approximately 50–70 kDa). The amino-terminal portion of each chain includes a variable region of approximately 100 to 110 or more amino acids primarily responsible for antigen recognition. This variable region is initially expressed as being linked to a cleavable signal peptide. Variable regions without a signal peptide are sometimes referred to as mature variable regions. Thus, for example, a light-chain mature variable region means a light-chain variable region without a light-chain signal peptide. The carboxyl-terminal portion of each chain defines a constant region primarily responsible for effector function.

[0462] The light chain is classified as κ or λ. The heavy chain is classified as γ, μ, α, δ, or ε, and antibody isotypes are defined as IgG, IgM, IgA, IgD, and IgE, respectively. Within both the light and heavy chains, variable and constant regions are linked by a “J” region of approximately 12 or more amino acids, with the heavy chain also including a “D” region of approximately 10 or more amino acids. See Fundamental Immunology, edited by Paul W., 2nd ed., Raven Press, NY, 1989, Chapter 7 (incorporated in its entirety for all purposes).

[0463] The variable regions of the immunoglobulin light or heavy chains (also referred to herein as the “light chain variable domain” (“VL domain”) or the “heavy chain variable domain” (“VH domain”) respectively) consist of “framework” regions separated by three “complementarity-determining regions” or “CDRs”. The framework regions are used to align the CDRs for epitope-specific binding to the antigen. The CDRs contain the amino acid residues of the antibody primarily responsible for antigen binding. From the amino terminus to the carboxyl terminus, both the VL and VH domains contain the following framework (FR) and CDR regions: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The CDRs 1, 2, and 3 of the VL domain are also referred to herein as CDR-L1, CDR-L2, and CDR-L3, respectively; and the CDRs 1, 2, and 3 of the VH domain are also referred to herein as CDR-H1, CDR-H2, and CDR-H3, respectively. When this application discloses a VL sequence with R as the C-terminal residue, R can also be considered as the N-terminal residue of the light chain constant region. Therefore, this application should also be understood as disclosing a VL sequence without a C-terminal R.

[0464] The amino acid assignment of each VL and VH domain is consistent with any conventional definition of CDR. Common definitions include the Kabat definition (Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, MD, 1987 and 1991), the Chothia definition (Chothia & Lesk, J. Mol. Biol. 196:901-917, 1987; Chothia et al., Nature 342:878-883, 1989); the ChothiaKabat CDR complex, where CDR-H1 is a complex of Chothia and Kabat CDRs; the AbM definition used by the Oxford Molecular antibody modeling software; and the contact definition by Martin et al. (bioinfo.org.uk / abs) (see Table 1). Kabat provides a widely used numbering convention (Kabat numbering) in which corresponding residues between different heavy chains or between different light chains are assigned the same number. When an antibody is said to contain a CDR by a definition of a CDR (e.g., Kabat), that definition indicates the presence of the CDR residues in the antibody (i.e., Kabat). The minimum number of CDRs. This does not preclude the presence of other residues belonging to another conventional CDR definition but outside the specified definition. For example, among other possibilities, antibodies containing a CDR defined by Kabat include antibodies in which the CDR contains Kabat CDR residues and no other CDR residues, and antibodies in which CDRH1 is the complex Chothia-Kabat CDR H1 and the other CDRs contain Kabat CDR residues and no additional CDR residues based on other definitions.

[0465] Table 1. General Definitions of CDRs Using Kabat Numbers

[0466]

[0467] Table 1. General Definitions of CDRs Using Kabat Numbers

[0468]

[0469] *According to Chothia's CDR-H1, the loop can end at H32, H33, or H34 (depending on the loop length). This is because the Kabat numbering scheme places the insertion of additional residues at 35A and 35B, while Chothia numbering places them at 31A and 31B. If neither H35A nor H35B (Kabat numbering) is present, then the Chothia CDR-H1 loop ends at H32. If only H35A is present, then it ends at H33. If both H35A and H35B are present, then it ends at H34.

[0470] The term "antibody" includes both intact antibodies and their binding fragments. Typically, fragments compete with the intact antibodies from which they are derived for specific binding to a target, including individual heavy chains, light chains Fab, Fab', F(ab')2, F(ab)c, Dab, nanobodies, and Fv. Fragments can be generated using recombinant DNA technology or through enzymatic or chemical separation of intact immunoglobulins. The term "antibody" also includes bispecific antibodies and / or humanized antibodies. Bispecific or bifunctional antibodies are artificial hybrid antibodies with two distinct heavy / light chain pairs and two distinct binding sites (see, for example, Songsivilai and Lachmann, Clin. Exp. Immunol., 79:315-321 (1990); Kostelny et al., J. Immunol., 148:1547-53 (1992)). In some bispecific antibodies, the two distinct heavy / light chain pairs include humanized 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3 heavy / light chain pairs and heavy / light chain pairs that are specific to epitopes on tau that are different from those bound by 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3.

[0471] In some bispecific antibodies, one heavy / light chain pair is a humanized 9F5 antibody, a humanized 10C12 antibody, a humanized 2D11 antibody, a humanized 12C4 antibody, a humanized 17C12 antibody, or a humanized 14H3 antibody, as further disclosed below, and the other heavy / light chain pair is derived from an antibody that binds to a receptor expressed on the blood-brain barrier, such as the insulin receptor, insulin-like growth factor (IGF) receptor, leptin receptor, or lipoprotein receptor or transferrin receptor (Friden et al., Proc. Natl. Acad. Sci. USA 88:4771-4775, 1991; Friden et al., Science 259:373-377, 1993). Such bispecific antibodies can be translocated across the blood-brain barrier via receptor-mediated transcellularity. Brain uptake of bispecific antibodies can be further enhanced by engineering them to reduce their affinity for blood-brain barrier receptors. Decreased affinity for the receptor leads to a wider distribution in the brain (see, for example, Atwal et al., Sci. Trans. Med. 3, 84ra43, 2011; Yu et al., Sci. Trans. Med. 3, 84ra44, 2011).

[0472] Exemplary bispecific antibodies may also be: (1) dual variable domain antibodies (DVD-Ig), wherein each light chain and heavy chain contains two variable domains tandemly linked by short peptide bonds (Wu et al., Generation and Characterization of a Dual Variable Domain Immunoglobulin (DVD-Ig)). TM(1) Molecule, Antibody Engineering, Springer Berlin Heidelberg (2010); (2) Tandab, which is a fusion of two single-chain biantibodies to obtain a tetravalent bispecific antibody with two binding sites for each target antigen; (3) Flexible body, which is a combination of scFv and biantibody to obtain a multivalent molecule; (4) So-called "dock and lock" molecule, which is based on the "dimerization and docking domain" in protein kinase A, and when applied to Fab, can obtain a trivalent bispecific binding protein composed of two identical Fab fragments linked to different Fab fragments; or (5) So-called Scorpion molecule, which contains, for example, two scFvs fused to the two ends of the human Fc region. Examples of platforms that can be used to prepare bispecific antibodies include BiTE (Micromet), DART (MacroGenics), Fcab and Mab2 (F-star), Fc engineered IgG1 (Xencor), or DuoBody (based on Fab arm exchange, Genmab).

[0473] The term "epitope" refers to a site on an antigen where an antibody binds. Epitopes can be formed from consecutive amino acids or from discontinuous amino acids juxtaposed through the ternary folding of one or more proteins. Epitopes formed from consecutive amino acids (also known as linear epitopes) are generally retained upon exposure to denaturing solvents, while epitopes formed through ternary folding (also known as conformational epitopes) are generally lost upon treatment with denaturing solvents. Epitopes typically consist of at least three, and more usually at least five or eight to ten amino acids in a unique spatial conformation. Methods for determining the spatial conformation of epitopes include, for example, X-ray crystallography and two-dimensional nuclear magnetic resonance. See, for example, Epitope Mapping Protocols, in Methods in Molecular Biology, Vol. 66, edited by Glenn E. Morris (1996).

[0474] Antibodies that recognize the same or overlapping epitopes can be identified in a simple immunoassay that shows the ability of one antibody to compete with another for binding to a target antigen. An antibody epitope can also be defined by X-ray crystallography to identify the contacting residues of an antibody by binding to its antigen. Alternatively, if all amino acid mutations in the antigen that reduce or eliminate the binding of one antibody reduce or eliminate the binding of another antibody, then the two antibodies have the same epitope. If some amino acid mutations that reduce or eliminate the binding of one antibody reduce or eliminate the binding of another antibody, then the two antibodies have overlapping epitopes.

[0475] Competition between antibodies is determined by an assay in which the test antibody inhibits the specific binding of a reference antibody to a common antigen (see, for example, Junghans et al., Cancer Res. 50:1495, 1990). The test antibody competes with the reference antibody if an excess of the test antibody (e.g., at least 2x, 5x, 10x, 20x, or 100x) inhibits at least 50% of the binding of the reference antibody (as measured in a competitive binding assay). Some test antibodies inhibit at least 75%, 90%, or 99% of the binding of the reference antibody. Antibodies identified by the competitive assay (competitive antibodies) include antibodies that bind to the same epitope as the reference antibody and antibodies that bind to adjacent epitopes sufficiently close to the epitope bound to the reference antibody to create steric hindrance.

[0476] The term "pharmaceutically acceptable" means that the carrier, diluent, excipient, or adjuvant is compatible with the other components of the formulation and is substantially harmless to the recipient.

[0477] The term "patient" includes people and other mammalian subjects who receive preventative or therapeutic treatment.

[0478] If a subject has at least one known risk factor (e.g., genetic, biochemical, family history, and situational exposure) that places an individual with said risk factor at a statistically significantly greater risk of developing the disease compared to an individual without that risk factor, then the individual’s risk of developing the disease is increased.

[0479] The term "biological sample" refers to a sample of biological material from or derived from a biological source, such as a human or mammalian subject. Such samples can be organs, organelles, tissues, tissue sections, body fluids, peripheral blood, plasma, serum, cells, molecules (such as proteins and peptides), and any parts or combinations derived therefrom. The term "biological sample" may also include any material derived by processing the sample. Derived material may include cells or their progeny. Processing of biological samples may include one or more of the following: filtration, distillation, extraction, concentration, fixation, inactivation of interfering components, etc.

[0480] The term "control sample" refers to a biological sample that is not known or suspected of including areas affected by tau-related disease, or at least not known or suspected of including diseased areas of a given type. Control samples can be obtained from individuals who have never had tau-related disease. Alternatively, control samples can be obtained from patients who have tau-related disease. Such samples can be obtained simultaneously with or on different occasions from biological samples believed to contain tau-related disease. Both the biological sample and the control sample can be obtained from the same tissue. Preferably, the control sample consists substantially or entirely of normal, healthy areas and can be used for comparison with biological samples believed to contain areas affected by tau-related disease. Preferably, the tissue in the control sample is of the same type as the tissue in the biological sample. Preferably, the tau-related disease-affected cells believed to be in the biological sample are of the same cell type (e.g., neurons or glial cells) as the cell types in the control sample.

[0481] The term "disease" refers to any abnormal condition that impairs physiological function. This term is used broadly to encompass any symptom, ailment, abnormality, pathology, discomfort, disorder, or syndrome that impairs physiological function, regardless of the nature of its cause.

[0482] The term "symptoms" refers to subjective evidence of disease perceived by the subject, such as changes in gait. "Signs" refers to objective evidence of disease observed by the physician.

[0483] The term "positive response to treatment" refers to a more favorable response in an individual patient or in a patient population than the average response in a control group that did not receive treatment.

[0484] To classify amino acid substitutions as conserved or non-conserved, amino acids are grouped as follows: Group I (hydrophobic side chains): met, ala, val, leu, ile; Group II (neutral hydrophilic side chains): cys, ser, thr; Group III (acidic side chains): asp, glu; Group IV (basic side chains): asn, gln, his, lys, arg; Group V (residues affecting chain orientation): gly, pro; and Group VI (aromatic side chains): trp, tyr, phe. Conservative substitutions include substitutions between amino acids of the same category. Non-conservative substitutions involve exchanging a member of one category for a member of another category.

[0485] The percentage of sequence identity is determined using antibody sequences that are aligned to the maximum extent possible using the Kabat numbering convention. After alignment, if a subject antibody region (e.g., the entire mature variable region of the heavy or light chain) is compared with the same region of a reference antibody, the percentage of sequence identity between the subject antibody region and the reference antibody region is calculated by dividing the number of positions occupied by the same amino acids in the subject antibody region and the reference antibody region by the total number of aligned positions in the two regions, excluding vacancies, and multiplying by 100 to convert to a percentage.

[0486] Compositions or methods that "comprise" or "include" one or more of the listed elements may include other elements not specifically listed. For example, a composition that "comprises" or "includes" an antibody may contain an antibody, alone or in combination with other components.

[0487] The specification of a range of values ​​includes all integers within or defining that range, as well as all subranges defined by integers within that range.

[0488] Unless otherwise apparent from the context, the term “about” covers non-substantial variations, such as values ​​within the standard tolerance of measurement error (e.g., SEM) of a specified value.

[0489] Statistical significance means p≤0.05.

[0490] Unless the context clearly specifies otherwise, the singular forms of the articles “a,” “an,” and “the” include multiple referents. For example, the terms “compound” or “at least one compound” can include multiple compounds, including mixtures thereof. Detailed Implementation

[0491] I. General Provisions

[0492] This invention provides antibodies that bind to tau. Some antibodies specifically bind to an epitope within (Q / E)IVYK(S / P) (SEQ ID NO:56). Some antibodies specifically bind to a peptide containing the amino acid sequence QIVYKP (SEQ ID NO:57, corresponding to residues 307-312 of the tau isoform in SEQ ID NO:1). Some antibodies specifically bind to a peptide containing the amino acid sequence EIVYKSP (SEQ ID NO:58, corresponding to residues 391-397 of the tau isoform in SEQ ID NO:1). These antibodies differ from 3D6 and other antibodies characterized by binding to the microtubule-binding region (MTBR) of human tau in that they have an additional epitope near the C-terminus of tau. The additional C-terminal specificity of the epitope provides a basis for antibody binding to an increased number of pathologically associated conformations of tau. Some antibodies specifically bind to a peptide containing the amino acid sequence EIVYKS (SEQ ID NO:277, corresponding to residues 391-396 of the tau isoform in SEQ ID NO:1). Exemplary antibodies of the present invention are 9F5, 10C12, 2D11, 12C4, 17C12, and 14H3. Some antibodies of the present invention are used to inhibit or delay the progression of tau-related pathology and associated symptoms. Although practice of the present invention does not require an understanding of the mechanisms, toxicity reduction can occur through mechanisms such as antibody-induced tau phagocytosis, inhibition of intermolecular or intramolecular aggregation of tau, or binding to other molecules, by stabilizing nontoxic conformations, by inhibiting intercellular or intracellular transmission of pathogenic tau forms, by blocking tau phosphorylation, by preventing tau binding to cells, or by inducing tau proteolytic cleavage. Some antibodies of the present invention can be used to increase tau aggregation, thereby reducing toxicity / cellular uptake and / or increasing clearance by increasing the molecular weight of certain aggregated tau substances. Large aggregates of tau molecules can reduce uptake by neuronal cells. Some antibodies of the present invention, by binding divalently to tau, bring dissociated tau molecules closer together, promoting aggregation into large tau aggregates that cannot be taken up by neuronal cells. Furthermore, Fc-mediated phagocytosis requires several tau-binding antibodies to be very close together. Large aggregates of tau molecules may bind to numerous anti-tau antibodies and provide the clusters required for Fc-mediated phagocytosis by macrophages. The antibodies of the present invention, or agents that induce such antibodies, may be used in methods for treating Alzheimer's disease and other tau-related diseases or for achieving prevention of said diseases.

[0493] II. Target Molecules

[0494] Unless otherwise apparent from the context, references to tau refer to the native human form of tau, including all isoforms, regardless of the presence of post-translational modifications (e.g., phosphorylation, glycosylation, or acetylation). There are six major tau isoforms (splice variants) present in the human brain. The longest of these variants has 441 amino acids, with the starting met residue cleaved. Residues are numbered according to the 441 isoform. Thus, for example, a reference to phosphorylation at position 404 means position 404 of the 441 isoform, or the corresponding position of any other isoform when maximally aligned with the 441 isoform. Amino acid sequences and Swiss-Prot numbers of the isoforms are indicated below.

[0495]

[0496] P10636-6(4R0N human tau)(SEQ ID NO:3)

[0497]

[0498] References to tau include known natural variants, of which approximately 30 are listed in the Swiss-Prot database and its alignments, as well as mutations associated with tauopathies such as dementia, Pick's disease, supranuclear palsy, etc. (see, e.g., the Swiss-Prot database and Poorkaj et al., Ann Neurol. 43:815-825 (1998)). Some examples of tau mutations numbered according to the 441 isoform are the lysine to threonine mutation (K257T) at amino acid residue 257; the isoleucine to valine mutation (I260V) at amino acid position 260; the glycine to valine mutation (G272V) at amino acid position 272; the asparagine to lysine mutation (N279K) at amino acid position 279; the asparagine to histidine mutation (N296H) at amino acid position 296; the proline to serine mutation (P301S) at amino acid position 301; the proline to leucine mutation (P301L) at amino acid 301; the glycine to valine mutation (G303V) at amino acid position 303; the serine to asparagine mutation (S305N) at position 305; the glycine to serine mutation (G335S) at amino acid position 335; the valine to methionine mutation (V337M) at position 337; the glutamate to valine mutation (E342V) at position 342; the lysine to isoleucine mutation (K369I) at amino acid position 369; the glycine to arginine mutation (G389R) at amino acid position 389; and the arginine to tryptophan mutation (R406W) at amino acid position 406.

[0499] Tau can be phosphorylated at one or more amino acid residues, including tyrosine at amino acid positions 18, 29, 97, 310, and 394; serine at amino acid positions 184, 185, 198, 199, 202, 208, 214, 235, 237, 238, 262, 293, 324, 356, 396, 400, 404, 409, 412, 413, and 422; and threonine at amino acid positions 175, 181, 205, 212, 217, 231, and 403.

[0500] Unless otherwise apparent from the context, references to tau or fragments thereof include natural human amino acid sequences, including their isotypes, mutants, and allele variants.

[0501] III. Antibodies

[0502] A. Combining specificity and functional characteristics

[0503] This invention provides antibodies that specifically bind to tau. Some antibodies specifically bind to tau at epitopes formed by amino acids from one or both regions of a common core motif of tau, namely IVYK (SEQ ID NO: 276). These regions are defined by residues 307-312 and 391-397 or 391-396 of SEQ ID NO: 1, respectively. Thus, antibodies such as scFv with one tau binding site can specifically bind to tau at an epitope formed solely by amino acids from either of these regions or to a hybrid epitope formed by amino acids from both regions. Antibodies with two tau binding sites can also simultaneously bind to epitopes within 307-312 and 391-397 or within 391-396 from both binding sites. Epitopes can be located on the same or different tau molecules. Some antibodies of this invention specifically bind to peptides composed of tau residues 307-312, namely residues QIVYKP (SEQ ID NO: 57). Some antibodies of the present invention specifically bind to peptides composed of tau residues 391-397, namely EIVYKSP (SEQ ID NO: 58). Some antibodies of the present invention specifically bind to peptides composed of tau residues 391-396, namely EIVYKS (SEQ ID NO: 277). Some antibodies of the present invention specifically bind to peptides composed of the common motif (Q / E)IVYK(S / P) (SEQ ID NO: 56). These antibodies can be obtained by immunization with tau polypeptides purified from natural sources or recombinantly expressed. Antibodies binding to tau in its unphosphorylated form and in forms where one or more readily phosphorylated residues are phosphorylated can be screened. The present invention also provides antibodies that bind to the same epitopes, such as 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3, as any of the aforementioned antibodies. This also includes antibodies that compete with any of the aforementioned antibodies for binding to tau, such as those competing with 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3. In one embodiment, the antibody that binds to the same epitope as the reference antibody (e.g., 9F5) or that competes with the reference antibody shares one or more of its functional properties, such as the ability to inhibit tau internalization into neuronal cells. Optionally, this property is of the same or greater extent than that of the reference antibody within the experimental error range.

[0504] This invention provides an antibody that competitively binds to tau with 9F5 and reduces tau-induced neurotoxicity. This invention also provides an antibody that competitively binds to tau with 9F5 and has enhanced resistance to agitation stress. Exemplary humanized antibodies against 9F5 that competitively bind to tau and have enhanced resistance to agitation stress are L27cS / L37Q / M51G / L54R(DIM2) (also known as hu9F5VHv9 / hu9F5VLv8_DIM2SEQ ID NO:127 / SEQ ID NO:133), L27cG / L37G / M51G / L54T(DIM14) (also known as hu9F5VHv9 / hu9F5VLv8_DIM14SEQ ID NO:127 / SEQ ID NO:145) and L27cG / L37G / M51G / L54R(DIM13) (also known as hu9F5VHv9 / hu9F5VLv8_DIM13SEQ ID NO:127 / SEQ ID NO:144). This invention provides an antibody that competitively binds to tau against 9F5 and has enhanced resistance to low pH stress. Exemplary humanized antibodies that compete with 9F5 for binding to tau and have enhanced resistance to low pH stress are L27cS / L37Q / M51G / L54R(DIM2) (also known as hu9F5VHv9 / hu9F5VLv8_DIM2SEQ ID NO:127 / SEQ ID NO:133), L27cD / L37Q / M51G / L54R(DIM6) (also known as hu9F5VHv9 / hu9F5VLv8_DIM6, SEQ ID NO:127 / SEQ ID NO:137) and L27cG / L37G / M51G / L54R(DIM13) (also known as hu9F5VHv9 / hu9F5VLv8_DIM13SEQ ID NO:127 / SEQ ID NO:144).Exemplary humanized antibodies against 9F5 that compete with tau binding and exhibit enhanced resistance to agitation stress and enhanced resistance to low pH stress are DIM2 [hu9F5VHv9 / hu9F5VLv8_DIM2SEQ ID NO:127 / SEQ ID NO:133], DIM6 [hu9F5VHv9 / hu9F5VLv8_DIM6SEQ ID NO:127 / SEQ ID NO:137], DIM7 [hu9F5VHv9 / hu9F5VLv8_DIM7SEQ ID NO:127 / SEQ ID NO:138], DIM8 [hu9F5VHv9 / hu9F5VLv8_DIM8SEQ ID NO:127 / SEQ ID NO:139], and DIM13 [hu9F5VHv9 / hu9F5VLv8_DIM13SEQ ID NO:127 / SEQ ID NO:139]. ID NO:144], DIM18[hu9F5VHv9 / hu9F5VLv8_DIM18SEQ ID NO:127 / SEQ ID NO:149], DIM28[hu9F5VHv9 / hu9F5VLv8_DIM28SEQ ID NO:127 / SEQ ID NO:159 and DIM30 [hu9F5VHv9 / hu9F5VLv8_DIM30SEQ ID NO:127 / SEQ ID NO:161].

[0505] This invention provides an antibody that competitively binds to tau with 9F5 and exhibits a reduced aggregation tendency under high concentration conditions. An exemplary antibody that competitively binds to tau with 9F5 and exhibits a reduced aggregation tendency under high concentration conditions is a combination of L37Q / M51G / L54R and the original leucine at position L27c (DIM27), also known as hu9F5VHv9 / hu9F5VLv8_DIM27SEQ ID NO:127 / SEQ ID NO:158.

[0506] The antibodies described above can be regenerated by immunization with a tau polypeptide comprising the amino acid sequence QIVYKP (SEQ ID NO:57) or composed thereof, comprising the amino acid sequence EIVYKSP (SEQ ID NO:58), comprising the amino acid sequence EIVYKS (SEQ ID NO:277), or comprising the amino acid sequence (Q / E)IVYK(S / P) (SEQ ID NO:56) or composed thereof, or by immunization with a full-length tau polypeptide or a fragment thereof containing such residues, followed by screening for specific binding to peptides containing such residues. Such tau peptides are preferably attached to heteroconjugate molecules that facilitate the initiation of an antibody response against the peptide. Attachment can be direct or via spacer peptides or amino acids. Cysteine ​​is used as the spacer amino acid because its free SH group facilitates attachment of the carrier molecule. Polyglycine linkers (e.g., 2-6 glycine residues) with or without cysteine ​​residues between glycine and the peptide can also be used. The carrier molecule serves to provide T-cell epitopes that facilitate the initiation of an antibody response against the peptide. Several carriers are commonly used, specifically key-foraminifera hemocyanin (KLH), ovalbumin, and bovine serum albumin (BSA). Peptide spacers can be added to the peptide immunogen as part of solid-phase peptide synthesis. The carriers are typically added via chemical cross-linking. Some examples of usable chemical crosslinking agents include crosslinked-N-maleimide-6-aminohexanoyl ester or m-maleimide-benzoyl-N-hydroxysuccinimide ester (MBS) (see, for example, Harlow, E. et al., Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY 1988; Sinigaglia et al., Nature, 336:778-780 (1988); Chicz et al., J. Exp. Med., 178:27-47 (1993); Hammer et al., Cell 74:197-203 (1993); Falk K. et al., Immunogenetics, 39:230-242 (1994); WO 98 / 23635; and Southwood et al. J. Immunology, 160:3363-3373 (1998)). If present, the vector and spacer can be attached to either end of the immunogen.

[0507] Peptides with optional spacers and carriers can be used to immunize laboratory animals or B cells, as described in more detail below. The ability of hybridoma supernatants to bind to the following can be tested: tau peptides containing the amino acid sequence QIVYKP (SEQ ID NO:57) or composed thereof, peptides containing the amino acid sequence EIVYKSP (SEQ ID NO:58) or composed thereof, peptides containing the amino acid sequence EIVYKS (SEQ ID NO:277) or composed thereof, or peptides containing the amino acid sequence (Q / E)IVYK(S / P) (SEQ ID NO:56) or composed thereof, and / or phosphorylated and non-phosphorylated forms of tau, such as the full-length isotype of tau having position 404 in the phosphorylated form. Peptides can be attached to carriers or other tags to facilitate screening assays. In this case, the carrier or tag is preferably different from the combination of spacer and carrier molecules used for immunization to eliminate antibodies specific to the spacer or carrier rather than the tau peptide. Any tau isotype can be used.

[0508] The antibody named 9F5 is an exemplary antibody that specifically binds to tau. Unless otherwise apparent from the context, references to 9F5 should be understood as referring to any of the mouse, chimeric, veneered, and humanized forms of the antibody. The antibody has been deposited under [accession number]. This antibody specifically binds to peptides comprising the amino acid sequence QIVYKP (SEQ ID NO:57) or thereof, peptides comprising the amino acid sequence EIVYKSP (SEQ ID NO:58) or thereof, or peptides comprising the amino acid sequence (Q / E)IVYK(S / P) (SEQ ID NO:56) or thereof. The Kabat / Chothia complex CDRs of the heavy chain of 9F5 are named SEQ ID NO:8, 9, and 10, respectively, and the Kabat CDRs of the light chain of 9F5 are named SEQ ID NO:12, 13, and 14, respectively.

[0509] Other antibodies that compete with 9F5 for binding to tau and / or bind to the same or overlapping epitopes as 9F5 have been isolated and named 10C12, 2D11, 12C4, 17C12, and 14H3, and generated from hybridomas of the same names. 10C12 has mature variable heavy and light regions (after signal peptide cleavage), characterized by SEQ ID NO:7 and SEQ ID NO:11, respectively. Unless otherwise apparent from the context, references to 10C12 should be understood as referring to any of the mouse, chimeric, veneered, and humanized forms of the antibody. 10C12 has been deposited with [accession number]. 10C12 is further characterized by its ability to bind to non-pathological and pathological forms and conformations of tau, as well as misfolded / aggregated forms of tau. 10C12 binds to structural features such as tau tangles and dystrophic neurites in Alzheimer's disease tissue and precipitates monomeric and aggregated tau from Alzheimer's disease extracts.

[0510] 2D11 possesses mature variable heavy and light regions (after signal peptide cleavage), characterized by SEQ ID NO:7 and SEQ ID NO:11, respectively. Unless otherwise apparent from the context, references to 2D11 should be understood as referring to any of the mouse, chimeric, veneered, and humanized forms of the antibody. 2D11 has been deposited under [accession number]. 2D11 is further characterized by its ability to bind non-pathological and pathological forms and conformations of tau, as well as misfolded / aggregated forms of tau. 2D11 binds to structural features such as tau tangles and dystrophic neurites in Alzheimer's disease tissue and precipitates monomeric and aggregated tau from Alzheimer's disease extracts.

[0511] 12C4 possesses mature variable heavy and light regions (after signal peptide cleavage), characterized by SEQ ID NO:219 and SEQ ID NO:11, respectively. Unless otherwise apparent from the context, references to 12C4 should be understood as referring to any of the mouse, chimeric, veneered, and humanized forms of the antibody. 12C4 has been deposited under [accession number]. A further characteristic of 12C4 is its ability to bind non-pathological and pathological forms and conformations of tau, as well as misfolded / aggregated forms of tau. 12C4 binds structural features such as tau tangles and dystrophic neurites in Alzheimer's disease tissue and precipitates monomeric and aggregated tau from Alzheimer's disease extracts.

[0512] 17C12 possesses mature variable heavy and light regions (after signal peptide cleavage), characterized by SEQ ID NO:225 and SEQ ID NO:228, respectively. Unless otherwise apparent from the context, references to 17C12 should be understood as referring to any of the mouse, chimeric, veneered, and humanized forms of the antibody. 17C12 is deposited under [accession number]. A further characteristic of 17C12 is its ability to bind to both non-pathological and pathological forms and conformations of tau, as well as misfolded / aggregated forms of tau.

[0513] 14H3 possesses mature variable heavy and light regions (after signal peptide cleavage), characterized by SEQ ID NO:240 and SEQ ID NO:244, respectively. Unless otherwise apparent from the context, references to 14H3 should be understood as referring to any of the mouse, chimeric, veneered, and humanized forms of the antibody. 14H3 has been deposited under [accession number]. A further characteristic of 14H3 is its ability to bind to both non-pathological and pathological forms and conformations of tau, as well as misfolded / aggregated forms of tau. 14H3 binds to structural features such as tau tangles and dystrophic neurites in Alzheimer's tissue.

[0514] The alignment of the mature heavy chain variable regions of mouse 9F5, 10C12, 2D11, 12C4, 14H3, and 17C12 antibodies is depicted on... Figure 24 In the study, the mature light chain variable regions of mouse 9F5, 10C12, 2D11, 12C4, 14H3, and 17C12 antibodies were compared and depicted. Figure 25The amino acid sequences of the mature heavy chain variable region of the mouse 10C12 antibody and the mature heavy chain variable region of the mouse 9F5 antibody share 100% sequence identity. The mature light chain variable region of the mouse 10C12 antibody also shares 100% sequence identity with the mature light chain variable region of the mouse 9F5 antibody. The amino acid sequences of the mature heavy chain variable region of the mouse 2D11 antibody and the mature light chain variable region of the mouse 9F5 antibody share 100% sequence identity. The mature light chain variable region of the mouse 12C4 antibody and the mature heavy chain variable region of the mouse 9F5 antibody share 96.6% sequence identity. The mature light chain variable region of the mouse 12C4 antibody and the mature light chain variable region of the mouse 9F5 antibody share 100% sequence identity. The amino acid sequence of the mature heavy chain variable region of the mouse 17C12 antibody shares 95.9% sequence identity with that of the mature heavy chain variable region of the mouse 9F5 antibody, and the amino acid sequence of the mature light chain variable region of the mouse 17C12 antibody shares 70.5% sequence identity with that of the mouse 9F5 antibody. Similarly, the amino acid sequence of the mature heavy chain variable region of the mouse 14H3 antibody shares 35.0% sequence identity with that of the mouse 9F5 antibody, and the amino acid sequence of the mature light chain variable region of the mouse 14H3 antibody shares 73.2% sequence identity with that of the mouse 9F5 antibody.

[0515] Optionally, the antibodies of the present invention do not include 10C12 antibodies. Optionally, the antibodies of the present invention do not include 2D11 antibodies. Optionally, the antibodies of the present invention do not include 12C4 antibodies. Optionally, the antibodies of the present invention do not include 17C12 antibodies. Optionally, the antibodies of the present invention do not include 14H3 antibodies.

[0516] Some antibodies of the present invention bind to the same or overlapping epitopes with antibodies named 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3. The sequences of the heavy and light chain maturation variable regions of 9F5 are named SEQ ID NO:7 and 11, respectively. The sequences of the heavy and light chain maturation variable regions of 10C12 are named SEQ ID NO:7 and 11, respectively. The sequences of the heavy and light chain maturation variable regions of 2D11 are named SEQ ID NO:7 and 11, respectively. The sequences of the heavy and light chain maturation variable regions of 12C4 are named SEQ ID NO:219 and 11, respectively. The sequences of the heavy and light chain maturation variable regions of 17C12 are named SEQ ID NO:225 and 228, respectively. The sequences of the heavy and light chain maturation variable regions of 14H3 are named SEQ ID NO:240 and 244, respectively. Other antibodies with this binding specificity can be generated by immunizing mice with tau or a portion thereof comprising the desired epitope (e.g., a tau peptide comprising the amino acid sequence QIVYKP (SEQ ID NO:57) or thereof, a tau peptide comprising the amino acid sequence EIVYKSP (SEQ ID NO:58) or thereof, a tau peptide comprising the amino acid sequence EIVYKS (SEQ ID NO:277) or thereof, or a tau peptide comprising the amino acid sequence (Q / E)IVYK(S / P) (SEQ ID NO:56) or thereof) and screening the resulting antibodies for competitive binding to tau against antibodies optionally having a variable region comprising mouse 9F5 (IgG1 / κ), 10C12 (IgG2a / κ), 2D11 (IgG2a / κ), 12C4 (IgG2a / κ), 17C12 (IgG2a / κ), or 14H3 (IgG2a / κ). A tau fragment containing the desired epitope can be linked to a vector that facilitates an antibody response to the fragment and / or combined with an adjuvant that facilitates such a response. Antibodies of this type can be screened for differential binding to tau or fragments thereof, compared to mutants of specific residues. Screening for such mutants more precisely defines binding specificity, allowing the identification of antibodies whose binding is inhibited by mutagenesis of specific residues and which may share functional properties with other exemplified antibodies. Mutations can be systematic substitutions of alanine (or serine, if alanine is already present) with a single residue at a time, or with wider intervals, throughout the entire target or the entire segment in which the known epitope resides. If the same set of mutations significantly reduces the binding of two antibodies, then the two antibodies bind to the same epitope.

[0517] Antibodies with binding specificity to the selected mouse antibody (e.g., 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3) can also be generated using variants of the phage display method. See Winter, WO 92 / 20791. This method is particularly suitable for generating human antibodies. In this method, the heavy or light chain variable region of the selected mouse antibody is used as the starting material. If, for example, the light chain variable region is chosen as the starting material, a phage library is constructed in which members exhibit the same light chain variable region (i.e., the mouse starting material) and different heavy chain variable regions. The heavy chain variable region can be obtained, for example, from a library of rearranged human heavy chain variable regions. Selection is made based on the strong specific binding (e.g., at least 10) to tau or fragments thereof. 8 And preferably at least 10 9 M -1 The phage is then used. The heavy chain variable region from this phage is then used as starting material to construct another phage library. In this library, each phage exhibits the same heavy chain variable region (i.e., the region identified from the first display library) and different light chain variable regions. The light chain variable regions can be obtained, for example, from libraries of rearranged human variable light chain regions. Again, phages showing strong specific binding to tau or fragments thereof are selected. The resulting antibodies typically have the same or similar epitope specificity as the mouse starting material.

[0518] The CDRs of the Kabat / Chothia complex of the heavy chain of 9F5 are named SEQ ID NO:8-10, and the CDRs of the Kabat complex of the light chain of 9F5 are named SEQ ID NO:12-14.

[0519] The CDRs of the Kabat / Chothia complexes of the heavy chain of 10C12 are named SEQ ID NO:8-10, and the CDRs of the Kabat complexes of the light chain of 10C12 are named SEQ ID NO:12-14.

[0520] The CDRs of the Kabat / Chothia complex of the heavy chain of 2D11 are named SEQ ID NO:8-10, and the CDRs of the Kabat complex of the light chain of 2D11 are named SEQ ID NO:12-14.

[0521] Table 2 indicates 9F5, 10C12, and 2D11CDR as defined by the complex of Kabat, Chothia, Chothia, and Kabat (also referred to herein as the "Kabat / Chothia complex"), AbM, and Contact.

[0522] Table 2: 9F5, 10C12, and 2D11CDR as defined by Kabat, Chothia, the complex of Chothia and Kabat, AbM, and Contact.

[0523]

[0524] The CDRs of the Kabat / Chothia complex of the heavy chain of 12C4 are named SEQ ID NO:8, 220 and 10, respectively, and the CDRs of the Kabat complex of the light chain of 12C4 are named SEQ ID NO:12-14, respectively.

[0525] Table 3 shows the 12C4CDR as defined by the complex of Kabat, Chothia, Chothia and Kabat (also referred to herein as the "Kabat / Chothia complex"), AbM and Contact.

[0526] Table 3: 12C4CDR as defined by Kabat, Chothia, the complex of Chothia and Kabat, AbM, and Contact

[0527]

[0528] The CDRs of the Kabat / Chothia complex of the heavy chain of 17C12 are named SEQ ID NO:226, 227 and 10, respectively, and the Kabat CDRs of the light chain of 17C12 are named SEQ ID NO:229-231, respectively.

[0529] Table 4 shows the 17C12CDR as defined by the complex of Kabat, Chothia, Chothia and Kabat (also referred to herein as the "Kabat / Chothia complex"), AbM and Contact.

[0530] Table 4: 17C12CDR as defined by Kabat, Chothia, the complex of Chothia and Kabat, AbM, and Contact

[0531]

[0532] The CDRs of the Kabat / Chothia complexes of the heavy chain of 14H3 were named SEQ ID NO:241-243, and the CDRs of the Kabat complexes of the light chain of 14H3 were named SEQ ID NO:245-247.

[0533] Table 5 shows the 14H3CDR as defined by the complex of Kabat, Chothia, Chothia and Kabat (also referred to herein as the "Kabat / Chothia complex"), AbM and Contact.

[0534] Table 5: 14H3CDR defined as by Kabat, Chothia, the complex of Chothia and Kabat, AbM, and Contact

[0535]

[0536] Table 5: 14H3CDR defined as by Kabat, Chothia, the complex of Chothia and Kabat, AbM, and Contact

[0537]

[0538] Other antibodies can be obtained by mutagenesis of the cDNA encoding the heavy and light chains of exemplary antibodies (such as 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3). Monoclonal antibodies that are at least 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identical in amino acid sequence to 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3 in the variable region of the mature heavy and / or light chains and maintain their functional properties, and / or differ from the corresponding antibodies in that they have a few functionally insignificant amino acid substitutions (e.g., conserved substitutions), deletions, or insertions are also included in this invention. It also includes monoclonal antibodies having at least one or all six CDRs, said CDRs as defined by any conventional definition, but preferably Kabat, which have 90%, 95%, 99% or 100% identity with the corresponding CDRs of 9F5, 10C12, 2D11, 12C4, 17C12 or 14H3.

[0539] The present invention also provides antibodies having some or all (e.g., 3, 4, 5, and 6) CDRs that are wholly or substantially derived from 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3. Such antibodies may include a heavy chain variable region having at least two and typically all three CDRs that are wholly or substantially derived from 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3; and / or a light chain variable region having at least two and typically all three CDRs that are wholly or substantially derived from 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3. The antibody may include both a heavy chain and a light chain. When a CDR contains no more than 4, 3, 2, or 1 substitution, insertion, or deletion, the CDR is substantially derived from the corresponding 9F5CDR, except that CDR-H2 (as defined by Kabat) may have no more than 6, 5, 4, 3, 2, or 1 substitution, insertion, or deletion. Such antibodies may have at least 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity with 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3 in the amino acid sequence of the mature heavy chain and / or light chain variable region while maintaining their functional properties, and / or differ from 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3 by a few functionally insignificant amino acid substitutions (e.g., conserved substitutions), deletions, or insertions.

[0540] Some antibodies identified by such assays can bind to tau monomers, misfolded forms, aggregates, phosphorylated or unphosphorylated forms, or other forms. Similarly, some antibodies are immunoreactive to both non-pathological and pathological forms and conformations of tau.

[0541] The present invention also provides a means for specifically binding to a peptide composed of residues (Q / E)IVYK(S / P) (SEQ ID NO: 56), residue QIVYKP (SEQ ID NO: 57), residue EIVYKSP (SEQ ID NO: 58), or residue EIVYKS (SEQ ID NO: 277). An exemplary means is an antibody comprising the heavy chain CDRs of SEQ ID NO: 8-10 and the light chain CDRs of SEQ ID NO: 12-14. An exemplary means is an antibody comprising the heavy chain CDRs of SEQ ID NO: 8, 220, and 10 and the light chain CDRs of SEQ ID NO: 12-14. An exemplary means is an antibody comprising the heavy chain CDRs of SEQ ID NO: 226, 227, and 10 and the light chain CDRs of SEQ ID NO: 229-231. An exemplary means is an antibody comprising the heavy chain CDRs of SEQ ID NO: 241-243 and the light chain CDRs of SEQ ID NO: 245-247.

[0542] B. Non-human antibodies

[0543] The generation of other non-human antibodies (e.g., mouse, guinea pig, primate, rabbit, or rat) against tau or fragments thereof (e.g., peptides containing the amino acid sequence of QIVYKP (SEQ ID NO:57), EIVYKSP (SEQ ID NO:58), EIVYKS (SEQ ID NO:277), or (Q / E)IVYK(S / P) (SEQ ID NO:56)) can be achieved, for example, by immunizing the animal with tau or fragments thereof. See Harlow & Lane, Antibodies, A Laboratory Manual (CSHP NY, 1988) (incorporated by reference for all purposes). Optionally, the immunogen may be 383 amino acid human tau (4R0N). Optionally, the immunogen may be human tau containing the P301S mutation. Optionally, the immunogen may be human tau, wherein the human tau is recombinant with an N-terminal His tag. Optionally, animals are immunized with a tau fragment linked to a vector, comprising a peptide represented by (Q / E)IVYK(S / P) (SEQ ID NO:56). Optionally, the peptide is QIVYKP (SEQ ID NO:57), EIVYKSP (SEQ ID NO:58), or EIVYKS (SEQ ID NO:277). This immunogen can be obtained from natural sources, through peptide synthesis, or through recombinant expression. Optionally, the immunogen can be administered by fusion with or otherwise compounding with a carrier protein. Optionally, the immunogen can be administered with an adjuvant. Several types of adjuvants can be used as described below. For immunization of laboratory animals, complete Freund's adjuvant can be used, followed by incomplete adjuvants. Rabbits or guinea pigs are commonly used for the preparation of polyclonal antibodies. Mice are commonly used for the preparation of monoclonal antibodies. Antibodies are screened for specific binding to tau or tau epitopes (e.g., QIVYKP (SEQ ID NO:57), EIVYKSP (SEQ ID NO:58), EIVYKS (SEQ ID NO:277), or (Q / E)IVYK(S / P) (SEQ ID NO:56)). Optionally, screening may be performed for 15-amino acid peptides comprising QIVYKP (SEQ ID NO:57), EIVYKSP (SEQ ID NO:58), or EIVYKS (SEQ ID NO:277), or any other shared motif represented by (Q / E)IVYK(S / P) (SEQ ID NO:56). Optionally, the peptide comprises QIVYKP (SEQ ID NO:57), EIVYKSP (SEQ ID NO:58), or EIVYKS (SEQ ID NO:277).This screening can be achieved by: identifying the binding of the antibody to a set of tau variants, such as tau variants containing amino acid residues 307-312, 391-397, or 391-396 of SEQ ID NO: 1, or tau variants composed of or containing mutations within these residues; and identifying which tau variants bind to the antibody. Binding can be assessed, for example, by Western blotting, FACS, or ELISA.

[0544] C. Humanized antibodies

[0545] Humanized antibodies are genetically engineered antibodies in which a CDR from a non-human "donor" antibody is grafted into a human "recipient" antibody sequence (see, for example, Queen, US 5,530,101 and 5,585,089; Winter, US 5,225,539; Carter, US 6,407,213; Adair, US 5,859,205; and Foote, US 6,881,557). The recipient antibody sequence can be, for example, a mature human antibody sequence, a complex of such sequences, a common sequence of a human antibody sequence, or a germline region sequence. Thus, a humanized antibody is an antibody having at least three, four, five, or all of the CDRs, wholly or substantially derived from the donor antibody, and a variable region framework sequence and a constant region (if present), wholly or substantially derived from the human antibody sequence. Similarly, the humanized heavy chain has at least one, two, and usually all three CDRs that are entirely or substantially derived from the donor antibody heavy chain, and a heavy chain variable region framework sequence and a heavy chain constant region (if present) that are substantially derived from the human heavy chain variable region framework and constant region sequences. Similarly, the humanized light chain has at least one, two, and usually all three CDRs that are entirely or substantially derived from the donor antibody light chain, and a light chain variable region framework sequence and a light chain constant region (if present) that are substantially derived from the human light chain variable region framework and constant region sequences. Humanized antibodies, excluding nanobodies and dAbs, comprise humanized heavy chains and humanized light chains. When at least 85%, 90%, 95%, or 100% of the corresponding residues (as defined by any conventional definition, but preferably by Kabat) are identical between the corresponding CDRs, the CDRs in the humanized antibody are substantially derived from the corresponding CDRs in the non-human antibody. When at least 85%, 90%, 95%, or 100% of the corresponding residues as defined by Kabat are identical, the variable region frame sequence or the constant region of the antibody chain is substantially derived from the human variable region frame sequence or the human constant region, respectively. To be classified as humanized according to the 2014 World Health Organization (WHO) International Non-Proprietary Name (INN) definition, an antibody must have at least 85% identity with a human germline antibody sequence (i.e., prior to somatic hypermutation). A chimeric antibody is an antibody in which one antibody chain (e.g., the heavy chain) meets a threshold but another chain (e.g., the light chain) does not. If neither chain meets the threshold, the antibody is classified as chimeric, even if the variable frame regions of both chains are substantially human and have some mouse reversion mutations. See Jones et al. (2016) The INNs and outs of antibody nonproprietary names, mAbs8:1,1-9, DOI:10.1080 / 19420862.2015.1114320.See also “WHO-INN: International nonproprietary names (INN) for biological and biotechnological substances (a review)” (Internet) 2014. Available at: http: / / www.who.int / medicines / services / inn / BioRev2014.pdf, which is incorporated herein by reference. For the avoidance of ambiguity, the term “humanized” as used herein is not intended to limit the 2014 WHO INN definition of humanized antibodies. Some humanized antibodies described herein have at least 85% sequence identity with human germline sequences, and some humanized antibodies described herein have less than 85% sequence identity with human germline sequences. Some heavy chains of humanized antibodies described herein have approximately 60% to 100% sequence identity with human germline sequences, for example, in the range of approximately 60% to 69%, 70% to 79%, 80% to 84%, or 85% to 89%. Some heavy chains do not meet the 2014 WHO INN definition and have sequence identity with human germline sequences, for example, approximately 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, or 82%, 83%, or 84%, while other heavy chains meet the 2014 WHO INN definition and have sequence identity with human germline sequences of approximately 85%, 86%, 87%, 88%, 89%, or higher. Some light chains of the humanized antibodies presented herein have sequence identity with human germline sequences of approximately 60% to 100%, for example, in the range of approximately 80% to 84% or 85% to 89%. Some light chains do not meet the 2014 WHO INN definition and have, for example, about 81%, 82%, 83%, or 84% sequence identity with human germline sequences, while other light chains meet the 2014 WHO INN definition and have about 85%, 86%, 87%, 88%, 89%, or higher sequence identity with human germline sequences. Some humanized antibodies provided herein, defined as “chimeric” according to the 2014 WHO INN definition, have a heavy chain with less than 85% sequence identity with a human germline sequence, paired with a light chain with less than 85% sequence identity with a human germline sequence. Some humanized antibodies provided herein, defined as “mixed” according to the 2014 WHO INN definition, have, for example, a heavy chain with at least 85% sequence identity with a human germline sequence, paired with a light chain with less than 85% sequence identity with a human germline sequence, or vice versa.Some of the humanized antibodies described herein meet the 2014 WHO INN definition of “humanized” and have a heavy chain with at least 85% sequence identity to a human germline sequence, said heavy chain being paired with a light chain having at least 85% sequence identity to a human germline sequence. An exemplary 12C4 antibody conforming to the 2014 WHO INN definition of “humanized” includes an antibody having a mature heavy chain with the amino acid sequence of SEQ ID NO:221 or SEQ ID NO:222, said mature heavy chain being paired with a mature light chain with the amino acid sequence of SEQ ID NO:223 or SEQ ID NO:224. An exemplary 14H3 antibody conforming to the 2014 WHO INN definition of “humanized” includes an antibody having a mature heavy chain with the amino acid sequence of SEQ ID NO:248 or SEQ ID NO:249, said mature heavy chain being paired with a mature light chain with the amino acid sequence of SEQ ID NO:251 or SEQ ID NO:252. Some of the humanized antibodies described herein meet the 2014 WHO INN definition of “mixed”. Exemplary 9F5 antibodies conforming to the 2014 WHO INN definition of “mixed” include antibodies having a mature heavy chain having an amino acid sequence of any one of SEQ ID NO:15-22 and SEQ ID NO:127-128, said mature heavy chain being paired with a mature light chain sequence having an amino acid sequence of any one of SEQ ID NO:26-29 and SEQ ID NO:130-131. Exemplary 10C12 antibodies conforming to the 2014 WHO INN definition of “humanized” include antibodies having a mature heavy chain having an amino acid sequence of SEQ ID NO:214 or SEQ ID NO:215, said mature heavy chain being paired with a mature light chain sequence having an amino acid sequence of SEQ ID NO:216 or SEQ ID NO:217. Exemplary 17C12 antibodies conforming to the 2014 WHO INN definition of “humanization” include antibodies having a mature heavy chain having the amino acid sequence of SEQ ID NO:232 or SEQ ID NO:233, said mature heavy chain being paired with a mature light chain having the amino acid sequence of SEQ ID NO:235. Further humanized 9F5 antibodies of the present invention include antibodies having a mature heavy chain having the amino acid sequence of any one of SEQ ID NO:15-22 and SEQ ID NO:127-128, said mature heavy chain being paired with a mature light chain having the amino acid sequence of any one of SEQ ID NO:23-25.Another humanized 17C12 antibody of the present invention includes an antibody having a mature heavy chain having an amino acid sequence of SEQ ID NO:232 or SEQ ID NO:233, said mature heavy chain being paired with a mature light chain having an amino acid sequence of SEQ ID NO:234.

[0546] Although humanized antibodies are typically incorporated with all six CDRs from mouse antibodies (as defined by any conventional definition, but preferably by Kabat), they can also be prepared with fewer than all the CDRs from mouse antibodies (e.g., at least 3, 4, or 5 CDRs) (e.g., Pascalis et al., J. Immunol. 169:3076, 2002; Vajdos et al., J. of Mol. Biol., 320:415-428, 2002; Iwahashi et al., Mol. Immunol. 36:1079-1091, 1999; Tamura et al., J. Immunol., 164:1432-1441, 2000).

[0547] In some antibodies, only a subset of the CDRs, i.e., the set of CDR residues required for binding, called the SDR, is needed to maintain binding in the humanized antibody. CDR residues that do not contact the antigen and are not in the SDR can be identified from the Kabat CDR region located outside the Chothia hypervariable ring (Chothia, J. Mol. Biol. 196:901, 1987) based on previous research (e.g., residues H60-H65 in CDR H2 are generally not required), through molecular modeling and / or empirically, or as described in Gonzales et al., Mol. Immunol. 41:863, 2004. In such humanized antibodies, at locations where one or more donor CDR residues are absent or the entire donor CDR is omitted, the amino acid occupying that position can be the amino acid occupying the corresponding position (by Kabat number) in the receptor antibody sequence. The number of such substitutions of the receptor for the donor amino acid that will be included in the CDR reflects a balance of competitive considerations. Such substitutions may benefit by reducing the number of mouse amino acids in humanized antibodies, and thus decreasing potential immunogenicity and / or meeting the WHO INN definition of “humanization.” However, substitutions can also cause alterations in affinity, and a significant reduction in affinity is preferred to be avoided. The position within the CDR for substitution and the amino acid used for substitution can also be selected empirically.

[0548] The human receptor antibody sequence may optionally be selected from many known human antibody sequences to provide a high degree of sequence identity (e.g., 65%–85% identity) between the variable region frame of the human receptor sequence and the corresponding variable region frame of the donor antibody chain.

[0549] Some humanized and chimeric antibodies possess the same (within the experimental error range) or improved functional properties, such as binding affinity for human tau, as described in the examples, similar to the inhibition of tau internalization into neurons by the mouse antibodies that derived from them. For example, the binding affinity of some humanized and chimeric antibodies is 3, 2, or 1 times that of the mouse antibodies that derived from them, or the affinity is indistinguishable within the experimental error range. Some humanized and chimeric antibodies inhibit tau internalization into neurons to the extent that, as described in the examples, it is 3, 2, or 1 times that of the mouse antibodies that derived from them, or the extent of inhibition is the same as that of the mouse antibodies that derived from them within the experimental error range.

[0550] An example of a receptor sequence for the 9F5 heavy chain is the human mature heavy chain variable region of 48G7Fab with PDB accession code 2RCS-VH_huFrwk (SEQ ID NO:32). Another example of a receptor sequence for the 9F5 heavy chain is the human mature heavy chain GenBank AAN16432-VH_huFrwk (SEQ ID NO:31). The CDR-H1 and H2 loops of the variable domains of 9F5 and 48G7Fab also share the same length. An example of a receptor sequence for the 9F5 heavy chain is the human mature heavy chain variable region IMGT#IGHV1-69-2*01 (SEQ ID NO:33). The Chothia CDR-H1 of IMGT#IGHV1-69-2*01 (SEQ ID NO:33) is canonical class 1, while the Chothia CDR-H2 is canonical class 2. IMGT#IGHV1-69-2*01 (SEQ ID NO:33) belongs to subgroup 1 of the human heavy chain. An example of a receptor sequence for the 9F5 light chain is the human mature light chain variable region 1911357B-VL_huFrwk (SEQ ID NO:36). An example of a receptor sequence for the 9F5 light chain is the human mature light chain variable region CAB51297-VL_huFrwk (SEQ ID NO:35). The CDR-L1, L2, and L3 loops of the light chain variable domains of the 9F5 and CAB51297 & 1911357B antibodies share the same length. An example of a receptor sequence for the 9F5 light chain is the human mature light chain variable region IGKV2-28*01 & IGKJ2*01 (SEQ ID NO:37). Chothia CDR-L1 of IGKV2-28*01 & IGKJ2*01 (SEQ ID NO:37) is normative category 4. Chothia CDR-L2 of IGKV2-28*01 & IGKJ2*01 (SEQ ID NO:37) is normative category 1. Chothia CDR-L3 of IGKV2-28*01 & IGKJ2*01 (SEQ ID NO:37) is normative category 1. IGKV2-28*01 & IGKJ2*01 (SEQ ID NO:37) belongs to human κ subgroup 2.

[0551] An example of a receptor sequence for the 10C12 heavy chain is the human mature CAC20421 (SEQ ID NO: 218). The variable domain of 10C12 and the CDR-H1 and H2 loops of CAC20421VH also share the same length. An example of a receptor sequence for the 10C12 heavy chain is the human mature heavy chain variable region IMGT#IGHV1-69-2*01 (SEQ ID NO: 33). The Chothia CDR-H1 of IMGT#IGHV1-69-2*01 (SEQ ID NO: 33) is canonical class 1, while the Chothia CDR-H2 is canonical class 2. IMGT#IGHV1-69-2*01 (SEQ ID NO: 33) belongs to subgroup 1 of the human heavy chain. An example of a receptor sequence for the 10C12 light chain is the human mature light chain variable region CAB51297-VL_huFrwk (SEQ ID NO: 35). The variable domain of the 10C12 light chain and the CDR-L1, L2, and L3 loops of CAB51297VL share the same length. An example of a receptor sequence for the 10C12 light chain is the human mature light chain variable region IGKV2-28*01 & IGKJ2*01 (SEQ ID NO:37). The Chothia CDR-L1 of IGKV2-28*01 & IGKJ2*01 (SEQ ID NO:37) is canonical class 4. The Chothia CDR-L2 of IGKV2-28*01 & IGKJ2*01 (SEQ ID NO:37) is canonical class 1. The Chothia CDR-L3 of IGKV2-28*01 & IGKJ2*01 (SEQ ID NO:37) is canonical class 1. IGKV2-28*01 & IGKJ2*01 (SEQ ID NO:37) belong to human κ subgroup 2.

[0552] An example of a receptor sequence for the 12C4 heavy chain is the human mature heavy chain variable region CAC20421-VH_huFrwk (SEQ ID NO:218). The variable domain of 12C4 and the CDR-H1 and H2 loops of CAC20421VH also share the same length. An example of a receptor sequence for the 12C4 heavy chain is the human mature heavy chain variable region IMGT#IGHV1-69-2*01 (SEQ ID NO:33). Chothia CDR-H1 of IMGT#IGHV1-69-2*01 (SEQ ID NO:33) is canonical class 1, while Chothia CDR-H2 is canonical class 2. IMGT#IGHV1-69-2*01 (SEQ ID NO:33) belongs to subgroup 1 of the human heavy chain. An example of a receptor sequence for the 12C4 light chain is the human mature light chain variable region CAB51297-VL_huFrwk (SEQ ID NO:35). The 12C4 light chain variable domain and the CDR-L1, L2, and L3 loops of CAB51297VL share the same length. An example of a receptor sequence for the 12C4 light chain is the human mature light chain variable region IGKV2-28*01&IGKJ2*01 (SEQ ID NO:37). The Chothia CDR-L1 of IGKV2-28*01&IGKJ2*01 (SEQ ID NO:37) is canonical class 4. The Chothia CDR-L2 of IGKV2-28*01&IGKJ2*01 (SEQ ID NO:37) is canonical class 1. Chothia CDR-L3 of IGKV2-28*01 & IGKJ2*01 (SEQ ID NO:37) is a normative category 1. IGKV2-28*01 & IGKJ2*01 (SEQ ID NO:37) belong to human κ subgroup 2.

[0553] An example of a receptor sequence for the 17C12 heavy chain is the human mature heavy chain variable region CAC20421-VH_huFrwk (SEQ ID NO:218). The CDR-H1 and H2 loops of the heavy chain variable domains of 17C12 and CAC20421 also share the same length. An example of a receptor sequence for the 17C12 heavy chain is the human mature heavy chain variable region IMGT#IGHV1-69-2*01 (SEQ ID NO:33). The Chothia CDR-H1 of IMGT#IGHV1-69-2*01 (SEQ ID NO:33) is canonical class 1, while the Chothia CDR-H2 is canonical class 2. IMGT#IGHV1-69-2*01 (SEQ ID NO:33) belongs to subgroup 1 of the human heavy chain. An example of a receptor sequence for the 17C12 light chain is the human mature light chain variable region QDO16713-VL_huFrwk (SEQ ID NO:238). The CDR-L1, L2, and L3 loops of the light chain variable domains of the 17C12 and QDO16713 antibodies also share the same length. An example of a receptor sequence for the 17C12 light chain is the human mature light chain variable region IGKV2-29*02&IGKJ4*01 (SEQ ID NO:239).

[0554] An example of a receptor sequence for the 14H3 heavy chain is the human mature heavy chain variable region QDJ57937-VH_huFrwk (SEQ ID NO:253). The variable domain of 14H3 and the CDR-H1 and H2 loops of QDJ57937VH also share the same length. An example of a receptor sequence for the 14H3 heavy chain is the human mature heavy chain variable region IGHV1-70*04&IGHJ4*01 (SEQ ID NO:254). An example of a receptor sequence for the 14H3 light chain is the human mature light chain variable region ABC66914-VL_huFrwk (SEQ ID NO:256). The variable domain of the 14H3 light chain and the CDR-L1, L2, and L3 loops of ABC66914VL share the same length. An example of a receptor sequence for the 14H3 light chain is the human mature light chain variable region IGKV2-28*01 & IGKJ2*01 (SEQ ID NO:37). Chothia CDR-L1 of IGKV2-28*01 & IGKJ2*01 (SEQ ID NO:37) is canonical class 4. Chothia CDR-L2 of IGKV2-28*01 & IGKJ2*01 (SEQ ID NO:37) is canonical class 1. Chothia CDR-L3 of IGKV2-28*01 & IGKJ2*01 (SEQ ID NO:37) is canonical class 1. IGKV2-28*01 & IGKJ2*01 (SEQ ID NO:37) belongs to human κ subgroup 2.

[0555] If more than one human receptor antibody sequence is chosen, then complexes or hybrids of those receptors can be used, and the amino acids used at different positions in the humanized light and heavy chain variable regions can be derived from any human receptor antibody sequence used. For example, the human mature heavy chain variable region of humanized 48G7Fab (SEQ ID NO:32) with PDB accession code 2RCS-VH_huFrwk (SEQ ID NO:31) and humanized 48G7Fab (SEQ ID NO:32) with PDB accession code 2RCS-VH_huFrwk can be used as the receptor sequence for the humanization of the 9F5 mature heavy chain variable region. Examples of locations where the two receptors differ include positions H1 (E or Q), H5 (V or Q), H11 (V or L), H12 (K or V), H20 (V or L), H23 (K or T), H28 (T or N), H38 (R or K), H40 (A or R), H42 (G or E), H43 (K or Q), H48 (M or I), H54 (D or N), H66 (R or K), H69 (M or I), H75 (T or S), H76 (D or N), H80 (M or L), H81 (E or Q), H83 (R or T), H108 (L or T), or H109 (V or L). Humanized versions of the 9F5 heavy chain variable region can include any amino acid at any of these positions. The germline sequence IMGT#IGHV1-69-2*01 (SEQ ID NO:25) was also used as the receptor sequence for the humanization of the 9F5 mature heavy chain variable region. For example, the human mature light chain variable regions of CAB51297-VL_huFrwk (SEQ ID NO:35) and 1911357B-VL_huFrwk (SEQ ID NO:36) were used as receptor sequences for the humanization of the 9F5 mature light chain variable region. Examples of positions where these two receptors differ include positions L7 (S or A), L8 (P or A), L9 (L or F), L11 (L or N), L15 (P or L), L17 (E or T), L18 (P or S), L30 (Y or I), L31 (N or T), L54 (R or L), L60 (D or N), L66 (G or E), or L74 (K or R). The humanized form of the 9F5 light chain variable region can include any amino acid at any of these positions. The germline sequences IGKV2-28*01&IGKJ2*01 (SEQ ID NO:37) were also used as the receptor sequences for the humanization of the mature 9F5 light chain variable region.

[0556] Substitutions can be selected based on the potential impact of certain amino acids from human variable region framework residues on CDR conformation and / or binding to antigens. Studies of such potential impacts are conducted by modeling, examining the characterization of amino acids at specific positions, or empirically observing the effects of substitution or mutagenesis of specific amino acids.

[0557] For example, when the amino acids between mouse variable region framework residues and selected human variable region framework residues differ, human framework amino acids can be substituted by equivalent framework amino acids from mouse antibodies when the following characteristics are reasonably expected:

[0558] (1) Directly non-covalently bound antigens;

[0559] (2) Adjacent to a CDR area or within a CDR defined by Chothia but not Kabat;

[0560] (3) Or interact with the CDR region (e.g., in the CDR region approximately (e.g., by modeling the light or heavy chains on the resolved structures of homologous known immunoglobulin chains); or

[0561] (4) are residues involved in the VL-VH interface.

[0562] In one implementation, a two-stage PCR protocol is used to generate humanized sequences, which allows for the introduction of multiple mutations, deletions, and insertions using QuikChange site-directed mutagenesis [Wang, W. and Malcolm, BA (1999) BioTechniques 26:680-682].

[0563] Framework residues from categories (1) to (3) as defined in Queen, US 5,530,101 are sometimes alternatively referred to as canonical residues and vernier residues. Framework residues that help define the conformation of the CDR ring are sometimes referred to as canonical residues (Chothia & Lesk, J. Mol. Biol. 196:901-917 (1987); Thornton & Martin, J. Mol. Biol. 263:800-815 (1996)). Framework residues that support the conformation of the antigen-binding ring and play a role in finely regulating the interaction between the antibody and the antigen are sometimes referred to as vernier residues (Foote & Winter, J. Mol. Biol. 224:487-499 (1992)).

[0564] Other framework residues that are candidates for substitution are residues that generate potential glycosylation sites. Other substitution candidates are recipient human framework amino acids that are uncommon for human immunoglobulins at that position. These amino acids can be substituted with amino acids from mouse donor antibodies at equivalent positions or from more typical human immunoglobulins.

[0565] Other framework residues that could be considered for substitution are the N-terminal glutamine residue (Q), which could be replaced with glutamate (E) to minimize the possibility of pyroglutamate conversion [Y. Diana Liu et al., 2011, J. Biol. Chem., 286: 11211-11217]. The conversion from glutamate (E) to pyroglutamate (pE) occurs more slowly than from glutamine (Q). Due to the loss of primary amine in the glutamine-to-pE conversion, the antibody becomes more acidic. Incomplete conversion results in antibody heterogeneity, which can be observed as multiple peaks using charge-based analytical methods. Differences in heterogeneity may indicate a lack of process control.Exemplary 9F5 humanized heavy chain variable regions having N-terminal glutamine-to-glutamic acid substitutions are SEQ ID NO:16 (hu9F5VHv2), SEQ ID NO:17 (hu9F5VHv3), SEQ ID NO:18 (hu9F5VHv4), SEQ ID NO:19 (hu9F5VHv5), SEQ ID NO:20 (hu9F5VHv6), SEQ ID NO:21 (hu9F5VHv7), SEQ ID NO:22 (hu9F5VHv8), SEQ ID NO:109 (hu9F5VHv4_L80P), SEQ ID NO:110 (hu9F5VHv4_L80D), SEQ ID NO:111 (hu9F5VHv4_L82cG), SEQ ID NO:112 (hu9F5VHv4_L82cD), and SEQ ID NO:109 (hu9F5VHv4_L80P). NO: 113 (hu9F5VHv4_L82P), SEQ ID NO: 114 (hu9F5VHv4_L80G), SEQ ID NO: 115 (hu9F5VHv4_L82K), SEQ ID NO: 116 (hu9F5VHv4_L82R), SEQ ID NO:117(hu9F5VHv4_L82E), SEQ ID NO:118(hu9F5VHv4_L82N), SEQ ID NO:119(hu9F5VHv4_Y79D), SEQ ID NO:120(hu9F5VHv4_Y79N), SEQ ID NO:121(hu9F5VHv4_Y79G), SEQ ID NO:122(hu9F5VHv5_M80E), SEQ ID NO:123(hu9F5VHv5_M80G), SEQ SEQ ID NO:124 (hu9F5VHv4_L82cS), SEQ ID NO:125 (hu9F5VHv4_Y79Q), SEQ ID NO:126 (hu9F5VHv4_S82aG), SEQ ID NO:127 (hu9F5VHv9), SEQ ID NO:128 (hu9F5VHv10), SEQ ID NO:129 (hu9F5VHv10_L82cG). An exemplary 10C12 humanized heavy chain variable region having an N-terminal glutamine-to-glutamic acid substitution is SEQ ID NO:215 (hu10C12VHv2). An exemplary 12C4 humanized heavy chain variable region having an N-terminal glutamine-to-glutamic acid substitution is SEQ ID NO:222 (hu12C4VHv2). An exemplary 17C12 humanized heavy chain variable region having an N-terminal glutamine-to-glutamic acid substitution is SEQ ID NO:233 (hu17C12VHv2).

[0566] An exemplary humanized antibody includes a humanized form of mouse 9F5, named Hu9F5.

[0567] Mouse antibody 9F5 includes mature heavy chain and light chain variable regions, the mature heavy chain and light chain variable regions having amino acid sequences comprising SEQ ID NO:7 and SEQ ID NO:11, respectively. This invention provides 29 exemplified humanized mature heavy chain variable regions: hu9F5VHv1 (SEQ ID NO:15), hu9F5VHv2 (SEQ ID NO:16), hu9F5VHv3 (SEQ ID NO:17), hu9F5VHv4 (SEQ ID NO:18), hu9F5VHv5 (SEQ ID NO:19), hu9F5VHv6 (SEQ ID NO:20), hu9F5VHv7 (SEQ ID NO:21), hu9F5VHv8 (SEQ ID NO:22), hu9F5VHv4_L80P (SEQ ID NO:109), hu9F5VHv4_L80D (SEQ ID NO:110), hu9F5VHv4_L82cG (SEQ ID NO:111), hu9F5VHv4_L82cD (SEQ ID NO:15). NO:112), hu9F5VHv4_L82P (SEQ ID NO:113), hu9F5VHv4_L80G (SEQ ID NO:114), hu9F5VHv4_L82K (SEQ ID NO 115:), hu9F5VHv4_L82R (SEQ ID NO:116), hu9F5VHv4_L82E (SEQ ID NO:117), hu9F5VHv4_L82N (SEQ ID NO:118), hu9F5VHv4_Y79D (SEQ ID NO:119), hu9F5VHv4_Y79N (SEQ ID NO:120), hu9F5VHv4_Y79G (SEQ ID NO:121), hu9F5VHv5_M80E (SEQ ID NO:122), hu9F5VHv5_M80G (SEQ ID The invention also provides 95 exemplified mature light chain variable regions: hu9F5VHv4_L82cS (SEQ ID NO:123), hu9F5VHv4_Y79Q (SEQ ID NO:125), hu9F5VHv4_S82aG (SEQ ID NO:126), hu9F5VHv9 (SEQ ID NO:127), hu9F5VHv10 (SEQ ID NO:128), and hu9F5VHv10_L82cG (SEQ ID NO:129).NO:25)、hu9F5VLv4(SEQ ID NO:26)、hu9F5VLv5(SEQ IDNO:27)、hu9F5VLv6(SEQ ID NO:28)、hu9F5VLv7(SEQ ID NO:29)、hu9F5VLv8(SEQ ID NO:130)、hu9F5VLv9(SEQ ID NO:131)、hu9F5VLv2_M51E(SEQ ID NO:61)、hu9F5VLv2_M51D(SEQIDNO:62)、hu9F5VLv2_L27cD(SEQ ID NO:63)、hu9F5VLv2_L27cG(SEQ ID NO:64)、hu9F5VLv2_L27cS(SEQ ID NO:65)、hu9F5VLv2_L27cE(SEQ ID NO:66)、hu9F5VLv2_I30E(SEQ IDNO:67)、hu9F5VLv2_I30K(SEQ ID NO:68)、hu9F5VLv2_L27cT(SEQ ID NO:69)、hu9F5VLv2_L27cN(SEQ ID NO:70、hu9F5VLv2_L27bD(SEQ ID NO:71)、hu9F5VLv2_I30G(SEQID NO:72)、hu9F5VLv2_L33N(SEQ ID NO:73)、hu9F5VLv2_L27cA(SEQ IDNO:74)、hu9F5VLv2_L33T(SEQ ID NO:75)、hu9F5VLv2_L33S(SEQID NO:76)、hu9F5VLv2_L33R(SEQID NO:77)、hu9F5VLv2_I30Q(SEQ ID NO:78)、hu9F5VLv2_L27bT(SEQ ID NO:79、hu9F5VLv2_T31G(SEQ ID NO:80)、hu9F5VLv2_L27bQ(SEQ IDNO:81)、hu9F5VLv2_L33G(SEQID NO:82)、hu9F5VLv2_L27cP(SEQID NO:83)、hu9F5VLv2_V78R(SEQ ID NO:84)、hu9F5VLv2_I75D(SEQ ID NO:85)、hu9F5VLv2_V78D(SEQ ID NO:86)、hu9F5VLv2_V78E(SEQID NO:87)、hu9F5VLv2_V78P(SEQ IDNO:88)、hu9F5VLv2_V78K(SEQ IDNO:89)、hu9F5VLv2_R77D(SEQID NO:90)、hu9F5VLv2_V78G(SEQ ID NO:91)、hu9F5VLv2_S76P(SEQ ID NO:92)、hu9F5VLv2_I75P(SEQ ID NO:93)、hu9F5VLv2_I75Q(SEQ ID NO:94)、hu9F5VLv2_I75G(SEQID NO:95)、hu9F5VLv2_L73P(SEQ ID NO:96)、hu9F5VLv2_L73G(SEQ IDNO:97)、hu9F5VLv2_V78Q(SEQ ID NO:98)、hu9F5VLv2_S76G(SEQID NO:99)、hu9F5VLv2_L92D(SEQ ID NO:100)、hu9F5VLv2_Y86T(SEQ ID NO:101)、hu9F5VLv2_L92E(SEQ ID NO:102)、hu9F5VLv2_L92G(SEQ ID NO:103)、hu9F5VLv2_L92Q(SEQ IDNO:104)、hu9F5VLv2_L93G(SEQ ID NO:105)、hu9F5VLv2_V85G(SEQ ID NO:106)、hu9F5VLv2_L92T(SEQ ID NO:107)、hu9F5VLv2_A89G(SEQ ID NO:108)、hu9F5VLv8_DIM1(SEQ IDNO:132)、hu9F5VLv8_DIM2(SEQ ID NO:133)、hu9F5VLv8_DIM3(SEQ ID NO:134)、hu9F5VLv8_DIM4(SEQ ID NO:135)、hu9F5VLv8_DIM5(SEQ ID NO:136)、hu9F5VLv8_DIM6(SEQ IDNO:137)、hu9F5VLv8_DIM7(SEQ ID NO:138、hu9F5VLv8_DIM8(SEQ ID NO:139)、hu9F5VLv8_DIM9(SEQ ID NO:140)、hu9F5VLv8_DIM10(SEQ ID NO:141)、hu9F5VLv8_DIM11(SEQ IDNO:142)、hu9F5VLv8_DIM12(SEQ ID NO:143)、hu9F5VLv8_DIM13(SEQ ID NO:144)、hu9F5VLv8_DIM14(SEQ IDNO:145), hu9F5VLv8_DIM15 (SEQ ID NO:146), hu9F5VLv8_DIM16 (SEQ ID NO:147), hu9F5VLv8_DIM17 (SEQ ID NO:148), hu9F5VLv8_DIM18 (SEQ ID NO:149), hu9F5VLv8_DIM19 (SEQ ID NO:150), hu9F5VLv8_DIM20 (SEQ ID NO:151), hu9F5VLv8_DIM21 (SEQ ID NO:152), hu9F5VLv8_DIM22 (SEQ ID NO:153), hu9F5VLv8_DIM23 (SEQ ID NO:154), hu9F5VLv8_DIM24 (SEQ ID NO:155), hu9F5VLv8_DIM25 (SEQ ID NO:156), hu9F5VLv8_DIM26 (SEQ ID NO:157), hu9F5VLv8_DIM27 (SEQ ID NO:158), hu9F5VLv8_DIM28 (SEQ ID NO:159), hu9F5VLv8_DIM29 (SEQ ID NO:160), hu9F5VLv8_DIM30 (SEQ ID NO:161), hu9F5VLv9_DIM1 (SEQ ID NO:162), hu9F5VLv9_DIM2 (SEQ ID NO:163), hu9F5VLv9_DIM4 (SEQ ID NO:164), hu9F5VLv9_DIM5 (SEQ ID NO:165), hu9F5VLv9_DIM8) (SEQ ID NO:166), hu9F5VLv9_DIM10 (SEQ ID NO:167), hu9F5VLv9_DIM11 (SEQ ID NO:168), (hu9F5VLv9_DIM13 (SEQ ID NO:169), hu9F5VLv9_DIM19 (SEQ ID NO:170), and hu9F5VLv9_DIM20 (SEQ ID NO:171). Figures 1A to 1B and Figures 4A to 4B The comparison of the heavy chain variable regions of mouse 9F5 and various humanized antibodies is shown. Figures 2A to 2B and Figures 5A to 5B The comparison of the light chain variable regions of mouse 9F5 and various humanized antibodies is shown. Figures 6A to 6C The comparison of the light chain variable regions of the humanized variant hu9F5VLv8 and various humanized antibodies is shown.

[0568] For reasons such as potentially affecting CDR conformation and / or binding to antigens, mediating interactions between heavy and light chains, interactions with constant regions, being a site of desired or undesirable post-translational modifications, being an uncommon residue in its position within the human variable region sequence and therefore potentially immunogenic, reducing aggregation potential, and other reasons, the following 48 variable region framework positions were considered as substitution candidates for the 95 illustrated human mature light chain variable regions and 29 illustrated human mature heavy chain variable regions, as further specified in the examples: L3 (V3Q), L7 (A7S), L8 (A... 8P), L9(F9L), L11(N11L), L15(L15P), L17(T17E), L18(S18P), L37(L37Q, L37G, L37I), L39(R39K), L60(N60D), L64(G64S) , L66(E66G), L73(L73P, L73G), L74(R74K), L75(I75D, I75P, I75Q, I75G), L76(S76P, S76G), L77(R77D), L78(V78R, V78D, V7 8E, V78P, V78K, V78G, V78Q), L85(V85G), L86(Y86T), L100(G100Q), H1(Q1E), H5(Q5V), H11(L11V), H12(V12K), H17(S17T) , H20(L20I), H23(T23K), H38(K38R, K38Q), H40(R40A), H42(E42G), H43(Q43K), H48(I48M), H66(K66R), H69(I69M), H75(S7 5T), H76(N76D), H79(Y79Q, Y79D, Y79N, Y79G), H80(L80M, L80P, L80D, L80G, L80E), H81(Q81E), H82(L82P, L82K, L82R, L82R, L82E, L82N), H82a(S82aG), H82c(L82cG, L82cD, L82cS), H83(T83R), H93(A93T), H94(S94T), H108(T108L), and H109(L109V).The following 14 variable region CDR positions are considered as substitution candidates for the 95 illustrated human mature light chain variable regions and the 29 illustrated human mature heavy chain variable regions, as further specified in the examples: L27b (L27bD, L27bT, L27bQ), L27c (L27cD, L27cG, L27cS, L27cE, L27cT, L27cN, L27cA, L27cP, L27cI), L30 (I30Y, I30E, I30K, I30G, I30Q), L31 (T31N, T31G), L33 (L33N, L33T, L33S, L33R, L33G), L51 (M51G, M51E, M51D, M51K, M51I), L54 (L54R, L54G, L54T), L89 (A89G), L92 (L92D, L92E, L92G, L92Q, L92T, L92I), L93 (E93G), H28 (N28T), H51 (I48M), H54 (N54D), and H56 (D56E). In some humanized 9F5 antibodies, the Kabat-Chothia complex CDR-H1 has an amino acid sequence containing SEQ ID NO:50. In some humanized 9F5 antibodies, the Kabat-Chothia complex CDR-H1 has the amino acid sequence containing SEQ ID NO:50, and the Kabat CDR-H2 has the amino acid sequence containing SEQ ID NO:51. In some humanized 9F5 antibodies, the Kabat CDR-H2 has the amino acid sequence containing SEQ ID NO:51. In some humanized 9F5 antibodies, the Kabat CDR-H2 has the amino acid sequence containing SEQ ID NO:52. In some humanized 9F5 antibodies, the Kabat CDR-L2 has the amino acid sequence containing SEQ ID NO:55. In some humanized 9F5 antibodies, the Kabat CDR-L1 has the amino acid sequence containing SEQ ID NO:53, and the Kabat CDR-L2 has the amino acid sequence containing SEQ ID NO:55. In some humanized 9F5 antibodies, the Kabat CDR-L1 has the amino acid sequence containing SEQ ID NO:54, and the Kabat CDR-L2 has the amino acid sequence containing SEQ ID NO:55. Among some humanized 9F5 antibodies, Kabat CDR-L1 has an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NO:172-193. Among some humanized 9F5 antibodies, Kabat CDR-L2 has an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NO:194-205.Among some humanized 9F5 antibodies, Kabat CDR-L3 has an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NO:206-213.

[0569] Here, as elsewhere, the first mentioned residue is the residue of the humanized antibody formed by grafting Kabat CDR or the complex Chothia-Kabat CDR (in the case of CDR-H1) into the human receptor framework, and the second mentioned residue is the residue that is considered to replace this residue. Thus, within the variable region framework, the first mentioned residue is human, and within the CDR, the first mentioned residue is mouse.

[0570] The exemplified antibodies include any permutation or combination of the exemplified mature heavy and light chain variable regions: hu9F5VHv1 / hu9F5VLv1, hu9F5VHv1 / hu9F5VLv2, hu9F5VHv1 / hu9F5VLv3, hu9F5VHv1 / hu9F5VLv4, hu9F5VLv1 / hu9F5VLv5, hu9F5VHv1 / hu9F5VLv6, hu9F5VHv1 / hu9F5VLv7, hu9F5VHv2 / hu9F5VLv1, hu9F5VHv2 / hu9F5VLv2, hu9F5VHv2 / hu9F5VLv3, hu9F5VHv2 / hu9F5VLv4, hu9F5VLv2 / hu9F5VLv5, hu9F5VHv2 / hu9F5VLv6, hu9F5VHv2 / hu9F5VLv7, hu9F5VHv3 / hu9F5VLv1, hu9F5VHv3 / hu9F5VLv2, hu9F5VHv3 / hu9F5VLv3, hu9F5VHv3 / hu9F5VLv4, hu9F5VLv3 / hu9F5VLv5, hu9F5VHv3 / hu9F5VLv6, hu9F5VHv3 / hu9F5VLv7, hu9F5VHv4 / hu9F5VLv1, hu9F5VHv4 / hu9F5VLv2, hu9F5VHv4 / hu9F5VLv3, hu9F5VHv4 / hu9F5VLv4, hu9F5VLv4 / hu9F5VLv5, hu9F5VHv4 / hu9F5VLv6, hu9F5VHv4 / hu9F5VLv7, hu9F5VHv5 / hu9F5VLv1, hu9F5VHv5 / hu9F5VLv2, hu9F5VHv5 / hu9F5VLv3, hu9F5VHv5 / hu9F5VLv4, hu9F5VLv5 / hu9F5VLv5, hu9F5VHv5 / hu9F5VLv6, hu9F5VHv5 / hu9F5VLv7, hu9F5VHv6 / hu9F5VLv1, hu9F5VHv6 / hu9F5VLv2, hu9F5VHvhu9F5VHv7 / hu9F5VLv7, hu9F5VHv8 / hu9F5VLv1, hu9F5VHv8 / hu9F5VLv2, hu9F5VHv8 / hu9F5VLv3, hu9F5VHv8 / hu9F5VLv4, hu9F5VLv8 / hu9F5VLv5, hu9F5VHv8 / hu9F5VLv6, hu9F5VHv8 / hu9F5VLv7

[0571] The illustrated antibodies include the illustrated mature heavy chain variable regions hu9F5VHv1 (SEQ ID NO:15), hu9F5VHv2 (SEQ ID NO:16), hu9F5VHv3 (SEQ ID NO:17), hu9F5VHv4 (SEQ ID NO:18), hu9F5VHv5 (SEQ ID NO:19), hu9F5VHv6 (SEQ ID NO:20), hu9F5VHv7 (SEQ ID NO:21), hu9F5VHv8 (SEQ ID NO:22), hu9F5VHv9 (SEQ ID NO:127), hu9F5VHv10 (SEQ ID NO:128), hu9F5VHv10_L82cG (SEQ ID NO:129), hu9F5VHv4_L80P (SEQ ID NO:109), and hu9F5VHv4_L80D (SEQ ID NO:15). NO:110), hu9F5VHv4_L82cG (SEQ ID NO:111), hu9F5VHv4_L82cD (SEQ ID NO:112), hu9F5VHv4_L82P (SEQ ID NO:113), hu9F5VHv4_L80G (SEQ ID NO:114), hu9F5VHv4_L82K (SEQ ID NO:115), hu9F5VHv4_L82R (SEQ ID NO:116), hu9F5VHv4_L82E (SEQ ID NO:117), hu9F5VHv4_L82N (SEQ ID NO:118), hu9F5VHv4_Y79D (SEQ ID NO:119), hu9F5VHv4_Y79N (SEQ ID NO:120), hu9F5VHv4_Y79G (SEQ ID NO:121), hu9F5VHv5_M80E (SEQ ID NO:122), hu9F5VHv5_M80G (SEQ ID NO:123), hu9F5VHv4_L82Cs (SEQ ID NO:124), hu9F5VHv4_Y79Q (SEQ ID NO:125) and hu9F5VHv4_S82aG (SEQ ID NO:126) and the illustrated mature light chain variable regions hu9F5VLv1 (SEQ ID NO:23), hu9F5VLv2 (SEQ ID NO:24), hu9F5VLv3 (SEQ ID NO:25), hu9F5VLv4 (SEQ ID NO:26), hu9F5VLv5 (SEQ ID NO:27), hu9F5VLv6 (SEQ ID NO:28), hu9F5VLv7 (SEQ ID NO:126).ID NO:29)、hu9F5VLv8(SEQ ID NO:130)、hu9F5VLv9(SEQ IDNO:131)、hu9F5VLv2_M51E(SEQ ID NO:61)、hu9F5VLv2_M51D(SEQ ID NO:62)、hu9F5VLv2_L27cD(SEQID NO:63)、hu9F5VLv2_L27cG(SEQ ID NO:64)、hu9F5VLv2_L27cS(SEQ IDNO:65)、hu9F5VLv2_L27cE(SEQ ID NO:66)、hu9F5VLv2_I30E(SEQ ID NO:67)、hu9F5VLv2_I30K(SEQID NO:68)、hu9F5VLv2_L27cT(SEQ ID NO:69)、hu9F5VLv2_L27cN(SEQ ID NO:70)、hu9F5VLv2_L27bD(SEQ ID NO:71)、hu9F5VLv2_I30G、(SEQ IDNO:72)、hu9F5VLv2_L33N(SEQID NO:73)、hu9F5VLv2_L27cA(SEQID NO:74)、hu9F5VLv2_L33T(SEQ ID NO:75)、hu9F5VLv2_L33S(SEQ ID NO:76)、hu9F5VLv2_L33R(SEQ ID NO:77)、hu9F5VLv2_I30Q(SEQID NO:78)、hu9F5VLv2_L27bT(SEQ IDNO:79)、hu9F5VLv2_T31G(SEQ ID NO:80)、hu9F5VLv2_L27bQ(SEQID NO:81)、hu9F5VLv2_L33G(SEQ ID NO:82)、hu9F5VLv2_L27cP(SEQID NO:83)、hu9F5VLv2_V78R(SEQ ID NO:84)、hu9F5VLv2_I75D(SEQ ID NO:85)、hu9F5VLv2_V78D(SEQ IDNO:86)、hu9F5VLv2_V78E(SEQ ID NO:87)、hu9F5VLv2_V78P(SEQIDNO:88)、hu9F5VLv2_V78K(SEQ ID NO:89)、hu9F5VLv2_R77D(SEQ ID NO:90)、hu9F5VLv2_V78G(SEQ ID NO:91)、hu9F5VLv2_S76P(SEQ IDNO:92)、hu9F5VLv2_I75P(SEQ IDNO:93)、hu9F5VLv2_I75Q(SEQ ID NO:94)、hu9F5VLv2_I75G(SEQID NO:95)、hu9F5VLv2_L73P(SEQID NO:96)、hu9F5VLv2_L73G(SEQ ID NO:97)、hu9F5VLv2_V78Q(SEQ ID NO:98)、hu9F5VLv2_S76G(SEQ ID NO:99)、hu9F5VLv2_L92D(SEQ IDNO:100)、hu9F5VLv2_Y86T(SEQID NO:101)、hu9F5VLv2_L92E(SEQ ID NO:102)、hu9F5VLv2_L92G(SEQ ID NO:103)、hu9F5VLv2_L92Q(SEQ ID NO:104)、hu9F5VLv2_L93G(SEQ IDNO:105)、hu9F5VLv2_V85G(SEQID NO:106)、hu9F5VLv2_L92T(SEQ ID NO:107)、hu9F5VLv2_A89G(SEQ ID NO:108)、hu9F5VLv8_DIM1(SEQ ID NO:132)、hu9F5VLv8_DIM2(SEQ IDNO:133)、hu9F5VLv8_DIM3(SEQID NO:134)、hu9F5VLv8_DIM4(SEQ ID NO:135)、hu9F5VLv8_DIM5(SEQ ID NO:136)、hu9F5VLv8_DIM6(SEQ ID NO:137)、hu9F5VLv8_DIM7(SEQ IDNO:138)、hu9F5VLv8_DIM8(SEQID NO:139)、hu9F5VLv8_DIM9(SEQ ID NO:140)、hu9F5VLv8_DIM10(SEQ ID NO:141)、hu9F5VLv8_DIM11(SEQ ID NO:142)、hu9F5VLv8_DIM12(SEQ IDNO:143)、hu9F5VLv8_DIM13(SEQ ID NO:144)、hu9F5VLv8_DIM14(SEQ ID NO:145)、hu9F5VLv8_DIM15(SEQ ID NO:146)、hu9F5VLv8_DIM16(SEQ ID NO:147)、hu9F5VLv8_DIM17(SEQAny permutation or combination of any one of hu9F5VLv8_DIM1(SEQ ID NO:148), hu9F5VLv8_DIM2(SEQ ID NO:149), hu9F5VLv8_DIM3(SEQ ID NO:150), hu9F5VLv8_DIM4(SEQ ID NO:151), hu9F5VLv8_DIM5(SEQ ID NO:152), hu9F5VLv8_DIM6(SEQ ID NO:153), hu9F5VLv8_DIM7(SEQ ID NO:154), hu9F5VLv8_DIM8(SEQ ID NO:155), hu9F5VLv8_DIM9(SEQ ID NO:156), hu9F5VLv8_DIM10(SEQ ID NO:157), hu9F5VLv8_DIM11(SEQ ID NO:158), hu9F5VLv8_DIM12(SEQ ID NO:159), hu9F5VLv8_DIM13(SEQ ID NO:160), hu9F5VLv8_DIM14(SEQ ID NO:161), hu9F5VLv9_DIM1(SEQ ID NO:162), hu9F5VLv9_DIM2(SEQ ID NO:163), hu9F5VLv9_DIM4(SEQ ID NO:164), hu9F5VLv9_DIM5(SEQ ID NO:165), hu9F5VLv9_DIM8(SEQ ID NO:166), hu9F5VLv9_DIM10(SEQ ID NO:167), hu9F5VLv9_DIM11(SEQ ID NO:168), hu9F5VLv9_DIM13(SEQ ID NO:169), hu9F5VLv9_DIM19(SEQ ID NO:170), and hu9F5VLv9_DIM20(SEQ ID NO:171).

[0572] This invention provides an antibody in which the humanized heavy chain variable region hu9F5VHv9 (SEQ ID NO:127) is combined with the humanized light chain variable region hu9F5VLv8_DIM18 (also known as hu9F5VLv8_V3Q, L27cD, L37G, M51G, L54R, L92I, SEQ ID NO:149). This invention also provides an antibody in which the humanized heavy chain variable region hu9F5VHv9 (SEQ ID NO:127) is combined with the humanized light chain variable region hu9F5VLv8_DIM11 (also known as hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54R, L92I, SEQ ID NO:142). This invention provides an antibody in which the humanized heavy chain variable region hu9F5VHv9 (SEQ ID NO:127) is combined with the humanized light chain variable region hu9F5VLv8_DIM28 (also known as hu9F5VLv8_V3Q, L27cS, M51G, L54R, L92I, SEQ ID NO:159). This invention also provides an antibody in which the humanized heavy chain variable region hu9F5VHv9 (SEQ ID NO:127) is combined with the humanized light chain variable region hu9F5VLv8_DIM17 (also known as hu9F5VLv8_V3Q, L27cS, L37G, M51G, L54T, L92I, SEQ ID NO:148).

[0573] This invention provides an antibody in which the humanized heavy chain variable region hu9F5VHv9 (SEQ ID NO:127) is combined with the humanized light chain variable region hu9F5VLv8_DIM6 (also known as hu9F5VLv8_V3Q, L27cD, L37Q, M51G, L54R, L92I, SEQ ID NO:137). This invention also provides an antibody in which the humanized heavy chain variable region hu9F5VHv9 (SEQ ID NO:127) is combined with the humanized light chain variable region hu9F5VLv8_DIM14 (also known as hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54T, L92I, SEQ ID NO:145). This invention provides an antibody in which the humanized heavy chain variable region hu9F5VHv9 (SEQ ID NO:127) is combined with the humanized light chain variable region hu9F5VLv8_DIM5 (also known as hu9F5VLv8_V3Q, L27cG, L37Q, M51G, L54R, L92I, SEQ ID NO:136). This invention also provides an antibody in which the humanized heavy chain variable region hu9F5VHv9 (SEQ ID NO:127) is combined with the humanized light chain variable region hu9F5VLv8_DIM7 (also known as hu9F5VLv8_V3Q, L27cD, L37Q, M51K, L54R, L92I, SEQ ID NO:138).

[0574] This invention provides an antibody in which the humanized heavy chain variable region hu9F5VHv9 (SEQ ID NO:127) is combined with the humanized light chain variable region hu9F5VLv8_DIM27 (also known as hu9F5VLv8_V3Q, L37Q, M51G, L54R, L92I, SEQ ID NO:158). This invention also provides an antibody in which the humanized heavy chain variable region hu9F5VHv9 (SEQ ID NO:127) is combined with the humanized light chain variable region hu9F5VLv8_DIM12 (also known as hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54R, L92G, SEQ ID NO:143). This invention provides an antibody in which the humanized heavy chain variable region hu9F5VHv9 (SEQ ID NO:127) is combined with the humanized light chain variable region hu9F5VLv8_DIM13 (also known as hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54R, SEQ ID NO:144). This invention also provides an antibody in which the humanized heavy chain variable region hu9F5VHv9 (SEQ ID NO:127) is combined with the humanized light chain variable region hu9F5VLv8_DIM2 (also known as hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54R, L92I, SEQ ID NO:133).

[0575] This invention provides an antibody in which the humanized heavy chain variable region hu9F5VHv9 (SEQ ID NO:127) is combined with the humanized light chain variable region hu9F5VLv8_DIM29 (also known as hu9F5VLv8_V3Q, L27cS, L37Q, L54R, L92I, SEQ ID NO:160). This invention also provides an antibody in which the humanized heavy chain variable region hu9F5VHv9 (SEQ ID NO:127) is combined with the humanized light chain variable region hu9F5VLv8_DIM30 (also known as hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L92I, SEQ ID NO:161). The present invention provides an antibody in which the humanized heavy chain variable region hu9F5VHv9 (SEQ ID NO:127) is combined with the humanized light chain variable region hu9F5VLv8_DIM8 (also known as hu9F5VLv8_V3Q, L27cG, L37Q, M51K, L54R, L92I, SEQ ID NO:139).

[0576] The present invention provides an antibody in which the humanized heavy chain variable region hu9F5VHv10 (SEQ ID NO:128) is combined with the humanized light chain variable region hu9F5VLv9_DIM11 (also known as hu9F5VLv9_V3Q, L27cG, L37G, M51G, L54R, L92I, SEQ ID NO:168).

[0577] This invention provides variants of the 9F5 humanized antibody, wherein the humanized mature heavy chain variable region exhibits similarity to hu9F5VHv1 (SEQ ID NO:15), hu9F5VHv2 (SEQ ID NO:16), hu9F5VHv3 (SEQ ID NO:17), hu9F5VHv4 (SEQ ID NO:18), hu9F5VHv5 (SEQ ID NO:19), hu9F5VHv6 (SEQ ID NO:20), hu9F5VHv7 (SEQ ID NO:21), hu9F5VHv8 (SEQ ID NO:22), hu9F5VHv9 (SEQ ID NO:127), hu9F5VHv10 (SEQ ID NO:128), and hu9F5VHv10_L82cG (SEQ ID NO:15). NO:129), hu9F5VHv4_L80P (SEQ ID NO:109), hu9F5VHv4_L80D (SEQ ID NO:110), hu9F5VHv4_L82cG (SEQ ID NO:111), hu9F5VHv4_L82cD (SEQ ID NO:112), hu9F5VHv4_L82P (SEQ ID NO:113), hu9F5VHv4_L80G (SEQ ID NO:114), hu9F5VHv4_L82K (SEQ ID NO:115), hu9F5VHv4_L82R (SEQ ID NO:116), hu9F5VHv4_L82E (SEQ ID NO:117), hu9F5VHv4_L82N (SEQ ID NO:118), hu9F5VHv4_Y79D (SEQ hu9F5VHv4_Y79N (SEQ ID NO:120), hu9F5VHv4_Y79G (SEQ ID NO:121), hu9F5VHv5_M80E (SEQ ID NO:122), hu9F5VHv5_M80G (SEQ ID NO:123), hu9F5VHv4_L82Cs (SEQ ID NO:124), hu9F5VHv4_Y79Q (SEQ ID NO:125), or hu9F5VHv4_S82aG (SEQ ID NO:126) are at least 90%, 95%, 96%, 97%, 98%, or 99% identical to hu9F5VHv4_S82aG (SEQ ID NO:126), and the humanized mature light chain variable region shows at least 90%, 95%, 96%, 97%, 98%, or 99% identity with hu9F5VHv1 (SEQ ID NO:23), hu9F5VHv4_Y79G (SEQ ID NO:121), hu9F5VHv5_M80E (SEQ ID NO:122), hu9F5VHv5_M80G (SEQ ID NO:123), hu9F5VHv4_L82Cs (SEQ ID NO:124), hu9F5VHv4_Y79Q (SEQ ID NO:125), or hu9F5VHv4_S82aG (SEQ ID NO:126). NO:25), hu9F5VLv4 (SEQ ID)NO:26)、hu9F5VLv5(SEQ ID NO:27、hu9F5VLv6(SEQ ID NO:28)、hu9F5VLv7(SEQ ID NO:29)、hu9F5VLv8(SEQ IDNO:130、hu9F5VLv9(SEQ ID NO:131)、hu9F5VLv2_M51E(SEQ IDNO:61)、hu9F5VLv2_M51D(SEQ ID NO:62)、hu9F5VLv2_L27cD(SEQID NO:63)、hu9F5VLv2_L27cG(SEQ ID NO:64)、hu9F5VLv2_L27cS(SEQ ID NO:65)、hu9F5VLv2_L27cE(SEQ IDNO:66)、hu9F5VLv2_I30E(SEQ ID NO:67)、hu9F5VLv2_I30K(SEQID NO:68)、hu9F5VLv2_L27cT(SEQ ID NO:69)、hu9F5VLv2_L27cN(SEQ ID NO:70)、hu9F5VLv2_L27bD(SEQ ID NO:71)、hu9F5VLv2_I30G、(SEQ ID NO:72)、hu9F5VLv2_L33N(SEQ IDNO:73)、hu9F5VLv2_L27cA(SEQ ID NO:74)、hu9F5VLv2_L33T(SEQ ID NO:75)、hu9F5VLv2_L33S(SEQ ID NO:76)、hu9F5VLv2_L33R(SEQ ID NO:77)、hu9F5VLv2_I30Q(SEQID NO:78)、hu9F5VLv2_L27bT(SEQ ID NO:79)、hu9F5VLv2_T31G(SEQ IDNO:80)、hu9F5VLv2_L27bQ(SEQ ID NO:81)、hu9F5VLv2_L33G(SEQ ID NO:82)、hu9F5VLv2_L27cP(SEQ ID NO:83)、hu9F5VLv2_V78R(SEQ ID NO:84)、hu9F5VLv2_I75D(SEQ ID NO:85)、hu9F5VLv2_V78D(SEQ ID NO:86)、hu9F5VLv2_V78E(SEQ IDNO:87)、hu9F5VLv2_V78P(SEQID NO:88)、hu9F5VLv2_V78K(SEQ ID NO:89)、hu9F5VLv2_R77D(SEQ IDNO:90)、hu9F5VLv2_V78G(SEQ ID NO:91)、hu9F5VLv2_S76P(SEQ ID NO:92)、hu9F5VLv2_I75P(SEQID NO:93)、hu9F5VLv2_I75Q(SEQ IDNO:94)、hu9F5VLv2_I75G(SEQ ID NO:95)、hu9F5VLv2_L73P(SEQ ID NO:96)、hu9F5VLv2_L73G(SEQ ID NO:97)、hu9F5VLv2_V78Q(SEQ ID NO:98)、hu9F5VLv2_S76G(SEQ ID NO:99)、hu9F5VLv2_L92D(SEQ ID NO:100)、hu9F5VLv2_Y86T(SEQIDNO:101)、hu9F5VLv2_L92E(SEQ ID NO:102)、hu9F5VLv2_L92G(SEQ ID NO:103)、hu9F5VLv2_L92Q(SEQ ID NO:104)、hu9F5VLv2_L93G(SEQ ID NO:105)、hu9F5VLv2_V85G(SEQ IDNO:106)、hu9F5VLv2_L92T(SEQ ID NO:107)、hu9F5VLv2_A89G(SEQ ID NO:108)、hu9F5VLv8_DIM1(SEQ ID NO:132)、hu9F5VLv8_DIM2(SEQ ID NO:133)、hu9F5VLv8_DIM3(SEQ IDNO:134)、hu9F5VLv8_DIM4(SEQ ID NO:135)、hu9F5VLv8_DIM5(SEQ ID NO:136)、hu9F5VLv8_DIM6(SEQ ID NO:137)、hu9F5VLv8_DIM7(SEQ ID NO:138)、hu9F5VLv8_DIM8(SEQ IDNO:139)、hu9F5VLv8_DIM9(SEQ ID NO:140)、hu9F5VLv8_DIM10(SEQ ID NO:141)、hu9F5VLv8_DIM11(SEQ ID NO:142)、hu9F5VLv8_DIM12(SEQ ID NO:143)、hu9F5VLv8_DIM13(SEQ IDNO:144)、hu9F5VLv8_DIM14(SEQ ID NO:145)、hu9F5VLv8_DIM15(SEQ IDNO: 146), hu9F5VLv8_DIM16 (SEQ ID NO: 147), hu9F5VLv8_DIM17 (SEQ ID NO: 148), hu9F5VLv8_DIM18 (SEQ ID NO: 149), hu9F5VLv8_DIM19 (SEQ ID NO: 150), hu9F5VLv8_DIM20 (SEQ ID NO: 151), hu9F5VLv8_DIM21 (SEQ ID NO: 152), hu9F5VLv8_DIM22 (SEQ ID NO: 153), hu9F5VLv8_DIM23 (SEQ ID NO: 154), hu9F5VLv8_DIM24 (SEQ ID NO: 155), hu9F5VLv8_DIM25 (SEQ ID NO: 156), hu9F5VLv8_DIM26 (SEQ ID NO: 157), hu9F5VLv8_DIM27 (SEQ ID NO: 158), hu9F5VLv8_DIM28 (SEQ ID NO: 159), hu9F5VLv8_DIM29 (SEQ ID NO: 160), hu9F5VLv8_DIM30 (SEQ ID NO: 161), hu9F5VLv9_DIM1 (SEQ ID NO: 162), hu9F5VLv9_DIM2 (SEQ ID NO: 163), hu9F5VLv9_DIM4 (SEQ ID NO: 164), hu9F5VLv9_DIM5 (SEQ ID NO: 165), hu9F5VLv9_DIM8 (SEQ ID NO: 166), hu9F5VLv9_DIM10 (SEQ ID NO: 167), hu9F5VLv9_DIM11 (SEQ ID NO: 168), hu9F5VLv9_DIM13 (SEQ ID NO: 169), hu9F5VLv9_DIM19 (SEQ ID NO: 170), or hu9F5VLv9_DIM20 (SEQ ID NO: 171) have at least 90%, 95%, 96%, 97%, 98%, or 99% identity. In some such antibodies, SEQ ID NOs: 15 - 22, 109 - 129, SEQ ID NOs: 23 - 29, SEQ ID NO: NO: 61 - 108, and SEQ IDAt least 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, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, or all 62 of the positions that have undergone reversion mutations or other mutations are otherwise reverted to mutations or otherwise mutated.

[0578] In some humanized 9F5 antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H1 is occupied by E, H17 by T, H20 by I, H69 by M, H75 by T, H93 by T, H94 by T, and H109 by V. In some humanized 9F5 antibodies, positions H1, H17, H20, H69, H75, H94, and H109 are occupied by E, T, I, M, T, T, T, and V, respectively.

[0579] In some humanized 9F5 antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H66 is occupied by R, and H81 is occupied by E. In some humanized 9F5 antibodies, positions H66 and H81 are occupied by R and E, respectively.

[0580] In some humanized 9F5 antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H23 is occupied by I, and H83 is occupied by R. In some humanized 9F5 antibodies, positions H23 and H83 are occupied by K and R, respectively.

[0581] In some humanized 9F5 antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H43 is occupied by K, H51 by V, H76 by D, M80 by M, and H108 by L. In some humanized 9F5 antibodies, positions H43, H51, H76, H80, and H108 are occupied by K, V, D, M, and L, respectively.

[0582] In some humanized 9F5 antibodies, the H28 position in the VH region is occupied by T.

[0583] In some humanized 9F5 antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H54 is occupied by D, and H56 is occupied by E. In some humanized 9F5 antibodies, positions H54 and H56 are occupied by D and E, respectively.

[0584] In some humanized 9F5 antibodies, position H40 in the VH region is occupied by A. In some humanized 9F5 antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H5 is occupied by V, H11 is occupied by V, H12 is occupied by K, H38 is occupied by R, and H42 is occupied by G.

[0585] In some humanized 9F5 antibodies, positions H5, H11, H12, H38, and H42 are occupied by V, V, K, R, and G, respectively.

[0586] In some humanized 9F5 antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H1 occupied by Q or E, H5 occupied by Q or V, H11 occupied by L or V, H12 occupied by V or K, H17 occupied by S or T, H20 occupied by L or I, H23 occupied by T or K, H28 occupied by N or T, H38 occupied by K, R, or Q, H40 occupied by R or A, H42 occupied by E or G, H43 occupied by Q or K, H48 occupied by I or M, H51 occupied by I or V, H54 occupied by N or D, H H66 is occupied by D or E, H66 by K or R, H69 by I or M, H75 by S or T, H76 by N or D, H79 by Y, Q, D, N or G, H80 by L, M, P, D, G or E, H81 by Q or E, H82 by L, P, K, R, E or N, H82a by S or G, H82c by L, G, D or S, H83 by T or R, H93 by A or T, H94 by S or T, H108 by T or L, and H109 by L or V.

[0587] In some humanized 9F5 antibodies, positions H1, H17, H20, H69, H75, H93, H94, and H109 in the VH region are occupied by E, T, I, M, T, T, T, and V, respectively, as in hu9F5VHv2. In some humanized 9F5 antibodies, positions H1, H17, H20, H66, H69, H75, H81, H93, H94, and H109 in the VH region are occupied by E, T, I, R, M, T, E, T, T, and V, respectively, as in hu9F5VHv3. In some humanized 9F5 antibodies, positions H1, H17, H20, H23, H28, H66, H69, H75, H81, H83, H93, H94 and H109 in the VH region are occupied by E, T, I, K, T, R, M, T, E, R, T, T and V, respectively, as in hu9F5VHv4. In some humanized 9F5 antibodies, positions H1, H17, H20, H23, H28, H43, H51, H54, H56, H66, H69, H75, H76, H80, H81, H83, H93, H94, H108, and H109 in the VH region are occupied by E, T, I, K, T, K, V, D, E, R, M, T, D, M, E, R, T, T, L, and V, respectively, as in hu9F5VHv5. In some humanized 9F5 antibodies, positions H1, H17, H20, H23, H28, H40, H43, H48, H51, H54, H56, H66, H69, H75, H76, H80, H81, H83, H93, H94, H108, and H109 in the VH region are occupied by E, T, I, K, T, A, K, M, V, D, E, R, M, T, D, M, E, R, T, T, L, and V, respectively, as in hu9F5VHv6. In some humanized 9F5 antibodies, positions H1, H5, H11, H12, H17, H20, H23, H38, H40, H42, H43, H51, H54, H56, H66, H69, H75, H76, H80, H81, H83, H93, H94, H108, and H109 in the VH region are occupied by E, V, V, K, T, I, K, R, A, G, K, V, D, E, R, M, T, D, M, E, R, T, T, L, and V, respectively, as in hu9F5VHv7. In some humanized 9F5 antibodies, positions H1, H5, H11, H12, H17, H20, H23, H38, H40, H42, H43, H51, H66, H69, H75, H76, H80, H81, H83, H93, H94, H108, and H109 in the VH region are occupied by E, V, V, K, T, I, K, R, A, G, K, V, R, M, T, D, M, E, R, T, T, L, and V, respectively, as in hu9F5VHv8.In some humanized 9F5 antibodies, positions H1, H5, H11, H12, H17, H20, H23, H38, H42, H43, H66, H69, H75, H80, H81, H83, H93, H94, H108, and H109 in the VH region are occupied by E, V, V, K, T, I, K, Q, G, K, R, M, T, M, E, R, T, T, L, and V, respectively, as in hu9F5VHv9. In some humanized 9F5 antibodies, the heavy chain variable region contains the amino acid sequence of SEQ ID NO:127.

[0588] In some humanized 9F5 antibodies, positions H1, H5, H11, H12, H17, H20, H23, H38, H42, H43, H66, H69, H75, H80, H81, H83, H93, H94, H108, and H109 in the VH region are occupied by E, V, V, K, T, I, K, K, E, K, R, M, T, M, E, R, T, T, L, and V, respectively, as in hu9F5VHv10. In some humanized 9F5 antibodies, the heavy chain variable region contains the amino acid sequence of SEQ ID NO:128.

[0589] In some humanized 9F5 antibodies, positions H1, H5, H11, H12, H17, H20, H23, H38, H42, H43, H66, H69, H75, H80, H81, H82c, H83, H93, H94, H108, and H109 in the VH region are occupied by E, V, V, K, T, I, K, K, E, K, R, M, T, M, E, G, R, T, T, L, and V, respectively, as in hu9F5VHv10_L82cG.

[0590] In some humanized 9F5 antibodies, at least one of the following positions in the VL region is occupied by a specified amino acid: L7 is occupied by S, L8 by P, L15 by P, and L100 by Q. In some humanized 9F5 antibodies, positions L7, L8, L15, and L100 are occupied by S, P, P, and Q, respectively.

[0591] In some humanized 9F5 antibodies, position L66 in the VL region is occupied by G. In some humanized 9F5 antibodies, position L64 in the VL region is occupied by S.

[0592] In some humanized 9F5 antibodies, position L17 in the VL region is occupied by E.

[0593] In some humanized 9F5 antibodies, at least one of the following positions in the VL region is occupied by a specified amino acid: L11 is occupied by L, L51 is occupied by G, and L54 is occupied by R. In some humanized 9F5 antibodies, positions L11, L51, and L54 are occupied by L, G, and R, respectively.

[0594] In some humanized 9F5 antibodies, position L30 in the VL region is occupied by Y.

[0595] In some humanized 9F5 antibodies, at least one of the following positions in the VL region is occupied by a specified amino acid: L3 is occupied by V or Q, L7 by A or S, L8 by A or P, L9 by F or L, L11 by N or L, L15 by L or P, L17 by T or E, L18 by S or P, L27b by L, D, T, or Q, L27c by L, D, G, S, E, T, N, A, P, or I, and L30 by I, Y, E, K, G, or Q. L31 is occupied by T, N, or G; L33 is occupied by L, N, T, S, R, or G; L37 is occupied by L, Q, G, or I; L39 is occupied by R or K; L51 is occupied by M, G, E, D, K, or I; L54 is occupied by L, R, G, or T; L60 is occupied by N or D; L64 is occupied by G or S; L66 is occupied by E or G; L73 is occupied by L, P, or G; L74 is occupied by R or K; L75 is occupied by I, D, P, Q, or G; L76 is occupied by S, P, or G; L77 is occupied by SEQ. ID NO:146 or D is occupied, L78 is occupied by V, R, D, E, P, K, G or Q, L85 is occupied by V or G, L86 is occupied by Y or T, L89 is occupied by A or G, L92 is occupied by L, D, E, G, Q, T or I, L93 is occupied by E or G, and L100 is occupied by G or Q.

[0596] In some humanized 9F5 antibodies, positions L64 and L66 in the VL region are occupied by S and G, respectively, as in hu9F5VLv1. In some humanized 9F5 antibodies, positions L7, L8, L15, L64, L66, and L100 in the VL region are occupied by S, P, P, S, G, and Q, respectively, as in hu9F5VLv2. In some humanized 9F5 antibodies, positions L7, L8, L15, L17, L66, and L100 in the VL region are occupied by S, P, P, E, G, and Q, respectively, as in hu9F5VLv3.

[0597] In some humanized 9F5 antibodies, positions L7, L8, L11, L15, L17, L51, L54, L66, and L100 in the VL region are occupied by S, P, L, P, E, G, R, G, and Q, respectively, as in hu9F5VLv4. In some humanized 9F5 antibodies, the light chain variable region contains the amino acid sequence of any one of SEQ ID NO:133, 135-137, 142-144, 149, 158, 159, and 168. In some humanized 9F5 antibodies, the light chain variable region contains the amino acid sequence of SEQ ID NO:133. In some humanized 9F5 antibodies, the light chain variable region contains the amino acid sequence of SEQ ID NO:137. In some humanized 9F5 antibodies, the light chain variable region contains the amino acid sequence of SEQ ID NO:149. In some humanized 9F5 antibodies, the light chain variable region contains the amino acid sequence of SEQ ID NO:159.

[0598] In some humanized 9F5 antibodies, positions L7, L8, L11, L15, L17, L30, L51, L54, L66, and L100 in the VL region are occupied by S, P, L, P, E, Y, G, R, G, and Q, respectively, as in hu9F5VLv5. In some humanized 9F5 antibodies, positions L7, L8, L11, L15, L17, L30, L51, L54, and L100 in the VL region are occupied by S, P, L, P, E, Y, G, R, and Q, respectively, as in hu9F5VLv6. In some humanized 9F5 antibodies, positions L7, L8, L9, L11, L15, L17, L18, L31, L39, L51, L54, L60, L66, L74, and L100 in the VL region are occupied by S, P, L, L, P, E, P, N, K, G, R, D, G, K, and Q, respectively, as in hu9F5VLv7.

[0599] In some humanized 9F5 antibodies, positions L7, L8, L11, L15, L17, L39, L64, L66, L74, and L100 in the VL region are occupied by S, P, L, P, E, K, S, G, K, and Q, respectively, as in hu9F5VLv8. In some humanized 9F5 antibodies, position L3 in the VL region is occupied by Q. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by D, G, I, L, or S; position L37 in the VL region is occupied by G, I, L, or Q; position L51 in the VL region is occupied by E, G, I, K, or M; position L54 in the VL region is occupied by G, L, R, or T; and position L92 in the VL region is occupied by G, I, or L. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by D or S, position L37 in the VL region is occupied by G, L or Q, position L51 in the VL region is occupied by G or K, position L54 in the VL region is occupied by R, and position L92 in the VL region is occupied by I.

[0600] In some humanized 9F5 antibodies, position L27c in the VL region is occupied by D, position L37 in the VL region is occupied by G, and position L51 in the VL region is occupied by G. In some humanized 9F5 antibodies, the heavy chain variable region has an amino acid sequence containing SEQ ID NO:127, and the light chain variable region has an amino acid sequence containing SEQ ID NO:149.

[0601] In some humanized 9F5 antibodies, position L27c in the VL region is occupied by D, position L37 in the VL region is occupied by Q, and position L51 in the VL region is occupied by G. In some humanized 9F5 antibodies, the heavy chain variable region has an amino acid sequence containing SEQ ID NO:127, and the light chain variable region has an amino acid sequence containing SEQ ID NO:137.

[0602] In some humanized 9F5 antibodies, position L27c in the VL region is occupied by S, position L37 in the VL region is occupied by L, and position L51 in the VL region is occupied by G. In some humanized 9F5 antibodies, the heavy chain variable region has an amino acid sequence containing SEQ ID NO:127, and the light chain variable region has an amino acid sequence containing SEQ ID NO:159.

[0603] In some humanized 9F5 antibodies, position L27c in the VL region is occupied by D, position L37 in the VL region is occupied by Q, and position L51 in the VL region is occupied by K. In some humanized 9F5 antibodies, the heavy chain variable region has an amino acid sequence containing SEQ ID NO:127, and the light chain variable region has an amino acid sequence containing SEQ ID NO:138.

[0604] In some humanized 9F5 antibodies, position L27c in the VL region is occupied by S, position L37 in the VL region is occupied by Q, and position L51 in the VL region is occupied by G. In some humanized 9F5 antibodies, the heavy chain variable region has an amino acid sequence containing SEQ ID NO:127, and the light chain variable region has an amino acid sequence containing SEQ ID NO:133.

[0605] In some humanized 9F5 antibodies, positions L7, L8, L11, L15, L17, L39, L60, L64, L66, L74, and L100 in the VL region are occupied by S, P, L, P, E, K, D, S, G, K, and Q, respectively, as in hu9F5VLv9. In some humanized 9F5 antibodies, position L3 in the VL region is occupied by Q.

[0606] In some humanized 9F5 antibodies, position L27c in the VL region is occupied by G or S, position L37 in the VL region is occupied by G, I, or Q, position L51 in the VL region is occupied by G, I, or K, position L54 in the VL region is occupied by G or R, and position L92 in the VL region is occupied by G, I, or L. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by G, position L37 in the VL region is occupied by G, position L51 in the VL region is occupied by G, and position L54 in the VL region is occupied by R. In some humanized 9F5 antibodies, position L92 in the VL region is occupied by I. In some humanized 9F5 antibodies, the heavy chain variable region has an amino acid sequence containing SEQ ID NO:129, and the light chain variable region has an amino acid sequence containing SEQ ID NO:168.

[0607] The light chain variable region of any of the antibodies mentioned above can be modified to further reduce immunogenicity. For example, in some humanized antibodies, the L27b position in the VL region is occupied by D, T, or Q; the L27c position in the VL region is occupied by D, G, S, E, T, N, A, I, or P; the L30 position in the VL region is occupied by E, K, G, or Q; the L31 position in the VL region is occupied by G; the L33 position in the VL region is occupied by N, T, S, R, or G; the L37 position in the VL region is occupied by Q, G, or I; the L51 position in the VL region is occupied by E, D, G, K, or I; the L54 position in the VL region is occupied by G, R, or T; and the L60 position in the VL region is occupied by D. Occupied; Position L73 in VL area is occupied by P or G; Position L75 in VL area is occupied by D, P, Q or G; Position L76 in VL area is occupied by P or G; Position L77 in VL area is occupied by D; Position L78 in VL area is occupied by R, D, E, P, K, G or Q; Position L85 in VL area is occupied by G; Position L86 in VL area is occupied by T; Position L89 in VL area is occupied by G; Position L92 in VL area is occupied by D, E, G, Q, I or T; and / or Position L93 in VL area is occupied by G (Kabat number).

[0608] In some humanized 9F5 antibodies, position L51 in the VL region is occupied by E, as in hu9F5VLv2_M51E. In some humanized 9F5 antibodies, position L51 in the VL region is occupied by D, as in hu9F5VLv2_M51D. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by D, as in hu9F5VLv2_L27cD. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by G, as in hu9F5VLv2_L27cG. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by S, as in hu9F5VLv2_L27cS. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by E, as in hu9F5VLv2_L27cE. In some humanized 9F5 antibodies, position L30 in the VL region is occupied by E, as in hu9F5VLv2_I30E. In some humanized 9F5 antibodies, position L30 in the VL region is occupied by K, as in hu9F5VLv2_I30K. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by T, as in hu9F5VLv2_L27cT. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by N, as in hu9F5VLv2_L27cN.

[0609] In some humanized 9F5 antibodies, position L27b in the VL region is occupied by D, as in hu9F5VLv2_L27bD. In some humanized 9F5 antibodies, position L30 in the VL region is occupied by G, as in hu9F5VLv2_I30G. In some humanized 9F5 antibodies, position L33 in the VL region is occupied by N, as in hu9F5VLv2_L33N. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by A, as in hu9F5VLv2_L27cA. In some humanized 9F5 antibodies, position L33 in the VL region is occupied by T, as in hu9F5VLv2_L33T. In some humanized 9F5 antibodies, position L33 in the VL region is occupied by S, as in hu9F5VLv2_L33S. In some humanized 9F5 antibodies, position L33 in the VL region is occupied by R, as in hu9F5VLv2_L33R. In some humanized 9F5 antibodies, position L30 in the VL region is occupied by Q, as in hu9F5VLv2_I30Q. In some humanized 9F5 antibodies, position L27b in the VL region is occupied by T, as in hu9F5VLv2_L27bT. In some humanized 9F5 antibodies, position L31 in the VL region is occupied by G, as in hu9F5VLv2_T31G.

[0610] In some humanized 9F5 antibodies, position L27b in the VL region is occupied by Q, as in hu9F5VLv2_L27bQ). In some humanized 9F5 antibodies, position L33 in the VL region is occupied by G, as in hu9F5VLv2_L33G. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by P, as in hu9F5VLv2_L27cP. In some humanized 9F5 antibodies, position L78 in the VL region is occupied by R, as in hu9F5VLv2_V78R. In some humanized 9F5 antibodies, position L75 in the VL region is occupied by D, as in hu9F5VLv2_I75D. In some humanized 9F5 antibodies, position L78 in the VL region is occupied by D, as in hu9F5VLv2_V78D. In some humanized 9F5 antibodies, position L78 in the VL region is occupied by E, as in hu9F5VLv2_V78E. In some humanized 9F5 antibodies, position L78 in the VL region is occupied by P, as in hu9F5VLv2_V78P. In some humanized 9F5 antibodies, position L78 in the VL region is occupied by K, as in hu9F5VLv2_V78K. In some humanized 9F5 antibodies, position L77 in the VL region is occupied by D, as in hu9F5VLv2_R77D.

[0611] In some humanized 9F5 antibodies, position L78 in the VL region is occupied by G, as in hu9F5VLv2_V78G. In some humanized 9F5 antibodies, position L76 in the VL region is occupied by P, as in hu9F5VLv2_S76P. In some humanized 9F5 antibodies, position L75 in the VL region is occupied by P, as in hu9F5VLv2_I75P. In some humanized 9F5 antibodies, position L75 in the VL region is occupied by Q, as in hu9F5VLv2_I75Q. In some humanized 9F5 antibodies, position L75 in the VL region is occupied by G, as in hu9F5VLv2_I75G. In some humanized 9F5 antibodies, position L73 in the VL region is occupied by P, as in hu9F5VLv2_L73P. In some humanized 9F5 antibodies, position L73 in the VL region is occupied by G, as in hu9F5VLv2_L73G. In some humanized 9F5 antibodies, position L78 in the VL region is occupied by Q, as in hu9F5VLv2_V78Q. In some humanized 9F5 antibodies, position L76 in the VL region is occupied by G, as in hu9F5VLv2_S76G. In some humanized 9F5 antibodies, position L92 in the VL region is occupied by D, as in hu9F5VLv2_L92D.

[0612] In some humanized 9F5 antibodies, position L86 in the VL region is occupied by T, as in hu9F5VLv2_Y86T. In some humanized 9F5 antibodies, position L92 in the VL region is occupied by E, as in hu9F5VLv2_L92E. In some humanized 9F5 antibodies, position L92 in the VL region is occupied by G, as in hu9F5VLv2_L92G. In some humanized 9F5 antibodies, position L92 in the VL region is occupied by Q, as in hu9F5VLv2_L92Q. In some humanized 9F5 antibodies, position L93 in the VL region is occupied by G, as in hu9F5VLv2_L93G. In some humanized 9F5 antibodies, position L85 in the VL region is occupied by G, as in hu9F5VLv2_V85G. In some humanized 9F5 antibodies, position L92 in the VL region is occupied by T, such as in hu9F5VLv2_L92T. In some humanized 9F5 antibodies, position L89 in the VL region is occupied by G, such as in hu9F5VLv2_A89G.

[0613] In some humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, G, G, and I, respectively, as in hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54G, and L92I (also known as hu9F5VLv8_DIM1). In other humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, G, R, and I, as in hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54R, and L92I (also known as hu9F5VLv8_DIM2). In some humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, G, T, and I, respectively, as in hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54T, and L92I (also known as hu9F5VLv8_DIM3). In other humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, G, R, and G, respectively, as in hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54R, and L92G (also known as hu9F5VLv8_DIM4). In some humanized 9F5 antibodies, the positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, Q, G, R, and I, respectively, as in hu9F5VLv8_V3Q, L27cG, L37Q, M51G, L54R, and L92I (also known as hu9F5VLv8_DIM5).

[0614] In some humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, D, Q, G, R, and I, respectively, as in hu9F5VLv8_V3Q, L27cD, L37Q, M51G, L54R, and L92I (also known as hu9F5VLv8_DIM6). In other humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, D, Q, K, R, and I, respectively, as in hu9F5VLv8_V3Q, L27cD, L37Q, M51K, L54R, and L92I (also known as hu9F5VLv8_DIM7). In some humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, Q, K, R, and I, respectively, as in hu9F5VLv8_V3Q, L27cG, L37Q, M51K, L54R, and L92I (also known as hu9F5VLv8_DIM8). In other humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, Q, K, G, and I, respectively, as in hu9F5VLv8_V3Q, L27cG, L37Q, M51K, L54G, and L92I (also known as hu9F5VLv8_DIM9). In some humanized 9F5 antibodies, the positions L3, L27c, L37, L51, L54 and L92 in the VL region are occupied by Q, S, Q, K, G and I, respectively, such as in hu9F5VLv8_V3Q, L27cS, L37Q, M51K, L54G, L92I (also known as hu9F5VLv8_DIM10).

[0615] In some humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, G, G, R, and I, respectively, as in hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54R, and L92I (also known as hu9F5VLv8_DIM11). In other humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, G, G, R, and G, respectively, as in hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54R, and L92G (also known as hu9F5VLv8_DIM12). In some humanized 9F5 antibodies, positions L3, L27c, L37, L51, and L54 in the VL region are occupied by Q, G, G, G, and R, respectively, as in hu9F5VLv8_V3Q, L27cG, L37G, M51G, and L54R (also known as hu9F5VLv8_DIM13). In other humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, G, G, T, and I, respectively, as in hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54T, and L92I (also known as hu9F5VLv8_DIM14). In some humanized 9F5 antibodies, the positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, G, G, T, and G, respectively, as in hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54T, and L92G (also known as hu9F5VLv8_DIM15).

[0616] In some humanized 9F5 antibodies, positions L3, L27c, L37, L51, and L54 in the VL region are occupied by Q, G, G, G, and T, respectively, as in hu9F5VLv8_V3Q, L27cG, L37G, M51G, and L54T (also known as hu9F5VLv8_DIM16). In other humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, G, G, T, and I, respectively, as in hu9F5VLv8_V3Q, L27cS, L37G, M51G, L54T, and L92I (also known as hu9F5VLv8_DIM17). In some humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, D, G, G, R, and I, respectively, as in hu9F5VLv8_V3Q, L27cD, L37G, M51G, L54R, and L92I (also known as hu9F5VLv8_DIM18). In other humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, I, I, R, and I, respectively, as in hu9F5VLv8_V3Q, L27cS, L37I, M51I, L54R, and L92I (also known as hu9F5VLv8_DIM19). In some humanized 9F5 antibodies, the positions L3, L27c, L37, L51, L54 and L92 in the VL region are occupied by Q, S, Q, I, G and I, respectively, such as in hu9F5VLv8_V3Q, L27cS, L37Q, M51I, L54G, L92I (also known as hu9F5VLv8_DIM20).

[0617] In some humanized 9F5 antibodies, positions L3, L27c, L37, L51, and L54 in the VL region are occupied by Q, S, Q, I, and G, respectively, as in hu9F5VLv8_V3Q, L27cS, L37Q, M51I, and L54G (also known as hu9F5VLv8_DIM21). In other humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, E, R, and I, respectively, as in hu9F5VLv8_V3Q, L27cS, L37Q, M51E, L54R, and L92I (also known as hu9F5VLv8_DIM22). In some humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, Q, E, G, and I, respectively, as in hu9F5VLv8_V3Q, L27cG, L37Q, M51E, L54G, and L92I (also known as hu9F5VLv8_DIM23). In other humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92L in the VL region are occupied by Q, G, I, E, R, and I, respectively, as in hu9F5VLv8_V3Q, L27cG, L37I, M51E, L54R, and L92I (also known as hu9F5VLv8_DIM24). In some humanized 9F5 antibodies, the positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, I, E, R, and G, respectively, as in hu9F5VLv8_V3Q, L27cG, L37I, M51E, L54R, and L92G (also known as hu9F5VLv8_DIM25).

[0618] In some humanized 9F5 antibodies, positions L3, L27c, L37, L51, and L54 in the VL region are occupied by Q, I, I, E, and R, respectively, as in hu9F5VLv8_V3Q, L27cI, L37I, M51E, and L54R (also known as hu9F5VLv8_DIM26). In other humanized 9F5 antibodies, positions L3, L37, L51, L54, and L92 in the VL region are occupied by Q, Q, G, R, and I, respectively, as in hu9F5VLv8_V3Q, L37Q, M51G, L54R, and L92I (also known as hu9F5VLv8_DIM27). In some humanized 9F5 antibodies, positions L3, L27c, L51, L54, and L92 in the VL region are occupied by Q, S, G, R, and I, respectively, as in hu9F5VLv8_V3Q, L27cS, M51G, L54R, and L92I (also known as hu9F5VLv8_DIM28). In other humanized 9F5 antibodies, positions L3, L27c, L37, L54, and L92 in the VL region are occupied by Q, S, Q, R, and I, respectively, as in hu9F5VLv8_V3Q, L27cS, L37Q, L54R, and L92I (also known as hu9F5VLv8_DIM29). In some humanized 9F5 antibodies, the positions L3, L27c, L37, L51, and L92 in the VL region are occupied by Q, S, Q, G, and I, respectively, such as in hu9F5VLv8_V3Q, L27cS, L37Q, M51G, and L92I (also known as hu9F5VLv8_DIM30).

[0619] In some humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, G, G, and I, respectively, as in hu9F5VLv9_V3Q, L27cS, L37Q, M51G, L54G, and L92I (also known as hu9F5VLv9_DIM1). In other humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, G, R, and I, respectively, as in hu9F5VLv9_V3Q, L27cS, L37Q, M51G, L54R, and L92I (also known as hu9F5VLv9_DIM2). In some humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, G, R, and G, respectively, as in hu9F5VLv9_V3Q, L27cS, L37Q, M51G, L54R, and L92G (also known as hu9F5VLv9_DIM4). In other humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, Q, G, R, and I, respectively, as in hu9F5VLv9_V3Q, L27cG, L37Q, M51G, L54R, and L92I (also known as hu9F5VLv9_DIM5). In some humanized 9F5 antibodies, the positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, Q, K, R, and I, respectively, as in hu9F5VLv9_V3Q, L27cG, L37Q, M51K, L54R, and L92I (also known as hu9F5VLv9_DIM8).

[0620] In some humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, K, G, and I, respectively, as in hu9F5VLv9_V3Q, L27cS, L37Q, M51K, L54G, and L92I (also known as hu9F5VLv9_DIM10). In other humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, G, G, R, and I, respectively, as in hu9F5VLv9_V3Q, L27cG, L37G, M51G, L54R, and L92I (also known as hu9F5VLv9_DIM11). In some humanized 9F5 antibodies, positions L3, L27c, L37, L51, and L54 in the VL region are occupied by Q, G, G, G, and R, respectively, as in hu9F5VLv9_V3Q, L27cG, L37G, M51G, and L54R (also known as hu9F5VLv9_DIM13). In other humanized 9F5 antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, I, I, R, and I, respectively, as in hu9F5VLv9_V3Q, L27cS, L37I, M51I, L54R, and L92I (also known as hu9F5VLv9_DIM19). In some humanized 9F5 antibodies, the positions L3, L27c, L37, L51, L54 and L92 in the VL region are occupied by Q, S, Q, I, G and I, respectively, such as in hu9F5VLv9_V3Q, L27cS, L37Q, M51I, L54G, L92I (also known as hu9F5VLv9_DIM20).

[0621] The heavy chain variable regions of any of the antibodies mentioned above can be modified to further reduce immunogenicity. For example, in some humanized antibodies, position H79 is occupied by D, N, G, or Q; position H80 is occupied by P, D, E, or G; position H82 is occupied by P, K, R, E, or N; position H82a is occupied by G; and / or position H82c is occupied by G, D, or S.

[0622] In some humanized 9F5 antibodies, position H80 is occupied by P, as in hu9F5VHv4_L80P. In some humanized 9F5 antibodies, position H80 is occupied by D, as in hu9F5VHv4_L80D. In some humanized 9F5 antibodies, position H82c is occupied by G, as in hu9F5VHv4_L82cG and hu9F5VHv10_L82cG. In some humanized 9F5 antibodies, position H82c is occupied by D, as in hu9F5VHv4_L82cD. In some humanized 9F5 antibodies, position H82 is occupied by P, as in hu9F5VHv4_L82P. In some humanized 9F5 antibodies, position H80 is occupied by G, as in hu9F5VHv4_L80G. In some humanized 9F5 antibodies, position H82 is occupied by K, as in hu9F5VHv4_L82K. In some humanized 9F5 antibodies, position H82 is occupied by R, as in hu9F5VHv4_L82R. In some humanized 9F5 antibodies, position H82 is occupied by E, as in hu9F5VHv4_L82E. In some humanized 9F5 antibodies, position H82 is occupied by N, as in hu9F5VHv4_L82N.

[0623] In some humanized 9F5 antibodies, position H79 is occupied by D, as in hu9F5VHv4_Y79D. In some humanized 9F5 antibodies, position H79 is occupied by N, as in hu9F5VHv4_Y79N. In some humanized 9F5 antibodies, position H79 is occupied by G, as in hu9F5VHv4_Y79G. ​​In some humanized 9F5 antibodies, position H80 is occupied by E, as in hu9F5VHv5_M80E. In some humanized 9F5 antibodies, position H80 is occupied by G, as in hu9F5VHv5_M80G. In some humanized 9F5 antibodies, position H82c is occupied by S, as in hu9F5VHv4_L82cS. In some humanized 9F5 antibodies, position H79 is occupied by Q, such as in hu9F5VHv4_Y79Q. In some humanized 9F5 antibodies, position H82a is occupied by G, such as in hu9F5VHv4_S82aG.

[0624] In some humanized 9F5 antibodies, the variable heavy chain has ≥85% identity with the human sequence. In some humanized 9F5 antibodies, the variable light chain has ≥85% identity with the human sequence. In some humanized 9F5 antibodies, each of the variable heavy chain and variable light chain has ≥85% identity with the human germline sequence. In some humanized 9F5 antibodies, the three heavy chain CDRs are as defined by the Kabat / Chothia complex (SEQ ID NO: 8, 9, and 10), and the three light chain CDRs are as defined by the Kabat / Chothia complex (SEQ ID NO: 8, 9, and 10). NO:12, 13 and 14); the conditions are: position H28 is occupied by N or T; position H51 is occupied by I or V; position H54 is occupied by N or D; position H56 is occupied by D or E; position L27b is occupied by L, D, T or Q; position L27c is occupied by L, D, G, S, E, T, N, A, P or I; position L30 is occupied by I, Y, E, K, G or Q; position L31 is occupied by T, N or G; position L33 is occupied by L, N, T, S, R or G; position L51 is occupied by M, G, E, D, K or I; position L54 is occupied by L, R, G or T; position L89 is occupied by A or G; position L92 is occupied by L, D, E, G, Q, T or I, and position L93 is occupied by E or G.

[0625] In some humanized 9F5 antibodies, the Kabat-Chothia complex CDR-H1 has the amino acid sequence comprising SEQ ID NO:50. In some humanized 9F5 antibodies, Kabat CDR-H2 has the amino acid sequence comprising SEQ ID NO:51 or SEQ ID NO:52. In some humanized 9F5 antibodies, Kabat CDR-L1 has the amino acid sequence comprising SEQ ID NO:53 or SEQ ID NO:54. In some humanized 9F5 antibodies, Kabat CDR-L2 has the amino acid sequence comprising SEQ ID NO:55. In some humanized 9F5 antibodies, Kabat CDR-L1 has the amino acid sequence selected from the group consisting of SEQ ID NO:172-193. In some humanized 9F5 antibodies, Kabat CDR-L2 has the amino acid sequence selected from the group consisting of SEQ ID NO:194-205. In some humanized 9F5 antibodies, Kabat CDR-L3 has the amino acid sequence selected from the group consisting of SEQ ID NO:206-213.

[0626] Exemplary humanized antibodies include a humanized form of mouse 10C12, named Hu10C12.

[0627] Mouse antibody 10C12 includes mature heavy and light chain variable regions, which have amino acid sequences comprising SEQ ID NO:7 and SEQ ID NO:11, respectively. This invention provides two exemplified humanized mature heavy chain variable regions: hu10C12VHv1 and hu10C12VHv2. This invention also provides two exemplified mature light chain variable regions: hu10C12VLv1 and hu10C12VLv2. Figure 7 and Figure 8 The comparisons of the heavy chain variable region and light chain variable region of mouse 10C12 and various humanized antibodies are shown respectively.

[0628] For reasons such as potentially affecting CDR conformation and / or binding to antigens, mediating interactions between heavy and light chains, interactions with constant regions, being a site of desired or undesirable post-translational modifications, being an uncommon residue in its position within the human variable region sequence and therefore potentially immunogenic, reducing aggregation potential, and other reasons, the following nine variable region framework positions were considered as substitution candidates for the two illustrated human mature light chain variable regions and the two illustrated human mature heavy chain variable regions, as further specified in the examples: L64 (G64S), L104 (V104L), H1 (Q1E), H24 (V24A), H48 (M48I), H67 (V67A), H69 (I69M), H93 (A93T), and H94 (R94T).

[0629] Here, as elsewhere, the first mentioned residue is the residue of the humanized antibody formed by grafting Kabat CDR or the complex Chothia-Kabat CDR (in the case of CDR-H1) into the human receptor framework, and the second mentioned residue is the residue that is considered to replace this residue. Thus, within the variable region framework, the first mentioned residue is human, and within the CDR, the first mentioned residue is mouse.

[0630] The illustrated antibodies include any arrangement or combination of the illustrated mature heavy chain and light chain variable regions: hu10C12VHv1 / hu10C12VLv1, hu10C12VHv1 / hu10C12VLv2, hu10C12VHv2 / hu10C12VLv1, hu10C12VHv2 / hu10C12VLv2.

[0631] The illustrated antibodies comprise any permutation or combination of the illustrated mature heavy chain variable regions hu10C12VHv1 (SEQ ID NO:214) and hu10C12VHv2 (SEQ ID NO:215) and any of the illustrated mature light chain variable regions hu10C12VLv1 (SEQ ID NO:216) and hu10C12VLv2 (SEQ ID NO:217).

[0632] This invention provides variants of 10C12 humanized antibodies, wherein the humanized mature heavy chain variable region exhibits at least 90%, 95%, 96%, 97%, 98%, or 99% identity with hu10C12VHv1 (SEQ ID NO: 214) or hu10C12VHv2 (SEQ ID NO: 215), and the humanized mature light chain variable region exhibits at least 90%, 95%, 96%, 97%, 98%, or 99% identity with hu10C12VLv1 (SEQ ID NO: 216) or hu10C12VLv2 (SEQ ID NO: 217). In some of these antibodies, at least 1, 2, 3, 4, 5, 6, 7, 8, or all nine reversion mutations or other mutations in SEQ ID NO: 214-215 and SEQ ID NO: 216-217 are retained.

[0633] In some humanized 10C12 antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H24 is occupied by A, H48 by I, H67 by A, H69 by M, H93 by T, and H94 by T. In some humanized 10C12 antibodies, positions H24, H48, H67, H69, H93, and H94 are occupied by A, I, A, M, T, and T, respectively.

[0634] In some humanized 10C12 antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H1 is occupied by Q or E, H24 is occupied by A, H48 is occupied by I, H67 is occupied by A, H69 is occupied by M, H93 is occupied by T, and H94 is occupied by T.

[0635] In some humanized 10C12 antibodies, positions H24, H48, H67, H69, H93, and H94 are substituted by A, I, A, M, T, and T, respectively, as in hu10C12VHv1. In some humanized 10C12 antibodies, positions H1, H24, H48, H67, H69, H93, and H94 are substituted by E, A, I, A, M, T, and T, respectively, as in hu10C12VHv2.

[0636] In some humanized 10C12 antibodies, position L64 in the VL region is occupied by S.

[0637] In some humanized 10C12 antibodies, at least one of the following positions in the VL region is occupied by a specified amino acid: L64 is occupied by S, and L104 is occupied by V or L.

[0638] In some humanized 10C12 antibodies, positions L64 and L104 in the VL region are occupied by S, as in hu10C12VLv1. In some humanized 10C12 antibodies, positions L64 and L104 in the VL region are occupied by S and L, respectively, as in hu10C12VLv2.

[0639] In some humanized 10C12 antibodies, the variable heavy chain has ≥85% identity with the human sequence. In some humanized 10C12 antibodies, the variable light chain has ≥85% identity with the human sequence. In some humanized 10C12 antibodies, each of the variable heavy chain and variable light chain has ≥85% identity with the human germline sequence. In some humanized 10C12 antibodies, the three heavy chain CDRs are as defined by the Kabat / Chothia complex (SEQ ID NO: 8, 9, and 10), and the three light chain CDRs are as defined by the Kabat / Chothia complex (SEQ ID NO: 12, 13, and 14).

[0640] An exemplary humanized antibody includes a humanized form of mouse 12C4, named Hu12C4.

[0641] Mouse antibody 12C4 includes mature heavy and light chain variable regions, which have amino acid sequences comprising SEQ ID NO:219 and SEQ ID NO:11, respectively. This invention provides two exemplified humanized mature heavy chain variable regions: hu12C4VHv1 and hu12C4VHv2. This invention also provides two exemplified mature light chain variable regions: hu12C4VLv1 and hu12C4VLv2. Figure 9 and Figure 10 The comparisons of the heavy chain variable region and light chain variable region of mouse 12C4 and various humanized antibodies are shown respectively.

[0642] For reasons such as potentially affecting CDR conformation and / or binding to antigens, mediating interactions between heavy and light chains, interactions with constant regions, being a site of desired or undesirable post-translational modifications, being an uncommon residue in its position within the human variable region sequence and therefore potentially immunogenic, gaining aggregation potential, and other reasons, the following six variable region framework positions are considered as substitution candidates for the two illustrated human mature light chain variable regions and the two illustrated human mature heavy chain variable regions, as further specified in the examples: L64 (G64S), L104 (V104L), H1 (Q1E), H48 (M48I), H93 (A93T), and H94 (R94T).

[0643] Here, as elsewhere, the first mentioned residue is the residue of the humanized antibody formed by grafting Kabat CDR or the complex Chothia-Kabat CDR (in the case of CDR-H1) into the human receptor framework, and the second mentioned residue is the residue that is considered to replace this residue. Thus, within the variable region framework, the first mentioned residue is human, and within the CDR, the first mentioned residue is mouse.

[0644] The illustrated antibodies include any arrangement or combination of the illustrated mature heavy chain and light chain variable regions: hu12C4VHv1 / hu12C4VLv1, hu12C4VHv1 / hu12C4VLv2, hu12C4VHv2 / hu12C4VLv1, hu12C4VHv2 / hu12C4VLv2.

[0645] The illustrated antibodies include any permutation or combination of the illustrated mature heavy chain variable regions hu12C4VHv1 (SEQ ID NO:221) and hu12C4VHv2 (SEQ ID NO:222) and any of the illustrated mature light chain variable regions hu12C4VLv1 (SEQ ID NO:223) and hu12C4VLv2 (SEQ ID NO:224).

[0646] This invention provides variants of 12C4 humanized antibodies, wherein the humanized mature heavy chain variable region exhibits at least 90%, 95%, 96%, 97%, 98%, or 99% identity with hu12C4VHv1 (SEQ ID NO:221) or hu12C4VHv2 (SEQ ID NO:222), and the humanized mature light chain variable region exhibits at least 90%, 95%, 96%, 97%, 98%, or 99% identity with hu12C4VLv1 (SEQ ID NO:223) or hu12C4VLv2 (SEQ ID NO:224). In some of these antibodies, at least 1, 2, 3, 4, 5, or all six reversion mutations or other mutations in SEQ ID NO:221-222 and SEQ ID NO:223-224 are retained.

[0647] In some humanized 12C4 antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H1 is occupied by Q or E, H48 is occupied by M or I, H93 is occupied by A or T, and H94 is occupied by R or T.

[0648] In some humanized 12C4 antibodies, positions H1, H48, H93, and H94 in the VH region are occupied by E, I, T, and T, respectively, as in hu12C4VHv2.

[0649] In some humanized 12C4 antibodies, at least one of the following positions in the VL region is occupied by a specified amino acid: L64 is occupied by G or S, and L104 is occupied by V or L.

[0650] In some humanized 12C4 antibodies, positions L64 and L104 in the VL region are occupied by S and L, respectively, as in hu12C4VLv2.

[0651] In some humanized 12C4 antibodies, the variable heavy chain has ≥85% identity with the human sequence. In some humanized 12C4 antibodies, the variable light chain has ≥85% identity with the human sequence. In some humanized 12C4 antibodies, each of the variable heavy chain and variable light chain has ≥85% identity with the human germline sequence. In some humanized 12C4 antibodies, the three heavy chain CDRs are as defined by the Kabat / Chothia complex (SEQ ID NO: 8, 220, and 10), and the three light chain CDRs are as defined by the Kabat / Chothia complex (SEQ ID NO: 12, 13, and 14).

[0652] An exemplary humanized antibody is a humanized form of mouse 17C12, named Hu17C12.

[0653] Mouse antibody 17C12 includes mature heavy and light chain variable regions, which have amino acid sequences comprising SEQ ID NO:225 and SEQ ID NO:228, respectively. This invention provides two exemplified humanized mature heavy chain variable regions: hu17C12VHv and hu17C12VHv2. This invention also provides two exemplified mature light chain variable regions: hu17C12VLv1 and hu17C12VLv2. Figure 11 and Figure 12 The comparisons of the heavy chain variable region and light chain variable region of mouse 17C12 and various humanized antibodies are shown respectively.

[0654] For reasons such as potentially affecting CDR conformation and / or binding to antigens, mediating interactions between heavy and light chains, interactions with constant regions, being a site of desired or undesirable post-translational modifications, being an uncommon residue in its position within the human variable region sequence and therefore potentially immunogenic, gaining aggregation potential, and other reasons, the following 13 variable region framework sites were considered as substitution candidates for the two illustrated human mature light chain variable regions and the two illustrated human mature heavy chain variable regions, as further specified in the examples: L2 (I2V), L36 (Y36L), L43 (P43S), H1 (Q1E), H2 (V2I), H24 (V24A), H48 (M48I), H67 (V67A), H69 (I69M), H93 (A93T), H94 (R94T), H108 (T108L), and H113 (R113S).

[0655] Here, as elsewhere, the first mentioned residue is the residue of the humanized antibody formed by grafting Kabat CDR or the complex Chothia-Kabat CDR (in the case of CDR-H1) into the human receptor framework, and the second mentioned residue is the residue that is considered to replace this residue. Thus, within the variable region framework, the first mentioned residue is human, and within the CDR, the first mentioned residue is mouse.

[0656] The illustrated antibodies include any arrangement or combination of the illustrated mature heavy chain and light chain variable regions: hu17C12VHv1 / hu17C12VLv1, hu17C12VHv1 / hu17C12VLv2, hu17C12VHv2 / hu17C12VLv1, hu17C12VHv2 / hu17C12VLv2.

[0657] The illustrated antibodies include any permutation or combination of the illustrated mature heavy chain variable regions hu17C12VHv1 (SEQ ID NO:232) and hu17C12VHv2 (SEQ ID NO:233) and any of the illustrated mature light chain variable regions hu17C12VLv1 (SEQ ID NO:234) and hu17C12VLv2 (SEQ ID NO:235).

[0658] This invention provides variants of 17C12 humanized antibodies, wherein the humanized mature heavy chain variable region exhibits at least 90%, 95%, 96%, 97%, 98%, or 99% identity with hu17C12VHv1 (SEQ ID NO:232) or hu17C12VHv2 (SEQ ID NO:233), and the humanized mature light chain variable region exhibits at least 90%, 95%, 96%, 97%, 98%, or 99% identity with hu17C12VLv1 (SEQ ID NO:234) or hu17C12VLv2 (SEQ ID NO:235). In some of these antibodies, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or all 13 reversion mutations or other mutations from SEQ ID NO:232-233 and SEQ ID NO:234-235 are retained.

[0659] In some humanized 17C12 antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H2 is occupied by I, H24 by A, H48 by I, H67 by A, H69 by M, H93 by T, and H94 by T. In some humanized 17C12 antibodies, positions H2, H24, H48, H67, H69, H93, and H94 are occupied by E, A, I, A, M, T, and T, respectively.

[0660] In some humanized 17C12 antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H1 is occupied by Q or E, H2 is occupied by I, H24 is occupied by A, H48 is occupied by I, H67 is occupied by A, H69 is occupied by M, H93 is occupied by T, H94 is occupied by T, H108 is occupied by T or L, and H113 is occupied by R or S.

[0661] In some humanized 17C12 antibodies, positions H2, H24, H48, H67, H69, H93, and H94 in the VH region are occupied by E, A, I, A, M, T, and T, respectively, as in hu17C12VHv1. In some humanized 17C12 antibodies, positions H1, H2, H24, H48, H67, H69, H93, H94, H108, and H113 in the VH region are occupied by E, I, A, I, A, M, T, T, L, and S, respectively, as in hu17C12VHv2.

[0662] In some humanized 17C12 antibodies, at least one of the following positions in the VL region is occupied by a specified amino acid: L2 is occupied by V, and L36 is occupied by L. In some humanized 17C12 antibodies, positions L2 and L36 are occupied by V and L, respectively.

[0663] In some humanized 17C12 antibodies, at least one of the following positions in the VL region is occupied by a specified amino acid: L2 is occupied by V, L36 is occupied by L, and L43 is occupied by P or S.

[0664] In some humanized 17C12 antibodies, positions L2 and L36 in the VL region are occupied by V and L, respectively, as in hu17C12VLv1. In some humanized 17C12 antibodies, positions L2, L36, and L43 in the VL region are occupied by V, L, and S, respectively, as in hu17C12VLv2.

[0665] In some humanized 17C12 antibodies, the variable heavy chain has ≥85% identity with the human sequence. In some humanized 17C12 antibodies, the variable light chain has ≥85% identity with the human sequence. In some humanized 17C12 antibodies, each of the variable heavy chain and variable light chain has ≥85% identity with the human germline sequence. In some humanized 9F5 antibodies, the three heavy chain CDRs are as defined by the Kabat / Chothia complex (SEQ ID NO: 226, 227, and 10), and the three light chain CDRs are as defined by the Kabat / Chothia complex (SEQ ID NO: 229-231).

[0666] An exemplary humanized antibody is a humanized form of mouse 14H3, named Hu14H3.

[0667] Mouse antibody 14H3 includes mature heavy and light chain variable regions, which have amino acid sequences comprising SEQ ID NO:240 and SEQ ID NO:244, respectively. This invention provides two exemplified humanized mature heavy chain variable regions: hu14H3VHv1 and hu14H3VHv2. This invention also provides two exemplified mature light chain variable regions: hu14H3VLv1 and hu14H3VLv2. Figure 13 and Figure 14 The comparisons of the heavy chain variable regions and light chain variable regions of mouse 14H3 and various humanized antibodies are shown respectively.

[0668] For reasons such as potentially affecting CDR conformation and / or binding to antigens, mediating interactions between heavy and light chains, interactions with constant regions, being a site of desired or undesirable post-translational modifications, being an uncommon residue in its position within the human variable region sequence and therefore potentially immunogenic, gaining aggregation potential, and other reasons, the following eight variable region framework sites are considered substitution candidates for the two illustrated human mature light chain variable regions and the two illustrated human mature heavy chain variable regions, as further specified in the examples: L2 (I2V), L7 (T7S), L37 (L37Q), L87 (Y87F), L100 (G100Q), L104 (V104L), H108 (M108L), and H113 (L113S). The following variable region CDR site is considered a substitution candidate for the two illustrated human mature light chain variable regions, as further specified in the examples: H35B (G35BS). Among some humanized 14H3 antibodies, the Kabat-Chothia complex CDR-H1 has an amino acid sequence containing SEQ ID NO:275.

[0669] Here, as elsewhere, the first mentioned residue is the residue of the humanized antibody formed by grafting Kabat CDR or the complex Chothia-Kabat CDR (in the case of CDR-H1) into the human receptor framework, and the second mentioned residue is the residue that is considered to replace this residue. Thus, within the variable region framework, the first mentioned residue is human, and within the CDR, the first mentioned residue is mouse.

[0670] The illustrated antibodies include any arrangement or combination of the illustrated mature heavy chain and light chain variable regions: hu14H3VHv1 / hu14H3VLv1, hu14H3VHv1 / hu14H3VLv2, hu14H3VHv2 / hu14H3VLv1, hu14H3VHv2 / hu14H3VLv2.

[0671] The illustrated antibodies include any permutation or combination of the illustrated mature heavy chain variable regions hu14H3VHv1 (SEQ ID NO:248) and hu14H3VHv2 (SEQ ID NO:249) and any of the illustrated mature light chain variable regions hu14H3VLv1 (SEQ ID NO:250) and hu14H3VLv2 (SEQ ID NO:251).

[0672] This invention provides variants of humanized antibodies against 14H3, wherein the humanized mature heavy chain variable region exhibits at least 90%, 95%, 96%, 97%, 98%, or 99% identity with hu14H3VHv1 (SEQ ID NO:249) or hu14H3VHv2 (SEQ ID NO:250), and the humanized mature light chain variable region exhibits at least 90%, 95%, 96%, 97%, 98%, or 99% identity with hu14H3VLv1 (SEQ ID NO:251) or hu14H3VLv2 (SEQ ID NO:252). In some of these antibodies, at least 1, 2, 3, 4, 5, 6, 7, 8, or all nine reversion mutations or other mutations from SEQ ID NO:249-250 and SEQ ID NO:251-252 are retained.

[0673] In some humanized 14H3 antibodies, the H35B position in the VH region is occupied by S.

[0674] In some humanized 14H3 antibodies, at least one of the following positions in the VH region is occupied by a specified amino acid: H35B is occupied by S, H108 is occupied by M or L, and H113 is occupied by L or S.

[0675] In some humanized 14H3 antibodies, position H35B in the VH region is occupied by S, such as in hu14H3VHv1. In some humanized 14H3 antibodies, positions H35B, H108, and H113 in the VH region are occupied by S, L, and S, respectively, such as in hu14H3VHv2.

[0676] In some humanized 14H3 antibodies, at least one of the following positions in the VL region is occupied by a specified amino acid: L2 is occupied by V and L87 is occupied by F. In some humanized 14H3 antibodies, positions L2 and L87 are occupied by V and F, respectively.

[0677] In some humanized 14H3 antibodies, at least one of the following positions in the VL region is occupied by a specified amino acid: L2 is occupied by V, L7 is occupied by T or S, L37 is occupied by L or Q, L87 is occupied by F, L100 is occupied by G or Q, and L104 is occupied by V or L.

[0678] In some humanized 14H3 antibodies, positions L2 and L87 in the VL region are occupied by V and F, respectively, as in hu14H3VLv1. In some humanized 14H3 antibodies, positions L2, L7, L37, L87, L100, and L104 in the VL region are occupied by V, S, Q, F, Q, and L, respectively, as in hu14H3VLv2.

[0679] In some humanized 14H3 antibodies, the variable heavy chain has ≥85% identity with the human sequence. In some humanized 14H3 antibodies, the variable light chain has ≥85% identity with the human sequence. In some humanized 14H3 antibodies, each of the variable heavy chain and variable light chain has ≥85% identity with the human germline sequence. In some humanized 14H3 antibodies, the three heavy chain CDRs are as defined by the Kabat / Chothia complex (SEQ ID NO:241-243), provided that position H35B is occupied by G or S; and the three light chain CDRs are as defined by the Kabat / Chothia complex (SEQ ID NO:245-247).

[0680] Among some humanized 14H3 antibodies, the Kabat-Chothia complex CDR-H1 has an amino acid sequence containing SEQ ID NO:275.

[0681] The CDR regions of these humanized 9F5, 10C12, 2D11, 12C4, 17C12, and 14H3 antibodies may be identical or substantially identical to the CDR regions of 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3. The CDR regions may be defined by any conventional definition (e.g., Chothia, or a complex of Chothia and Kabat), but are preferably defined as by Kabat.

[0682] Unless otherwise specified, the variable region frame positions conform to the Kabat numbering. Other such variants typically differ from the illustrated Hu9F5, Hu10C12, Hu12C4, Hu17C12, or Hu14H3 heavy and light chain sequences by a small number of substitutions, deletions, or insertions (e.g., usually no more than 1, 2, 3, 5, 10, or 15). Such differences are usually in frames but can also exist in CDRs.

[0683] Further variations of the humanized 9F5, 10C12, 2D11, 12C4, 17C12, and 14H3 variants are possible through additional reversion mutations in the variable region framework. Many framework residues in the humanized mAb that are not in contact with the CDR can be adapted for substitution of corresponding amino acids from donor mouse mAbs or other mouse or human antibodies, and even many potential CDR-contacting residues are permitted for substitution. Even amino acids within the CDR can be altered, for example, by using residues found at corresponding positions in the human receptor sequence used to provide the variable region framework. Alternatively, alternative human receptor sequences can be used, for example, for the heavy and / or light chains. If a different receptor sequence is used, then one or more of the reversion mutations recommended above may not be necessary, since the corresponding donor and receptor residues are already identical without reversion mutations.

[0684] Preferably, substitution or reversion mutations (whether conserved or not) in humanized 9F5, 10C12, 2D11, 12C4, 17C12 and 14H3 variants have no substantial effect on the binding affinity or potency of the humanized mAb, i.e. its ability to bind to tau.

[0685] Humanized 9F5 antibodies were further characterized by their ability to bind both phosphorylated and unphosphorylated tau, as well as misfolded / aggregated forms of tau.

[0686] D. Chimeric and decorative antibodies

[0687] The present invention also provides non-human antibodies, specifically chimeric and veneered forms of examples of 9F5, 10C12, 2D11, 12C4, 17C12 or 14H3 antibodies.

[0688] Chimeric antibodies are antibodies in which the mature variable regions of the light and heavy chains of a non-human antibody (e.g., mouse) are combined with the constant regions of the human light and heavy chains. Such antibodies retain essentially or completely the binding specificity of mouse antibodies and are approximately two-thirds human sequences.

[0689] Decorated antibodies are a type of humanized antibody that retains some, and typically all, of the core derivations (CDRs) and some nonhuman variable region framework residues of the nonhuman antibody, but replaces other variable region framework residues that can contribute to B or T cell epitopes, such as exposed residues (Padlan, Mol. Immunol. 28:489, 1991), with residues from corresponding positions in the human antibody sequence. The result is where the CDRs are entirely or substantially derived from the nonhuman antibody, and the variable region framework of the nonhuman antibody becomes more like a human antibody through substitution. Decorated forms of 9F5, 10C12, 2D11, 12C4, 17C12, and 14H3 antibodies are included in this invention.

[0690] E. Human antibodies

[0691] Human antibodies that specifically bind to tau or fragments thereof (e.g., amino acid sequences comprising QIVYKP (SEQ ID NO:57), EIVYKSP (SEQ ID NO:58), EIVYKS (SEQ ID NO:277), or (Q / E)IVYK(S / P) (SEQ ID NO:56) or peptides thereof) are provided by a variety of techniques described below. Some human antibodies are selected to have the same epitope specificity as a specific mouse antibody (such as one of the mouse monoclonal antibodies described in the examples) by competitive binding assays, by Winter's phage display method (above), or otherwise. Human antibodies can also be screened for specific epitope specificity by using only fragments of tau, such as tau fragments containing the amino acid sequence of QIVYKP (SEQ ID NO:57), EIVYKSP (SEQ ID NO:58), EIVYKS (SEQ ID NO:277) or (Q / E)IVYK(S / P) (SEQ ID NO:56) as target antigens, and / or by screening antibodies against a set of tau variants, such as tau variants containing various mutations in amino acid residues 307-312, 391-397, or 391-396 of SEQ ID NO:1.

[0692] Methods for generating human antibodies include the trisomy hybridoma method, as described in Oestberg et al., Hybridoma 2:361-367 (1983); Oestberg, U.S. Patent 4,634,664; and Engleman et al., U.S. Patent 4,634,666; using transgenic mice containing human immunoglobulin genes (see, for example, Lonberg et al., WO93 / 12227 (1993); US5,877,397; US5,874,299; US5,814,318; US5,789,650; US5,770,429; US5,661,016; US5,633,425; US5,625,126; US5,569,825; US 5,545,806; Neuberger, Nat. Biotechnol. 14:826 (1996); and Kucherlapati, WO91 / 10741 (1991); phage display methods (see, for example, Dower et al., WO91 / 17271; McCafferty et al., WO 92 / 01047; US 5,877,218; US5,871,907; US 5,858,657; US 5,837,242; US 5,733,743; and US5,565,332); and the methods described in WO 2008 / 081008 (e.g., immortalizing memory B cells isolated from humans, for example with EBV, screening for desired characteristics, and cloning and expressing recombinant forms).

[0693] F. Selection of the constant region

[0694] The variable regions of the heavy and light chains of chimeric, veneered, or humanized antibodies can be linked to at least a portion of the human constant region. The choice of the constant region depends in part on whether antibody-dependent cell-mediated cytotoxicity, antibody-dependent phagocytosis, and / or complement-dependent cytotoxicity are required. For example, human allotypes IgG1 and IgG3 exhibit complement-dependent cytotoxicity, while human allotypes IgG2 and IgG4 do not. Human IgG1 and IgG3 also induce stronger cell-mediated effector functions than human IgG2 and IgG4. The light chain constant region can be λ or κ. The numbering conventions for constant regions include EU numbers (Edelman, GM et al., Proc. Natl. Acad. USA, 63, 78-85 (1969)), Kabat numbers (Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, MD, 1991), IMGT unique numbers (Lefranc M.-P. et al., IMGT unique numbering for immunoglobulin and T cell receptor constant domains and Ig superfamily C-like domains, Dev. Comp. Immunol., 29, 185-203 (2005)), and IMGT exon numbers (Lefranc, ibid.).

[0695] One or more amino acids at the amino or carboxyl terminus of the light and / or heavy chains, such as the C-terminal lysine of the heavy chain, may be deleted or derived in part or all of the molecule. Substitutions may be made in the constant region to reduce or increase effector functions, such as complement-mediated cytotoxicity or ADCC (see, e.g., Winter et al., U.S. Patent No. 5,624,821; Tso et al., U.S. Patent No. 5,834,597; and Lazar et al., Proc. Natl. Acad. Sci. USA 103:4005, 2006), or to prolong the half-life in humans (see, e.g., Hinton et al., J. Biol. Chem. 279:6213, 2004). Exemplary substitutions include Gln at position 250 and / or Leu at position 428 (EU number used in the constant region in this section) to increase the half-life of the antibody. Substitutions at any or all of positions 234, 235, 236, and / or 237 reduce affinity for Fcγ receptors, particularly the FcγRI receptor (see, for example, US 6,624,821). Alanine substitutions at positions 234, 235, and 237 of human IgG1 can be used to reduce effector function. Some antibodies have alanine substitutions at positions 234, 235, and 237 of human IgG1 for the purpose of reducing effector function. Optionally, positions 234, 236, and / or 237 of human IgG2 are substituted with alanine, and position 235 is substituted with glutamine (see, for example, US 5,624,821). In some antibodies, mutations are used at one or more of positions 241, 264, 265, 270, 296, 297, 322, 329, and 331 (EU numbers) of human IgG1. In some antibodies, mutations are used at one or more of positions 318, 320, and 322 (EU number) of human IgG1. In some antibodies, positions 234 and / or 235 are substituted with alanine and / or position 329 is substituted with glycine. In some antibodies, positions 234 and 235 are substituted with alanine. In some antibodies, the isotype is human IgG2 or IgG4.

[0696] Antibodies can be expressed as tetramers containing two light chains and two heavy chains, single heavy chains, light chains, Fab, Fab', F(ab')2, and Fv, or single-chain antibodies in which the mature variable domains of the heavy and light chains are linked by spacers.

[0697] Human constant regions exhibit both allotropic and ethnotropic variations between individuals; that is, at one or more polymorphic locations, the constant region can differ among individuals. Ethnotropics differ from allotropics in that serum recognizing an ethnotropic binds to one or more other non-polymorphic regions of the same allotype. Thus, for example, another heavy chain constant region belongs to IgG1G1m3 with or without a C-terminal lysine. References to human constant regions include constant regions having any natural allotype or any arrangement of residues occupying a position within a natural allotype.

[0698] G. Expression of recombinant antibodies

[0699] Numerous methods are known for generating chimeric and humanized antibodies u...

Claims

1. An isolated monoclonal antibody that competes with antibody 9F5 for binding to human tau. 2 . The antibody of claim 1 , wherein the heavy chain CDR-H3 has an amino acid sequence comprising SEQ ID NO:

10. 3 . The antibody of claim 1 , wherein the heavy chain CDR-H1 has an amino acid sequence comprising SEQ ID NO:

8.

4. The antibody of claim 2, wherein the light chain CDRs CDR-L1, CDR-L2 and CDR-L3 have amino acid sequences comprising SEQ ID NOs: 12, 13 and 14, respectively). The antibody of claim 4 , wherein the heavy chain CDR-H1 has an amino acid sequence comprising SEQ ID NO:

8.

6. The antibody of claim 1, which binds to the same epitope on human tau as 9F5.

7. The antibody of claim 1, comprising three light chain CDRs and three heavy chain CDRs of monoclonal antibody 9F5, wherein 9F5 is a mouse antibody characterized by having a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 7 and a light chain variable region comprising an amino acid sequence of SEQ ID NO:

11.

8. The antibody of claim 7, wherein the three heavy chain CDRs CDR-H1, CDR-H2 and CDR-H3 are as defined by the Kabat / Chothia complex (SEQ ID NOs: 8, 9 and 10, respectively), except that position H28 may be occupied by N or T, position H51 may be occupied by I or V, position H54 may be occupied by N or D and position H56 may be occupied by D or E; and the three light chain CDRs CDR-L1, CDR-L2 and CDR-L3 are as defined by the Kabat / Chothia complex (SEQ ID NOs: 8, 9 and 10, respectively). NO:12, 13 and 14), except that position L27b is occupied by L, D, T or Q, position L27c is occupied by L, D, G, S, E, T, N, A, P or I, position L30 can be occupied by I, Y, E, K, G or Q, position L31 can be occupied by T, N or G, position L33 can be occupied by L, N, T, S, R or G, position L51 can be occupied by M, G, E, D, K or I, position L54 can be occupied by L, R, G or T, position L89 is occupied by A or G, position L92 is occupied by L, D, E, G, Q, T or I and position L93 is occupied by E or G. 9 . The antibody of claim 8 , wherein CDR-H1 has an amino acid sequence comprising SEQ ID NO:

50.

10. The antibody of claim 8, wherein CDR-H2 has an amino acid sequence comprising SEQ ID NO:

51.

11. The antibody of claim 8, wherein CDR-H2 has an amino acid sequence comprising SEQ ID NO:

52. 12 . The antibody of claim 8 , wherein CDR-L1 has an amino acid sequence comprising any one of SEQ ID NO: 53, SEQ ID NO: 54, and SEQ ID NOs: 172-193.

13. The antibody of claim 8, wherein CDR-L2 has an amino acid sequence comprising any one of SEQ ID NO: 55 and SEQ ID NO: 194-205.

14. The antibody of claim 8, wherein CDR-L3 has an amino acid sequence comprising any one of SEQ ID NOs: 206-213.

15. The antibody of claim 8, wherein CDR-H1 has an amino acid sequence comprising SEQ ID NO:50, and CDR-H2 has an amino acid sequence comprising SEQ ID NO:

51.

16. The antibody of claim 8, wherein CDR-L1 has an amino acid sequence comprising SEQ ID NO: 53, and CDR-L2 has an amino acid sequence comprising SEQ ID NO:

55.

17. The antibody of claim 8, wherein CDR-L1 has an amino acid sequence comprising SEQ ID NO: 54, and CDR-L2 has an amino acid sequence comprising SEQ ID NO:

55.

18. The antibody of any one of the preceding claims, wherein the antibody is a humanized antibody.

19. The humanized antibody of claim 18, wherein the humanized mature heavy chain variable region comprises the three Kabat heavy chain CDRs of 9F5 (SEQ ID NO:40, SEQ ID NO:9, and SEQ ID NO:10), and the humanized mature light chain variable region comprises the three Kabat light chain CDRs of 9F5 (SEQ ID NO:12-14).

20. The humanized antibody of any one of claims 1-19, comprising a humanized mature heavy chain variable region having an amino acid sequence at least 90% identical to any one of SEQ ID NOs: 15-22 and SEQ ID NOs: 109-129, and a humanized mature light chain variable region having an amino acid sequence at least 90% identical to any one of SEQ ID NOs: 23-29, SEQ ID NOs: 61-108, and SEQ ID NOs: 130-171.

21. The humanized antibody of claim 20, wherein at least one of the following positions in the VH region is occupied by a specified amino acid: H1 is occupied by Q or E, H5 is occupied by Q or V, H11 is occupied by L or V, H12 is occupied by V or K, H17 is occupied by S or T, H20 is occupied by L or I, H23 is occupied by T or K, H28 is occupied by N or T, H38 is occupied by K, R or Q, H40 is occupied by R or A, H42 is occupied by E or G, H43 is occupied by Q or K, H48 is occupied by I or M, H51 is occupied by I or V, and H54 is occupied by N or D. , H56 is occupied by D or E, H66 is occupied by K or R, H69 is occupied by I or M, H75 is occupied by S or T, H76 is occupied by N or D, H79 is occupied by Y, Q, D, N or G, H80 is occupied by L, M, P, D, G or E, H81 is occupied by Q or E, H82 is occupied by L, P, K, R, E or N, H82a is occupied by S or G, H82c is occupied by L, G, D or S, H83 is occupied by T or R, H93 is occupied by A or T, H94 is occupied by S or T, H108 is occupied by T or L, and H109 is occupied by L or V.

22. A humanized antibody as described in claim 21, with the proviso that positions H1, H5, H11, H12, H17, H20, H23, H38, H42, H43, H66, H69, H75, H80, H81, H83, H93, H94, H108 and H109 in the VH region are occupied by E, V, V, K, T, I, K, Q, G, K, R, M, T, M, E, R, T, T, L and V, respectively.

23. The humanized antibody of claim 22, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:

127.

24. A humanized antibody as described in claim 21, with the proviso that positions H1, H5, H11, H12, H17, H20, H23, H38, H42, H43, H66, H69, H75, H80, H81, H83, H93, H94, H108 and H109 in the VH region are occupied by E, V, V, K, T, I, K, K, E, K, R, M, T, M, E, R, T, T, L and V, respectively.

25. The humanized antibody of claim 24, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:

128.

26. The humanized antibody of claim 20, wherein at least one of the following positions in the VL region is occupied by a specified amino acid: L3 is occupied by V or Q, L7 is occupied by A or S, L8 is occupied by A or P, L9 is occupied by F or L, L11 is occupied by N or L, L15 is occupied by L or P, L17 is occupied by T or E, L18 is occupied by S or P, L27b is occupied by L, D, T or Q, L27c is occupied by L, D, G, S, E, T, N, A, P or I, L30 is occupied by I, Y, E, K, G, L31 is occupied by T, N or G, L33 is occupied by L, N, T, S, R or G, and L37 is occupied by L, Q, G or I , L39 is occupied by R or K, L51 is occupied by M, G, E, D, K or I, L54 is occupied by R, G or T, L60 is occupied by N or D, L64 is occupied by G or S, L66 is occupied by E or G, L73 is occupied by L, P or G, L74 is occupied by R or K, L75 is occupied by I, D, P, Q or G, L76 is occupied by S, P or G, L77 is occupied by R or D, L78 is occupied by V, R, D, E, P, K, G or Q, L85 is occupied by V or G, L86 is occupied by Y or T, L89 is occupied by A or G, L92 is occupied by L, D, E, G, Q, T or I, L93 is occupied by E or G, and L100 is occupied by G or Q.

27. The humanized antibody of claim 21, wherein positions L3Q, L27c, L37, L51, L54 and L92 in the VL region are occupied by Q, S, Q, G, R and I.

28. The humanized antibody of claim 21, wherein positions L3, L27c, L37, L51, L54 and L92 in the VL region are occupied by Q, S, Q, G, T and I, respectively.

29. The humanized antibody of claim 21, wherein positions L3, L27c, L37, L51, L54 and L92 in the VL region are occupied by Q, G, Q, G, R and I, respectively.

30. The humanized antibody of claim 21, wherein positions L3, L27c, L37, L51, L54 and L92 in the VL region are occupied by Q, D, Q, G, R and I, respectively.

31. A humanized antibody as described in claim 21, with the proviso that positions L3, L27c, L37, L51, L54 and L92 in the VL region are occupied by Q, D, Q, K, R and I, respectively.

32. A humanized antibody as described in claim 21, with the proviso that positions L3, L27c, L37, L51, L54 and L92 in the VL region are occupied by Q, G, Q, K, R and I, respectively.

33. A humanized antibody as described in claim 21, with the proviso that positions L3, L27c, L37, L51, L54 and L92 in the VL region are occupied by Q, G, G, G, R and I, respectively.

34. The humanized antibody of claim 21, wherein positions L3, L27c, L37, L51, L54 and L92 in the VL region are occupied by Q, G, G, G, R and G, respectively.

35. The humanized antibody of claim 21, wherein positions L3, L27c, L37, L51, L54 and L92 in the VL region are occupied by Q, G, G, G, T and I.

36. A humanized antibody as described in claim 21, with the proviso that positions L3, L27c, L37, L51, L54 and L92 in the VL region are occupied by Q, S, G, G, T and I, respectively.

37. A humanized antibody as described in claim 21, with the proviso that positions L3, L27c, L37, L51, L54 and L92 in the VL region are occupied by Q, D, G, G, R and I, respectively.

38. The humanized antibody of claim 21, wherein positions L3, L37, L51, L54 and L92 in the VL region are occupied by Q, Q, G, R and I, respectively.

39. The humanized antibody of claim 21, wherein positions L3, L27c, L51, L54 and L92 in the VL region are occupied by Q, S, G, R and I, respectively.

40. The humanized antibody of claim 21, wherein positions L3, L27c, L37, L54 and L92 in the VL region are occupied by Q, S, Q, R and I, respectively.

41. The humanized antibody of claim 21, wherein positions L3, L27c, L37, L51 and L92 in the VL region are occupied by Q, S, Q, G and I, respectively.

42. A humanized antibody as described in claim 21, with the proviso that positions L3, L27c, L37, L51, L54 and L92 in the VL region are occupied by Q, G, G, G, R and I, respectively.

43. The humanized antibody of claim 20, wherein the mature heavy chain variable region has the amino acid sequence of any one of SEQ ID NOs: 15-22 and SEQ ID NOs: 109-129, and the mature light chain variable region has the amino acid sequence of any one of SEQ ID NOs: 23-29, SEQ ID NOs: 61-108, and SEQ ID NOs: 130-171.

44. The humanized antibody of claim 43, wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:

149.

45. The humanized antibody of claim 437, wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:

142.

46. ​​The humanized antibody of claim 43, wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:

159.

47. The humanized antibody of claim 43, wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:

148.

48. The humanized antibody of claim 43, wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:

137.

49. The humanized antibody of claim 43, wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:

145.

50. The humanized antibody of claim 43, wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:

136.

51. The humanized antibody of claim 43, wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:

138.

52. The humanized antibody of claim 44, wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:

158.

53. The humanized antibody of claim 43, wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:

143.

54. The humanized antibody of claim 43, wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:

144.

55. The humanized antibody of claim 43, wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:

133.

56. The humanized antibody of claim 43, wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:

160.

57. The humanized antibody of claim 43, wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:

161.

58. The humanized antibody of claim 43, wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 127, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:

139.

59. The humanized antibody of claim 43, wherein the mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 128, and the mature light chain variable region has the amino acid sequence of SEQ ID NO:

168.

60. The antibody of claim 1, comprising three light chain CDRs and three heavy chain CDRs of monoclonal antibody 10C12, wherein 10C12 is a mouse antibody characterized by having a heavy chain variable region comprising an amino acid sequence of SEQ ID NO:7 and a light chain variable region comprising an amino acid sequence of SEQ ID NO:

11.

61. The antibody of claim 60, wherein the antibody is a humanized antibody.

62. A humanized antibody as described in claim 61, wherein the humanized mature heavy chain variable region comprises the three Kabat heavy chain CDRs of 10C12 (SEQ ID NO:40, SEQ ID NO:9 and SEQ ID NO:10), and the humanized mature light chain variable region comprises the three Kabat light chain CDRs of 10C12 (SEQ ID NO:12-14).

63. A humanized antibody as described in any one of claims 60-62, wherein the humanized antibody comprises a humanized mature heavy chain variable region having an amino acid sequence at least 90% identical to any one of SEQ ID NOs: 214-215 and a humanized mature light chain variable region having an amino acid sequence at least 90% identical to any one of SEQ ID NOs: 216-217.

64. A humanized antibody as described in claim 63, wherein at least one of the following positions in the VH region is occupied by a specified amino acid: H1 is occupied by Q or E, H24 is occupied by A, H48 is occupied by I, H67 is occupied by A, H69 is occupied by M, H93 is occupied by T, and H94 is occupied by T.

65. The humanized antibody of claim 64, wherein positions H1, H24, H48, H67, H69, H93 and H94 are occupied by E, A, I, A, M, T and T, respectively.

66. A humanized antibody as described in claim 63, wherein at least one of the following positions in the VL region is occupied by a specified amino acid: L64 is occupied by S, and L104 is occupied by V or L.

67. The humanized antibody of claim 66, wherein positions L64 and L104 in the VL region are occupied by S and L, respectively.

68. The humanized antibody of claim 63, wherein the mature heavy chain variable region has the amino acid sequence of any one of SEQ ID NOs: 214-215, and the mature light chain variable region has the amino acid sequence of any one of SEQ ID NOs: 216-217.

69. The antibody of claim 5, comprising three light chain CDRs and three heavy chain CDRs of monoclonal antibody 12C4, wherein 12C4 is a mouse antibody characterized by having a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 219 and a light chain variable region comprising an amino acid sequence of SEQ ID NO:

11.

70. The antibody of claim 69, wherein the antibody is a humanized antibody.

71. A humanized antibody as described in claim 70, wherein the humanized mature heavy chain variable region comprises the three Kabat heavy chain CDRs of 12C4 (SEQ ID NO:40, SEQ ID NO:220 and SEQ ID NO:10), and the humanized mature light chain variable region comprises the three Kabat light chain CDRs of 12C4 (SEQ ID NOs:12-14).

72. A humanized antibody as described in any one of claims 69-71, wherein the humanized antibody comprises a humanized mature heavy chain variable region having an amino acid sequence at least 90% identical to any one of SEQ ID NOs: 221-222 and a humanized mature light chain variable region having an amino acid sequence at least 90% identical to any one of SEQ ID NOs: 223-224.

73. A humanized antibody as described in claim 72, wherein at least one of the following positions in the VH region is occupied by a specified amino acid: H1 is occupied by Q or E, H48 is occupied by M or I, H93 is occupied by A or T, and H94 is occupied by R or T.

74. The humanized antibody of claim 73, wherein positions H1, H48, H93 and H94 in the VH region are occupied by E, I, T and T, respectively.

75. A humanized antibody as described in claim 72, wherein the positions in the VL region are occupied by specified amino acids: L64 is occupied by G or S, and L104 is occupied by V or L.

76. The humanized antibody of claim 75, wherein positions L64 and L104 in the VL region are occupied by S and L, respectively.

77. The humanized antibody of claim 72, wherein the mature heavy chain variable region has the amino acid sequence of any one of SEQ ID NOs: 221-222, and the mature light chain variable region has the amino acid sequence of any one of SEQ ID NOs: 223-224.

78. The antibody of claim 2, comprising three light chain CDRs and three heavy chain CDRs of monoclonal antibody 17C12, wherein 17C12 is a mouse antibody characterized by having a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 225 and a light chain variable region comprising an amino acid sequence of SEQ ID NO:

228.

79. The antibody of claim 78, wherein the antibody is a humanized antibody.

80. A humanized antibody as described in claim 79, wherein the humanized mature heavy chain variable region comprises the three Kabat heavy chain CDRs of 17C12 (SEQ ID NO:40, SEQ ID NO:227 and SEQ ID NO:10), and the humanized mature light chain variable region comprises the three Kabat light chain CDRs of 17C12 (SEQ ID NOs:229-231).

81. A humanized antibody as described in any one of claims 78-80, wherein the humanized antibody comprises a humanized mature heavy chain variable region having an amino acid sequence at least 90% identical to any one of SEQ ID NOs: 232-233 and a humanized mature light chain variable region having an amino acid sequence at least 90% identical to any one of SEQ ID NOs: 234-235.

82. A humanized antibody as described in claim 81, wherein at least one of the following positions in the VH region is occupied by a specified amino acid: H1 is occupied by Q or E, H2 is occupied by I, H24 is occupied by A, H48 is occupied by I, H67 is occupied by A, H69 is occupied by M, H93 is occupied by T, H94 is occupied by T, H108 is occupied by T or L, and H113 is occupied by R or S.

83. A humanized antibody as described in claim 82, with the proviso that positions H1, H2, H24, H48, H67, H69, H93, H94, H108 and H113 in the VH region are occupied by E, I, A, I, A, M, T, T, L and S, respectively.

84. A humanized antibody as described in claim 81, wherein at least one of the following positions in the VL region is occupied by a specified amino acid: L2 is occupied by V, L36 is occupied by L, and L43 is occupied by P or S.

85. The humanized antibody of claim 84, with the proviso that positions L2, L36 and L43 in the VL region are occupied by V, L and S, respectively.

86. The humanized antibody of claim 81, wherein the mature heavy chain variable region has the amino acid sequence of any one of SEQ ID NOs: 232-233, and the mature light chain variable region has the amino acid sequence of any one of SEQ ID NOs: 234-235.

87. An antibody as described in claim 1, wherein the antibody comprises three light chain CDRs and three heavy chain CDRs of monoclonal antibody 14H3, wherein 14H3 is a mouse antibody characterized by having a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 240 and a light chain variable region comprising an amino acid sequence of SEQ ID NO:

244.

88. The antibody of claim 87, wherein the three heavy chain CDRs CDR-H1, CDR-H2, and CDR-H3 are as defined by the Kabat / Chothia complex (SEQ ID NOs: 241, 242, and 243, respectively), except that position H35B may be occupied by G or S, and the three light chain CDRs CDR-L1, CDR-L2, and CDR-L3 are as defined by the Kabat / Chothia complex (SEQ ID NOs: 245, 246, and 247, respectively).

89. The antibody of claim 88, wherein CDR-H1 has an amino acid sequence comprising SEQ ID NO:

277.

90. The antibody of any one of claims 87-89, wherein the antibody is a humanized antibody.

91. A humanized antibody as described in claim 90, wherein the humanized mature heavy chain variable region comprises the three Kabat heavy chain CDRs of 14H3 (SEQ ID NO:265, SEQ ID NO:242 and SEQ ID NO:243), and the humanized mature light chain variable region comprises the three Kabat light chain CDRs of 14H3 (SEQ ID NO:245-247).

92. A humanized antibody as described in any one of claims 87-91, wherein the humanized antibody comprises a humanized mature heavy chain variable region having an amino acid sequence at least 90% identical to any one of SEQ ID NOs: 248-249 and a humanized mature light chain variable region having an amino acid sequence at least 90% identical to any one of SEQ ID NOs: 250-251.

93. A humanized antibody as described in claim 92, wherein at least one of the following positions in the VH region is occupied by a specified amino acid: H35B is occupied by S, H108 is occupied by M or L, and H113 is occupied by L or S.

94. The humanized antibody of claim 93, wherein positions H35B, H108 and H113 in the VH region are occupied by S, L and S, respectively.

95. A humanized antibody as described in claim 92, wherein at least one of the following positions in the VL region is occupied by a specified amino acid: L2 is occupied by V, L7 is occupied by T or S, L37 is occupied by L or Q, L87 is occupied by F, L100 is occupied by G or Q, and L104 is occupied by V or L.

96. A humanized antibody as described in claim 95, with the proviso that positions L2, L7, L37, L87, L100 and L104 in the VL region are occupied by V, S, Q, F, Q and L, respectively.

97. The humanized antibody of claim 92, wherein the mature heavy chain variable region has the amino acid sequence of any one of SEQ ID NOs: 248-249, and the mature light chain variable region has the amino acid sequence of any one of SEQ ID NOs: 250-251.

98. The antibody of any one of claims 1-97, which is a whole antibody.

99. The antibody of any one of claims 1-97, which is a binding fragment.

100. The antibody of any one of the preceding claims, wherein the isotype is human IgGl.

101. A humanized antibody as described in any one of claims 18-59, 61-68, 70-77, 79-86 and 90-100, wherein the mature light chain variable region is fused to the light chain constant region and the mature heavy chain variable region is fused to the heavy chain constant region.

102. The humanized antibody of claim 101, wherein the heavy chain constant region is a mutated form of a native human heavy chain constant region, and the mutated form of the native human heavy chain constant region has reduced binding to an Fcγ receptor relative to the native human heavy chain constant region.

103. The antibody of any one of claims 1-99 and 101-102, wherein the isotype is of the human IgG2 or IgG4 isotype.

104. A pharmaceutical composition comprising an antibody as defined in any one of claims 1-103 and a pharmaceutically acceptable carrier.

105. A nucleic acid encoding the heavy chain and / or light chain of the antibody of any one of claims 1-104.

106. A method for humanizing a mouse antibody, the method comprising: (a) selecting one or more receptor antibodies; (b) identifying amino acid residues of the mouse antibody to be retained; (c) synthesizing a nucleic acid encoding a humanized heavy chain comprising the CDRs of a mouse antibody heavy chain and a nucleic acid encoding a humanized light chain comprising the CDRs of a mouse antibody light chain; and (d) expressing the nucleic acid in a host cell to produce a humanized antibody; wherein the mouse antibody is 9F5, wherein 9F5 is characterized by a mature heavy chain variable region of SEQ ID NO: 7 and a mature light chain variable region of SEQ ID NO: 11, wherein the mouse antibody is 10C12, wherein 10C12 is characterized by a mature heavy chain variable region of SEQ ID NO:7 and a mature light chain variable region of SEQ ID NO:11, wherein the mouse antibody is 2D11, wherein 2D11 is characterized by a mature heavy chain variable region of SEQ ID NO: 7 and a mature light chain variable region of SEQ ID NO: 11, wherein the mouse antibody is 12C4, wherein 12C4 is characterized by a mature heavy chain variable region of SEQ ID NO: 219 and a mature light chain variable region of SEQ ID NO: 11, wherein the mouse antibody is 17C12, wherein 17C12 is characterized by a mature heavy chain variable region of SEQ ID NO: 225 and a mature light chain variable region of SEQ ID NO: 228, Wherein the mouse antibody is 14H3, wherein 14H3 is characterized by a mature heavy chain variable region of SEQ ID NO:240 and a mature light chain variable region of SEQ ID NO:

244.

107. A method of inhibiting or reducing tau aggregation in a subject having or at risk for developing tau-mediated amyloidosis, the method comprising administering to the subject an effective regimen of an antibody as described in any one of claims 1-104, thereby inhibiting or reducing tau aggregation in the subject.

108. A method for treating a tau-associated disease or achieving prevention of a tau-associated disease in a subject, the method comprising administering an effective regimen of an antibody as defined in any one of claims 1-104, and thereby treating the disease or achieving prevention of the disease.

109. The method of claim 108, wherein the tau-related disease is Alzheimer's disease, Down syndrome, mild cognitive impairment, primary age-related tauopathy, postencephalitic Parkinson's disease, post-traumatic dementia or dementia pugilistica, Pick's disease, Niemann-Pick disease type C, supranuclear palsy, frontotemporal dementia, frontotemporal lobar degeneration, argyrophilic grain disease, globular glial tauopathy, Guam amyotrophic lateral sclerosis / Parkinson's dementia complex, corticobasal degeneration (CBD), Lewy body dementia, Lewy body variant of Alzheimer's disease (LBVAD), chronic traumatic encephalopathy (CTE), globular glial tauopathy (GGT), Parkinson's disease, or progressive supranuclear palsy (PSP).

110. The method of claim 109, wherein the tau-related disease is Alzheimer's disease.

111. A method for detecting tau protein deposits in a subject having or at risk for a disease associated with tau aggregation or deposition, the method comprising administering to the subject an antibody as defined in any one of claims 1-104, and detecting the antibody that binds to tau in the subject.

112. An isolated monoclonal antibody that specifically binds to a peptide consisting of residues (Q / E)IVYK(S / P) (SEQ ID NO:56).

113. An isolated monoclonal antibody that specifically binds to a peptide consisting of residues QIVYKP (SEQ ID NO: 57).

114. An isolated monoclonal antibody that specifically binds to a peptide consisting of residues EIVYKSP (SEQ ID NO:58).

115. An isolated monoclonal antibody that specifically binds to a peptide consisting of residues EIVYKS (SEQ ID NO: 277).

116. A method for treating or preventing a tau-related disease in a subject, the method comprising administering an immunogen comprising a tau peptide of up to 20 consecutive amino acids of SEQ ID NO: 1 that specifically binds to antibody 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3, wherein the peptide induces the formation of an antibody that specifically binds to tau in the subject.

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