Antibodies that recognize tau

Abstract

The present invention provides antibodies that specifically bind to tau, which inhibit or slow the progression of tau-related pathologies and associated symptoms.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit under 35 U.S.C. §119(e) 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, each of which is incorporated by reference in its entirety for all purposes.

[0002] A reference to a sequence list The sequence listing in file 2020_02_07_542496WO_SEQLST.txt is 3560 kb, was created on February 7, 2020, and is incorporated herein by reference. [Background technology]

[0003] Tau is a well-known human protein that can exist in phosphorylated forms (see, e.g., 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 those in the central nervous system. It has been reported that t-tau (total tau, i.e., phosphorylated and non-phosphorylated forms), as well as phospho-tau (p-tau, i.e., phosphorylated tau), are released by the brain in response to neuronal injury and neurodegeneration and appear at elevated levels in the CSF of Alzheimer's patients compared to the general population (Jack et al., Lancet Neurol 9: 119-28 (2010)).

[0004] Tau is the main component of neurofibrillary tangles, which, together with plaques, are a hallmark of Alzheimer's disease. These tangles consist of abnormal fibrils measuring 10 nm in diameter that are in the form of paired coiled helices with a regular period of 80 nm. Tau in neurofibrillary tangles is abnormally phosphorylated (hyperphosphorylated), with phosphate groups attached to specific sites on the molecule. In Alzheimer's disease, significant involvement of neurofibrillary tangles is found in neurons of layer II of the entorhinal cortex, the CA1 and subiculum regions of the hippocampus, the amygdala, and in deeper layers of the neocortex (layers III, V, and superficial layer VI). Hyperphosphorylated tau has also been reported to interfere with microtubule assembly, which may promote the collapse of neuronal networks.

[0005] Tau inclusions are part of the defining neuropathology of several neurodegenerative diseases, including Alzheimer's disease, frontotemporal lobar degeneration, progressive supranuclear palsy, and Pick's disease. Summary of the Invention

[0006] In one embodiment, 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 chain CDRs 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 such antibodies bind to the same epitope on human tau as 9F5.

[0007] Some such antibodies comprise the three light chain CDRs and three heavy chain CDRs of monoclonal antibody 9F5, a murine antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:7 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:11.

[0008] In some such antibodies, the three heavy chain CDRs, CDR-H1, CDR-H2, and CDR-H3, are as defined by the Kabat / Chothia combination, except that N or T may occupy position H28, I or V may occupy position H51, N or D may occupy position H54, and D or E may occupy position H56 (SEQ ID NOs: 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 combination, except that N or T may occupy position H28, I or V may occupy position H51, N or D may occupy position H54, and D or E may occupy position H56 (SEQ ID NOs: 8, 9, and 10, respectively). as defined by the Kabat / Chothia complex, except that I, Y, E, K, G, or Q may occupy position L30, T, N, or G may occupy position L31, L, N, T, S, R, or G may occupy position L33, M, G, E, D, K, or I may occupy position L51, L, R, G, or T may occupy position L54, A or G may occupy position L89, L, D, E, G, Q, T, or I may occupy position L92, and E or G may occupy position L93 (SEQ ID NOs: 12, 13, and 14, respectively).

[0009] In some antibodies, CDR-H1 has an amino acid sequence comprising SEQ ID NO: 50. In some antibodies, CDR-H2 has an amino acid sequence comprising SEQ ID NO: 51. In some antibodies, CDR-H2 has an amino acid sequence comprising SEQ ID NO: 52. In some antibodies, CDR-L1 has an amino acid sequence comprising any of SEQ ID NO: 53, SEQ ID NO: 54, and SEQ ID NO: 172-193. In some antibodies, CDR-L2 has an amino acid sequence comprising any of SEQ ID NO: 55, and SEQ ID NO: 194-205. In some antibodies, CDR-L3 has an amino acid sequence comprising any of SEQ ID NO: 206-213. In some antibodies, 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. In some antibodies, 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. In some antibodies, 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.

[0010] Some antibodies are 9F5, or chimeric, veneered, or humanized forms thereof. In some antibodies, the variable heavy chain has 85% or more identity to a human sequence. In some antibodies, the variable light chain has 85% or more identity to a human sequence. In some antibodies, the variable heavy and variable light chains each have 85% or more identity to a human germline sequence. Some antibodies are humanized antibodies.

[0011] Some antibodies are humanized or chimeric 9F5 antibodies that specifically bind human tau, where 9F5 is a murine antibody 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. Some antibodies comprise a humanized mature heavy chain variable region comprising the three heavy chain CDRs of 9F5 and a humanized mature light chain variable region comprising the three light chain CDRs of 9F5. In some antibodies, the CDRs are defined CDRs selected from the group of Kabat, Chothia, Kabat / Chothia composite, AbM, and Contact.

[0012] In some antibodies, the humanized mature heavy chain variable region comprises the three Kabat / Chothia composite heavy chain CDRs of 9F5 (SEQ ID NO:8-10), and the humanized mature light chain variable region comprises the three Kabat / Chothia composite light chain CDRs of 9F5 (SEQ ID NO:12-14). In some antibodies, 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). In some antibodies, the humanized mature heavy chain variable region comprises the 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 the three Chothia light chain CDRs of 9F5 (SEQ ID NO:12-14). In some antibodies, the humanized mature heavy chain variable region comprises the 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 comprises the three AbM light chain CDRs of 9F5 (SEQ ID NO:12-14). In some antibodies, the humanized mature heavy chain variable region comprises the three Contact heavy chain CDRs of 9F5 (SEQ ID NO:44-46), and the humanized mature light chain variable region comprises the three Contact light chain CDRs of 9F5 (SEQ ID NO:47-49).

[0013] For example, the antibody is a humanized antibody, a veneered antibody, or a chimeric antibody.

[0014] Some such antibodies comprise a humanized mature heavy chain variable region having an amino acid sequence that is at least 90% identical to any one of SEQ ID NOs: 15-22 and 109-129, and a humanized mature light chain variable region having an amino acid sequence that is at least 90% identical to any one of SEQ ID NOs: 23-29, 61-108, and 130-171.

[0015] In some antibodies, at least one of the following positions in the VH region is occupied by the indicated amino acid: H1 is occupied by E, H17 is occupied by T, H20 is occupied by I, H69 is occupied by M, H75 is occupied by T, H93 is occupied by T, H94 is occupied by T, and H109 is occupied 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.

[0016] In some antibodies, at least one of the following positions in the VH region is occupied by the indicated 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.

[0017] In some antibodies, at least one of the following positions in the VH region is occupied by the indicated 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.

[0018] In some antibodies, at least one of the following positions in the VH region is occupied by the indicated amino acid: H43 is occupied by K, H51 is occupied by V, H76 is occupied by D, M80 is occupied by M, and H108 is occupied by L. In some antibodies, positions H43, H51, H76, H80, and H108 are occupied by K, V, D, M, and L, respectively.

[0019] In some antibodies, position H28 of the VH region is occupied by a T.

[0020] In some antibodies, at least one of the following positions in the VH region is occupied by the indicated 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.

[0021] In some antibodies, position H40 of the VH region is occupied by an A.

[0022] In some antibodies, at least one of the following positions in the VH region is occupied by the indicated 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. In some antibodies, positions H5, H11, H12, H38, and H42 are occupied by V, V, K, R, and G, respectively.

[0023] In some antibodies, at least one of the following positions in the VH region is occupied by the indicated 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, H54 is occupied by N or D, and H 56 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.

[0024] In some 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. In some 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. In some 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. In some antibodies, positions H1, H17, H20, H23, H28, H43, H51, H54, H56, H66, H69, H75, H76, H80, H81, H83, H93, H94, H108, and H109 of 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, positions H1, H17, H20, H23, H28, H40, H43, H48, H51, H54, H56, H66, H69, H75, H76, H80, H81, H83, H93, H94, H108, and H109 of 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, 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 of 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, 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 of 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.

[0025] 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 of the VH region are occupied, respectively, by E, V, V, K, T, I, K, Q, G, K, R, M, T, M, E, R, T, T, L, and V. In some antibodies, the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 127.

[0026] 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 of the VH region are occupied, respectively, by E, V, V, K, T, I, K, K, E, K, R, M, T, M, E, R, T, T, L, and V. In some antibodies, the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:128.

[0027] In some antibodies, positions H1, H5, H11, H12, H17, H20, H23, H38, H42, H43, H66, H69, H75, H80, H81, H82c, H83, H93, H94, H108, and H109 of 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.

[0028] 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.

[0029] 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.

[0030] In some antibodies, at least one of the following positions in the VL region is occupied by the indicated amino acid: L7 is occupied by S, L8 is occupied by P, L15 is occupied by P, and L100 is occupied by Q. In some antibodies, positions L7, L8, L15, and L100 are occupied by S, P, P, and Q, respectively.

[0031] In some antibodies, position L66 of the VL region is occupied by G. In some antibodies, position L64 of the VL region is occupied by S.

[0032] In some antibodies, position L17 of 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 designated amino acid: L11 is occupied by L, L51 is occupied by G, and L54 is occupied by R.

[0033] In some antibodies, positions L11, L51, and L54 are occupied by L, G, and R, respectively.

[0034] In some antibodies, at least one of the following positions in the VL region is occupied by the indicated amino acid: L3 is V or Q, L7 is A or S, L8 is A or P, L9 is F or L, L11 is N or L, L15 is L or P, L17 is T or E, L18 is S or P, L27b is L, D, T, or Q, L27c is L, D, G, S, E, T, N, A, P, or I, L30 is I, Y, E, K, G, L31 is T, N, or G, L33 is L, N, T, S, R, or G, L37 is L, Q, G, or I, and L39 is R or or K, L51 is M, G, E, D, K, or I, L54 is R, G, or T, L60 is N or D, L64 is G or S, L66 is E or G, L73 is L, P, or G, L74 is R or K, L75 is I, D, P, Q, or G, L76 is S, P, or G, L77 is R or D, L78 is V, R, D, E, P, K, G, or Q, L85 is V or G, L86 is Y or T, L89 is A or G, L92 is L, D, E, G, Q, T, or I, L93 is E or G, and L100 is G or Q.

[0035] 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.

[0036] In some antibodies, the light chain variable region comprises any of the amino acid sequences of SEQ ID NOs: 133, 135-137, 142-144, 149, 158, 159, and 168. In some antibodies, the light chain variable region comprises the amino acid sequence of SEQ ID NO: 133. In some antibodies, the light chain variable region comprises the amino acid sequence of SEQ ID NO: 137. In some antibodies, the light chain variable region comprises the amino acid sequence of SEQ ID NO: 149. In some antibodies, the light chain variable region comprises the amino acid sequence of SEQ ID NO: 159.

[0037] 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 of the VL region are occupied by S, P, L, L, P, E, P, N, K, G, R, D, G, K, and Q, respectively.

[0038] In some antibodies, positions L7, L8, L11, L15, L17, L39, L60, L64, L66, L74, and L100 of the VL region are occupied by S, P, L, P, E, K, N, S, G, K, and Q, respectively. In some antibodies, position L3 of the VL region is occupied by Q. In some antibodies, position L27c of the VL region is occupied by D, G, I, L, or S, position L37 of the VL region is occupied by G, I, L, or Q, position L51 of the VL region is occupied by E, G, I, K, or M, position L54 of the VL region is occupied by G, L, R, or T, and position L92 of 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.

[0039] In some antibodies, position L27c of the VL region is occupied by D, position L37 of the VL region is occupied by G, and position L51 of the VL region is occupied by G. In some antibodies, the heavy chain variable region has an amino acid sequence comprising SEQ ID NO: 127, and the light chain variable region has an amino acid sequence comprising SEQ ID NO: 149.

[0040] In some antibodies, position L27c of the VL region is occupied by D, position L37 of the VL region is occupied by Q, and position L51 of the VL region is occupied by G. In some antibodies, the heavy chain variable region has an amino acid sequence comprising SEQ ID NO:127, and the light chain variable region has an amino acid sequence comprising SEQ ID NO:137.

[0041] In some antibodies, position L27c of the VL region is occupied by S, position L37 of the VL region is occupied by L, and position L51 of the VL region is occupied by G. In some antibodies, the heavy chain variable region has an amino acid sequence comprising SEQ ID NO:127, and the light chain variable region has an amino acid sequence comprising SEQ ID NO:159.

[0042] In some antibodies, position L27c of the VL region is occupied by D, position L37 of the VL region is occupied by Q, and position L51 of the VL region is occupied by K. In some antibodies, the heavy chain variable region has an amino acid sequence comprising SEQ ID NO:127, and the light chain variable region has an amino acid sequence comprising SEQ ID NO:138.

[0043] In some antibodies, position L27c of the VL region is occupied by S, position L37 of the VL region is occupied by Q, and position L51 of the VL region is occupied by G. In some antibodies, the heavy chain variable region has an amino acid sequence comprising SEQ ID NO: 127, and the light chain variable region has an amino acid sequence comprising SEQ ID NO: 133.

[0044] 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.

[0045] In some antibodies, position L27c of the VL region is occupied by G, position L37 of the VL region is occupied by G, position L51 of the VL region is occupied by G, and position L54 of the VL region is occupied by R. In some antibodies, position L92 of the VL region is occupied by I. In some antibodies, the heavy chain variable region has an amino acid sequence comprising SEQ ID NO:129, and the light chain variable region has an amino acid sequence comprising SEQ ID NO:168.

[0046] 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.

[0047] 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 provided 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.

[0048] 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.

[0049] 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 provided 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.

[0050] 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 provided 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.

[0051] 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. In some antibodies, positions L3Q, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, G, R, and I. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, G, T, 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 of the VL region are occupied by Q, G, Q, G, R, and I, respectively.

[0052] 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. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, D, Q, K, 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. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, Q, K, G, and I, respectively. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 of the VL region are occupied by Q, S, Q, K, G, and I, respectively.

[0053] 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. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, G, G, R, and G, respectively. In some antibodies, positions L3, L27c, L37, L51, and L54 in the VL region are occupied by Q, G, G, G, and R, respectively. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, G, G, T, and I. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 of the VL region are occupied by Q, G, G, G, T, and G, respectively.

[0054] 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, I, I, R, and I, respectively. In some antibodies, positions L3, L27c, L37, L51, L54, and L92 of the VL region are occupied by Q, S, Q, I, G, and I, respectively.

[0055] 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, Q, E, G, 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 of the VL region are occupied by Q, G, I, E, R, and G, respectively.

[0056] 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, R, and I, respectively. In some antibodies, positions L3, L27c, L37, L51, and L92 in the VL region are occupied by Q, S, Q, R, and I, respectively.

[0057] 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 of the VL region are occupied by Q, G, Q, K, R, and I, respectively.

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

[0059] Some antibodies comprise a mature heavy chain variable region having an amino acid sequence that is at least 95% identical to any one of SEQ ID NOs: 15 - 22 and SEQ ID NOs: 109 - 129, and a mature light chain variable region having an amino acid sequence that is at least 95% identical to any one of SEQ ID NOs: 23 - 29, SEQ ID NOs: 61 - 108, and SEQ ID NOs: 130 - 171. Some antibodies comprise a mature heavy chain variable region having an amino acid sequence that is at least 98% identical to any one of SEQ ID NOs: 15 - 22 and SEQ ID NOs: 109 - 129, and a mature light chain variable region having an amino acid sequence that is at least 98% identical to any one of SEQ ID NOs: 23 - 29, SEQ ID NOs: 61 - 108, and SEQ ID NOs: 130 - 171. In some antibodies, 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.

[0060] 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.

[0061] 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.

[0062] 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.

[0063] 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.

[0064] 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.

[0065] 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.

[0066] 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.

[0067] 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.

[0068] 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.

[0069] 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.

[0070] 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.

[0071] 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.

[0072] Some such antibodies comprise the three light chain CDRs and three heavy chain CDRs of monoclonal antibody 10C12, a murine antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:7 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:11.

[0073] 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 NOs: 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 NOs: 12, 13, and 14, respectively).

[0074] Some antibodies are 10C12, or chimeric, veneered, or humanized forms thereof. In some antibodies, the variable heavy chain has 85% or more identity to human sequences. In some antibodies, the variable light chain has 85% or more identity to human sequences. In some antibodies, the variable heavy and variable light chains each have 85% or more identity to human germline sequences. Some antibodies are humanized antibodies.

[0075] Some antibodies are humanized or chimeric 10C12 antibodies that specifically bind to human tau, where 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. Some antibodies comprise a humanized mature heavy chain variable region comprising the three heavy chain CDRs of 10C12 and a humanized mature light chain variable region comprising the three light chain CDRs of 10C12. In some antibodies, the CDRs are defined CDRs selected from the group of Kabat, Chothia, Kabat / Chothia composite, AbM, and Contact.

[0076] In some antibodies, the humanized mature heavy chain variable region comprises the three Kabat / Chothia composite heavy chain CDRs of 10C12 (SEQ ID NO:8-10), and the humanized mature light chain variable region comprises the three Kabat / Chothia composite light chain CDRs of 10C12 (SEQ ID NO:12-14). In some antibodies, 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). In some antibodies, the humanized mature heavy chain variable region comprises the 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 the three Chothia light chain CDRs of 10C12 (SEQ ID NO:12-14). In some antibodies, the humanized mature heavy chain variable region comprises the 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 the three AbM light chain CDRs of 10C12 (SEQ ID NO:12-14). In some antibodies, the humanized mature heavy chain variable region comprises the three Contact heavy chain CDRs of 10C12 (SEQ ID NO:44-46), and the humanized mature light chain variable region comprises the three Contact light chain CDRs of 10C12 (SEQ ID NO:47-49).

[0077] For example, the antibody can be a humanized antibody, a veneered antibody, or a chimeric antibody.

[0078] Some such antibodies comprise a humanized mature heavy chain variable region having an amino acid sequence that is 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 that is at least 90% identical to any one of SEQ ID NOs: 216-217.

[0079] In some antibodies, at least one of the following positions in the VH region is occupied by the indicated amino acid: 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. In some antibodies, positions H24, H48, H67, H69, H93, and H94 are occupied by A, I, A, M, T, and T, respectively.

[0080] In some antibodies, at least one of the following positions in the VH region is occupied by the indicated 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.

[0081] In some antibodies, positions H24, H48, H67, H69, H93, and H94 are occupied by A, I, A, M, T, and T, respectively. In some antibodies, positions H1, H24, H48, H67, H69, H93, and H94 are occupied by E, A, I, A, M, T, and T, respectively.

[0082] In some antibodies, position L64 of the VL region is occupied by S.

[0083] In some antibodies, at least one of the following positions in the VL region is occupied by the indicated amino acid: L64 is S and L104 is 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.

[0084] Some antibodies comprise a mature heavy chain variable region having an amino acid sequence at least 95% identical to any one of SEQ ID NOs: 214-215, and a mature light chain variable region having an amino acid sequence at least 95% identical to any one of SEQ ID NOs: 216-217. Some antibodies comprise a mature heavy chain variable region having an amino acid sequence at least 98% identical to any one of SEQ ID NOs: 214-215, and a mature light chain variable region having an amino acid sequence at least 98% identical to any one of SEQ ID NOs: 216-217. In some antibodies, 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.

[0085] 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.

[0086] Some such antibodies comprise the three light chain CDRs and three heavy chain CDRs of monoclonal antibody 12C4, a murine antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:219 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:11.

[0087] 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 NOs: 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 NOs: 12, 13, and 14, respectively).

[0088] Some antibodies are 12C4 or chimeric, veneered, or humanized forms thereof. In some antibodies, the variable heavy chain has 85% or more identity to a human sequence. In some antibodies, the variable light chain has 85% or more identity to a human sequence. In some antibodies, the variable heavy and variable light chains each have 85% or more identity to a human germline sequence. Some antibodies are humanized antibodies.

[0089] Some antibodies are humanized or chimeric 12C4 antibodies that specifically bind to human tau, where 12C4 is a murine antibody 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. Some antibodies comprise a humanized mature heavy chain variable region that comprises the three heavy chain CDRs of 12C4 and a humanized mature light chain variable region that comprises the three light chain CDRs of 12C4. In some antibodies, the CDRs are defined CDRs selected from the group of Kabat, Chothia, Kabat / Chothia composite, AbM, and Contact.

[0090] In some antibodies, the humanized mature heavy chain variable region contains the three Kabat / Chothia combined heavy chain CDRs of 12C4 (SEQ ID NOs: 8, 220, and 10), and the humanized mature light chain variable region contains the three Kabat / Chothia combined light chain CDRs of 12C4 (SEQ ID NOs: 12 - 14). In some antibodies, the humanized mature heavy chain variable region contains 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 contains the three Kabat light chain CDRs of 12C4. In some antibodies, the humanized mature heavy chain variable region contains the three Chothia heavy chain CDRs of 12C4 (SEQ ID NOs: 41, 42, and SEQ ID NO: 10), and the humanized mature light chain variable region contains the three Chothia light chain CDRs of 12C4 (SEQ ID NOs: 12 - 14). In some antibodies, the humanized mature heavy chain variable region contains the 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 contains the three AbM light chain CDRs of 12C4 (SEQ ID NOs: 12 - 14). In some antibodies, the humanized mature heavy chain variable region contains the three Contact heavy chain CDRs of 12C4 (SEQ ID NOs: 44, 258, and 46), and the humanized mature light chain variable region contains the three Contact light chain CDRs of 12C4 (SEQ ID NOs: 47 - 49).

[0091] For example, the antibody can be a humanized antibody, a veneered antibody, or a chimeric antibody.

[0092] Some such antibodies include a humanized mature heavy chain variable region having an amino acid sequence that is 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 that is at least 90% identical to any one of SEQ ID NOs: 223 - 224.

[0093] In some antibodies, at least one of the following positions in the VH region is occupied by the indicated 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. In some antibodies, positions H1, H48, H93, and H94 in the VH region are occupied by E, I, T, and T, respectively.

[0094] In some antibodies, the VL region positions are occupied by the indicated amino acids: L64 is G or S, and L104 is V or L. In some antibodies, positions L64 and L104 in the VL region are occupied by S and L, respectively.

[0095] Some antibodies comprise a mature heavy chain variable region having an amino acid sequence at least 95% identical to any one of SEQ ID NOs: 221-222 and a mature light chain variable region having an amino acid sequence at least 95% identical to any one of SEQ ID NOs: 223-224. Some antibodies comprise a mature heavy chain variable region having an amino acid sequence at least 98% identical to any one of SEQ ID NOs: 221-222 and a mature light chain variable region having an amino acid sequence at least 98% identical to any one of SEQ ID NOs: 223-224. In some antibodies, 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.

[0096] 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.

[0097] Some such antibodies comprise the three light chain CDRs and three heavy chain CDRs of monoclonal antibody 17C12, a murine antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:225 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:228.

[0098] In some such antibodies, the three heavy chain CDRs, CDR-H1, CDR-H2, and CDR-H3, are as defined by the Kabat / Chothia composite (SEQ ID NOs: 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 composite (SEQ ID NOs: 229, 230, and 231, respectively).

[0099] Some antibodies are 17C12, or chimeric, veneered, or humanized forms thereof. In some antibodies, the variable heavy chain has 85% or more identity to human sequences. In some antibodies, the variable light chain has 85% or more identity to human sequences. In some antibodies, the variable heavy and variable light chains each have 85% or more identity to human germline sequences. Some antibodies are humanized antibodies.

[0100] Some antibodies are humanized or chimeric 17C12 antibodies that specifically bind human tau, where 17C12 is a murine antibody 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. Some antibodies comprise a humanized mature heavy chain variable region comprising the three heavy chain CDRs of 17C12 and a humanized mature light chain variable region comprising the three light chain CDRs of 17C12. In some antibodies, the CDRs are defined CDRs selected from the group of Kabat, Chothia, Kabat / Chothia composite, AbM, and Contact.

[0101] In some antibodies, the humanized mature heavy chain variable region comprises the three Kabat / Chothia composite heavy chain CDRs of 17C12 (SEQ ID NOs:226, 227, and 10) and the humanized mature light chain variable region comprises the three Kabat / Chothia composite light chain CDRs of 17C12 (SEQ ID NOs:229-231). In some antibodies, the humanized mature heavy chain variable region comprises the three Kabat heavy chain CDRs of 17C12 (SEQ ID NOs:40, 227, and 10) and the humanized mature light chain variable region comprises the three Kabat light chain CDRs of 17C12 (SEQ ID NOs:229-231). In some antibodies, the humanized mature heavy chain variable region comprises the 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 the three Chothia light chain CDRs of 17C12 (SEQ ID NO:229-231). In some antibodies, the humanized mature heavy chain variable region comprises the 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 the three AbM light chain CDRs of 17C12 (SEQ ID NO:229-231). In some antibodies, the humanized mature heavy chain variable region comprises the three Contact heavy chain CDRs of 17C12 (sequence numbers 44, 261, and 46), and the humanized mature light chain variable region comprises the three Contact light chain CDRs of 17C12 (sequence numbers 262-264).

[0102] For example, the antibody can be a humanized antibody, a veneered antibody, or a chimeric antibody.

[0103] Some such antibodies comprise a humanized mature heavy chain variable region having an amino acid sequence that is 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 that is at least 90% identical to any one of SEQ ID NOs: 234-235.

[0104] In some antibodies, at least one of the following positions in the VH region is occupied by the indicated amino acid: 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, and H94 is occupied 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.

[0105] In some antibodies, at least one of the following positions in the VH region is occupied by the indicated 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. 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 of the VH region are occupied by E, I, A, I, A, M, T, T, L, and S, respectively.

[0106] In some antibodies, at least one of the following positions in the VL region is occupied by the indicated 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.

[0107] In some antibodies, at least one of the following positions in the VL region is occupied by the indicated amino acid: L2 is V, L36 is L, and L43 is P or S. In some antibodies, positions L2 and 36 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.

[0108] Some antibodies comprise a mature heavy chain variable region having an amino acid sequence at least 95% identical to any one of SEQ ID NOs: 232 to 233, and a mature light chain variable region having an amino acid sequence at least 95% identical to any one of SEQ ID NOs: 234 to 235. Some antibodies comprise a mature heavy chain variable region having an amino acid sequence at least 98% identical to any one of SEQ ID NOs: 232 to 233, and a mature light chain variable region having an amino acid sequence at least 98% identical to any one of SEQ ID NOs: 234 to 235. In some antibodies, the mature heavy chain variable region has the amino acid sequence of any one of SEQ ID NOs: 232 to 233, and the mature light chain variable region has the amino acid sequence of any one of SEQ ID NOs: 234 to 235.

[0109] 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.

[0110] Some such antibodies comprise the three light chain CDRs and three heavy chain CDRs of monoclonal antibody 14H3, a murine antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:240 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:244.

[0111] In some such antibodies, the three heavy chain CDRs, CDR-H1, CDR-H2, and CDR-H3, are as defined by the Kabat / Chothia complex, except that G or S can occupy position H35B (SEQ ID NOs: 241, 242, and 243, respectively), 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). In some antibodies, CDR-H1 has an amino acid sequence comprising SEQ ID NO:277.

[0112] Some antibodies are 14H3 or chimeric, veneered, or humanized forms thereof. In some antibodies, the variable heavy chain has 85% or more identity to a human sequence. In some antibodies, the variable light chain has 85% or more identity to a human sequence. In some antibodies, the variable heavy and variable light chains each have 85% or more identity to a human germline sequence. Some antibodies are humanized antibodies.

[0113] Some antibodies are humanized or chimeric 14H3 antibodies that specifically bind to human tau, where 14H3 is a murine antibody 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. Some antibodies comprise a humanized mature heavy chain variable region that comprises the three heavy chain CDRs of 14H3 and a humanized mature light chain variable region that comprises the three light chain CDRs of 14H3. In some antibodies, the CDRs are defined CDRs selected from the group of Kabat, Chothia, Kabat / Chothia composite, AbM, and Contact.

[0114] In some antibodies, the humanized mature heavy chain variable region comprises the three Kabat / Chothia composite heavy chain CDRs of 14H3 (SEQ ID NOs:241-243) and the humanized mature light chain variable region comprises the three Kabat / Chothia composite light chain CDRs of 14H3 (SEQ ID NOs:245-247). In some antibodies, the humanized mature heavy chain variable region comprises the three Kabat heavy chain CDRs of 14H3 (SEQ ID NOs:265, 242, and 243) and the humanized mature light chain variable region comprises the three Kabat light chain CDRs of 14H3 (SEQ ID NOs:245-247). In some antibodies, the humanized mature heavy chain variable region comprises the 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 comprises the three Chothia light chain CDRs of 14H3 (SEQ ID NO:245-247). In some antibodies, the humanized mature heavy chain variable region comprises the 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 comprises the three AbM light chain CDRs of 14H3 (SEQ ID NO:245-247). In some antibodies, the humanized mature heavy chain variable region comprises the three Contact heavy chain CDRs of 14H3 (SEQ ID NO:269-271), and the humanized mature light chain variable region comprises the three Contact light chain CDRs of 14H3 (SEQ ID NO:272-274).

[0115] For example, the antibody can be a humanized antibody, a veneered antibody, or a chimeric antibody.

[0116] Some such antibodies comprise a humanized mature heavy chain variable region having an amino acid sequence that is 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 that is at least 90% identical to any one of SEQ ID NOs: 250-251.

[0117] In some antibodies, position H35B in the VH region is occupied by S.

[0118] In some antibodies, at least one of the following positions in the VH region is occupied by the indicated 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.

[0119] In some antibodies, at least one of the following positions in the VL region is occupied by the indicated 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.

[0120] In some antibodies, at least one of the following positions in the VL region is occupied by the indicated amino acid: L2 is V, L7 is T or S, L37 is L or Q, L87 is F, L100 is G or Q, and L104 is 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.

[0121] Some antibodies comprise a mature heavy chain variable region having an amino acid sequence at least 95% identical to any one of SEQ ID NOs: 248 to 249, and a mature light chain variable region having an amino acid sequence at least 95% identical to any one of SEQ ID NOs: 250 to 251. Some antibodies comprise a mature heavy chain variable region having an amino acid sequence at least 98% identical to any one of SEQ ID NOs: 248 to 249, and a mature light chain variable region having an amino acid sequence at least 98% identical to any one of SEQ ID NOs: 250 to 251. In some antibodies, the mature heavy chain variable region has the amino acid sequence of any one of SEQ ID NOs: 248 to 249, and the mature light chain variable region has the amino acid sequence of any one of SEQ ID NOs: 250 to 251.

[0122] 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.

[0123] For example, the antibody can be a chimeric antibody, for example, the antibody can be a veneered antibody.

[0124] The antibody may be an intact antibody. The antibody may be a binding fragment. In one embodiment, the binding fragment is a single chain antibody, a Fab, or a Fab'2 fragment. The antibody may be a Fab fragment or a single chain Fv. Some of the antibodies have a human IgG1 isotype, and others 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 version of the natural human heavy chain constant region that has reduced binding to Fcγ receptors compared to the natural human heavy chain constant region. In some antibodies, the heavy chain constant region is a heavy chain constant region of IgG1 isotype.

[0125] Some antibodies may have at least one mutation in the constant region, e.g., a mutation that reduces complement fixation or activation by the constant region, e.g., a mutation at one or more of positions 241, 264, 265, 270, 296, 297, 318, 320, 322, 329, and 331 according to EU numbering. Some antibodies have alanines at positions 318, 320, and 322. Some antibodies may be at least 95% (w / w) pure. The antibodies may be conjugated to a therapeutic, cytotoxic, cytostatic, neurotrophic, or neuroprotective agent.

[0126] In another aspect, the invention provides a pharmaceutical composition comprising any of the antibodies disclosed herein and a pharma- ceutically acceptable carrier.

[0127] In another aspect, the invention provides nucleic acids encoding the heavy and / or light chains of any of the antibodies disclosed herein, recombinant expression vectors comprising the nucleic acids, and host cells transformed with the recombinant expression vectors. Some nucleic acids have a sequence comprising any one of SEQ ID NOs: 38-39.

[0128] In another aspect, the invention provides a vector comprising a nucleic acid encoding a mature heavy chain variable region and a mature light chain variable region operably linked to one or more control sequences that affect the expression in a mammalian cell of any of the antibodies disclosed herein, a recombinant expression vector comprising the nucleic acid, a host cell transformed with the recombinant expression vector, and a host cell transformed with the 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 an scFv. In some vectors, the antibody is a Fab fragment. In some vectors, the one or more control sequences comprise one or more of a promoter, an enhancer, a ribosome binding site, and a transcription termination signal. In some vectors, the nucleic acid further encodes a signal peptide fused to the mature heavy chain variable region and the mature light chain variable region. In some vectors, the nucleic acid is codon-optimized for expression in a host cell. In some vectors, the one or more control sequences comprise a eukaryotic promoter. In some vectors, the nucleic acid further encodes a selectable gene.

[0129] In yet another aspect, the invention provides a method of expressing an antibody in a mammalian cell comprising the step of expressing said antibody by incorporating a nucleic acid disclosed herein into the genome of a transgenic animal.

[0130] In yet another aspect, the invention provides a first vector and a second vector comprising nucleic acid encoding a mature heavy chain variable region and a mature light chain variable region, respectively, each operably linked to one or more control sequences for affecting expression in a mammalian cell of any of the antibodies disclosed herein, and a host cell comprising said nucleic acid. In some first and second vectors, the nucleic acid further encodes 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.

[0131] In yet another aspect, the invention provides a method of expressing an antibody in a mammalian cell comprising the step of expressing said antibody by incorporating any of the nucleic acids disclosed herein into the genome of a transgenic animal.

[0132] In yet another aspect, the invention provides a method for humanizing any of the non-human antibodies described herein, such as murine antibody 9F5, 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, such as murine antibody 10C12, 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; such as murine antibody 2D11, 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; such as murine antibody 12C4, 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; such as murine antibody 17C12, 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; such as murine antibody 14H3, 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. Such methods may include the steps of selecting one or more acceptor antibodies and identifying the amino acid residues of the murine antibody to be retained; synthesizing a nucleic acid encoding a humanized heavy chain comprising the CDRs of the heavy chain of the murine antibody and a nucleic acid encoding a humanized light chain comprising the CDRs of the light chain of the murine antibody; and expressing the nucleic acids in a host cell to produce the humanized antibody.

[0133] Methods for producing humanized, chimeric, or veneered antibodies, such as 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 such that the cells secrete the antibody. The antibody can then be purified from the cell culture medium.

[0134] A cell line producing any of the antibodies disclosed herein can be produced by introducing into cells vectors encoding the antibody heavy and light chains and a selectable marker, growing the cells under conditions to select for cells with increased copy number of the vector, isolating a single cell from the selected cells; and depositing the cloned cells from the selected single cell based on antibody yield.

[0135] Some cells can grow under selective conditions and can be grown at least at 100 mg / L / 10 6 One can screen for cell lines that naturally express and secrete at 10 ...

[0136] The invention also provides methods of inhibiting or reducing tau aggregation in a subject having or at risk of developing tau-mediated amyloidosis, comprising administering to the subject an effective regime of an antibody disclosed herein, thereby inhibiting or reducing tau aggregation in the subject. Exemplary antibodies include humanized versions of 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3.

[0137] Also provided is a method of treating or affecting prevention of a tau-associated disease in a subject, comprising administering an effective regimen of an antibody disclosed herein to treat or affect prevention of the disease. Examples of such diseases are Alzheimer's disease, Down's syndrome, mild cognitive impairment, primary age-related tauopathy, postencephalitic parkinsonism, posttraumatic dementia or dementia pugilistica, Pick's disease, Niemann-Pick type C, supranuclear palsy, frontotemporal dementia, frontotemporal lobar degeneration, argyrophilic grain disease, globular glial tauopathy, Guam amyotrophic lateral sclerosis / Parkinsonism dementia complex, corticobasal degeneration (CBD), Lewy body dementia, Lewy body variant of Alzheimer's disease (LBVAD), chronic traumatic encephalopathy (CTE), globular glial tauopathy, Parkinson's disease, or progressive supranuclear palsy (PSP). In some methods, the tau-related disease is Alzheimer's disease. In some methods, the patient is a carrier of ApoE4.

[0138] Also provided is a method of reducing tau transmission abnormalities, comprising the step of reducing tau transmission by administering an effective regime of an antibody disclosed herein.

[0139] Also provided is a method of inducing phagocytosis of tau, comprising inducing phagocytosis of tau by administering an effective regime of the antibodies disclosed herein.

[0140] Also provided is a method of inhibiting tau aggregation or precipitation, comprising the step of inhibiting tau aggregation or precipitation by administering an effective regime of an antibody disclosed herein.

[0141] Also provided is a method of inhibiting the formation of tau fibrillary tangles comprising administering an effective regime of the antibodies disclosed herein.

[0142] The present invention also provides a method for detecting tau protein precipitates in a subject having or at risk for a disease associated with tau aggregation or precipitation, comprising administering to the subject an antibody disclosed herein and detecting said antibody bound to tau in the subject. Examples of such diseases are Alzheimer's disease, Down's syndrome, mild cognitive impairment, primary age-related tauopathy, postencephalitic parkinsonism, post-traumatic dementia or dementia pugilistica, Pick's disease, Niemann-Pick type C, supranuclear palsy, frontotemporal dementia, frontotemporal lobar degeneration, argyrophilic grain disease, globular glial tauopathy, Guam amyotrophic lateral sclerosis / Parkinsonism dementia complex, corticobasal degeneration (CBD), dementia with Lewy bodies, Lewy body variant of Alzheimer's disease (LBVAD), chronic traumatic encephalopathy (CTE), globular glial tauopathy, Parkinson's disease, or progressive supranuclear palsy (PSP). In some embodiments, the antibody is administered into the subject's body by intravenous injection. In some embodiments, the antibody is administered directly to the brain by intravenous injection or by drilling a hole in the subject's skull. In some embodiments, the antibody is labeled. In some embodiments, the antibody is labeled with a fluorescent label, a paramagnetic label, or a radioactive label. In some embodiments, the radioactive label is detected using positron emission tomography (PET) or single photon emission computed tomography (SPECT).

[0143] The present invention also provides a method for measuring the effectiveness of a treatment in a subject being treated for a disease associated with tau aggregation or precipitation, comprising the steps of measuring a first level of tau protein precipitates in the subject prior to treatment by administering to the subject an antibody disclosed herein, detecting a first amount of the antibody bound to tau in the subject, administering the treatment to the subject, measuring a second level of tau protein precipitates in the subject after treatment by administering to the subject the antibody, and detecting the antibody bound to tau in the subject, wherein a reduction in the level of tau protein precipitates indicates a positive response to the treatment.

[0144] The present invention also provides a method for measuring the effectiveness of a treatment in a subject being treated for a disease associated with tau aggregation or precipitation, comprising the steps of measuring a first level of tau protein precipitates in the subject prior to treatment by administering an antibody disclosed herein to the subject, detecting a first amount of the antibody bound to tau in the subject, administering the treatment to the subject, measuring a second level of tau protein precipitates in the subject after treatment by administering the antibody to the subject, and detecting the second amount of antibody bound to tau in the subject, wherein no change in the level of tau protein precipitates or a small increase in tau protein precipitates indicates a positive response to the treatment.

[0145] The invention also provides an isolated monoclonal antibody that specifically binds to a peptide consisting of the residues (Q / E)IVYK(S / P) (SEQ ID NO: 56). The invention also provides an isolated monoclonal antibody that specifically binds to a peptide consisting of the residues QIVYKP (SEQ ID NO: 57). The invention also provides an isolated monoclonal antibody that specifically binds to a peptide consisting of the residues EIVYKSP (SEQ ID NO: 58). The invention also provides an isolated monoclonal antibody that specifically binds to a peptide consisting of the residues EIVYKS (SEQ ID NO: 277).

[0146] The invention also provides isolated monoclonal antibodies that specifically bind to a polypeptide of SEQ ID NO:1 at an epitope that includes at least one residue from 307-312 of SEQ ID NO:1. Some such antibodies bind to an epitope from 307-312 of SEQ ID NO:1. The invention also provides isolated monoclonal antibodies that specifically bind to a polypeptide of SEQ ID NO:1 at an epitope that includes at least one residue from 391-397 of SEQ ID NO:1. Some such antibodies bind to an epitope from 391-397 of SEQ ID NO:1. The invention also provides isolated monoclonal antibodies that specifically bind to a polypeptide of SEQ ID NO:1 at an epitope that includes at least one residue from 391-396 of SEQ ID NO:1. Some such antibodies bind to an epitope from 391-396 of SEQ ID NO:1. The invention also provides isolated monoclonal antibodies that specifically bind to a polypeptide of SEQ ID NO:1 at an epitope that includes at least one residue from 307-312 of SEQ ID NO:1 and at least one residue from 391-397 of SEQ ID NO:1. The present invention also provides isolated monoclonal antibodies that specifically bind to a polypeptide of SEQ ID NO:1 at an epitope comprising at least one residue derived from among residues 307-312 of SEQ ID NO:1 and at least one residue derived from among residues 391-396 of SEQ ID NO:1.

[0147] The present invention also provides a method of treating or preventing a tau-related disease in a subject, comprising administering an immunogen comprising a tau peptide of up to 20 contiguous amino acids of SEQ ID NO:1 to which antibodies 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3 specifically bind, the immunogen inducing formation of antibodies that specifically bind to tau in the subject. In some such methods, the tau peptide consists of 4 to 7 contiguous amino acids from residues 307-312 of SEQ ID NO:1, or from residues 391-397 of SEQ ID NO:1, or from residues 391-396 of SEQ ID NO:1. In some such methods, the tau peptide consists of residues (Q / E)IVYK(S / P) (SEQ ID NO:56). In some such methods, the tau peptide consists of residues QIVYKP (SEQ ID NO:57). In some such methods, the tau peptide consists of residues EIVYKSP (SEQ ID NO:58). In some such methods, the tau peptide consists of residues EIVYKS (SEQ ID NO: 277). In some such methods, the tau peptide is attached to a heterologous conjugate molecule.

[0148] The invention also provides methods for producing antibodies that specifically bind to an epitope comprising (Q / E)IVYK(S / P) (SEQ ID NO:56), 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 comprises a P301S mutation. In some such methods, the human tau is recombinant and N-terminally his-tagged.

[0149] In some such methods, screening is performed against a 15 amino acid peptide that includes QIVYKP (SEQ ID NO:57), EIVYKSP (SEQ ID NO:58), EIVYKS (SEQ ID NO:277), or any other consensus motif represented by (Q / E)IVYK(S / P) (SEQ ID NO:56). In some such methods, the peptide includes QIVYKP (SEQ ID NO:57), or EIVYKSP (SEQ ID NO:58), or EIVYKS (SEQ ID NO:277).

[0150] In some such methods, animals are immunized with a tau fragment comprising a peptide presented by (Q / E)IVYK(S / P) (SEQ ID NO: 56) conjugated to a carrier. In some such methods, the peptide is QIVYKP (SEQ ID NO: 57) or EIVYKSP (SEQ ID NO: 58) or EIVYKS (SEQ ID NO: 277). [Brief description of the drawings]

[0151] [Figure 1A] FIG. 1A depicts an alignment of the human germline heavy chain variable region sequence IGHV1-69-2*01 (SEQ ID NO: 33), the human acceptor heavy chain variable region sequence AAN16432-VH_huFrwk (AAN16432_H; SEQ ID NO: 31), and the human acceptor heavy chain variable region sequence 2RCS-VH_huFrwk (2RCS_H; SEQ ID NO: 32) with the heavy chain variable region of the murine 9F5 antibody (SEQ ID NO: 7) and the heavy chain variable regions of the humanized versions of the 9F5 antibody (hu9F5VHv1, hu9F5VHv2, hu9F5VHv3, hu9F5VHv4, hu9F5VHv5, hu9F5VHv6, hu9F5VHv7, and hu9F5VHv8). 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 CDRs of mouse 9F5 VH are in bold, as defined by the Kabat / Chothia composite. [Figure 1B] FIG. 1B depicts an alignment of the human germline heavy chain variable region sequence IGHV1-69-2*01 (SEQ ID NO: 33), the human acceptor heavy chain variable region sequence AAN16432-VH_huFrwk (AAN16432_H; SEQ ID NO: 31), and the human acceptor heavy chain variable region sequence 2RCS-VH_huFrwk (2RCS_H; SEQ ID NO: 32) with the heavy chain variable region of the murine 9F5 antibody (SEQ ID NO: 7) and the heavy chain variable regions of the humanized versions of the 9F5 antibody (hu9F5VHv1, hu9F5VHv2, hu9F5VHv3, hu9F5VHv4, hu9F5VHv5, hu9F5VHv6, hu9F5VHv7, and hu9F5VHv8). 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 CDRs of mouse 9F5 VH are in bold, as defined by the Kabat / Chothia composite. [Figure 2A]FIG. 2A depicts an alignment of the human germline light chain variable region sequence IGKV2-28*01&_IGKJ2*01 (IGKV2-28*01_IGKJ2*01; SEQ ID NO: 37), human acceptor CAB51297-VL_huFrwk (CAB51297_L; SEQ ID NO: 35), and human acceptor 1911357B-VL_huFRwk (1911357B_L; SEQ ID NO: 36) with the light chain variable region of the murine 9F5 antibody (SEQ ID NO: 11) and the light chain variable regions of the humanized versions of the 9F5 antibody (hu9F5VLv1, hu9F5VLv2, hu9F5VLv3, hu9F5VLv4, hu9F5VLv5, hu9F5VLv6, and hu9F5VLv7). 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 CDRs of mouse 9F5 VL are in bold as defined by Kabat. [Figure 2B] FIG. 2B represents an alignment of the human germline light chain variable region sequence IGKV2-28*01&_IGKJ2*01 (IGKV2-28*01_IGKJ2*01; SEQ ID NO: 37), human acceptor CAB51297-VL_huFrwk (CAB51297_L; SEQ ID NO: 35), and human acceptor 1911357B-VL_huFRwk (1911357B_L; SEQ ID NO: 36) with the light chain variable region of the murine 9F5 antibody (SEQ ID NO: 11) and the light chain variable regions of the humanized versions of the 9F5 antibody (hu9F5VLv1, hu9F5VLv2, hu9F5VLv3, hu9F5VLv4, hu9F5VLv5, hu9F5VLv6, and hu9F5VLv7). 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 CDRs of mouse 9F5 VL are in bold as defined by Kabat. [Diagram 3] Figure 3 represents the results of an assay showing that the mouse 9F5 antibody blocks the internalization of tau into neurons. [Figure 4A] Figure 4A represents the alignment of the human germline heavy chain variable region sequence IGHV1-69-2*01 (SEQ ID NO: 33), the human acceptor heavy chain variable region sequences AAN16432-VH_huFrwk (AAN16432_H; SEQ ID NO: 31), and 2RCS-VH_huFrwk (2RCS_H; SEQ ID NO: 32), with the heavy chain variable region of the mouse 9F5 antibody (SEQ ID NO: 7) and the heavy chain variable regions of humanized versions of the 9F5 antibody (hu9F5VHv1, hu9F5VHv2, hu9F5VHv3, hu9F5VHv4, hu9F5VHv5, hu9F5VHv6, hu9F5VHv7, hu9F5VHv8, hu9F5VHv9, and hu9F5VHv10). 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 CDRs of the mouse 9F5 VH are in bold as defined by the Kabat / Chothia composite. Residues identical to those of the mouse 9F5 VH are represented by "". [Figure 4B]FIG. 4B depicts an alignment of the human germline heavy chain variable region sequence IGHV1-69-2*01 (SEQ ID NO: 33), the human acceptor heavy chain variable region sequence AAN16432-VH_huFrwk (AAN16432_H; SEQ ID NO: 31), and the human acceptor heavy chain variable region sequence 2RCS-VH_huFrwk (2RCS_H; SEQ ID NO: 32) with the heavy chain variable region of the murine 9F5 antibody (SEQ ID NO: 7) and the heavy chain variable regions of the humanized versions of the 9F5 antibody (hu9F5VHv1, hu9F5VHv2, hu9F5VHv3, hu9F5VHv4, hu9F5VHv5, hu9F5VHv6, hu9F5VHv7, hu9F5VHv8, hu9F5VHv9, and hu9F5VHv10). 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 CDRs of mouse 9F5 VH are in bold as defined by the Kabat / Chothia composite. Residues identical to those in mouse 9F5 VH are represented by "". [Figure 5A]FIG. 5A shows the human germline light chain variable region sequence IGKV2-28*01&_IGKJ2*01 (IGKV2-28*01_IGKJ2*01; SEQ ID NO: 37), human acceptor CAB51297-VL_huFrwk (CAB51297_L; SEQ ID NO: 35), and human acceptor 1911357B-VL_huFRwk (1911357B_L; SEQ ID NO: 36). FIG. 36) represents an alignment of the light chain variable region of the murine 9F5 antibody (sequence number 11) and the light chain variable regions of humanized versions of the 9F5 antibody (hu9F5VLv1, hu9F5VLv2, hu9F5VLv3, hu9F5VLv4, hu9F5VLv5, hu9F5VLv6, hu9F5VLv7, hu9F5VLv8, and hu9F5VLv9). 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 CDRs of mouse 9F5 VL are in bold as defined by Kabt. Residues identical to those in mouse 9F5 VL are represented by "". [Figure 5B]FIG. 5B shows the human germline light chain variable region sequence IGKV2-28*01&_IGKJ2*01 (IGKV2-28*01_IGKJ2*01; SEQ ID NO: 37), human acceptor CAB51297-VL_huFrwk (CAB51297_L; SEQ ID NO: 35), and human acceptor 1911357B-VL_huFRwk (1911357B_L; SEQ ID NO: 36). FIG. 36) represents an alignment of the light chain variable region of the murine 9F5 antibody (sequence number 11) and the light chain variable regions of humanized versions of the 9F5 antibody (hu9F5VLv1, hu9F5VLv2, hu9F5VLv3, hu9F5VLv4, hu9F5VLv5, hu9F5VLv6, hu9F5VLv7, hu9F5VLv8, and hu9F5VLv9). 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 CDRs of mouse 9F5 VL are in bold as defined by Kabt. Residues identical to those in mouse 9F5 VL are represented by "". [Figure 6A]FIG. 6A shows the light chain variable region of the humanized version 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), hu9F 5VLv8_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), h u9F5VLv8_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), h u9F5VLv9_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), hu9F5VLv9_DIM20 (SEQ ID NO: 171), hu9F5VLv8_DIM27 (SEQ ID NO: 158), hu9F5VLv8_DIM28 (SEQ ID NO: 159), hu9F5VLv8_DIM29 (SEQ ID NO: 160),FIG. 1 shows an alignment of the light chain variable region of hu9F5VLv8 with hu9F5VLv8_DIM30 (SEQ ID NO: 161). The CDRs of hu9F5VLv8 are in bold as defined by Kabat. [Figure 6B]FIG. 6B shows the light chain variable region of the humanized version 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), hu9F 5VLv8_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), h u9F5VLv8_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), h u9F5VLv9_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), hu9F5VLv9_DIM20 (SEQ ID NO: 171), hu9F5VLv8_DIM27 (SEQ ID NO: 158), hu9F5VLv8_DIM28 (SEQ ID NO: 159), hu9F5VLv8_DIM29 (SEQ ID NO: 160),FIG. 1 shows an alignment of the light chain variable region of hu9F5VLv8 with hu9F5VLv8_DIM30 (SEQ ID NO: 161). The CDRs of hu9F5VLv8 are in bold as defined by Kabat. [Figure 6C]FIG. 6C shows the light chain variable regions of the humanized versions 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), hu9F 5VLv8_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), h u9F5VLv8_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), h u9F5VLv9_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), hu9F5VLv9_DIM20 (SEQ ID NO: 171), hu9F5VLv8_DIM27 (SEQ ID NO: 158), hu9F5VLv8_DIM28 (SEQ ID NO: 159), hu9F5VLv8_DIM29 (SEQ ID NO: 160),FIG. 1 shows an alignment of the light chain variable region of hu9F5VLv8 with hu9F5VLv8_DIM30 (SEQ ID NO: 161). The CDRs of hu9F5VLv8 are in bold as defined by Kabat. [Figure 7] Figure 7 depicts an alignment of the human germline heavy chain variable region sequence IGHV1-69-2*01 (SEQ ID NO: 33) and the human acceptor heavy chain variable region sequence CAC20421 VH (SEQ ID NO: 218) with the heavy chain variable region of the murine 10C12 antibody (SEQ ID NO: 7, labeled m10C12 VH in Figure 7) and the heavy chain variable region of the humanized version of the 10C12 antibody (hu10C12VHv1 and hu10C12VHv2). hu10C12VHv1 is SEQ ID NO: 214 and hu10C12VHv2 is SEQ ID NO: 215. The CDRs of the murine 10C12 VH are in bold, as defined by the Kabat / Chothia composite. [Figure 8] Figure 8 depicts an alignment of the human germline light chain variable region sequence IGKV2-28*01 &_IGKJ2*01 (SEQ ID NO: 37) and human acceptor CAB51297-VL_huFrwk (SEQ ID NO: 35) with the light chain variable region of murine 10C12 (SEQ ID NO: 11) and the light chain variable region of the humanized version of the 10C12 antibody (hu10C12VLv1 and hu10C12VLv2). hu10C12VLv1 is SEQ ID NO: 216 and hu10C12VLv2 is SEQ ID NO: 217. The CDRs of murine 10C12 VL are in bold as defined by Kabat. [Figure 9] Figure 9 depicts an alignment of the human germline heavy chain variable region sequence IGHV1-69-2*01 (SEQ ID NO: 33) and the human acceptor heavy chain variable region sequence CAC20421 VH (SEQ ID NO: 218) with the heavy chain variable region of the murine 12C4 antibody (SEQ ID NO: 219) and the heavy chain variable region of the humanized version of the 12C4 antibody (hu12C4VHv1 and hu12C4VHv2). hu12C4VHv1 is SEQ ID NO: 221 and hu12C4VHv2 is SEQ ID NO: 222. The CDRs of the murine 12C4 VH are in bold, as defined by the Kabat / Chothia composite. [Figure 10]Figure 10 depicts an alignment of the human germline light chain variable region sequence IGKV2-28*01 &_IGKJ2*01 (SEQ ID NO:37) and human acceptor CAB51297 (SEQ ID NO:35) with the light chain variable region of the murine 12C4 antibody (SEQ ID NO:11) and the light chain variable region of a humanized version of the 12C4 antibody (hu12C4VLv1 and huvVLv2). hu12C4VLv1 is SEQ ID NO:223 and hu12C4VLv2 is SEQ ID NO:224. The CDRs of the murine 12C4 VL are in bold as defined by Kabat. [Figure 11] Figure 11 depicts an alignment of the human germline heavy chain variable region sequence IGHV1-69-2*01 (SEQ ID NO:33) and the human acceptor heavy chain variable region sequence CAC20421 VH (SEQ ID NO:218) with the heavy chain variable region of the murine 17C12 antibody (SEQ ID NO:225) and the heavy chain variable region of the humanized version of the 17C12 antibody (hu17C12VHv1 and hu17C12VHv2). hu17C12VHv1 is SEQ ID NO:232 and hu17C12VHv2 is SEQ ID NO:233. The CDRs of the murine 17C12 VH are in bold, as defined by the Kabat / Chothia composite. [Figure 12] Figure 12 depicts an alignment of the human germline light chain variable region sequences IGKV2-29*02 & IGKJ4*01 (SEQ ID NO: 239) and human acceptor QDO16713 VL (SEQ ID NO: 238) with the light chain variable region of the murine 17C12 antibody (SEQ ID NO: 228) and the light chain variable region of the humanized version of the 17C12 antibody (hu17C12VLv1 and hu17C12VLv2). hu17C12VLv1 is SEQ ID NO: 234 and hu17C12VLv2 is SEQ ID NO: 235. The CDRs of the murine 17C12 VL are in bold as defined by Kabat. [Figure 13]Figure 13 depicts an alignment of the human germline heavy chain variable region sequences IGHV2-70*04 & IGHJ4*01 (SEQ ID NO: 254) and the human acceptor heavy chain variable region sequence QDJ57937VH hFrwk (SEQ ID NO: 253) with the heavy chain variable region of the murine 14H3 antibody (SEQ ID NO: 240) and the heavy chain variable region of the humanized version of the 14H3 antibody (hu14H3VHv1 and hu14H3VHv2). hu14H3VHv1 is SEQ ID NO: 248 and hu14H3VHv2 is SEQ ID NO: 249. The CDRs of the murine 14H3 VH are in bold, as defined by the Kabat / Chothia composite. [Figure 14] Figure 14 depicts an alignment of the human germline light chain variable region sequence IGKV2-28*01 &_IGKJ2*01 (IGKV2-28*01_IGKJ2*01; SEQ ID NO: 37) and human acceptor ABC66914VL_hFwrk (SEQ ID NO: 256) with the light chain variable region of the murine 14H3 antibody (SEQ ID NO: 244) and the light chain variable region of the humanized version of the 14H3 antibody (hu14H3VLv1 and hu14H3VLv2). hu14H3VLv1 is SEQ ID NO: 250 and hu14H3VLv2 is SEQ ID NO: 251. The CDRs of the murine 14H3 VL are in bold as defined by Kabat. [Figure 15] FIG. 15 depicts the results of an assay showing that the mouse 10C12, 12C4, 2D11, 17C12, 14H3, and 9F5 antibodies block internalization of tau into neurons. [Figure 16] FIG. 16 depicts the results of an assay showing that mouse 10C12, 12C4, 2D11, and 9F5 antibodies prevent tau toxicity in primary neurons (neuronal viability). [Figure 17] FIG. 17 depicts the results of an assay showing that mouse 10C12, 12C4, 2D11, and 9F5 antibodies prevent tau toxicity in primary neurons (LDH release). [Figure 18]FIG. 18 depicts the results of a Western blot assay showing that mouse 10C12, 12C4, 2D11, 17C12, 14H3, and 9F5 antibodies detect tau in samples derived from the brains of Alzheimer's disease patients. [Figure 19] FIG. 19 shows the results of an immunoprecipitation assay using mouse 10C12, 12C4, 2D11, 17C12, 14H3, and 9F5 antibodies, as well as a sample derived from the brain of an Alzheimer's disease patient. [Figure 20] FIG. 20 depicts the results of an assay to measure the ability of 9F5 humanized variants to resist agitation stress-induced aggregation. [Figure 21] FIG. 21 depicts the results of an assay to measure the ability of 9F5 humanized variants to withstand exposure to low pH. [Figure 22] FIG. 22 depicts the results of an assay to measure the propensity of 9F5 humanized variants to aggregate under simulated high concentration conditions. [Figure 23A] FIG. 23A depicts the results of an immunohistochemistry assay using a control, mouse 2D11, mouse 9F5, mouse 12C4, mouse 14H3, and mouse 17C12. [Figure 23B] FIG. 23B depicts the results of an immunohistochemistry assay using a control, mouse 2D11, mouse 9F5, mouse 12C4, mouse 14H3, and mouse 17C12. [Figure 23C] FIG. 23C depicts the results of an immunohistochemistry assay using a control, mouse 2D11, mouse 9F5, mouse 12C4, mouse 14H3, and mouse 17C12. [Figure 23D] FIG. 23D depicts the results of an immunohistochemistry assay using control, mouse 2D11, mouse 9F5, mouse 12C4, mouse 14H3, and mouse 17C12. [Figure 23E] FIG. 23E depicts the results of an immunohistochemistry assay using control, mouse 2D11, mouse 9F5, mouse 12C4, mouse 14H3, and mouse 17C12. [Figure 23F] FIG. 23F depicts the results of an immunohistochemistry assay using control, mouse 2D11, mouse 9F5, mouse 12C4, mouse 14H3, and mouse 17C12. [Figure 24] Figure 24 represents an alignment of the heavy chain variable region of the murine 9F5 antibody (SEQ ID NO: 7), the murine 10C12 antibody (SEQ ID NO: 7), the murine 2D11 antibody (SEQ ID NO: 7), the murine 12C4 antibody (SEQ ID NO: 219), the murine 14H3 antibody (SEQ ID NO: 240), and the murine 17C12 antibody (SEQ ID NO: 225). The CDRs of the murine 9F5 VH are in bold, as defined by the Kabat / Chothia composite. [Diagram 25] Figure 25 represents an alignment of the light chain variable region of the murine 9F5 antibody (SEQ ID NO: 11), the murine 10C12 antibody (SEQ ID NO: 11), the murine 2D11 antibody (SEQ ID NO: 11), the murine 12C4 antibody (SEQ ID NO: 11), the murine 14H3 antibody (SEQ ID NO: 244), and the murine 17C12 antibody (SEQ ID NO: 228). The CDRs of the murine 9F5 VL are in bold as defined by Kabat. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0152] A brief description of the sequence SEQ ID NO:1 represents the amino acid sequence of a human tau isoform (Swiss-Prot P10636-8).

[0153] SEQ ID NO:2 represents the amino acid sequence of a human tau isoform (Swiss-Prot P10636-7).

[0154] SEQ ID NO:3 represents the amino acid sequence of a human tau isoform (Swiss-Prot P10636-6), (4R0N human tau).

[0155] SEQ ID NO: 4 represents the amino acid sequence of a human tau isoform (Swiss-Prot P10636-5).

[0156] SEQ ID NO:5 represents the amino acid sequence of a human tau isoform (Swiss-Prot P10636-4).

[0157] SEQ ID NO:6 represents the amino acid sequence of a human tau isoform (Swiss-Prot P10636-2).

[0158] SEQ ID NO:7 represents the amino acid sequence of the heavy chain variable region of the murine 9F5 antibody.

[0159] SEQ ID NO:8 represents the amino acid sequence of the composite Kabat / Chothia CDR-H1 of the murine 9F5 antibody.

[0160] SEQ ID NO: 9 represents the amino acid sequence of the Kabat CDR-H2 of the murine 9F5 antibody.

[0161] SEQ ID NO: 10 represents the amino acid sequence of the Kabat CDR-H3 of the murine 9F5 antibody.

[0162] SEQ ID NO: 11 represents the amino acid sequence of the light chain variable region of the murine 9F5 antibody.

[0163] SEQ ID NO: 12 represents the amino acid sequence of the Kabat CDR-L1 of the murine 9F5 antibody.

[0164] SEQ ID NO: 13 represents the amino acid sequence of the Kabat CDR-L2 of the murine 9F5 antibody.

[0165] SEQ ID NO: 14 represents the amino acid sequence of the Kabat CDR-L3 of the murine 9F5 antibody.

[0166] SEQ ID NO: 15 represents the amino acid sequence of the humanized heavy chain variable region hu9F5VHv1.

[0167] SEQ ID NO: 16 represents the amino acid sequence of the humanized heavy chain variable region hu9F5VHv2.

[0168] SEQ ID NO: 17 represents the amino acid sequence of the humanized heavy chain variable region hu9F5VHv3.

[0169] SEQ ID NO: 18 represents the amino acid sequence of the humanized heavy chain variable region hu9F5VHv4.

[0170] SEQ ID NO: 19 represents the amino acid sequence of the humanized heavy chain variable region hu9F5VHv5.

[0171] SEQ ID NO:20 represents the amino acid sequence of the humanized heavy chain variable region hu9F5VHv6.

[0172] SEQ ID NO: 21 represents the amino acid sequence of the humanized heavy chain variable region hu9F5VHv7.

[0173] SEQ ID NO: 22 represents the amino acid sequence of the humanized heavy chain variable region hu9F5VHv8.

[0174] SEQ ID NO: 23 represents the amino acid sequence of the humanized light chain variable region hu9F5VLv1.

[0175] SEQ ID NO:24 represents the amino acid sequence of the humanized light chain variable region hu9F5VLv2.

[0176] SEQ ID NO:25 represents the amino acid sequence of the humanized light chain variable region hu9F5VLv3.

[0177] SEQ ID NO: 26 represents the amino acid sequence of the humanized light chain variable region hu9F5VLv4.

[0178] SEQ ID NO: 27 represents the amino acid sequence of the humanized light chain variable region hu9F5VLv5 of the humanized 9F5 antibody.

[0179] SEQ ID NO: 28 represents the amino acid sequence of the humanized light chain variable region hu9F5VLv6.

[0180] SEQ ID NO:29 represents the amino acid sequence of the humanized light chain variable region hu9F5VLv7.

[0181] SEQ ID NO: 30 represents the amino acid sequence of the heavy chain variable region structural model PDB.# 5OBF-VH_mSt.

[0182] SEQ ID NO: 31 represents the amino acid sequence of the heavy chain variable region acceptor GenBank Acc.# AAN16432-VH_huFrwk.

[0183] SEQ ID NO: 32 represents the amino acid sequence of the heavy chain variable region acceptor PDB # 2RCS-VH_huFrwk.

[0184] SEQ ID NO: 33 is the heavy chain variable region germline sequence IMGT# IGHV1-69-2 * This represents the amino acid sequence of 01.

[0185] SEQ ID NO: 34 represents the amino acid sequence of the light chain variable region structural model PDB # 5OBF-VL_mSt.

[0186] SEQ ID NO: 35 represents the amino acid sequence of the light chain variable region acceptor GenBank Acc. # CAB51297-VL_huFrwk.

[0187] SEQ ID NO: 36 represents the amino acid sequence of the light chain variable region acceptor GenBank Acc. # 1911357B-VL_huFrwk.

[0188] SEQ ID NO: 37 is the light chain variable region germline sequence IMGT# IGKV2-28 * 01 & IGKJ2 * This represents the amino acid sequence of 01.

[0189] SEQ ID NO:38 represents the nucleic acid sequence encoding the heavy chain variable region of the murine 9F5 antibody.

[0190] SEQ ID NO:39 represents the nucleic acid sequence encoding the light chain variable region of the murine 9F5 antibody.

[0191] SEQ ID NO: 40 represents the amino acid sequence of the Kabat CDR-H1 of the murine 9F5 antibody.

[0192] SEQ ID NO: 41 represents the amino acid sequence of the Chothia CDR-H1 of the murine 9F5 antibody.

[0193] SEQ ID NO: 42 represents the amino acid sequence of the Chothia CDR-H2 of the murine 9F5 antibody.

[0194] SEQ ID NO: 43 represents the amino acid sequence of AbM CDR-H2 of the murine 9F5 antibody.

[0195] SEQ ID NO: 44 represents the amino acid sequence of Contact CDR-H1 of mouse 9F5 antibody.

[0196] SEQ ID NO: 45 represents the amino acid sequence of Contact CDR-H2 of mouse 9F5 antibody.

[0197] SEQ ID NO: 46 represents the amino acid sequence of Contact CDR-H3 of mouse 9F5 antibody.

[0198] SEQ ID NO: 47 represents the amino acid sequence of Contact CDR-L1 of mouse 9F5 antibody.

[0199] SEQ ID NO: 48 represents the amino acid sequence of Contact CDR-L2 of mouse 9F5 antibody.

[0200] SEQ ID NO: 49 represents the amino acid sequence of Contact CDR-L3 of mouse 9F5 antibody.

[0201] SEQ ID NO:50 represents the amino acid sequence of another Kabat / Chothia composite CDR-H1 of the humanized 9F5 antibody (present in hu9F5VHv4, hu9F5VHv5, and hu9F5VHv6).

[0202] SEQ ID NO:51 represents the amino acid sequence of another Kabat CDR-H2 of the humanized 9F5 antibody (present in hu9F5VHv5, hu9F5VHv6, and hu9F5VHv7).

[0203] SEQ ID NO: 52 represents the amino acid sequence of another Kabat CDR-H2 of the humanized 9F5 antibody (present in hu9F5VHv8).

[0204] SEQ ID NO:53 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv5 and hu9F5VLv6).

[0205] SEQ ID NO: 54 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv7).

[0206] SEQ ID NO: 55 identifies another Kabat CDR-L2 of the humanized 9F5 antibody (hu9F5VLv4, hu9F5VLv5, hu9F5VLv6, hu9F5VLv7, hu9F5VLv8_DIM2, hu9F5VLv8_DIM4, hu9F5VLv8_DIM5, hu9F5VLv8_DIM6, hu9F5VLv8_DIM11, hu9F5VLv8_DIM12, hu9F5VLv8 hu9F5VLv9_DIM13, hu9F5VLv8_DIM18, hu9F5VLv8_DIM27, hu9F5VLv8_DIM28, hu9F5VLv9_DIM2, hu9F5VLv9_DIM4, hu9F5VLv9_DIM5, hu9F5VLv9_DIM11, and hu9F5VLv9_DIM13).

[0207] SEQ ID NO: 56 represents the amino acid sequence of the epitope of antibody 9F5.

[0208] SEQ ID NO:57 represents the amino acid sequence of the consensus motif of the peptide to which antibody 9F5 binds.

[0209] SEQ ID NO:58 represents the amino acid sequence of the consensus motif of the peptide to which antibody 9F5 binds.

[0210] SEQ ID NO: 59 represents the amino acid sequence of the linker.

[0211] SEQ ID NO: 60 represents the amino acid sequence of the HA control peptide.

[0212] SEQ ID NO: 61 represents the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_M51E).

[0213] SEQ ID NO: 62 represents the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_M51D).

[0214] SEQ ID NO: 63 represents the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27cD).

[0215] SEQ ID NO: 64 represents the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27cG).

[0216] SEQ ID NO: 65 represents the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27cS).

[0217] SEQ ID NO: 66 represents the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27cE).

[0218] SEQ ID NO: 67 represents the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_I30E).

[0219] SEQ ID NO: 68 represents the amino acid sequence of a variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_I30K).

[0220] SEQ ID NO: 69 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27cT).

[0221] SEQ ID NO: 70 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27cN).

[0222] SEQ ID NO: 71 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27bD).

[0223] SEQ ID NO: 72 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_I30G).

[0224] SEQ ID NO: 73 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L33N).

[0225] SEQ ID NO: 74 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27cA).

[0226] SEQ ID NO: 75 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L33T).

[0227] SEQ ID NO: 76 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L33S).

[0228] SEQ ID NO: 77 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L33R).

[0229] SEQ ID NO: 78 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_I30Q).

[0230] SEQ ID NO: 79 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27bT).

[0231] SEQ ID NO: 80 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_T31G).

[0232] SEQ ID NO: 81 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27bQ).

[0233] SEQ ID NO: 82 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L33G).

[0234] SEQ ID NO: 83 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L27cP).

[0235] SEQ ID NO: 84 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_V78R).

[0236] SEQ ID NO: 85 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_I75D).

[0237] SEQ ID NO: 86 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_V78D).

[0238] SEQ ID NO: 87 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_V78E).

[0239] SEQ ID NO: 88 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_V78P).

[0240] SEQ ID NO: 89 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_V78K).

[0241] SEQ ID NO: 90 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_R77D).

[0242] SEQ ID NO: 91 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_V78G).

[0243] SEQ ID NO: 92 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_S76P).

[0244] SEQ ID NO: 93 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_I75P).

[0245] SEQ ID NO: 94 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_I75Q).

[0246] SEQ ID NO: 95 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_I75G).

[0247] SEQ ID NO: 96 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L73P).

[0248] SEQ ID NO: 97 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L73G).

[0249] SEQ ID NO: 98 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_V78Q).

[0250] SEQ ID NO: 99 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_S76G).

[0251] SEQ ID NO: 100 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L92D).

[0252] SEQ ID NO: 101 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_Y86T).

[0253] SEQ ID NO: 102 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L92E).

[0254] SEQ ID NO: 103 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L92G).

[0255] SEQ ID NO: 104 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L92Q).

[0256] SEQ ID NO: 105 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L93G).

[0257] SEQ ID NO: 106 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_V85G).

[0258] SEQ ID NO: 107 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_L92T).

[0259] SEQ ID NO: 108 represents the amino acid sequence of the variant of the hu9F5VLv2 light chain variable region (also known as hu9F5VLv2_A89G).

[0260] SEQ ID NO: 109 represents the amino acid sequence of the variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L80P).

[0261] SEQ ID NO: 110 represents the amino acid sequence of the variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L80D).

[0262] SEQ ID NO:111 represents the amino acid sequence of the variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L82cG).

[0263] SEQ ID NO: 112 represents the amino acid sequence of the variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L82cD).

[0264] SEQ ID NO: 113 represents the amino acid sequence of the variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L82P).

[0265] SEQ ID NO: 114 represents the amino acid sequence of the variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L80G).

[0266] SEQ ID NO: 115 represents the amino acid sequence of the variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L82K).

[0267] SEQ ID NO: 116 represents the amino acid sequence of the variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L82R).

[0268] SEQ ID NO: 117 represents the amino acid sequence of the variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L82E).

[0269] SEQ ID NO: 118 represents the amino acid sequence of the variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L82N).

[0270] SEQ ID NO: 119 represents the amino acid sequence of the variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_Y79D).

[0271] SEQ ID NO: 120 represents the amino acid sequence of the variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_Y79N).

[0272] SEQ ID NO: 121 represents the amino acid sequence of the variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_Y79G).

[0273] SEQ ID NO: 122 represents the amino acid sequence of the variant of the hu9F5VHv5 heavy chain variable region (also known as hu9F5VHv5_M80E).

[0274] Sequence number 123 represents the amino acid sequence of a variant of the hu9F5VHv5 heavy chain variable region (also known as hu9F5VHv5_M80G).

[0275] Sequence number 124 represents the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_L82cS).

[0276] Sequence number 125 represents the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_Y79Q).

[0277] Sequence number 126 represents the amino acid sequence of a variant of the hu9F5VHv4 heavy chain variable region (also known as hu9F5VHv4_S82aG).

[0278] Sequence number 127 represents the amino acid sequence of the heavy chain variable region hu9F5VHv9.

[0279] Sequence number 128 represents the amino acid sequence of the heavy chain variable region hu9F5VHv10 (also known as hu9F5VHv9_Q38K_G42E).

[0280] Sequence number 129 represents the amino acid sequence of the heavy chain variable region hu9F5VHv10_L82cG.

[0281] Sequence number 130 represents the amino acid sequence of the light chain variable region hu9F5VLv8.

[0282] Sequence number 131 represents the amino acid sequence of a variant of the light chain variable region hu9F5VLv9 (also known as hu9F5VLv8_N60D).

[0283] Sequence number 132 represents the amino acid sequence of a variant of the hu9F5VLv8 light chain variable region (also known as hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54G, L92I, hu9F5VLv8_DIM1).

[0284] SEQ ID NO: 133 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54R, L92I, also known as hu9F5VLv8_DIM2).

[0285] SEQ ID NO: 134 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54T, L92I, also known as hu9F5VLv8_DIM3).

[0286] SEQ ID NO: 135 represents the amino acid sequence of the hu9F5VLv8 light chain variable region variant (hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54R, L92G, also known as hu9F5VLv8_DIM4).

[0287] SEQ ID NO: 136 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37Q, M51G, L54R, L92I, also known as hu9F5VLv8_DIM5).

[0288] SEQ ID NO: 137 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cD, L37Q, M51G, L54R, L92I, also known as hu9F5VLv8_DIM6).

[0289] SEQ ID NO: 138 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cD, L37Q, M51K, L54R, L92I, also known as hu9F5VLv8_DIM7).

[0290] SEQ ID NO: 139 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37Q, M51K, L54R, L92I, also known as hu9F5VLv8_DIM8).

[0291] SEQ ID NO: 140 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37Q, M51K, L54G, L92I, also known as hu9F5VLv8_DIM9).

[0292] SEQ ID NO: 141 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, M51K, L54G, L92I, also known as hu9F5VLv8_DIM10).

[0293] SEQ ID NO: 142 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54R, L92I, also known as hu9F5VLv8_DIM11).

[0294] SEQ ID NO: 143 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54R, L92G, also known as hu9F5VLv8_DIM12).

[0295] SEQ ID NO: 144 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54R, also known as hu9F5VLv8_DIM13).

[0296] SEQ ID NO: 145 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54T, L92I, also known as hu9F5VLv8_DIM14).

[0297] SEQ ID NO: 146 represents the amino acid sequence of a variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54T, L92G, also known as hu9F5VLv8_DIM15).

[0298] SEQ ID NO: 147 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54T, also known as hu9F5VLv8_DIM16).

[0299] SEQ ID NO: 148 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37G, M51G, L54T, L92I, also known as hu9F5VLv8_DIM17).

[0300] SEQ ID NO: 149 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cD, L37G, M51G, L54R, L92I, also known as hu9F5VLv8_DIM18).

[0301] SEQ ID NO: 150 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37I, M51I, L54R, L92I, also known as hu9F5VLv8_DIM19).

[0302] SEQ ID NO: 151 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, M51I, L54G, L92I, also known as hu9F5VLv8_DIM20).

[0303] SEQ ID NO: 152 represents the amino acid sequence of a variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, M51I, L54G, also known as hu9F5VLv8_DIM21).

[0304] SEQ ID NO: 153 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, M51E, L54R, L92I, also known as hu9F5VLv8_DIM22).

[0305] SEQ ID NO: 154 Variant of Hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37Q, M51E, L54G, L92I, also known as hu9F5VLv8_DIM23).

[0306] SEQ ID NO: 155 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37I, M51E, L54R, L92I, also known as hu9F5VLv8_DIM24).

[0307] SEQ ID NO: 156 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cG, L37I, M51E, L54R, L92G, also known as hu9F5VLv8_DIM25).

[0308] SEQ ID NO: 157 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cI, L37I, M51E, L54R, also known as hu9F5VLv8_DIM26).

[0309] SEQ ID NO: 158 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L37Q, M51G, L54R, L92I, also known as hu9F5VLv8_DIM27).

[0310] SEQ ID NO: 159 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, M51G, L54R, L92I, also known as hu9F5VLv8_DIM28).

[0311] SEQ ID NO: 160 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, L54R, L92I, also known as hu9F5VLv8_DIM29).

[0312] SEQ ID NO: 161 represents the amino acid sequence of the variant of the hu9F5VLv8 light chain variable region (hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L92I, also known as hu9F5VLv8_DIM30).

[0313] SEQ ID NO: 162 represents the amino acid sequence of a variant of the hu9F5VLv9 light chain variable region (hu9F5VLv9_V3Q, L27cS, L37Q, M51G, L54G, L92I, also known as hu9F5VLv9_DIM1).

[0314] SEQ ID NO: 163 represents the amino acid sequence of the variant of the hu9F5VLv9 light chain variable region (hu9F5VLv9_V3Q, L27cS, L37Q, M51G, L54R, L92I, also known as hu9F5VLv9_DIM2).

[0315] SEQ ID NO: 164 represents the amino acid sequence of a variant of the hu9F5VLv9 light chain variable region (hu9F5VLv9_V3Q, L27cS, L37Q, M51G, L54R, L92G, also known as hu9F5VLv9_DIM4).

[0316] SEQ ID NO: 165 represents the amino acid sequence of the hu9F5VLv9 light chain variable region variant (hu9F5VLv9_V3Q, L27cG, L37Q, M51G, L54R, L92I, also known as hu9F5VLv9_DIM5).

[0317] SEQ ID NO: 166 represents the amino acid sequence of the variant of the hu9F5VLv9 light chain variable region (hu9F5VLv9_V3Q, L27cG, L37Q, M51K, L54R, L92I, also known as hu9F5VLv9_DIM8).

[0318] SEQ ID NO: 167 represents the amino acid sequence of the hu9F5VLv9 light chain variable region variant (hu9F5VLv9_V3Q, L27cS, L37Q, M51K, L54G, L92I, also known as hu9F5VLv9_DIM10).

[0319] SEQ ID NO: 168 represents the amino acid sequence of the variant of the hu9F5VLv9 light chain variable region (hu9F5VLv9_V3Q, L27cG, L37G, M51G, L54R, L92I, also known as hu9F5VLv9_DIM11).

[0320] SEQ ID NO: 169 represents the amino acid sequence of the hu9F5VLv9 light chain variable region variant (hu9F5VLv9_V3Q, L27cG, L37G, M51G, L54R, also known as hu9F5VLv9_DIM13).

[0321] SEQ ID NO: 170 represents the amino acid sequence of the hu9F5VLv9 light chain variable region variant (hu9F5VLv9_V3Q, L27cS, L37I, M51I, L54R, L92I, also known as hu9F5VLv9_DIM19).

[0322] SEQ ID NO: 171 represents the amino acid sequence of the hu9F5VLv9 light chain variable region variant (hu9F5VLv9_V3Q, L27cS, L37Q, M51I, L54G, L92I, also known as hu9F5VLv9_DIM20).

[0323] SEQ ID NO: 172 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_L27bD).

[0324] SEQ ID NO: 173 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_L27bT).

[0325] SEQ ID NO: 174 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_L27bQ).

[0326] SEQ ID NO: 175 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_L27cD, hu9F5VLv8_DIM6, hu9F5VLv8_DIM7, and hu9F5VLv8_DIM18).

[0327] SEQ ID NO: 176 identifies another Kabat CDR-L1 of the humanized 9F5 antibody (hu9F5VLv2_L27cG, hu9F5VLv8_DIM5, hu9F5VLv8_DIM8, hu9F5VLv8_DIM9, hu9F5VLv8_DIM11, hu9F5VLv8_DIM12, hu9F5VLv8_DIM13, hu9F5VLv8_DIM14, hu9F5VLv8_DIM 15, hu9F5VLv8_DIM16, hu9F5VLv8_DIM23, hu9F5VLv8_DIM24, hu9F5VLv8_DIM25, hu9F5VLv9_DIM5, hu9F5VLv9_DIM8, hu9F5VLv9_DIM11, and hu9F5VLv9_DIM13).

[0328] SEQ ID NO: 177 is another Kabat sequence for the humanized 9F5 antibody. This represents 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).

[0329] SEQ ID NO: 178 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_L27cE).

[0330] SEQ ID NO: 179 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_L27cT).

[0331] SEQ ID NO: 180 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_L27cN).

[0332] SEQ ID NO: 181 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_L27cA).

[0333] SEQ ID NO: 182 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_L27cP).

[0334] SEQ ID NO: 183 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv8_DIM26).

[0335] SEQ ID NO: 184 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_I30E).

[0336] SEQ ID NO: 185 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_I30K).

[0337] SEQ ID NO: 186 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_I30G).

[0338] SEQ ID NO: 187 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_I30Q).

[0339] SEQ ID NO: 188 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_T31G).

[0340] SEQ ID NO: 189 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_L33N).

[0341] SEQ ID NO: 190 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_L33T).

[0342] SEQ ID NO: 191 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_L33S).

[0343] SEQ ID NO: 192 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_L33R).

[0344] SEQ ID NO: 193 represents the amino acid sequence of another Kabat CDR-L1 of the humanized 9F5 antibody (present in hu9F5VLv2_L33G).

[0345] SEQ ID NO: 194 represents the amino acid sequence of another Kabat CDR-L2 of the humanized 9F5 antibody (present in hu9F5VLv2_M51E).

[0346] SEQ ID NO: 195 represents the amino acid sequence of another Kabat CDR-L2 of the humanized 9F5 antibody (present in hu9F5VLv2_M51D).

[0347] SEQ ID NO: 196 represents the amino acid sequence of another Kabat CDR-L2 of the humanized 9F5 antibody (present in hu9F5VLv8_DIM30).

[0348] SEQ ID NO: 197 represents the amino acid sequence of another Kabat CDR-L2 of the humanized 9F5 antibody (present in hu9F5VLv8_DIM29).

[0349] SEQ ID NO: 198 represents the amino acid sequence of another Kabat CDR-L2 of the humanized 9F5 antibody (present in hu9F5VLv8_DIM1, and hu9F5VLv9_DIM1).

[0350] SEQ ID NO: 199 represents the amino acid sequence of another Kabat CDR-L2 of the humanized 9F5 antibody (present in hu9F5VLv8_DIM3, hu9F5VLv8_DIM14, hu9F5VLv8_DIM15, hu9F5VLv8_DIM16, and hu9F5VLv8_DIM17).

[0351] SEQ ID NO: 200 represents the amino acid sequence of another Kabat CDR-L2 of the humanized 9F5 antibody (present in hu9F5VLv8_DIM7, hu9F5VLv8_DIM8, and hu9F5VLv9_DIM8).

[0352] SEQ ID NO: 201 represents the amino acid sequence of another Kabat CDR-L2 of the humanized 9F5 antibody (present in hu9F5VLv8_DIM9, hu9F5VLv8_DIM10, and hu9F5VLv9_DIM10).

[0353] SEQ ID NO: 202 represents the amino acid sequence of another Kabat CDR-L2 of the humanized 9F5 antibody (present in hu9F5VLv8_DIM19, and hu9F5VLv9_DIM19).

[0354] SEQ ID NO: 203 represents the amino acid sequence of another Kabat CDR-L2 of the humanized 9F5 antibody (present in hu9F5VLv8_DIM20, hu9F5VLv8_DIM21, and hu9F5VLv9_DIM20).

[0355] SEQ ID NO: 204 represents the amino acid sequence of another Kabat CDR-L2 of the humanized 9F5 antibody (present in hu9F5VLv8_DIM22, hu9F5VLv8_DIM24, hu9F5VLv8_DIM25, and hu9F5VLv8_DIM26).

[0356] SEQ ID NO: 205 represents the amino acid sequence of another Kabat CDR-L2 of the humanized 9F5 antibody (present in hu9F5VLv8_DIM23).

[0357] SEQ ID NO: 206 represents the amino acid sequence of another Kabat CDR-L3 of the humanized 9F5 antibody (present in hu9F5VLv2_A89G).

[0358] SEQ ID NO: 207 represents the amino acid sequence of another Kabat CDR-L3 of the humanized 9F5 antibody (present in hu9F5VLv2_L92D).

[0359] SEQ ID NO: 208 represents the amino acid sequence of another Kabat CDR-L3 of the humanized 9F5 antibody (present in hu9F5VLv2_L92E).

[0360] SEQ ID NO: 209 represents the amino acid sequence of another Kabat CDR-L3 of the humanized 9F5 antibody (present in hu9F5VLv8_DIM4, hu9F5VLv8_DIM12, hu9F5VLv8_DIM15, hu9F5VLv8_DIM25, and hu9F5VLv9_DIM4).

[0361] SEQ ID NO: 210 represents the amino acid sequence of another Kabat CDR-L3 of the humanized 9F5 antibody (present in hu9F5VLv2_L92Q).

[0362] SEQ ID NO: 211 represents the amino acid sequence of another Kabat CDR-L3 of the humanized 9F5 antibody (present in hu9F5VLv2_L92T).

[0363] SEQ ID NO: 212 identifies another Kabat CDR-L3 of humanized 9F5 antibody (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, hu9F5VLv8_DIM21, hu9F5VLv8_DIM22, hu9F5VLv8_DIM23, hu9F5VLv8_DIM24, hu9F5VLv8_DIM25, hu9F5VLv8_DIM26, hu9F5VLv8_DIM27, hu9F5VLv8_DIM28, hu9F5VLv8_DIM29, hu9F5VLv8_DIM30, hu9F5VLv8_DIM31, hu9F5VLv8_DIM32, hu9F5VLv8_DIM33, hu9F5VLv8_DIM34, hu9F5VLv8_DIM35, hu9F5VLv8_DIM36, hu9F5VLv8_DIM37, hu9F5VLv8_DIM3 hu9F5VLv9_DIM30, hu9F5VLv9_DIM1, hu9F5VLv9_DIM2, hu9F5VLv9_DIM5, hu9F5VLv9_DIM8, hu9F5VLv9_DIM10, hu9F5VLv9_DIM11, hu9F5VLv9_DIM19, and hu9F5VLv9_DIM20).

[0364] SEQ ID NO: 213 represents the amino acid sequence of another Kabat CDR-L3 of the humanized 9F5 antibody (present in hu9F5VLv2_L93G).

[0365] SEQ ID NO: 214 represents the amino acid sequence of the humanized heavy chain variable region hu10C12VHv1.

[0366] SEQ ID NO: 215 represents the amino acid sequence of the humanized heavy chain variable region hu10C12VHv2.

[0367] SEQ ID NO: 216 represents the amino acid sequence of the humanized light chain variable region hu10C12VLv1.

[0368] SEQ ID NO: 217 represents the amino acid sequence of the humanized light chain variable region hu10C12VLv2.

[0369] SEQ ID NO: 218 represents the amino acid sequence of the heavy chain variable region acceptor CAC20421-VH_huFrwk.

[0370] SEQ ID NO: 219 represents the amino acid sequence of the heavy chain variable region of the murine 12C4 antibody.

[0371] SEQ ID NO: 220 represents the amino acid sequence of the Kabat CDR-H2 of the murine 12C4 antibody.

[0372] SEQ ID NO: 221 represents the amino acid sequence of the humanized heavy chain variable region hu12C4VHv1.

[0373] SEQ ID NO: 222 represents the amino acid sequence of the humanized heavy chain variable region hu12C4VHv2.

[0374] SEQ ID NO: 223 represents the amino acid sequence of the humanized light chain variable region hu12C4VLv1.

[0375] SEQ ID NO: 224 represents the amino acid sequence of the humanized light chain variable region hu12C4VLv2.

[0376] SEQ ID NO: 225 represents the amino acid sequence of the heavy chain variable region of the murine 17C12 antibody.

[0377] SEQ ID NO: 226 represents the amino acid sequence of the composite Kabat / Chothia CDR H1 of the murine 17C12 antibody.

[0378] SEQ ID NO: 227 represents the amino acid sequence of the Kabat CDR H2 of the murine 17C12 antibody.

[0379] SEQ ID NO: 228 represents the amino acid sequence of the light chain variable region of the murine 17C12 antibody.

[0380] SEQ ID NO: 229 represents the amino acid sequence of the Kabat CDR-L1 of the murine 17C12 antibody.

[0381] SEQ ID NO: 230 represents the amino acid sequence of the Kabat CDR-L2 of the murine 17C12 antibody.

[0382] SEQ ID NO: 231 represents the amino acid sequence of the Kabat CDR-L3 of the murine 17C12 antibody.

[0383] SEQ ID NO: 232 represents the amino acid sequence of the humanized heavy chain variable region hu17C12VHv1.

[0384] SEQ ID NO: 233 represents the amino acid sequence of the humanized heavy chain variable region hu17C12VHv2.

[0385] SEQ ID NO: 234 represents the amino acid sequence of the humanized light chain variable region hu17C12VLv1.

[0386] SEQ ID NO: 235 represents the amino acid sequence of the humanized light chain variable region hu17C12VLv2.

[0387] SEQ ID NO: 236 represents the amino acid sequence of the heavy chain variable region structural model 3PP3-VH_mSt.

[0388] SEQ ID NO: 237 represents the amino acid sequence of the light chain variable region structural model 3PP3-VL_mSt.

[0389] SEQ ID NO: 238 represents the amino acid sequence of the light chain variable region acceptor QDO16713-VL_huFrwk.

[0390] SEQ ID NO: 239 is the light chain variable region germline sequence IGKV2-29 * 02 & IGKJ4 * This represents the amino acid sequence of 01.

[0391] SEQ ID NO: 240 represents the amino acid sequence of the heavy chain variable region of the murine 14H3 antibody.

[0392] SEQ ID NO: 241 represents the amino acid sequence of the composite Kabat / Chothia CDR H1 of the murine 14H3 antibody.

[0393] SEQ ID NO: 242 represents the amino acid sequence of the Kabat CDR H2 of the murine 14H3 antibody.

[0394] SEQ ID NO: 243 represents the amino acid sequence of the Kabat CDR H3 of the murine 14H3 antibody.

[0395] SEQ ID NO: 244 represents the amino acid sequence of the light chain variable region of the murine 14H3 antibody.

[0396] SEQ ID NO: 245 represents the amino acid sequence of the Kabat CDR L1 of the murine 14H3 antibody.

[0397] SEQ ID NO: 246 represents the amino acid sequence of the Kabat CDR L2 of the murine 14H3 antibody.

[0398] SEQ ID NO: 247 represents the amino acid sequence of the Kabat CDR L3 of the murine 14H3 antibody.

[0399] SEQ ID NO: 248 represents the amino acid sequence of the humanized heavy chain variable region hu14H3VHv1.

[0400] SEQ ID NO: 249 represents the amino acid sequence of the humanized heavy chain variable region hu14H3VHv2.

[0401] SEQ ID NO: 250 represents the amino acid sequence of the humanized light chain variable region hu14H3VLv1.

[0402] SEQ ID NO: 251 represents the amino acid sequence of the humanized light chain variable region hu14H3VLv2.

[0403] SEQ ID NO: 252 represents the amino acid sequence of the heavy chain variable region structural model 2VQ1-VH_mSt.

[0404] SEQ ID NO: 253 represents the amino acid sequence of the heavy chain variable region acceptor QDJ57937-VH_huFrwk.

[0405] SEQ ID NO: 254 is the heavy chain variable region germline sequence IGHV1-70 * 04 & IGHJ4 * This represents the amino acid sequence of 01.

[0406] SEQ ID NO: 255 represents the amino acid sequence of the light chain variable region structural model 2VQ1-VL_mSt.

[0407] SEQ ID NO: 256 represents the amino acid sequence of the light chain variable region acceptor ABC66914-VL_huFrwk.

[0408] SEQ ID NO: 257 represents the amino acid sequence of AbM CDR-H2 of the murine 12C4 antibody.

[0409] SEQ ID NO: 258 represents the amino acid sequence of Contact CDR-H2 of mouse 12C4 antibody.

[0410] SEQ ID NO: 259 represents the amino acid sequence of the Chothia CDR-H1 of the murine 17C12 antibody.

[0411] SEQ ID NO: 260 represents the amino acid sequence of AbM CDR-H2 of the murine 17C12 antibody.

[0412] SEQ ID NO: 261 represents the amino acid sequence of Contact CDR-H2 of mouse 17C12 antibody.

[0413] SEQ ID NO: 262 represents the amino acid sequence of Contact CDR-L1 of mouse 17C12 antibody.

[0414] SEQ ID NO: 263 represents the amino acid sequence of Contact CDR-L2 of mouse 17C12 antibody.

[0415] SEQ ID NO: 264 represents the amino acid sequence of Contact CDR-L3 of mouse 17C12 antibody.

[0416] SEQ ID NO: 265 represents the amino acid sequence of the Kabat CDR-H1 of the murine 14H3 antibody.

[0417] SEQ ID NO: 266 represents the amino acid sequence of the Chothia CDR-H1 of the murine 14H3 antibody.

[0418] SEQ ID NO: 267 represents the amino acid sequence of the Chothia CDR-H2 of the murine 14H3 antibody.

[0419] SEQ ID NO: 268 represents the amino acid sequence of AbM CDR-H2 of the murine 14H3 antibody.

[0420] SEQ ID NO: 269 represents the amino acid sequence of Contact CDR-H1 of mouse 14H3 antibody.

[0421] SEQ ID NO: 270 represents the amino acid sequence of Contact CDR-H2 of mouse 14H3 antibody.

[0422] SEQ ID NO: 271 represents the amino acid sequence of Contact CDR-H3 of mouse 14H3 antibody.

[0423] SEQ ID NO: 272 represents the amino acid sequence of Contact CDR-L1 of mouse 14H3 antibody.

[0424] SEQ ID NO: 273 represents the amino acid sequence of Contact CDR-L2 of mouse 14H3 antibody.

[0425] SEQ ID NO: 274 represents the amino acid sequence of Contact CDR-L3 of mouse 14H3 antibody.

[0426] SEQ ID NO:275 represents the amino acid sequence of another composite Kabat / Chothia CDR H1 of the humanized 14H3 antibody (present in hu14H3VHv1 and hu14H3VHv2).

[0427] SEQ ID NO:276 represents the amino acid sequence of the consensus motif of the peptide to which antibodies 9F5, 10C12, 2D11, 12C4, 17C12, and 14H3 bind.

[0428] SEQ ID NO:277 represents the amino acid sequence of the consensus motif of the peptide to which antibody 2D11 binds.

[0429] definition Monoclonal antibodies or other biological entities are usually provided in isolated form. This means that the antibody or other biological entity usually contains interfering proteins and other contaminants resulting from its production or purification with a purity of at least 50 (w / w)%, but does not exclude the possibility that the monoclonal antibody may be combined with an excess of a pharma- ceutical acceptable carrier or other vehicle intended to facilitate its use. In some cases, the monoclonal antibody is at least 60 (w / w)%, 70 (w / w)%, 80 (w / w)%, 90 (w / w)%, 95 (w / w)%, or 99 (w / w)% pure of interfering proteins and contaminants resulting from its production or purification. In many cases, an isolated monoclonal antibody or other biological entity is the major macromolecular species remaining after its purification.

[0430] Specific binding of an antibody to its target antigen is at least 10 6 , 10 7, 10 8 , 10 9 , 10 10 , 10 11 , or 10 12 M -1 Specific binding refers to the affinity and / or avidity of an antibody. Specific binding is detectably significant in magnitude and is comparable to non-specific binding occurring to at least one unrelated target. Specific binding can be the result of the formation of bonds between specific functional groups or specific spatial matches (e.g., lock and key types), whereas non-specific binding is usually the result of van der Waals forces. However, specific binding does not necessarily imply that an antibody binds to one and only one target.

[0431] The basic antibody structural unit is a tetramer of subunits. Each tetramer contains two identical pairs of polypeptide chains, each pair having one "light" chain (about 25 kDa) and one "heavy" chain (about 50-70 kDa). The amino-terminal portion of each chain contains a variable region of about 100-110 or more amino acids primarily responsible for antigen recognition. This variable region is initially expressed linked to a cleavable signal peptide. The variable region without the signal peptide is sometimes referred to as a mature variable region. Thus, for example, light chain mature variable region refers to the light chain variable region without the light chain signal peptide. The carboxy terminus of each chain defines a constant region primarily responsible for effector function.

[0432] Light chains are classified as either kappa or lambda. Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, and define the antibody's isotype as IgG, IgM, IgA, IgD, and IgE, respectively. Within light and heavy chains, the variable and constant regions are joined by a "J" region of about 12 or more amino acids, with heavy chains also including a "D" region of about 10 more amino acids. See generally, Fundamental Immunology, Paul, W., ed., 2nd ed. Raven Press, NY, 1989, Ch. 7 (incorporated by reference in its entirety for all purposes).

[0433] An immunoglobulin light or heavy chain variable region (also referred to herein as a "light chain variable domain" ("VL domain") or a "heavy chain variable domain" ("VH domain"), respectively) consists of a "framework" region interrupted by three "complementarity determining regions" or "CDRs". The framework regions act to align the CDRs for specific binding to an epitope of an antigen. The CDRs contain the amino acid residues of an antibody that primarily contribute to antigen binding. From the amino-terminus to the carboxyl-terminus, the VL and VH domains comprise the following framework regions (FR) and CDR regions: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. CDR1, 2, and 3 of the VL domain are also referred to herein as CDR-L1, CDR-L2, and CDR-L3, respectively; CDR1, 2, and 3 of the VH domain are also referred to herein as CDR-H1, CDR-H2, and CDR-H3, respectively. Where the present application discloses a VL sequence with R as the C-terminal residue, R may alternatively be considered as the N-terminal residue of the light chain constant region, and thus, it should be understood that the present application also discloses a VL sequence without a C-terminal R.

[0434] The amino acid assignments for each VL and VH domain follow one of the conventional CDR definitions, including 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 Chothia Kabat CDR combination, in which CDR-H1 is a combination of the Chothia and Kabat CDRs; The AbM definition used by antibody modeling software from Molecular; and the contact definition from Martin et al. (bioinfo.org.uk / abs) (see Table 1). Kabat provides a widely used numbering scheme (Kabat numbering) in which corresponding residues between different heavy chains or different light chains are assigned the same number. When an antibody is said to contain a CDR according to a particular definition of CDR (e.g. Kabat), this definition specifies the minimum number of CDR residues present in the antibody (i.e. Kabat CDRs). This does not exclude that other residues that are within the alternative conventional CDR definitions but outside the scope of the specified definition may be present as well. For example, an antibody that contains a CDR as defined by Kabat includes, among other possibilities, an antibody in which the CDR contains Kabat CDR residues and no other CDR residues, and an antibody in which the CDR H1 is a composite Chothia-Kabat CDR H1 and the other CDRs contain Kabat CDR residues and no further CDR residues according to other definitions. [Table 1]

[0435] The term "antibody" includes intact antibodies and binding fragments thereof, typically consisting of separate heavy and light chains, Fab, Fab', F(ab') 2 Fragments, including F(ab)c, Dabs, Nanobodies, and Fv, compete with the intact antibody from which they were derived for specific binding to a target. Fragments can be produced by recombinant DNA techniques or by enzymatic or chemical separation of intact immunoglobulins. The term "antibody" also includes bispecific and / or humanized antibodies. Bispecific or bifunctional antibodies are artificial hybrid antibodies having two different heavy / light chain pairs and two different binding sites (see, e.g., 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 heavy / light chain pairs include a humanized 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3 heavy / light chain pair and a heavy / light chain pair specific for a different epitope on tau other than that bound by 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3.

[0436] In some bispecific antibodies, one heavy chain / light chain pair is the humanized 9F5 antibody, humanized 10C12 antibody, humanized 2D11 antibody, humanized 12C4 antibody, humanized 17C12 antibody, or humanized 14H3 antibody further disclosed below, and the other heavy chain / light chain pair is derived from an antibody that binds to a receptor expressed at the blood-brain barrier, such as the insulin receptor, insulin-like growth factor (IGF) receptor, leptin receptor, 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 delivered across the blood-brain barrier by receptor-mediated transcytosis. Brain uptake of the bispecific antibody can be further enhanced by engineering the bispecific antibody to reduce its affinity for the blood-brain barrier receptor. Reduction of the affinity for the receptor resulted in a broader distribution in the brain (see, e.g., Atwal et al., Sci. Trans. Med. 3, 84ra43, 2011; Yu et al., Sci. Trans. Med. 3, 84ra44, 2011).

[0437] Also exemplary bispecific antibodies include: (1) dual variable domain antibodies (DVD-Ig), in which each light and heavy chain contains two variable domains in tandem via a short peptide bond (Wu et al., Generation and Characterization of a Dual Variable Domain Immunoglobulin (DVD-Ig™) Molecule, In: Antibody Engineering, Springer Berlin Heidelberg (2010)); (2) Tandbab, a fusion of two single-chain diabodies resulting in a tetravalent bispecific antibody with two binding sites for each of the target antigens; (3) flexibodies, a combination of diabody and scFv resulting in a multivalent molecule; (4) so-called "dock and dock antibodies" based on the "D / D domain" of protein kinase A, which when applied to Fabs, can result in a trivalent bispecific binding protein consisting of two identical Fab fragments bound to different Fab fragments. or (5) so-called Scorpion molecules, which comprise, for example, two scFvs fused to either end of a human Fc region. Examples of platforms useful for the preparation of bispecific antibodies include BiTE (Micromet), DART (MacroGenics), Fcab and Mab2 (F-star), Fc-engineered IgG1 (Xencor), or DuoBody (based on the exchange of Fab arms, Genmab).

[0438] The term "epitope" refers to a site on an antigen to which an antibody binds. Epitopes can be formed from contiguous or non-contiguous amino acids juxtaposed by tertiary folding of one or more proteins. Epitopes formed from contiguous amino acids (also known as linear epitopes) are usually retained after exposure to denaturing solvents, whereas epitopes formed by tertiary folding (conformational epitopes) are usually lost after treatment with denaturing solvents. An epitope comprises at least three, usually at least five or eight to ten amino acids in a unique spatial conformation. Methods for determining the spatial conformation of an epitope 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, Glenn E. Morris, Ed. (1996).

[0439] Antibodies that recognize the same or overlapping epitopes can be identified in a simple immunoassay that shows the property of one antibody competing with the binding of another antibody to a target antigen. The epitope of an antibody can also be defined by X-ray crystallography of the antibody bound to its antigen to identify the exact residues. Alternatively, two antibodies have the same epitope if all amino acid mutations in the antigen that reduce or eliminate binding of one antibody reduce or eliminate binding of the other antibody. Two antibodies have overlapping epitopes if some amino acid mutations that reduce or eliminate binding of one antibody reduce or eliminate binding of the other antibody.

[0440] Competition between antibodies is determined by an assay in which the antibody under test inhibits specific binding of a reference antibody to a common antigen (see, e.g., Junghans et al., Cancer Res. 50:1495, 1990). A test antibody competes with a reference antibody if an excess of the test antibody (e.g., at least 2-fold, 5-fold, 10-fold, 20-fold, or 100-fold) inhibits binding of the reference antibody by at least 50% as measured in a competitive binding assay. Some test antibodies inhibit binding of the reference antibody by at least 75%, 90%, or 99%. Antibodies identified by competitive assays (competitor antibodies) include antibodies that bind to the same epitope as the reference antibody and antibodies that bind to adjacent epitopes that are sufficiently close to the epitope bound by the reference antibody due to steric hindrance that occurs.

[0441] The term "pharmaceutically acceptable" means the carrier, diluent, excipient or auxiliary must be compatible with the other ingredients of the formulation and not substantially deleterious to the recipient thereof.

[0442] The term "patient" includes human and other mammalian subjects receiving either prophylactic or therapeutic treatment.

[0443] An individual is at high risk for a disease if the subject has at least one known risk factor (genetic factors, biochemical factors, familial, and situational exposures) that places the individual with the risk factor at a statistically significantly higher risk of developing the disease than individuals without the risk factor.

[0444] The term "biological sample" refers to a sample of biological material available in or from a biological source, such as a human or mammalian subject. Such samples may be organs, organelles, tissues, tissue sections, bodily 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 encompass any material derived by processing the sample. Derived materials may include cells or their passages. Processing of biological samples may include one or more of filtration, distillation, extraction, concentration, fixation, inactivation of interfering components, and the like.

[0445] The term "control sample" refers to a biological sample that is not known or suspected to contain a region affected by a tau-related disease, or at least not known or suspected to contain a disease region of a given type. A control sample may be obtained from an individual not affected by a tau-related disease. Alternatively, a control sample may be obtained from a patient affected by a tau-related disease. Such a biological sample may be obtained at the same time as a biological sample suspected to contain a tau-related disease, or in a different context. Both the biological sample and the control sample may be obtained from the same tissue. Preferably, the control sample consists of an essentially or grossly normal, healthy region, and may be used for comparison with a biological sample suspected to contain a region affected by a tau-related disease. Preferably, the tissue of the control sample is of the same type as the tissue of the biological sample. Preferably, the cells suspected to be in the biological sample affected by a tau-related disease arise from the same cell type (e.g., neurons or glia) as the cell type in the control sample.

[0446] The term "disease" refers to any abnormal condition that impairs physiological function. The term is used broadly to encompass any disorder, disease, disorder, pathology, disorder, condition, or syndrome in which physiological function is impaired, regardless of the nature of the etiology.

[0447] The term "symptom" refers to subjective evidence of disease, such as a change in gait, as perceived by the subject. "Sign" refers to objective evidence of disease as observed by a physician.

[0448] The term "positive response to a treatment" refers to a more favorable individual patient response or the average response of a population of patients compared to the average response of a control population that does not receive the treatment.

[0449] For the purposes of classifying amino acid substitutions as conservative or non-conservative, 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 that affect chain orientation): gly, pro; and Group VI (aromatic side chains): trp, tyr, phe. Conservative substitutions involve substitutions between amino acids of the same class. Non-conservative substitutions constitute the exchange of a member of one of these classes for another.

[0450] Percentage sequence identity is determined with maximally aligned antibody sequences according to the Kabat numbering convention. After alignment, when a region of an antibody of interest (e.g., the entire mature variable region of a heavy or light chain) is compared to the same region of a reference antibody, the percentage sequence identity between the subject and reference antibody regions is the number of positions occupied by the same amino acid in the subject and reference antibodies, divided by the total number of aligned positions of the two regions, not accounting for gaps, multiplied by 100 to convert to a percentage.

[0451] A composition or method "comprising or including" one or more recited elements may include other elements not specifically recited. For example, a composition "comprising" an antibody may include the antibody alone or may include the antibody in combination with other components.

[0452] The representation of a range of values ​​includes every integer within or defining that range, and every subrange defined by the integers within that range.

[0453] Unless otherwise clear from the context, the term "about" includes definite values, such as values ​​within the standard error (e.g., SEM) of measurement of the stated value.

[0454] Statistical significance means p≦0.05.

[0455] The singular forms of articles "a," "an," and "the" include the plural forms unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" may include a plurality of compounds, including mixtures thereof.

[0456] Detailed Description I. General Provisions The present invention provides antibodies that bind to tau. Some antibodies specifically bind to an epitope in (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 of 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 of SEQ ID NO:1). The antibodies differ from 3D6 and other antibodies that are 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 specificity of the additional C-terminus of the epitope provides the basis for the antibodies to bind to an increasing number of conformational forms of tau associated with pathology. 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 of SEQ ID NO:1). Exemplary antibodies of the invention are 9F5, 10C12, 2D11, 12C4, 17C12, and 14H3. Some antibodies of the invention act to inhibit or slow the worsening of tau-related pathology and associated symptoms. Although understanding of the mechanism is not necessary for the practice of the invention, reduced toxicity may occur as a result of the antibody stabilizing a non-toxic conformation, inhibiting intercellular or intracellular transmission of pathogenic tau forms, inducing phagocytosis of tau by blocking tau phosphorylation, preventing tau binding to cells, or inducing proteolytic cleavage of tau, preventing tau from inter- or intramolecularly aggregating or binding to other molecules, among other mechanisms. Some antibodies of the invention are useful for increasing tau aggregation by increasing the molecular weight of certain aggregated tau species to reduce increased toxicity / cellular uptake and / or clearance. Large aggregates of tau molecules may exhibit reduced uptake into neuronal cells. Some antibodies of the invention, by bivalent binding to tau, bring other tau molecules into closer proximity and encourage their aggregation into tau aggregates that are too large to be internalized within neurons. Furthermore, Fc-mediated phagocytosis requires some tau-binding antibodies to be brought into closer proximity.Large aggregates of tau molecules may have many anti-tau antibodies bound to them, providing the clusters necessary for Fc-mediated phagocytosis by macrophages. The antibodies of the invention or agents that induce such antibodies may be used in methods to treat or affect the prevention of Alzheimer's disease and other diseases associated with tau.

[0457] II.Target molecule Unless otherwise clear from the context, reference to tau refers to the native human form of tau, including all isoforms, whether or not post-translational modifications (e.g., phosphorylation, glycosylation, or acetylation) are present. There are six major isoforms (splice variants) of tau present in the human brain. The longest of these variants has 441 amino acids, the first met residue of which is truncated. Residues are numbered according to the 441 isoform. Thus, for example, reference to phosphorylation at position 404 refers to position 404 in the 441 isoform, or the corresponding position in any other isoform when maximally aligned with the 441 isoform. The amino acid sequences and Swiss-Prot numbers of the isoforms are set forth below. [ka] JPEG0007681316000003.jpg178162

[0458] Reference to tau includes known naturally occurring variations and permutations thereof, about 30 of which are listed in the Swiss-Prot database, as well as mutations associated with tau pathologies, 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 by the 441 isoforms, are a lysine to threonine mutation at amino acid residue 257 (K257T); an isoleucine to valine mutation at amino acid position 260 (I260V); a glycine to valine mutation at amino acid position 272 (G272V); an asparagine to lysine mutation at amino acid position 279 (N279K); an asparagine to histidine mutation at amino acid position 296 (N296H); a proline to serine mutation at amino acid position 301 (P301S); a proline to leucine mutation at amino acid position 301 (P301L). ); a glycine to valine mutation at amino acid position 303 (G303V); a serine to asparagine mutation at position 305 (S305N); a glycine to serine mutation at amino acid position 335 (G335S); a valine to methionine mutation at position 337 (V337M); a glutamic acid to valine mutation at position 342 (E342V); a lysine to isoleucine mutation at amino acid position 369 (K3691); a glycine to arginine mutation at amino acid position 389 (G389R); and an arginine to tryptophan mutation at amino acid position 406 (R406W).

[0459] 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.

[0460] Unless otherwise clear from the context, reference to tau or a fragment thereof includes the naturally occurring human amino acid sequence, including its isoforms, mutants, and allelic variants.

[0461] III. Antibodies A. Binding Specificity and Functional Properties The present invention provides antibodies that specifically bind to tau. Some antibodies specifically bind to tau at an epitope formed by amino acids from either or both of two regions of tau that have a common core motif of 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, an antibody that includes one binding site for tau, such as an scFv, may specifically bind to tau at an epitope formed by amino acids in either of these regions individually, or may specifically bind to a hybrid epitope formed by amino acids from both of these regions. An antibody that includes two binding sites for tau may also bind to an epitope within the ranges 307-312 and 391-397, or within the range 391-396, simultaneously from its two binding sites. The epitopes may be on the same or different molecules of tau. Some antibodies of the invention specifically bind to a peptide consisting of residues 307-312 of tau, i.e., residues QIVYKP (SEQ ID NO: 57). Some antibodies of the invention specifically bind to a peptide consisting of residues 391-397 of tau, i.e., residues EIVYKSP (SEQ ID NO: 58). Some antibodies of the invention specifically bind to a peptide consisting of residues 391-396 of tau, i.e., EIVYKS (SEQ ID NO: 277). Some antibodies of the invention specifically bind to a peptide consisting of the consensus motif (Q / E)IVYK(S / P) (SEQ ID NO: 56). These antibodies may be obtained by immunization with tau polypeptides purified from natural sources or expressed recombinantly. Antibodies may be screened for binding to unphosphorylated forms of tau and to forms in which one or more phosphorylation-prone residues are phosphorylated. The present invention also provides antibodies that bind to the same epitope as any of the above antibodies, e.g., 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3, and also includes antibodies that compete with any of the above antibodies, e.g., 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3, for binding to tau.In one embodiment, an antibody that binds to the same epitope as a reference antibody, such as 9F5, or competes with the reference antibody, shares one or more of its functional properties, such as inhibiting internalization of tau into neuronal cells, and optionally retains such properties to the same extent, or to a greater extent than the reference antibody, within experimental error.

[0462] The present invention provides antibodies that compete with 9F5 for binding to tau and reduce tau-induced toxicity in neurons. The present invention provides antibodies that compete with 9F5 for binding to tau and have increased resistance to agitation stress. Exemplary 9F5 humanized antibodies that compete with 9F5 for binding to tau and have increased resistance to agitation stress are L27cS / L37Q / M51G / L54R (DIM2), also known as hu9F5VHv9 / hu9F5VLv8_DIM2 SEQ ID NO: 127 / SEQ ID NO: 133, L27cG / L37G / M51G / L54T (DIM14), also known as hu9F5VHv9 / hu9F5VLv8_DIM14 SEQ ID NO: 127 / SEQ ID NO: 145, and L27cG / L37G / M51G / L54R (DIM13), also known as hu9F5VHv9 / hu9F5VLv8_DIM13 SEQ ID NO: 127 / SEQ ID NO: 144. The present invention provides antibodies that compete with 9F5 for binding to tau and have increased resistance to low pH stress. Exemplary 9F5 humanized antibodies that compete with 9F5 for binding to tau and have increased resistance to low pH stress are L27cS / L37Q / M51G / L54R (DIM2), also known as hu9F5VHv9 / hu9F5VLv8_DIM2 SEQ 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_DIM13 SEQ ID NO:127 / SEQ ID NO:144.Exemplary 9F5 humanized antibodies that compete with 9F5 for binding to tau, have increased resistance to agitation stress, and have increased resistance to low pH stress are DIM 2 [hu9F5VHv9 / hu9F5VLv8_DIM2 SEQ ID NO:127 / SEQ ID NO:133], DIM6 [hu9F5VHv9 / hu9F5VLv8_DIM6 SEQ ID NO:127 / SEQ ID NO:137], DIM7 [hu9F5VHv9 / hu9F5VLv8_DIM7 SEQ ID NO:127 / SEQ ID NO:138], DIM8 [hu9F5VHv9 / hu9F5VLv8_DIM8 SEQ ID NO:127 / SEQ ID NO:139], DIM13 [hu9F5VHv9 / hu9F5VLv8_DIM13 SEQ ID NO:127 / SEQ ID NO:144], DIM18 [hu9F5VHv9 / hu9F5VLv8_DIM18 SEQ ID NO:127 / SEQ ID NO:149], DIM28 [hu9F5VHv9 / hu9F5VLv8_DIM28 SEQ ID NO:127 / SEQ ID NO:159], and DIM30 [hu9F5VHv9 / hu9F5VLv8_DIM30 SEQ ID NO:127 / SEQ ID NO:161].

[0463] The present invention provides antibodies that compete with 9F5 for binding to tau and have a reduced tendency to aggregate under high concentration conditions. An exemplary antibody that competes with 9F5 for binding to tau and has a reduced tendency to aggregate under high concentration conditions is L37Q / M51G / L54R (DIM27) combined with the original leucine at position L27c, also known as hu9F5VHv9 / hu9F5VLv8_DIM27 SEQ ID NO: 127 / SEQ ID NO: 158.

[0464] The above-mentioned antibodies can be produced de novo by immunizing with a tau peptide comprising or consisting of the amino acid sequence QIVYKP (SEQ ID NO:57), a tau peptide comprising the amino acid sequence EIVYKSP (SEQ ID NO:58), a tau peptide comprising the amino acid sequence EIVYKS (SEQ ID NO:277), or a tau peptide comprising or consisting of the amino acid sequence (Q / E)IVYK(S / P) (SEQ ID NO:56), or by immunizing with a full-length tau peptide or a fragment thereof comprising said residues, and screening for specific binding to a peptide comprising said residues. Preferably, such tau peptides are conjugated to a heterologous conjugate molecule that helps elicit an antibody response against said peptide. Conjugation can be direct or through a spacer peptide or amino acid. Cysteine ​​is used as a spacer amino acid because its free SH group facilitates the attachment of a carrier molecule. Polyglycine linkers (e.g. 2-6 glycines) with or without a cysteine ​​residue between the glycine and the peptide can also be used. Carrier molecules act to provide T cell epitopes that help elicit an antibody response to the peptide. Several carriers are commonly used, notably keyhole limpet hemocyanin (KLH), ovalbumin, and bovine serum albumin (BSA). Peptide spacers can be added to the peptide immunogen as part of solid-phase peptide synthesis. Carriers are usually added by chemical cross-linking.Some examples of chemical cross-linking agents that can be used include cross-N-maleimido-6-aminocaproyl ester or m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS) (see, e.g., 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); International Patent Publication No. 98 / 23635; and Southwood et al. J. Immunology, 160:3363-3373). (1998)). Carriers and spacers, when present, can be attached to either end of the immunogen.

[0465] The peptides, including optional spacers and carriers, may be used to immunize experimental animals or B cells, as described in more detail below. Hybridoma supernatants may be tested for the property of binding to tau peptides comprising or consisting of the amino acid sequence QIVYKP (SEQ ID NO: 57), peptides comprising or consisting of the amino acid sequence EIVYKSP (SEQ ID NO: 58), peptides comprising or consisting of the amino acid sequence EIVYKS (SEQ ID NO: 277), or peptides comprising or consisting of the amino acid sequence (Q / E)IVYK(S / P) (SEQ ID NO: 56), and / or phosphorylated and non-phosphorylated forms of tau, such as the full-length isoform of tau with position 404 in phosphorylated form. The peptides may be bound to a carrier or other tag to facilitate screening assays. In this case, the carrier or tag is selectively different from the combination of spacer and carrier molecule used in immunization, to exclude antibodies specific for the spacer or carrier rather than the tau peptide.

[0466] The antibody designated 9F5 is an exemplary antibody that specifically binds to tau. Unless otherwise clear from the context, reference to 9F5 should be understood to refer to any of the murine, chimeric, veneered, and humanized forms of this antibody. The antibody has been deposited under [DEPOSIT NUMBER]. This antibody specifically binds to a peptide comprising or consisting of the amino acid sequence QIVYKP (SEQ ID NO:57), a peptide comprising or consisting of the amino acid sequence EIVYKSP (SEQ ID NO:58), or a peptide comprising or consisting of the amino acid sequence (Q / E)IVYK(S / P) (SEQ ID NO:56). The Kabat / Chothia composite CDRs of the heavy chain of 9F5 are designated SEQ ID NOs:8, 9, and 10, respectively, and the Kabat / Chothia composite CDRs of the light chain of 9F5 are designated SEQ ID NOs:12, 13, and 14, respectively.

[0467] Additional antibodies that compete with 9F5 for binding to tau and / or bind to the same or overlapping epitopes as 9F5 have been isolated, designated 10C12, 2D11, 12C4, 17C12, and 14H3, and are produced by hybridomas of the same name. 10C12 has mature variable heavy and light chain regions (after cleavage of the signal peptide) characterized by SEQ ID NO:7 and SEQ ID NO:11, respectively. Unless otherwise clear from the context, reference to 10C12 should be understood to represent either the murine, chimeric, veneered, or humanized form of this antibody. 10C12 has been deposited under [deposit number]. 10C12 is further characterized by the property of binding to both non-pathological and pathological forms and conformations of tau, as well as misfolded / aggregated forms of tau. 10C12 is associated with structural properties such as tau fibrillar tangles and degenerated neurites in tissue from Alzheimer's disease, and precipitates monomeric and aggregated tau from Alzheimer's disease extracts.

[0468] 2D11 has mature variable heavy and light chain regions (after cleavage of the signal peptide) characterized by SEQ ID NO: 7 and SEQ ID NO: 11, respectively. Unless otherwise clear from the context, reference to 2D11 should be understood to represent any of the murine, chimeric, veneered, and humanized forms of this antibody. 2D11 has been deposited under [deposit number]. 2D11 is further characterized by its ability to bind both non-pathological and pathological forms and conformations of tau, as well as misfolded / aggregated forms of tau. 2D11 has been associated with structural properties such as fibrillar tangles and degenerated neurites of tau in tissues from Alzheimer's disease, and precipitates monomeric and aggregated tau from Alzheimer's disease extracts.

[0469] 12C4 has mature variable heavy and light chain regions (after cleavage of the signal peptide) characterized by SEQ ID NO: 219 and SEQ ID NO: 11, respectively. Unless otherwise clear from the context, reference to 12C4 should be understood to represent any of the murine, chimeric, veneered, and humanized forms of this antibody. 12C4 has been deposited under [deposit number]. 12C4 is further characterized by its ability to bind both non-pathological and pathological forms and conformations of tau, as well as misfolded / aggregated forms of tau. 12C4 has been associated with structural properties such as fibrillar tangles and degenerated neurites of tau in tissues from Alzheimer's disease, and precipitates monomeric and aggregated tau from Alzheimer's disease extracts.

[0470] 17C12 has mature variable heavy and light chain regions (after cleavage of the signal peptide) characterized by SEQ ID NO: 225 and SEQ ID NO: 228, respectively. Unless otherwise clear from the context, reference to 17C12 should be understood to represent any of the murine, chimeric, veneered, and humanized forms of this antibody. 17C12 has been deposited under [deposit number]. 17C12 is further characterized by its ability to bind to both non-pathological and pathological forms and conformations of tau, as well as misfolded / aggregated forms of tau.

[0471] 14H3 has mature variable heavy and light chain regions (after cleavage of the signal peptide) characterized by SEQ ID NOs: 240 and 244, respectively. Unless otherwise clear from the context, reference to 14H3 should be understood to represent any of the murine, chimeric, veneered, and humanized forms of this antibody. 14H3 is deposited under [deposit number]. 14H3 is further characterized by its ability to bind both non-pathological and pathological forms and conformations of tau, as well as misfolded / aggregated forms of tau. 14H3 has been associated with structural properties such as fibrillary tangles and degenerated neurites of tau in tissues from Alzheimer's disease.

[0472] An alignment of the mature heavy chain variable regions of the murine 9F5, 10C12, 2D11, 12C4, 14H3, and 17C12 antibodies is presented in Figure 24, and the mature light chain variable regions of the murine 9F5, 10C12, 2D11, 12C4, 14H3, and 17C12 antibodies is presented in Figure 25. The amino acid sequence of the mature heavy chain variable region of the murine 10C12 antibody has 100% sequence identity to that region of the murine 9F5 antibody, and the mature light chain variable region of the murine 10C12 antibody has 100% sequence identity to that region of the murine 9F5 antibody. The amino acid sequence of the mature heavy chain variable region of the mouse 2D11 antibody has 100% sequence identity to the corresponding region of the mouse 9F5 antibody, and the mature light chain variable region of the mouse 2D11 antibody has 100% sequence identity to the corresponding region of the mouse 9F5 antibody. The amino acid sequence of the mature heavy chain variable region of the mouse 12C4 antibody has 96.6% sequence identity to the corresponding region of the mouse 9F5 antibody, and the mature light chain variable region of the mouse 12C4 antibody has 100% sequence identity to the corresponding region of the mouse 9F5 antibody. The amino acid sequence of the mature heavy chain variable region of the mouse 17C12 antibody has 95.9% sequence identity to the corresponding region of the mouse 9F5 antibody, and the mature light chain variable region of the mouse 17C12 antibody has 70.5% sequence identity to the corresponding region of the mouse 9F5 antibody. The amino acid sequence of the mature heavy chain variable region of the mouse 14H3 antibody has 35.0% sequence identity to the corresponding region of the mouse 9F5 antibody, and the mature light chain variable region of the mouse 14H3 antibody has 73.2% sequence identity to the corresponding region of the mouse 9F5 antibody.

[0473] Optionally, the antibodies of the present invention do not include a 10C12 antibody. Optionally, the antibodies of the present invention do not include a 2D11 antibody. Optionally, the antibodies of the present invention do not include a 12C4 antibody. Optionally, the antibodies of the present invention do not include a 17C12 antibody. Optionally, the antibodies of the present invention do not include a 14H3 antibody.

[0474] Some antibodies of the present invention bind to the same or overlapping epitopes as the antibodies designated 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3. The sequences of the heavy and light chain mature variable regions of 9F5 are designated SEQ ID NOs: 7 and 11, respectively. The sequences of the heavy and light chain mature variable regions of 10C12 are designated SEQ ID NOs: 7 and 11, respectively. The sequences of the heavy and light chain mature variable regions of 2D11 are designated SEQ ID NOs: 7 and 11, respectively. The sequences of the heavy and light chain mature variable regions of 12C4 are designated SEQ ID NOs: 219 and 11, respectively. The sequences of the heavy and light chain mature variable regions of 17C12 are designated SEQ ID NOs: 225 and 228, respectively. The sequences of the heavy and light chain mature variable regions of 14H3 are designated SEQ ID NOs: 240 and 244, respectively. Other antibodies having such binding specificity can be produced by immunizing mice with tau or a portion thereof comprising the desired epitope (e.g., a tau peptide comprising or consisting of the amino acid sequence QIVYKP (SEQ ID NO:57), a tau peptide comprising or consisting of the amino acid sequence EIVYKSP (SEQ ID NO:58), a tau peptide comprising or consisting of the amino acid sequence EIVYKS (SEQ ID NO:277), or a tau peptide comprising or consisting of the amino acid sequence (Q / E)IVYK(S / P) (SEQ ID NO:56)) and screening the resulting antibodies for binding to tau, optionally in competition with antibodies having the variable regions of murine 9F5 (IgG1 / κ), 10C12 (IgG2a / κ), 2D11 (IgG2a / κ), 12C4 (IgG2a / κ), 17C12 (IgG2a / κ), or 14H3 (IgG2a / κ). Fragments of tau containing the desired epitope may be coupled to a carrier and / or combined with an adjuvant that helps elicit an antibody response against the fragment. Such antibodies may be screened for differential binding to tau or its fragments compared to mutants of specified residues.Screening against such mutants allows for the identification of antibodies whose binding is inhibited by mutagenesis of specific residues, possibly sharing functional properties of the other exemplified antibodies, by defining the binding specificity more precisely. The mutations can be systematic substitutions with alanine (or serine, if alanine is already present), one residue at a time or at more widely spaced intervals, across the target or across a section of it where the epitope is known to remain. If the same set of mutations significantly reduces binding of the two antibodies, then the two antibodies bind to the same epitope.

[0475] Antibodies having the binding specificity of a selected murine antibody (e.g., 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3) can also be produced using a variant of the phage display method. See Winter, International Patent Publication No. 92 / 20791. This method is particularly suitable for producing human antibodies. In this method, the heavy or light chain variable region of a selected murine antibody is used as the starting material. For example, if a light chain variable region is selected as the starting material, a phage library is constructed in which members display the same light chain variable region (i.e., the murine 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. Phages (e.g., at least 10 8 , preferably at least 10 9 M -1 ) are selected. The heavy chain variable region from this phage then serves as starting material for constructing a further phage library, in which each phage displays the same heavy chain variable region (i.e., the region identified from the first display library) and a different light chain variable region. The light chain variable region may be obtained from a library of rearranged human variable light chain regions. Again, phage that show strong specific binding to tau or a fragment thereof are selected. The resulting antibodies usually have the same or similar epitope specificity as the murine starting material.

[0476] The composite Kabat / Chothia CDRs of the heavy chain of 9F5 are represented as SEQ ID NOs:8 to 10, respectively, and the composite Kabat / Chothia CDRs of the light chain of 9F5 are represented as SEQ ID NOs:12 to 14, respectively.

[0477] The composite Kabat / Chothia CDRs of the heavy chain of 10C12 are represented as SEQ ID NOs:8 to 10, respectively, and the composite Kabat / Chothia CDRs of the light chain of 10C12 are represented as SEQ ID NOs:12 to 14, respectively.

[0478] The composite Kabat / Chothia CDRs of the heavy chain of 2D11 are represented as SEQ ID NOs: 8 to 10, respectively, and the composite Kabat / Chothia CDRs of the light chain of 2D11 are represented as SEQ ID NOs: 12 to 14, respectively.

[0479] Table 2 presents the CDRs of 9F5, 10C12, and 2D11 as defined by Kabat, Chothia, Chothia and Kabat composite (also referred to herein as "Kabat / Chothia composite"), AbM, and Contact. [Table 2]

[0480] The composite Kabat / Chothia CDRs of the heavy chain of 12C4 are designated SEQ ID NOs: 8, 220, and 10, respectively, and the Kabat CDRs of the light chain of 12C4 are designated SEQ ID NOs: 12-14, respectively.

[0481] Table 3 presents the 12C4 CDRs as defined by Kabat, Chothia, Chothia and Kabat composite (also referred to herein as the "Kabat / Chothia composite"), AbM, and Contact. [Table 3]

[0482] The composite Kabat / Chothia CDRs of the heavy chain of 17C12 are represented as SEQ ID NOs: 226, 227, and 10, respectively, and the Kabat CDRs of the light chain of 17C12 are represented as SEQ ID NOs: 229-231, respectively.

[0483] Table 4 presents the 17C12 CDRs as defined by Kabat, Chothia, Chothia and Kabat composite (also referred to herein as the "Kabat / Chothia composite"), AbM, and Contact. [Table 4]

[0484] The composite Kabat / Chothia CDRs of the heavy chain of 14H3 are represented as SEQ ID NOs: 241-243, respectively, and the Kabat CDRs of the light chain of 14H3 are represented as SEQ ID NOs: 245-247, respectively.

[0485] Table 5 presents the 14H3 CDRs defined by Kabat, Chothia, Chothia and Kabat composite (also referred to herein as the "Kabat / Chothia composite"), AbM, and Contact. [Table 5]

[0486] Other antibodies can be obtained by mutagenesis of cDNA encoding the heavy and light chains of exemplary antibodies such as 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3. Also included in the invention are 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 mature heavy and / or light chain variable region, retain their functional properties, and / or differ from the respective antibodies by a small number of functionally minor amino acid substitutions (e.g., conservative substitutions), deletions, or insertions. Also included are monoclonal antibodies having at least one or all six CDRs, as defined by any conventional definition, but preferably as defined by Kabat, that are 90%, 95%, 99% or 100% identical to the corresponding CDRs of 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3.

[0487] The invention also provides antibodies having some or all (e.g., 3, 4, 5, and 6) CDRs derived completely or substantially from 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3. Such antibodies may comprise a heavy chain variable region having at least two, usually three, CDRs derived completely or substantially from the heavy chain variable region of 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3, and / or a light chain variable region having at least two, usually three, CDRs derived completely or substantially from the light chain variable region of 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3. An antibody may comprise both a heavy chain and a light chain. A CDR is substantially derived from the corresponding 9F5 CDR if it contains no more than 4, 3, 2, or 1 substitutions, insertions, or deletions, except that CDR-H2 (as defined by Kabat) may have no more than 6, 5, 4, 3, 2, or 1 substitutions, insertions, or deletions. Such antibodies may have at least 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% identity to 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3 in the amino acid sequence of the mature heavy chain variable region and / or the mature light chain variable region, and may retain its functional properties, and / or may differ from 9F5, 10C12, 2D11, 12C4, 17C12, or 14H3 by a small number of functionally minor amino acid substitutions (e.g., conservative substitutions), deletions, or insertions.

[0488] Some antibodies identified by such assays may bind to monomeric, misfolded, aggregated, phosphorylated, or nonphosphorylated forms of tau, or other forms. Similarly, some antibodies are immunoreactive with nonpathological and pathological forms and conformations of tau.

[0489] The invention further provides means for specifically binding to a peptide consisting of residues (Q / E)IVYK(S / P) (SEQ ID NO:56), residues QIVYKP (SEQ ID NO:57), residues EIVYKSP (SEQ ID NO:58), or residues EIVYKS (SEQ ID NO:277). Exemplary means are antibodies comprising a heavy chain CDR of SEQ ID NO:8-10 and a light chain CDR of SEQ ID NO:12-14. Exemplary means are antibodies comprising a heavy chain CDR of SEQ ID NO:8, 220, and 10 and a light chain CDR of SEQ ID NO:12-14. Exemplary means are antibodies comprising a heavy chain CDR of SEQ ID NO:226, 227, and 10 and a light chain CDR of SEQ ID NO:229-231. Exemplary means are antibodies comprising a heavy chain CDR of SEQ ID NO:241-243 and a light chain CDR of SEQ ID NO:245-247.

[0490] B. Non-human antibodies Production of other non-human antibodies, e.g., mouse, guinea pig, primate, rabbit, or rat antibodies, against tau, or a fragment thereof (e.g., a peptide comprising 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 a fragment thereof. See Harlow & Lane, Antibodies, A Laboratory Manual (CSHP NY, 1988) (incorporated by reference for all purposes). Optionally, the immunogen can be 383 amino acid human tau (4R0N). Optionally, the immunogen can be human tau containing the P301S mutation. Optionally, the immunogen can be human tau that is recombinantly N-terminally His-tagged. Optionally, the animal is immunized with a tau fragment containing a peptide represented by (Q / E)IVYK(S / P) (SEQ ID NO:56) conjugated to a carrier. Optionally, the peptide is QIVYKP (SEQ ID NO:57), EIVYKSP (SEQ ID NO:58), or EIVYKS (SEQ ID NO:277). Such immunogens can be obtained from natural sources, by peptide synthesis, or by recombinant expression. Optionally, the immunogens can be administered fused to or otherwise complexed with a carrier protein. Optionally, the immunogens can be administered with an adjuvant. Several types of adjuvants can be used, as described below. Complete Freund's adjuvant, followed by incomplete adjuvant, can be used to immunize laboratory animals. Rabbits or guinea pigs are usually used to generate polyclonal antibodies. Mice are usually used to generate monoclonal antibodies. Antibodies are screened for specific binding to tau or an epitope within tau (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 against 15 amino acid peptides comprising QIVYKP (SEQ ID NO:57), EIVYKSP (SEQ ID NO:58), EIVYKS (SEQ ID NO:277), or any other consensus motif presented by (Q / E)IVYK(S / P) (SEQ ID NO:56). Optionally, the peptide comprises QIVYKP (SEQ ID NO:57) EIVYKSP (SEQ ID NO:58), EIVYKS (SEQ ID NO:277). Such screening may be accomplished by determining binding of the antibody to a collection of tau variants, such as tau variants comprising or consisting of amino acid residues 307-312 or 391-397 or 391-396 of SEQ ID NO:1, or mutations therein, and determining whether the tau variants bind to the antibody. Binding may be assessed, for example, by Western blot, FACS, or ELISA.

[0491] C. Humanized Antibodies A humanized antibody is a genetically engineered antibody in which CDRs from a non-human "donor" antibody are grafted into the sequences of a human "acceptor" antibody (see, e.g., U.S. Pat. Nos. 5,530,101 and 5,585,089 (Queen); U.S. Pat. Nos. 5,225,539 (Winter); 6,407,213 (Carter); 5,859,205 (Adair); and 6,881,557 (Foote)). The sequences of the acceptor antibody can be, for example, the sequences of a mature human antibody, a composite of such sequences, a consensus sequence of human antibody sequences, or a sequence of a germline region. Thus, a humanized antibody is an antibody having at least three, four, five, or all of the CDRs, completely or substantially, derived from the donor antibody, and variable region framework sequences and constant regions, if present, completely or substantially, derived from human antibody sequences. Similarly, a humanized heavy chain has at least one, two, and usually all three CDRs entirely or substantially from the heavy chain of a donor antibody, and a heavy chain variable region framework sequence and a heavy chain constant region, if present, substantially from the human heavy chain variable region framework and constant region sequences. Similarly, a humanized light chain has at least one, two, and usually all three CDRs entirely or substantially from the donor antibody light chain, and a light chain variable region framework sequence and a light chain constant region, if present, substantially from the human light chain variable region framework and constant region sequences. Other than nanobodies and dAbs, a humanized antibody comprises a humanized heavy chain and a humanized light chain. The CDRs of a humanized antibody are derived from substantially the corresponding CDRs in a non-human antibody when at least 85%, 90%, 95%, or 100% of the corresponding residues (defined by any conventional definition, but preferably defined by Kabat) are identical between each CDR. The framework sequence of a variable region of an antibody chain or a constant region of an antibody chain is substantially derived from a framework sequence of a human variable region or a human constant region, respectively, if at least 85%, 90%, 95%, or 100% of the corresponding residues as defined by Kabat are identical.For a humanized antibody to be classified as humanized under the 2014 World Health Organization (WHO) International Nonproprietary Name (INN) definition, the antibody must have at least 85% identity with the human germline antibody sequence (i.e., prior to somatic mutations). A mixed antibody is one in which one antibody chain (e.g., the heavy chain) matches the threshold, but the other chain (e.g., the light chain) does not. If neither chain matches the threshold, even though the variable framework regions in both chains are substantially human with some mouse back mutations, the antibody is classified as chimeric. See Jones et al. (2016) The INNs and outs of antibody nonproprietary names, mAbs 8:1, 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) (hereby incorporated by reference). For the avoidance of doubt, the term "humanized" as used herein is not intended to be limited to the 2014 WHO INN definition of a humanized antibody. Some of the humanized antibodies provided herein have at least 85% sequence identity with human germline sequences, and some of the humanized antibodies provided herein have less than 85% sequence identity with human germline sequences. Some of the heavy chains of the humanized antibodies provided herein have about 60-100% sequence identity with human germline sequences, e.g., within the range of about 60%-69%, 70%-79%, 80%-84%, or 85%-89% sequence identity.Some heavy chains fall below the 2014 WHO INN definition, e.g., have about 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, or 82%, 83%, or 84% sequence identity to human germline sequences, and other heavy chains meet the 2014 WHO INN definition, have about 85%, 86%, 87%, 88%, 89% or more sequence identity to human germline sequences. Some of the light chains of the humanized antibodies provided herein have about 60%-100% sequence identity to human germline sequences, e.g., within the range of about 80%-84% or 85%-89% sequence identity. Some light chains fall below the 2014 WHO INN definition, e.g., have about 81%, 82%, 83%, or 84% sequence identity to human germline sequences, while other light chains meet the 2014 WHO INN definition and have about 85%, 86%, 87%, 88%, 89%, or more sequence identity to human germline sequences. Some humanized antibodies provided herein that are "chimeric" under the 2014 WHO INN definition have a heavy chain that has less than 85% identity to human germline sequences paired with a light chain that has less than 85% identity to human germline sequences. For example, some humanized antibodies provided herein that have a heavy chain that has less than 85% identity to human germline sequences paired with a light chain that has less than 85% identity to human germline sequences, or vice versa, are "mixed" under the 2014 WHO INN definition. Some humanized antibodies provided herein meet the 2014 WHO INN definition of "humanized" and have a heavy chain having at least 85% sequence identity to a human germline sequence paired with a light chain having at least 85% sequence identity to a human germline sequence. Exemplary 12C4 antibodies that meet the 2014 WHO INN definition of "humanized" include antibodies having a mature heavy chain having the amino acid sequence of SEQ ID NO:221 or SEQ ID NO:222 paired with a mature light chain sequence having the amino acid sequence of SEQ ID NO:223 or SEQ ID NO:224.Exemplary 14H3 antibodies that meet the 2014 WHO INN definition of "humanized" include antibodies having a mature heavy chain having the amino acid sequence of SEQ ID NO: 248 or SEQ ID NO: 249 paired with a mature light chain sequence having the amino acid sequence of SEQ ID NO: 251 or SEQ ID NO: 252. Some humanized antibodies provided herein meet the 2014 WHO INN definition of "mixed." Exemplary 9F5 antibodies that meet the 2014 WHO INN definition of "mixed" include antibodies having a mature heavy chain having the amino acid sequence of any of SEQ ID NOs: 15-22 and SEQ ID NOs: 127-128 paired with a mature light chain sequence having the amino acid sequence of any of SEQ ID NOs: 26-29 and SEQ ID NOs: 130-131. Exemplary 10C12 antibodies that meet the 2014 WHO INN definition of "mixed" include antibodies having a mature heavy chain having the amino acid sequence of SEQ ID NO: 214 or SEQ ID NO: 215 paired with a mature light chain sequence having the amino acid sequence of SEQ ID NO: 216 or SEQ ID NO: 217. Exemplary 17C12 antibodies that meet the 2014 WHO INN definition of "mixed" include antibodies having a mature heavy chain having the amino acid sequence of SEQ ID NO: 232 or SEQ ID NO: 233 paired with a mature light chain having the amino acid sequence of SEQ ID NO: 235. Further humanized 9F5 antibodies of the invention include antibodies having a mature heavy chain having the amino acid sequence of any of SEQ ID NOs: 15-22 and 127-128 paired with a mature light chain having the amino acid sequence of any of SEQ ID NOs: 23-25. Further humanized 17C12 antibodies of the invention include antibodies having a mature heavy chain having the amino acid sequence of SEQ ID NO: 232 or SEQ ID NO: 233 paired with a mature light chain having the amino acid sequence of SEQ ID NO: 234.

[0492] Humanized antibodies often incorporate all six CDRs (as defined by any conventional definition, but preferably as defined by Kabat) from a murine antibody, although they can also be made with fewer than all of the CDRs from a murine antibody (e.g., at least three, four, or five 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).

[0493] In some antibodies, only a portion of the CDRs, called SDRs, i.e., a subset of CDR residues required for binding, are required to retain binding of the humanized antibody. CDR residues that do not contact the antigen and are not in the SDRs can be identified from regions of the Kabat CDRs other than the Chothia hypervariable loops (Chothia, J. Mol. Biol. 196:901, 1987), by molecular modeling and / or experience, or based on previous testing as described in Gonzales et al., Mol. Immunol. 41: 863, 2004 (e.g., residues H60-H65 of CDR H2 are often not required). In such humanized antibodies, where one or more donor CDR residues are absent or the entire donor CDR is omitted, the amino acid residue that occupies that position can be the amino acid that occupies the corresponding position (according to Kabat numbering) in the acceptor antibody sequence. The number of substitutions of acceptor donor amino acids in such included CDRs reflects a balance of competing considerations. Such substitutions are potentially advantageous in reducing the number of mouse amino acids in the humanized antibody and thus reducing potential immunogenicity, and / or to meet the WHO INN definition of "humanization". However, substitutions may also result in changes in affinity, and significant reductions in affinity are preferably avoided. The position of substitution in the CDR and the amino acid to be substituted may also be empirically selected.

[0494] The sequence of the human acceptor antibody may optionally be selected from the many known human antibodies to provide a high degree of sequence identity (e.g., 65-85% identity) between the framework of the human acceptor sequence variable region and the framework of the corresponding variable region of the donor antibody chain. Some humanized and chimeric antibodies have the same or improved functional properties (within experimental error), such as binding affinity for human tau, inhibition of internalization of tau into neurons, as described in the Examples, as the mouse antibodies from which they are derived. For example, some humanized and chimeric antibodies have binding affinities within a factor of 3, 2, or 1 of the mouse antibodies from which they are derived, or affinities that are indistinguishable within experimental error. Some humanized and chimeric antibodies inhibit internalization of tau into neurons as described in the Examples, within a factor of 3, 2, or 1 of the mouse antibodies from which they are derived, or inhibit as well as the mouse antibodies from which they are derived, within experimental error.

[0495] An example of an acceptor sequence for the 9F5 heavy chain is the human mature heavy chain variable region of humanized 48G7 Fab with PDB accession code 2RCS-VH_huFrwk (SEQ ID NO: 32). An example of an acceptor sequence for the 9F5 heavy chain is the human mature heavy chain GenBank AAN16432-VH_huFrwk (SEQ ID NO: 31). The variable domains of 9F5 and 48G7 Fab also share the same length for the CDR-H1, H2 loops. An example of an acceptor sequence for the 9F5 heavy chain is the human mature heavy chain variable region IMGT# IGHV1-69-2 * 01 (SEQ ID NO: 33). IMGT# IGHV1-69-2 * The Chothia CDR-H1 of 01 (SEQ ID NO: 33) is in canonical class 1, and the Chothia CDR-H2 is in canonical class 2. IMGT# IGHV1-69-2 *01 (SEQ ID NO: 33) belongs to human heavy chain subgroup 1. An example of an acceptor sequence for the 9F5 light chain is the human mature light chain variable region 1911357B-VL_huFrwk (SEQ ID NO: 36). An example of an acceptor sequence for the 9F5 light chain is the human mature light chain variable region CAB51297-VL_huFrwk (SEQ ID NO: 35). The variable light chain domains of the 9F5 antibody and CAB51297 & 1911357B antibodies also share the same length for the CDR-L1, L2, and L3 loops. An example of an acceptor sequence for the 9F5 light chain is IGKV2-28 * 01 & IGKJ2 * 01 (SEQ ID NO: 37). IGKV2-28 * 01 & IGKJ2 * The Chothia CDR-L1 of IGKV2-28 (SEQ ID NO: 37) is in canonical class 4. * 01 & IGKJ2 * The Chothia CDR-L2 of IGKV2-28 (SEQ ID NO: 37) is in canonical class 1. * 01 & IGKJ2 * The Chothia CDR-L3 of IGKV2-28 (SEQ ID NO: 37) is canonical class 1. * 01 & IGKJ2 * 01 (SEQ ID NO:37) belongs to the human kappa subgroup 2.

[0496] An example of an acceptor sequence for the 10C12 heavy chain is the human mature CAC20421 (SEQ ID NO: 218). The variable domains of 10C12 and CAC20421 VH also share the same length for the CDR-H1, H2 loops. An example of an acceptor sequence for the 10C12 heavy chain is the human mature heavy chain variable region IMGT# IGHV1-69-2 * 01 (SEQ ID NO: 33). IMGT# IGHV1-69-2 * The Chothia CDR-H1 of 01 (SEQ ID NO: 33) is in canonical class 1, and the Chothia CDR-H2 is in canonical class 2. IMGT# IGHV1-69-2 *01 (SEQ ID NO: 33) belongs to human heavy chain subgroup 1. An example of an acceptor sequence for the 10C12 light chain is the human mature light chain variable region CAB51297-VL_huFrwk (SEQ ID NO: 35). The variable light chain domains of 10C12 and CAB51297 VL also share identical lengths for the CDR-L1, L2, and L3 loops. An example of an acceptor sequence for the 10C12 light chain is IGKV2-28 * 01 & IGKJ2 * 01 (SEQ ID NO: 37). IGKV2-28 * 01 & IGKJ2 * The Chothia CDR-L1 of IGKV2-28 (SEQ ID NO: 37) is in canonical class 4. * 01 & IGKJ2 * The Chothia CDR-L2 of IGKV2-28 (SEQ ID NO: 37) is in canonical class 1. * 01 & IGKJ2 * The Chothia CDR-L3 of IGKV2-28 (SEQ ID NO: 37) is canonical class 1. * 01 & IGKJ2 * 01 (SEQ ID NO:37) belongs to the human kappa subgroup 2.

[0497] An example of an acceptor sequence for the 12C4 heavy chain is the human mature heavy chain variable region of CAC20421-VH_huFrwk (SEQ ID NO: 218). The variable domains of 12C4 and CAC20421 VH also share the same length for the CDR-H1, H2 loops. An example of an acceptor sequence for the 12C4 heavy chain is the human mature heavy chain variable region IMGT# IGHV1-69-2 * 01 (SEQ ID NO: 33). IMGT# IGHV1-69-2 * The Chothia CDR-H1 of 01 (SEQ ID NO: 33) is in canonical class 1, and the Chothia CDR-H2 is in canonical class 2. IMGT# IGHV1-69-2 *01 (SEQ ID NO: 33) belongs to human heavy chain subgroup 1. An example of an acceptor sequence for the 12C4 light chain is the human mature light chain variable region CAB51297-VL_huFrwk (SEQ ID NO: 35). The variable light chain domains of 12C4 and CAB51297 VL also share identical lengths for the CDR-L1, L2, and L3 loops. An example of an acceptor sequence for the 12C4 light chain is IGKV2-28 * 01 & IGKJ2 * 01 (SEQ ID NO: 37). IGKV2-28 * 01 & IGKJ2 * The Chothia CDR-L1 of IGKV2-28 (SEQ ID NO: 37) is in canonical class 4. * 01 & IGKJ2 * The Chothia CDR-L2 of IGKV2-28 (SEQ ID NO: 37) is in canonical class 1. * 01 & IGKJ2 * The Chothia CDR-L3 of IGKV2-28 (SEQ ID NO: 37) is canonical class 1. * 01 & IGKJ2 * 01 (SEQ ID NO:37) belongs to the human kappa subgroup.

[0498] An example of an acceptor sequence for the 17C12 heavy chain is the human mature heavy chain variable region of CAC20421-VH_huFrwk (SEQ ID NO: 218). The variable heavy chain domains of 17C12 and CAC20421 also share the same length for the CDR-H1, H2 loops. An example of an acceptor sequence for the 17C12 heavy chain is the human mature heavy chain variable region IMGT# IGHV1-69-2 * 01 (SEQ ID NO: 33). IMGT# IGHV1-69-2 * The Chothia CDR-H1 of 01 (SEQ ID NO: 33) is in canonical class 1, and the Chothia CDR-H2 is in canonical class 2. IMGT# IGHV1-69-2 *01 (SEQ ID NO: 33) belongs to human heavy chain subgroup 1. An example of an acceptor sequence for the 17C12 light chain is the human mature light chain variable region QDO16713-VL_huFrwk (SEQ ID NO: 238). The variable light chain domains of the 17C12 and QDO16713 antibodies also share identical lengths for the CDR-L1, L2, and L3 loops. An example of an acceptor sequence for the 17C12 light chain is IGKV2-29 * 02 & IGKJ4 * 01 (SEQ ID NO: 239).

[0499] An example of an acceptor sequence for the 14H3 heavy chain is the human mature heavy chain variable region of QDJ57937-VH_huFrwk (SEQ ID NO: 253). The variable domains of 14H3 and QDJ57937 VH share the same length of the CDR-H1, H2 loops. An example of an acceptor 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 an acceptor sequence for the 14H3 light chain is the human mature light chain variable region ABC66914-VL_huFrwk (SEQ ID NO: 256). The variable light chain domains of 14H3 and ABC66914 VL also share identical lengths for the CDR-L1, L2, and L3 loops. An example of an acceptor sequence for the 14H3 light chain is IGKV2-28 * 01 & IGKJ2 * 01 (SEQ ID NO: 37). IGKV2-28 * 01 & IGKJ2 * The Chothia CDR-L1 of IGKV2-28 (SEQ ID NO: 37) is in canonical class 4. * 01 & IGKJ2 * The Chothia CDR-L2 of IGKV2-28 (SEQ ID NO: 37) is in canonical class 1. * 01 & IGKJ2 * The Chothia CDR-L3 of IGKV2-28 (SEQ ID NO: 37) is canonical class 1. * 01 & IGKJ2 *01 (SEQ ID NO:37) belongs to the human kappa subgroup 2.

[0500] If more than one human acceptor antibody sequence is selected, a composite or hybrid of these acceptors can be used, and the amino acids used at different positions in the humanized light and heavy chain variable regions can be taken from any of the human acceptor antibody sequences used. For example, the human mature heavy chain variable region of AAN16432-VH_huFrwk (SEQ ID NO: 31) and humanized 48G7 Fab with PDB accession code 2RCS-VH_huFrwk (SEQ ID NO: 32) were used as hybrid acceptor sequences for humanization of the 9F5 mature heavy chain variable region. Examples of positions where the two acceptors differ are 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). A humanized version of the 9F5 heavy chain variable region may include any of the amino acids at any of these positions. Human germline sequence IMGT# IGHV1-69-2 *01 (SEQ ID NO: 25) was also used as an acceptor sequence for 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 hybrid acceptor sequences for humanization of the 9F5 mature light chain variable region. Examples of positions where these two acceptors differ are 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). A humanized version of the 9F5 light chain variable region can include any of the amino acids at any of these positions. * 01 & IGKJ2 * 01 (SEQ ID NO:37) was also used as the acceptor sequence for humanization of the 9F5 mature light chain variable region.

[0501] Particular amino acids from the human variable region framework residues may be selected for substitution based on their possible effect on the CDR conformation and / or binding to antigen, which may be examined by modeling, testing the characteristics of the amino acid at a particular position, or by experimental observation of the effect of substitution or mutagenesis of a particular amino acid.

[0502] For example, if an amino acid differs between a residue in the mouse variable region framework and a residue in the selected human variable region framework, the amino acid in the human framework may be (1) Direct non-covalent binding to the antigen, (2) adjacent to or within the CDR regions, as defined by Chothia but not by Kabat; (3) interact with the CDR region in some other way (e.g., within about 6 Å of the CDR region) (e.g., identified by modeling the light or heavy chain on the resolved structure of a homologous known immunoglobulin chain); or (4) Residues involved in the VL-VH interface Where it is reasonably predicted that this will be the case, amino acids from the equivalent framework from the mouse antibody may be substituted.

[0503] In one embodiment, humanized sequences are generated using QuikChange site-directed mutagenesis, a two-step PCR protocol that allows for the introduction of multiple mutations, deletions, and insertions [Wang, W. and Malcolm, BA (1999) BioTechniques 26:680-682].

[0504] Framework residues of classes (1)-(3) as defined by Queen et al., U.S. Patent No. 5,530,101, are sometimes alternatively referred to as canonical and vernier residues. Framework residues that help define the conformation of the CDR loops 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 help define the conformation of the antigen-binding loops and play a role in fine-tuning the fit of the antibody to the antigen are sometimes referred to as veneer residues (Foote & Winter, J. Mol. Biol 224:487-499 (1992)).

[0505] Other framework residues that are candidates for substitution are those that create potential glycosylation sites. Further candidates for substitution are amino acids in the acceptor human framework that are rare in human immunoglobulins at that position. These amino acids can be replaced with amino acids from the equivalent position in the mouse donor antibody or from the equivalent position in a more typical human immunoglobulin.

[0506] Another framework residue that is a candidate for substitution is the N-terminal glutamine residue (Q), which can be replaced with glutamic acid (E) to minimize the possibility of conversion to pyroglutamic acid [Y. Diana Liu, et al., 2011, J. Biol. Chem., 286: 11211-11217]. Conversion of glutamic acid (E) to pyroglutamic acid (pE) is slower than conversion from glutamine (Q). Due to the loss of a primary amine in the conversion of glutamine to pE, the antibody becomes more acidic. Incomplete conversion leads to antibody heterogeneity, which can be observed as multiple peaks using charge-based analytical methods. Differences in heterogeneity may represent loss of process control. Exemplary 9F5 humanized heavy chain variable regions with N-terminal glutamine to glutamate 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), SEQ ID 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 ), sequence number 123 (hu9F5VHv5_M80G), sequence number 124 (hu9F5VHv4_L82cS), sequence number 125 (hu9F5VHv4_Y79Q), sequence number 126 (hu9F5VHv4_S82aG), sequence number 127 (hu9F5VHv9), sequence number 128 (hu9F5VHv10), sequence number 129 (hu9F5VHv10_L82cG).An exemplary 10C12 humanized heavy chain variable region with an N-terminal glutamine to glutamate substitution is SEQ ID NO: 215 (hu10C12VHv2). An exemplary 12C4 humanized heavy chain variable region with an N-terminal glutamine to glutamate substitution is SEQ ID NO: 222 (hu12C4VHv2). An exemplary 17C12 humanized heavy chain variable region with an N-terminal glutamine to glutamate substitution is SEQ ID NO: 233 (hu17C12VHv2).

[0507] Exemplary humanized antibodies include the humanized form of murine 9F5, designated Hu9F5.

[0508] The murine antibody 9F5 comprises a mature heavy chain variable region and a mature light chain variable region having amino acid sequences comprising SEQ ID NO: 7 and SEQ ID NO: 11, respectively. The present 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: 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), Provided herein are u9F5VHv5_M80G (SEQ ID NO: 123), hu9F5VHv4_L82cS (SEQ ID NO: 124), 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). The present invention further provides 95 exemplified 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:29), hu9F5VLv8 (SEQ ID NO:130), hu9F5VLv9 (SEQ ID NO:131), hu9F5VLv2_M51E (SEQ ID NO:61), hu9F5VLv2_M51D (SEQ ID NO: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 ID NO: 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 (SEQ ID NO: 72), hu9F5VLv2_L33N (SEQ ID NO: 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 (SEQ ID NO: 78), hu9F5VLv2_L27bT (SEQ ID NO: 79), hu9F5VLv2_T31G (SEQ ID NO: 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 ID NO: 87), hu9F5VLv2_V78P (SEQ ID NO: 88), hu9F5VLv2_V78K (SEQ ID NO: 89), hu9F5VLv2_R77D (SEQ ID 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_I75 G (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 (SEQ ID NO: 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 ID NO: 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 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), hu9F5V Lv8_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), hu9F 5VLv8_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) are provided. Figures 1A-1B and 4A-4B show alignments of the heavy chain variable regions of mouse 9F5 and various humanized antibodies. Figures 2A-2B and 5A-5B show alignments of the light chain variable regions of mouse 9F5 and various humanized antibodies. Figures 6A-6C show alignments of the light chain variable regions of humanized variant hu9F5VLv8 and various humanized antibodies.

[0509] For reasons such as possible effects on CDR conformation and / or binding to antigen, interactions between heavy and light chains, mediating interactions with the constant region, being sites of desirable or undesirable post-translational modifications, being rare residues at that position in human variable region sequences and thus potentially immunogenic, reducing the likelihood of aggregation, and other reasons, the following 48 variable region framework positions were selected from the 95 illustrated human mature light chain possible residues, as further detailed in the Examples: Variable regions and 29 exemplified substitutions in human mature light chain variable regions were considered: L3 (V3Q), L7 (A7S), L8 (A8P), 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 (I76D, I76P, I76Q, I76G), L77 (I77D, I77P, I77Q, I77G), L78 (I78D, I78P, I78Q, I78G), L79 (I79D, I79P, I79Q, I79G), L80 (I79D, I79P, I79Q, I79G), L81 (I79D, I79P, I79Q, I79G), L82 (I79D, I79P, I79Q, I79G), L83 (I79D, I79P, I79Q, I79G), L84 (I79D, I79P, I79Q, I79G), L85 (I79D, I79P, I79Q, I79G), L86 (I79D, I79D, I79Q, I79G), L87 (I79D, I79D, I79Q, I79G), L88 (I79D, I79D, I79Q, I79G), L89 (I79D, I79 6(S76P, S76G), L77(R77D), L78(V78R, V78D, V78E, V78P, V78K, V78G, V78Q), L85(V85G), L86(Y86T), L100(G100Q), H1(Q1E), H5(Q5V), H 11(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(S75T), 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 were considered as candidates for substitution in the 95 exemplified human mature light chain variable regions and the 29 exemplified human mature heavy chain variable regions, as further detailed 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 composite CDR-H1 has an amino acid sequence comprising SEQ ID NO:50. In some humanized 9F5 antibodies, the Kabat / Chothia composite CDR-H1 has an amino acid sequence comprising SEQ ID NO: 50, and the Kabat CDR-H2 has an amino acid sequence comprising SEQ ID NO: 51. In some humanized 9F5 antibodies, the Kabat CDR-H2 has an amino acid sequence comprising SEQ ID NO: 51. In some humanized 9F5 antibodies, the Kabat CDR-H2 has an amino acid sequence comprising SEQ ID NO: 52. In some humanized 9F5 antibodies, the Kabat CDR-L2 has an amino acid sequence comprising SEQ ID NO: 55. In some humanized 9F5 antibodies, the Kabat CDR-L1 has an amino acid sequence comprising SEQ ID NO: 53, and the Kabat CDR-L2 has an amino acid sequence comprising SEQ ID NO: 55. In some humanized 9F5 antibodies, the Kabat CDR-L1 has an amino acid sequence comprising SEQ ID NO: 54, and the Kabat CDR-L2 has an amino acid sequence comprising SEQ ID NO: 55. In some humanized 9F5 antibodies, the Kabat CDR-L1 has an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NOs: 172-193. In some humanized 9F5 antibodies, the Kabat CDR-L2 has an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NOs: 194-205.In some humanized 9F5 antibodies, the Kabat CDR-L3 has an amino acid sequence comprising a sequence selected from the group consisting of SEQ ID NOs: 206-213.

[0510] Here, as elsewhere, the first mentioned residue is the residue of a humanized antibody formed by grafting a Kabat CDR, or in the case of CDR-H1, a composite Chothia-Kabat CDR, onto a human acceptor framework, and the second mentioned residue is the residue proposed to replace that residue. Thus, within the variable region frameworks the first mentioned residue is human and within the CDRs the first mentioned residue is murine.

[0511] The exemplified antibodies include any permutation or combination of the exemplified mature heavy chain variable regions and mature light chain variable regions: hu9F5VHv1 / hu9F5VLv1, hu9F5VHv1 / hu9F5VLv2, hu9F5VHv1 / hu9F5VLv3, hu9F5VHv1 / hu9F5VLv4, hu9F5VLv1 / hu9F5VLv5, hu9F5VHv1 / hu9F5VLv6, hu9F5VHv1 / hu9F5VLv7, hu9F5VHv2 / hu9F5VLv1, hu9F5VHv2 / hu9F5VLv2, hu9F5VHv2 ...2 / hu9F5VLv4, hu9F5VL Lv3, hu9F5VHv2 / hu9F5VLv4, hu9F5VLv2 / hu9F5VLv5, hu9F5VHv2 / hu9F5VLv6, hu9F5VHv2 / hu9F5VLv7, hu9F5VHv3 / hu9F5VLv1, hu9F5VHv3 / hu9F5VLv2, hu9F5VHv3 / hu9F5VLv3, hu9F5VHv3 / hu9F5VLv4, hu9F5VLv3 / hu9F5VLv5, hu9F5VHv3 / hu9F5VLv6, hu9F5VHv3 / hu9F5VLv7, hu9F5VHv4 / hu9F5VLv1, hu9F 5VHv4 / hu9F5VLv2, hu9F5VHv4 / hu9F5VLv3, hu9F5VHv4 / hu9F5VLv4, hu9F5VLv4 / hu9F5VLv5, hu9F5VHv4 / hu9F5VLv6, hu9F5VHv4 / hu9F5VLv7, hu9F5VHv 5 / hu9F5VLv1, hu9F5VHv5 / hu9F5VLv2, hu9F5VHv5 / hu9F5VLv3, hu9F5VHv5 / hu9F5VLv4, hu9F5VLv5 / hu9F5VLv5, hu9F5VHv5 / hu9F5VLv6, hu9F5VHv5 / hu 9F5VLv7, hu9F5VHv6 / hu9F5VLv1, hu9F5VHv6 / hu9F5VLv2, hu9F5VHv6 / hu9F5VLv3, hu9F5VHv6 / hu9F5VLv4, hu9F5VHv6 / hu9F5VLv5, hu9F5VHv6 / hu9F5V Lv6, hu9F5VHv6 / hu9F5VLv7, hu9F5VHv7 / hu9F5VLv1, hu9F5VHv7 / hu9F5VLv2, hu9F5VHv7 / hu9F5VLv3, hu9F5VHv7 / hu9F5VLv4, hu9F5VLv7 / hu9F5VLv5,Includes hu9F5VHv7 / hu9F5VLv6, hu9F5VHv7 / hu9F5VLv7, hu9F5VHv8 / hu9F5VLv1, hu9F5VHv8 / hu9F5VLv2, hu9F5VHv8 / hu9F5VLv3, hu9F5VHv8 / hu9F5VLv4, hu9F5VLv8 / hu9F5VLv5, hu9F5VHv8 / hu9F5VLv6, and hu9F5VHv8 / hu9F5VLv7.

[0512] Exemplary antibodies include the 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: 29), hu9F5VLv8 (SEQ ID NO: 130), hu9F5VLv9 (SEQ ID NO: 131), hu9F5VLv2_M51E (SEQ ID NO: 61), hu9F5VLv2_M51D (SEQ ID NO: 62), hu9F5VLv2_L 27cD (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 ID NO: 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 (SEQ ID NO: 72), hu9F5VLv2_L33N (SEQ ID NO: 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 (SEQ ID NO: 78), hu9F5VLv2_L27bT (SEQ ID NO: 79), hu9F5VLv2_T31G (SEQ ID NO: 80), hu9F5VLv2_L27bQ (SEQ ID NO: 81), hu9F5VLv2_L33G (SEQ ID NO: 82), hu9F5VLv2_L27cP (SEQ ID NO: 83), hu9F5VL v2_V78R (SEQ ID NO: 84), hu9F5VLv2_I75D (SEQ ID NO: 85), hu9F5VLv2_V78D (SEQ ID NO: 86), hu9F5VLv2_V78E (SEQ ID NO: 87), hu9F5VLv2_V78P (SEQ ID NO: 88), hu9F5VLv2_V78K (SEQ ID NO: 89), hu9F5VLv2_R77D (SEQ ID 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 (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 (SEQ ID NO: 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 ID NO: 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 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), 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: 1 68), hu9F5VLv9_DIM13 (SEQ ID NO: 169), 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_DIM13 (SEQ ID NO: 169), hu9F5VLv9_DIM19 (SEQ ID NO: 170), and hu9F5VLv9_DIM20 (SEQ ID NO: 171). No. 22), hu9F5VHv9 (SEQ ID NO: 127), hu9F5VHv10 (SEQ ID NO: 128), hu9F5VHv10_L82cG (SEQ ID 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 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).

[0513] The 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 (hu9F5VLv8_V3Q, L27cD, L37G, M51G, L54R, L92I, also known as SEQ ID NO: 149). The 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_DIM11 (hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54R, L92I, also known as SEQ ID NO: 142). The 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 (hu9F5VLv8_V3Q, L27cS, M51G, L54R, L92I, also known as SEQ ID NO: 159). The 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_DIM17 (hu9F5VLv8_V3Q, L27cS, L37G, M51G, L54T, L92I, also known as SEQ ID NO: 148).

[0514] The 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 (hu9F5VLv8_V3Q, L27cD, L37Q, M51G, L54R, L92I, also known as SEQ ID NO: 137). The 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_DIM14 (hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54T, L92I, also known as SEQ ID NO: 145). The 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 (hu9F5VLv8_V3Q, L27cG, L37Q, M51G, L54R, L92I, also known as SEQ ID NO: 136). The 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_DIM7 (hu9F5VLv8_V3Q, L27cD, L37Q, M51K, L54R, L92I, also known as SEQ ID NO: 138).

[0515] The 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 (hu9F5VLv8_V3Q, L37Q, M51G, L54R, L92I, also known as SEQ ID NO: 158). The 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_DIM12 (hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54R, L92G, also known as SEQ ID NO: 143). The 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 (hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54R, also known as SEQ ID NO: 144). The 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_DIM2 (hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54R, L92I, also known as SEQ ID NO: 133).

[0516] The 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 (hu9F5VLv8_V3Q, L27cS, L37Q, L54R, L92I, also known as SEQ ID NO: 160). The 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_DIM30 (hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L92I, also known as 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).

[0517] 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 (hu9F5VLv9_V3Q, L27cG, L37G, M51G, L54R, L92I, also known as SEQ ID NO: 168).

[0518] The present invention relates to humanized mature heavy chain variable regions, including 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: 130), hu9F5VHv5_L82cG (SEQ ID NO: 131), hu9F5VHv5_L82cG (SEQ ID NO: 132), hu9F5VHv5_L82cG (SEQ ID NO: 133), hu9F5VHv5_L82cG (SEQ ID NO: 134), hu9F5VHv5_L82cG (SEQ ID NO: 135), hu9F5VHv5_L82cG (SEQ ID NO: 136), hu9F5VHv5_L82cG (SEQ ID NO: 137), hu9F5VHv5_L82cG (SEQ ID NO: 138), hu9F5VHv5_L82cG (SEQ ID NO: 139), hu9F5VHv5_L82cG (SEQ 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 ID NO: 1 19), 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), and the humanized mature light chain variable region is selected from the group consisting of hu9F5VLv1 (SEQ ID NO: 127), 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). 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: 29), hu9F5VLv8 (SEQ ID NO: 130), hu9F5VLv9 (SEQ ID NO: 131), hu9F5VLv2_M51E (SEQ ID NO: 61), hu9F5VLv2_M51D (SEQ ID NO: 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 ID NO: 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 (SEQ ID NO: 72), hu9F5VLv2_L33N (SEQ ID NO: 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 (SEQ ID NO: 78), hu9F5VLv2_L27bT (SEQ ID NO: 79), hu9F5VLv2_T31G (SEQ ID NO: 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 ID NO: 87), hu9F5VLv2_V78P (SEQ ID NO: 88), hu9F5VLv2_V78K (SEQ ID NO: 89), hu9F5VLv2_R77D (SEQ ID 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 (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 (SEQ ID NO: 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 ID NO: 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 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), hu9F5V Lv8_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), hu9 F5VLv8_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),Variants of the 9F5 humanized antibody are provided that exhibit at least 90%, 95%, 96%, 97%, 98%, or 99% identity to hu9F5VLv9_DIM19 (SEQ ID NO: 170), or hu9F5VLv9_DIM20 (SEQ ID NO: 171). In some such antibodies, at 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 are subject to back mutation or other mutation in SEQ ID NOs: 15-22, 109-129, SEQ ID NOs: 23-29, SEQ ID NOs: 61-108, and SEQ ID NOs: 130-171 have likewise been back mutated or otherwise mutated. ,

[0519] In some humanized 9F5 antibodies, at least one of the following positions in the VH region is occupied by the indicated amino acid: H1 is occupied by E, H7 is occupied by T, H20 is occupied by I, H69 is occupied by M, H75 is occupied by T, H93 is occupied by T, H94 is occupied by T, and H109 is occupied 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.

[0520] In some humanized 9F5 antibodies, at least one of the following positions in the VH region is occupied by the indicated 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.

[0521] In some humanized 9F5 antibodies, at least one of the following positions in the VH region is occupied by the indicated 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.

[0522] In some humanized 9F5 antibodies, at least one of the following positions in the VH region is occupied by the indicated amino acid: H43 is occupied by K, H51 is occupied by V, H76 is occupied by D, M80 is occupied by M, and H108 is occupied by L. In some humanized 9F5 antibodies, positions H43, H51, H76, H80, and H108 are occupied by K, V, D, M, and L, respectively.

[0523] In some humanized 9F5 antibodies, position H28 of the VH region is occupied by a T.

[0524] In some humanized 9F5 antibodies, at least one of the following positions in the VH region is occupied by the indicated 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.

[0525] In some humanized 9F5 antibodies, position H40 of 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 the indicated 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.

[0526] In some humanized 9F5 antibodies, positions H5, H11, H12, H38, and H42 are occupied by V, V, K, R, and G, respectively.

[0527] In some humanized 9F5 antibodies, at least one of the following positions in the VH region is occupied by the indicated 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.

[0528] In some humanized 9F5 antibodies, such as hu9F5VHv2, 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. In some humanized 9F5 antibodies, such as hu9F5VHv3, 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. In some humanized 9F5 antibodies, such as hu9F5VHv4, 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. In some humanized 9F5 antibodies, such as hu9F5VHv5, 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 humanized 9F5 antibodies, such as hu9F5VHv6, 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 humanized 9F5 antibodies, such as hu9F5VHv7, 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 humanized 9F5 antibodies, such as hu9F5VHv8, 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. In some humanized 9F5 antibodies, such as hu9F5VHv9, positions H1, H5, H11, H12, H17, H20, H23, H38, H42, H43, H66, H69, H75, H80, H81, H83, H93, H94, H108, and H109 of the VH region are occupied, respectively, by E, V, V, K, T, I, K, Q, G, K, R, M, T, M, E, R, T, T, L, and V. In some humanized 9F5 antibodies, the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:127.

[0529] 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 of 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, such as in the case of hu9F5VHv10. In some humanized 9F5 antibodies, the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:128.

[0530] 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, such as in hu9F5VHv10_L82cG.

[0531] In some humanized 9F5 antibodies, at least one of the following positions in the VL region is occupied by the indicated amino acid: L7 is occupied by S, L8 is occupied by P, L15 is occupied by P, and L100 is occupied by Q. In some humanized 9F5 antibodies, positions L7, L8, L15, and L100 are occupied by S, P, P, and Q, respectively.

[0532] In some humanized 9F5 antibodies, position L66 of the VL region is occupied by G. In some humanized 9F5 antibodies, position L64 of the VL region is occupied by S.

[0533] In some humanized 9F5 antibodies, position L17 of the VL region is occupied by E.

[0534] In some humanized 9F5 antibodies, at least one of the following positions in the VL region is occupied by the indicated 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.

[0535] In some humanized 9F5 antibodies, position L30 of the VL region is occupied by Y.

[0536] In some humanized 9F5 antibodies, at least one of the following positions in the VL region is occupied by the indicated amino acid: L3 is V or Q, L7 is A or S, L8 is A or P, L9 is F or L, L11 is N or L, L15 is L or P, L17 is T or E, L18 is S or P, L27b is L, D, T, or Q, L27c is L, D, G, S, E, T, N, A, P, or I, L30 is I, Y, E, K, G, or Q, L31 is T, N, or G, L33 is L, N, T, S, R, or G, L37 is L, Q, G, or I, and L39 is R. or K, L51 is M, G, E, D, K, or I, L54 is L, R, G, or T, L60 is N or D, L64 is G or S, L66 is E or G, L73 is L, P, or G, L74 is R or K, L75 is I, D, P, Q, or G, L76 is S, P, or G, L77 is SEQ ID NO: 146 or D, L78 is V, R, D, E, P, K, G, or Q, L85 is V or G, L86 is Y or T, L89 is A or G, L92 is L, D, E, G, Q, T, or I, L93 is E or G, and L100 is G or Q.

[0537] In some humanized 9F5 antibodies, such as in hu9F5VLv1, positions L64 and L66 in the VL region are occupied by S and G, respectively. In some humanized 9F5 antibodies, such as in hu9F5VLv2, 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 humanized 9F5 antibodies, such as in hu9F5VLv3, positions L7, L8, L15, L17, L66, and L100 in the VL region are occupied by S, P, P, E, G, and Q, respectively, such as in hu9F5VLv3.

[0538] In some humanized 9F5 antibodies, such as hu9F5VLv4, positions L7, L8, L11, L15, L17, L51, L54, L66, and L100 of the VL region are occupied by S, P, L, P, E, G, R, G, and Q, respectively. In some humanized 9F5 antibodies, the light chain variable region comprises any of the amino acid sequences of SEQ ID NOs: 133, 135-137, 142-144, 149, 158, 159, and 168. In some humanized 9F5 antibodies, the light chain variable region comprises the amino acid sequence of SEQ ID NO: 133. In some humanized 9F5 antibodies, the light chain variable region comprises the amino acid sequence of SEQ ID NO: 137. In some humanized 9F5 antibodies, the light chain variable region comprises the amino acid sequence of SEQ ID NO: 149. In some humanized 9F5 antibodies, the light chain variable region comprises the amino acid sequence of SEQ ID NO: 159.

[0539] In some humanized 9F5 antibodies, such as hu9F5VLv5, 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 humanized 9F5 antibodies, such as hu9F5VLv6, 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 humanized 9F5 antibodies, such as hu9F5VLv7, positions L7, L8, L9, L11, 15, 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.

[0540] In some humanized 9F5 antibodies, such as hu9F5VLv8, positions L7, L8, L11, L15, L17, L39, L64, L66, L74, and L100 of the VL region are occupied by S, P, L, P, E, K, S, G, K, and Q, respectively. In some humanized 9F5 antibodies, position L3 of the VL region is occupied by Q. In some humanized 9F5 antibodies, position L27c of the VL region is occupied by D, G, I, L, or S, position L37 of the VL region is occupied by G, I, L, or Q, position L51 of the VL region is occupied by E, G, I, K, or M, position L54 of the VL region is occupied by G, L, R, or T, and position L92 of 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.

[0541] In some humanized 9F5 antibodies, position L27c of the VL region is occupied by D, position L37 of the VL region is occupied by G, and position L51 of the VL region is occupied by G. In some humanized 9F5 antibodies, the heavy chain variable region has an amino acid sequence comprising SEQ ID NO: 127 and the light chain variable region has an amino acid sequence comprising SEQ ID NO: 149.

[0542] In some humanized 9F5 antibodies, position L27c of the VL region is occupied by D, position L37 of the VL region is occupied by Q, and position L51 of the VL region is occupied by G. In some humanized 9F5 antibodies, the heavy chain variable region has an amino acid sequence comprising SEQ ID NO:127, and the light chain variable region has an amino acid sequence comprising SEQ ID NO:137.

[0543] In some humanized 9F5 antibodies, position L27c of the VL region is occupied by S, position L37 of the VL region is occupied by L, and position L51 of the VL region is occupied by G. In some humanized 9F5 antibodies, the heavy chain variable region has an amino acid sequence comprising SEQ ID NO:127, and the light chain variable region has an amino acid sequence comprising SEQ ID NO:159.

[0544] In some humanized 9F5 antibodies, position L27c of the VL region is occupied by D, position L37 of the VL region is occupied by Q, and position L51 of the VL region is occupied by K. In some humanized 9F5 antibodies, the heavy chain variable region has an amino acid sequence comprising SEQ ID NO:127, and the light chain variable region has an amino acid sequence comprising SEQ ID NO:138.

[0545] In some humanized 9F5 antibodies, position L27c of the VL region is occupied by S, position L37 of the VL region is occupied by Q, and position L51 of the VL region is occupied by G. In some humanized 9F5 antibodies, the heavy chain variable region has an amino acid sequence comprising SEQ ID NO:127, and the light chain variable region has an amino acid sequence comprising SEQ ID NO:133.

[0546] In some humanized 9F5 antibodies, such as hu9F5VLv9, positions L7, L8, L11, L15, L17, L39, L60, L64, L66, L74, and L100 of the VL region are occupied by S, P, L, P, E, K, D, S, G, K, and Q, respectively. In some humanized 9F5 antibodies, position L3 of the VL region is occupied by Q.

[0547] In some humanized 9F5 antibodies, position L27c of the VL region is occupied by G or S, position L37 of the VL region is occupied by G, I or Q, position L51 of the VL region is occupied by G, I or K, position L54 of the VL region is occupied by Q or R, and position L92 of the VL region is occupied by G, I or L. In some humanized 9F5 antibodies, position L27c of the VL region is occupied by G, position L37 of the VL region is occupied by G, position L51 of the VL region is occupied by G, and position L54 of the VL region is occupied by R. In some humanized 9F5 antibodies, position L92 of the VL region is occupied by I. In some humanized 9F5 antibodies, the heavy chain variable region has an amino acid sequence comprising SEQ ID NO: 129 and the light chain variable region has an amino acid sequence comprising SEQ ID NO: 168.

[0548] The light chain variable region of any of the above-mentioned antibodies may be modified to further reduce immunogenicity. For example, in some humanized antibodies, position L27b of the VL region is occupied by D, T, or Q; position L27c of the VL region is occupied by D, G, S, E, T, N, A, I, or P; position L30 of the VL region is occupied by E, K, G, or Q; position L31 of the VL region is occupied by G; position L33 of the VL region is occupied by N, T, S, R, or G; position L37 of the VL region is occupied by Q, G, or I; position L51 of the VL region is occupied by E, D, G, K, or I; position L54 of the VL region is occupied by G, R, or T; and position L60 of the VL region is occupied by D. position L73 in the VL region is occupied by P or G; position L75 in the VL region is occupied by D, P, Q, or G; position L76 in the VL region is occupied by P or G; position L77 in the VL region is occupied by D; position L78 in the VL region is occupied by R, D, E, P, K, G, or Q; position L85 in the VL region is occupied by G; position L86 in the VL region is occupied by T; position L89 in the VL region is occupied by G; position L92 in the VL region is occupied by D, E, G, Q, I, or T; and / or position L93 in the VL region is occupied by G (Kabat numbering).

[0549] In some humanized 9F5 antibodies, position L51 in the VL region is occupied by E, such as in the case of hu9F5VLv2_M51E. In some humanized 9F5 antibodies, position L51 in the VL region is occupied by D, such as in the case of hu9F5VLv2_M51D. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by D, such as in the case of hu9F5VLv2_L27cD. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by G, such as in the case of hu9F5VLv2_L27cG. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by S, such as in the case of hu9F5VLv2_L27cS. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by E, such as in the case of hu9F5VLv2_L27cE. In some humanized 9F5 antibodies, position L30 in the VL region is occupied by E, such as in hu9F5VLv2_I30E. In some humanized 9F5 antibodies, position L30 in the VL region is occupied by K, such as in hu9F5VLv2_I30K. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by T, such as in hu9F5VLv2_L27cT. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by N, such as in hu9F5VLv2_L27cN.

[0550] In some humanized 9F5 antibodies, position L27b in the VL region is occupied by D, such as in hu9F5VLv2_L27bD. In some humanized 9F5 antibodies, position L30 in the VL region is occupied by G, such as in hu9F5VLv2_I30G. In some humanized 9F5 antibodies, position L33 in the VL region is occupied by N, such as in hu9F5VLv2_L33N. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by A, such as in hu9F5VLv2_L27cA. In some humanized 9F5 antibodies, position L33 in the VL region is occupied by T, such as in hu9F5VLv2_L33T. In some humanized 9F5 antibodies, position L33 in the VL region is occupied by S, such as in hu9F5VLv2_L33S. In some humanized 9F5 antibodies, position L33 in the VL region is occupied by R, such as in hu9F5VLv2_L33R. In some humanized 9F5 antibodies, position L30 in the VL region is occupied by Q, such as in hu9F5VLv2_I30Q. In some humanized 9F5 antibodies, position L27b in the VL region is occupied by T, such as in hu9F5VLv2_L27bT. In some humanized 9F5 antibodies, position L31 in the VL region is occupied by G, such as in hu9F5VLv2_T31G.

[0551] In some humanized 9F5 antibodies, position L27b in the VL region is occupied by Q, such as in hu9F5VLv2_L27bQ. In some humanized 9F5 antibodies, position L33 in the VL region is occupied by G, such as in hu9F5VLv2_L33G. In some humanized 9F5 antibodies, position L27c in the VL region is occupied by P, such as in hu9F5VLv2_L27cP. In some humanized 9F5 antibodies, position L78 in the VL region is occupied by R, such as in hu9F5VLv2_V78R. In some humanized 9F5 antibodies, position L75 in the VL region is occupied by D, such as in hu9F5VLv2_I75D. In some humanized 9F5 antibodies, position L78 in the VL region is occupied by D, such as in hu9F5VLv2_V78D. In some humanized 9F5 antibodies, position L78 in the VL region is occupied by E, such as in hu9F5VLv2_V78E. In some humanized 9F5 antibodies, position L78 in the VL region is occupied by P, such as in hu9F5VLv2_V78P. In some humanized 9F5 antibodies, position L78 in the VL region is occupied by K, such as in hu9F5VLv2_V78K. In some humanized 9F5 antibodies, position L77 in the VL region is occupied by D, such as in hu9F5VLv2_R77D.

[0552] In some humanized 9F5 antibodies, position L78 in the L region is occupied by G, such as in hu9F5VLv2_V78G. In some humanized 9F5 antibodies, position L76 in the VL region is occupied by P, such as in hu9F5VLv2_S76P. In some humanized 9F5 antibodies, position L75 in the VL region is occupied by P, such as in hu9F5VLv2_I75P. In some humanized 9F5 antibodies, position L75 in the VL region is occupied by Q, such as in hu9F5VLv2_I75Q. In some humanized 9F5 antibodies, position L75 in the VL region is occupied by G, such as in hu9F5VLv2_I75G. In some humanized 9F5 antibodies, position L73 in the VL region is occupied by P, such as in hu9F5VLv2_L73P. In some humanized 9F5 antibodies, position L73 in the VL region is occupied by G, such as in hu9F5VLv2_L73G. In some humanized 9F5 antibodies, position L78 in the VL region is occupied by Q, such as in hu9F5VLv2_V78Q. In some humanized 9F5 antibodies, position L76 in the VL region is occupied by G, such as in hu9F5VLv2_S76G. In some humanized 9F5 antibodies, position L92 in the VL region is occupied by D, such as in hu9F5VLv2_L92D.

[0553] In some humanized 9F5 antibodies, position L86 in the VL region is occupied by T, such as in hu9F5VLv2_Y86T. In some humanized 9F5 antibodies, position L92 in the VL region is occupied by E, such as in hu9F5VLv2_L92E. In some humanized 9F5 antibodies, position L92 in the VL region is occupied by G, such as in hu9F5VLv2_L92G. In some humanized 9F5 antibodies, position L92 in the VL region is occupied by Q, such as in hu9F5VLv2_L92Q. In some humanized 9F5 antibodies, position L93 in the VL region is occupied by G, such as in hu9F5VLv2_L93G. In some humanized 9F5 antibodies, position L85 in the VL region is occupied by G, such as in hu9F5VLv2_V85G. In some humanized 9F5 antibodies, position L92 in the VL region is occupied by a T, such as in hu9F5VLv2_L92T. In some humanized 9F5 antibodies, position L89 in the VL region is occupied by a G, such as in hu9F5VLv2_A89G.

[0554] In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54G, L92I, also known as hu9F5VLv8_DIM1, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, G, G, and I, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54R, L92I, also known as hu9F5VLv8_DIM2, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, G, R, and I. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54T, L92I, also known as hu9F5VLv8_DIM3, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, G, T, and I, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L54R, L92G, also known as hu9F5VLv8_DIM4, 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 humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cG, L37Q, M51G, L54R, L92I, also known as hu9F5VLv8_DIM5, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, Q, G, R, and I, respectively.

[0555] In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cD, L37Q, M51G, L54R, L92I, also known as hu9F5VLv8_DIM6, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, D, Q, G, R, and I, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cD, L37Q, M51K, L54R, L92I, also known as hu9F5VLv8_DIM7, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, D, Q, K, R, and I, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cG, L37Q, M51K, L54R, L92I, also known as hu9F5VLv8_DIM8, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, Q, K, R, and I, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cG, L37Q, M51K, L54G, L92I, also known as hu9F5VLv8_DIM9, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, Q, K, G, and I, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cS, L37Q, M51K, L54G, L92I, also known as hu9F5VLv8_DIM10, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, K, G, and I, respectively.

[0556] In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54R, L92I, also known as hu9F5VLv8_DIM11, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, G, G, R, and I, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54R, L92G, also known as hu9F5VLv8_DIM12, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, G, G, R, and G, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54R, also known as hu9F5VLv8_DIM13, positions L3, L27c, L37, L51, and L54 in the VL region are occupied by Q, G, G, G, and R, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54T, L92I, also known as hu9F5VLv8_DIM14, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, G, G, T, and I. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54T, L92G, also known as hu9F5VLv8_DIM15, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, G, G, T, and G, respectively.

[0557] In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cG, L37G, M51G, L54T, also known as hu9F5VLv8_DIM16, positions L3, L27c, L37, L51, and L54 in the VL region are occupied by Q, G, G, G, and T, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cS, L37G, M51G, L54T, L92I, also known as hu9F5VLv8_DIM17, 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 humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cD, L37G, M51G, L54R, L92I, also known as hu9F5VLv8_DIM18, 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 humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cS, L37I, M51I, L54R, L92I, also known as hu9F5VLv8_DIM19, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, I, I, R, and I, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cS, L37Q, M51I, L54G, L92I, also known as hu9F5VLv8_DIM20, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, I, G, and I, respectively.

[0558] In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cS, L37Q, M51I, L54G, also known as hu9F5VLv8_DIM21, positions L3, L27c, L37, L51, and L54 in the VL region are occupied by Q, S, Q, I, and G, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cS, L37Q, M51E, L54R, L92I, also known as hu9F5VLv8_DIM22, 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 humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cG, L37Q, M51E, L54G, L92I, also known as hu9F5VLv8_DIM23, positions L3, L27c, L37, L51, L54, and L92, respectively, in the VL region are occupied by Q, G, Q, E, G, and I. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cG, L37I, M51E, L54R, L92I, also known as hu9F5VLv8_DIM24, positions L3, L27c, L37, L51, L54, and L92L, respectively, in the VL region are occupied by Q, G, I, E, R, and I. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cG, L37I, M51E, L54R, L92G, also known as hu9F5VLv8_DIM25, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, I, E, R, and G, respectively.

[0559] In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cI, L37I, M51E, L54R, also known as hu9F5VLv8_DIM26, positions L3, L27c, L37, L51, and L54 in the VL region are occupied by Q, I, I, E, and R, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L37Q, M51G, L54R, L92I, also known as hu9F5VLv8_DIM27, positions L3, L37, L51, L54, and L92 in the VL region are occupied by Q, Q, G, R, and I, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cS, M51G, L54R, L92I, also known as hu9F5VLv8_DIM28, positions L3, L27c, L51, L54, and L92 in the VL region are occupied by Q, S, G, R, and I, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cS, L37Q, L54R, L92I, also known as hu9F5VLv8_DIM29, positions L3, L27c, L37, L54, and L92 in the VL region are occupied by Q, S, Q, R, and I, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv8_V3Q, L27cS, L37Q, M51G, L92I, also known as hu9F5VLv8_DIM30, positions L3, L27c, L37, L51, and L92 in the VL region are occupied by Q, S, Q, G, and I, respectively.

[0560] In some humanized 9F5 antibodies, such as hu9F5VLv9_V3Q, L27cS, L37Q, M51G, L54G, L92I, also known as hu9F5VLv9_DIM1, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, G, G, and I, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv9_V3Q, L27cS, L37Q, M51G, L54R, L92I, also known as hu9F5VLv9_DIM2, 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 humanized 9F5 antibodies, such as hu9F5VLv9_V3Q, L27cS, L37Q, M51G, L54R, L92G, also known as hu9F5VLv9_DIM4, 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 humanized 9F5 antibodies, such as hu9F5VLv9_V3Q, L27cG, L37Q, M51G, L54R, L92I, also known as hu9F5VLv9_DIM5, 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 humanized 9F5 antibodies, such as hu9F5VLv9_V3Q, L27cG, L37Q, M51K, L54R, L92I, also known as hu9F5VLv9_DIM8, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, Q, K, R, and I, respectively.

[0561] In some humanized 9F5 antibodies, such as hu9F5VLv9_V3Q, L27cS, L37Q, M51K, L54G, L92I, also known as hu9F5VLv9_DIM10, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, K, G, and I, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv9_V3Q, L27cG, L37G, M51G, L54R, L92I, also known as hu9F5VLv9_DIM11, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, G, G, G, R, and I, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv9_V3Q, L27cG, L37G, M51G, L54R, also known as hu9F5VLv9_DIM13, positions L3, L27c, L37, L51, and L54 in the VL region are occupied by Q, G, G, G, and R, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv9_V3Q, L27cS, L37I, M51I, L54R, L92I, also known as hu9F5VLv9_DIM19, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, I, I, R, and I, respectively. In some humanized 9F5 antibodies, such as hu9F5VLv9_V3Q, L27cS, L37Q, M51I, L54G, L92I, also known as hu9F5VLv9_DIM20, positions L3, L27c, L37, L51, L54, and L92 in the VL region are occupied by Q, S, Q, I, G, and I, respectively.

[0562] The heavy chain variable region of any of the above-mentioned antibodies may also 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.

[0563] In some humanized 9F5 antibodies, position H80 is occupied by P, such as in hu9F5VHv4_L80P. In some humanized 9F5 antibodies, position H80 is occupied by D, such as in hu9F5VHv4_L80D. In some humanized 9F5 antibodies, position H82c is occupied by G, such as in hu9F5VHv4_L82cG and hu9F5VHv10_L82cG. In some humanized 9F5 antibodies, position H82c is occupied by D, such as in hu9F5VHv4_L82cD. In some humanized 9F5 antibodies, position H82 is occupied by P, such as in hu9F5VHv4_L82P. In some humanized 9F5 antibodies, position H80 is occupied by G, such as in hu9F5VHv4_L80G. In some humanized 9F5 antibodies, position H82 is occupied by K, such as in hu9F5VHv4_L82K. In some humanized 9F5 antibodies, position H82 is occupied by R, such as in hu9F5VHv4_L82R. In some humanized 9F5 antibodies, position H82 is occupied by E, such as in hu9F5VHv4_L82E. In some humanized 9F5 antibodies, position H82 is occupied by N, such as in hu9F5VHv4_L82N.

[0564] In some humanized 9F5 antibodies, position H79 is occupied by D, such as in hu9F5VHv4_Y79D. In some humanized 9F5 antibodies, position H79 is occupied by N, such as in hu9F5VHv4_Y79N. In some humanized 9F5 antibodies, position H79 is occupied by G, such as in hu9F5VHv4_Y79G. ​​In some humanized 9F5 antibodies, position H80 is occupied by E, such as in hu9F5VHv5_M80E. In some humanized 9F5 antibodies, position H80 is occupied by G, such as in hu9F5VHv5_M80G. In some humanized 9F5 antibodies, position H82c is occupied by S, such 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.

[0565] In some humanized 9F5 antibodies, the variable heavy chain has 85% or more identity to the human sequence. In some humanized 9F5 antibodies, the variable light chain has 85% or more identity to the human sequence. In some humanized 9F5 antibodies, the variable heavy chain and the variable light chain each have 85% or more identity to the human germline sequence. In some humanized 9F5 antibodies, the three heavy chain CDRs are as defined by the Kabat / Chothia combination (SEQ ID NOs: 8, 9, and 10, respectively) and the three light chain CDRs are as defined by the Kabat / Chothia combination (SEQ ID NOs: 12, 13, and 14); except that 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, and position L27c 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.

[0566] In some humanized 9F5 antibodies, the Kabat / Chothia composite CDR-H1 has an amino acid sequence comprising SEQ ID NO: 50. In some humanized 9F5 antibodies, the Kabat CDR-H2 has an amino acid sequence comprising SEQ ID NO: 51 or SEQ ID NO: 52. In some humanized 9F5 antibodies, the Kabat CDR-L1 has an amino acid sequence comprising SEQ ID NO: 53 or SEQ ID NO: 54. In some humanized 9F5 antibodies, the Kabat CDR-L2 has an amino acid sequence comprising SEQ ID NO: 55. In some humanized 9F5 antibodies, the Kabat CDR-L1 has an amino acid sequence selected from the group consisting of SEQ ID NOs: 172 to 193. In some humanized 9F5 antibodies, the Kabat CDR-L2 has an amino acid sequence selected from the group consisting of SEQ ID NOs: 194 to 205. In some humanized 9F5 antibodies, the Kabat CDR-L3 has an amino acid sequence selected from the group consisting of SEQ ID NOs: 206 to 213.

[0567] Exemplary humanized antibodies include a humanized form of mouse 10C12 designated Hu10C12.

[0568] Mouse antibody 10C12 includes a mature heavy chain variable region and a mature light chain variable region having amino acid sequences comprising SEQ ID NO: 7 and SEQ ID NO: 11, respectively. The present invention provides two exemplified humanized mature heavy chain variable regions: hu10C12VHv1 and hu10C12VHv2. Further, the present invention provides two exemplified mature light chain variable regions hu10C12VLv1 and hu10C12VLv2. Figures 7 and 8 show the alignment of the respective heavy chain variable regions and light chain variable regions of mouse 10C12 and various humanized antibodies.

[0569] For reasons such as possible effects on CDR conformation and / or binding to antigen, interactions between heavy and light chains, mediating interactions with the constant region, being sites of desirable or undesirable post-translational modifications, being rare residues at that position in human variable region sequences and therefore potentially immunogenic, reducing the likelihood of aggregation, and other reasons, the following nine variable region framework positions were considered as candidates for substitution in the two exemplified human mature light chain variable regions and the two exemplified human mature light chain variable regions, as further detailed in the Examples: L64 (G64S), L104 (V104L), H1 (Q1E), H24 (V24A), H48 (M48I), H67 (V67A), H69 (I69M), H93 (A93T), and H94 (R94T).

[0570] Here, as elsewhere, the first mentioned residue is the residue of a humanized antibody formed by grafting a Kabat CDR, or in the case of CDR-H1, a composite Chothia-Kabat CDR, onto a human acceptor framework, and the second mentioned residue is the residue proposed to replace that residue. Thus, within the variable region frameworks the first mentioned residue is human and within the CDRs the first mentioned residue is murine.

[0571] The exemplified antibodies include any permutation or combination of the exemplified mature heavy chain variable regions and mature light chain variable regions: hu10C12VHv1 / hu10C12VLv1, hu10C12VHv1 / hu10C12VLv2, hu10C12VHv2 / hu10C12VLv1, hu10C12VHv2 / hu10C12VLv2.

[0572] The exemplified antibodies include any of the exemplified mature light chain variable regions hu10C12VLv1 (SEQ ID NO: 216) and hu10C12VLv2 (SEQ ID NO: 217) permutations or combinations of any of the exemplified mature heavy chain variable regions hu10C12VHv1 (SEQ ID NO: 214) and hu10C12VHv2 (SEQ ID NO: 215).

[0573] The invention provides variants of the 10C12 humanized antibody in which the humanized mature heavy chain variable region exhibits at least 90%, 95%, 96%, 97%, 98%, or 99% identity to 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 to hu10C12VLv1 (SEQ ID NO:216) or hu10C12VLv2 (SEQ ID NO:217). Some such antibodies retain at least one, two, three, four, five, six, seven, eight, or all nine of the back mutations or other mutations in SEQ ID NOs:214-215 and SEQ ID NOs:216-217.

[0574] In some humanized 10C12 antibodies, at least one of the following positions in the VH region is occupied by the indicated amino acid: 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. In some humanized 10C12 antibodies, positions H24, H48, H67, H69, H93, and H94 are occupied by A, I, A, M, T, and T, respectively.

[0575] In some humanized 10C12 antibodies, at least one of the following positions in the VH region is occupied by the indicated 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.

[0576] In some humanized 10C12 antibodies, such as in hu10C12VHv1, positions H24, H48, H67, H69, H93, and H94 are occupied by A, I, A, M, T, and T, respectively. In some humanized 10C12 antibodies, such as in hu10C12VHv2, positions H1, H24, H48, H67, H69, H93, and H94 are occupied by E, A, I, A, M, T, and T, respectively.

[0577] In some humanized 10C12 antibodies, position L64 in the VL region is occupied by S.

[0578] In some humanized 10C12 antibodies, at least one of the following positions in the VL region is occupied by the indicated amino acid: L64 is S and L104 is V or L.

[0579] In some humanized 10C12 antibodies, positions L64 and L104 in the VL region are occupied by S, such as in hu10C12VLv1. In some humanized 10C12 antibodies, positions L64 and L104 in the VL region are occupied by S and L, respectively, such as in hu10C12VLv2.

[0580] In some humanized 10C12 antibodies, the variable heavy chain has 85% or more identity to the human sequence. In some humanized 10C12 antibodies, the variable light chain has 85% or more identity to the human sequence. In some humanized 10C12 antibodies, the variable heavy chain and the variable light chain each have 85% or more identity to the human germline sequence. In some humanized 10C12 antibodies, the three heavy chain CDRs are as defined by the Kabat / Chothia combination (SEQ ID NOs: 8, 9, and 10) and the three light chain CDRs are as defined by the Kabat / Chothia combination (SEQ ID NOs: 12, 13, and 14).

[0581] Exemplary humanized antibodies include a humanized form of the murine 12C4 antibody, designated Hu12C4.

[0582] Murine antibody 12C4 comprises a mature heavy chain variable region and a mature light chain variable region having amino acid sequences comprising SEQ ID NO:219 and SEQ ID NO:11, respectively. The present invention provides two exemplary humanized mature heavy chain variable regions: hu12C4VHv1 and hu12C4VHv2. The present invention further provides two exemplary mature light chain variable regions: hu12C4VLv1 and hu12C4VLv2. Figures 9 and 10 show alignments of the heavy and light chain variable regions of murine 12C4 and various humanized antibodies, respectively.

[0583] For reasons such as possible effects on CDR conformation and / or binding to antigen, interactions between heavy and light chains, mediating interactions with the constant region, being sites of desirable or undesirable post-translational modifications, being rare residues at that position in human variable region sequences and therefore potentially immunogenic, gaining potential for aggregation, and other reasons, the following six variable region framework positions were considered as candidates for substitution in the two exemplified human mature light chain variable regions and the two exemplified human mature light chain variable regions, as further detailed in the Examples: L64 (G64S), L104 (V104L), H1 (Q1E), H48 (M48I), H93 (A93T), and H94 (R94T).

[0584] Here, as elsewhere, the first mentioned residue is the residue of a humanized antibody formed by grafting a Kabat CDR, or in the case of CDR-H1, a composite Chothia-Kabat CDR, onto a human acceptor framework, and the second mentioned residue is the residue proposed to replace that residue. Thus, within the variable region frameworks the first mentioned residue is human and within the CDRs the first mentioned residue is murine.

[0585] The exemplified antibodies include any permutation or combination of the exemplified mature heavy chain variable regions and mature light chain variable regions: hu12C4VHv1 / hu12C4VLv1, hu12C4VHv1 / hu12C4VLv2, hu12C4VHv2 / hu12C4VLv1, hu12C4VHv2 / hu12C4VLv2.

[0586] The exemplified antibodies include any permutation or combination of the exemplified mature light chain variable regions hu12C4VLv1 (SEQ ID NO: 223) and hu12C4VLv2 (SEQ ID NO: 224) with any of the exemplified mature heavy chain variable regions hu12C4VHv1 (SEQ ID NO: 221) and hu12C4VHv2 (SEQ ID NO: 222).

[0587] The invention provides variants of the 12C4 humanized antibody in which the humanized mature heavy chain variable region exhibits at least 90%, 95%, 96%, 97%, 98%, or 99% identity to 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 to hu12C4VLv1 (SEQ ID NO:223) or hu12C4VLv2 (SEQ ID NO:224). Some such antibodies retain at least one, two, three, four, five, or all six of the back mutations or other mutations in SEQ ID NOs:221-222 and SEQ ID NOs:223-224.

[0588] In some humanized 12C4 antibodies, at least one of the following positions in the VH region is occupied by the indicated 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.

[0589] In some humanized 12C4 antibodies, such as hu12C4VHv2, positions H1, H48, H93, and H94 of the VH region are occupied by E, I, T, and T, respectively.

[0590] In some humanized 12C4 antibodies, at least one of the following positions in the VL region is occupied by the indicated amino acid: L64 is G or S, and L104 is V or L.

[0591] In some humanized 12C4 antibodies, such as hu12C4VLv2, positions L64 and L104 in the VL region are occupied by S and L, respectively.

[0592] In some humanized 12C4 antibodies, the variable heavy chain has 85% or more identity to human sequences. In some humanized 12C4 antibodies, the variable light chain has 85% or more identity to human sequences. In some humanized 12C4 antibodies, the variable heavy chain and the variable light chain each have 85% or more identity to human germline sequences. In some humanized 12C4 antibodies, the three heavy chain CDRs are as defined by the Kabat / Chothia composite (SEQ ID NOs: 8, 220, and 10) and the three light chain CDRs are as defined by the eKabat / Chothia composite (SEQ ID NOs: 12, 13, and 14).

[0593] An exemplary humanized antibody is a humanized form of murine 17C12, designated Hu17C12.

[0594] Murine antibody 17C12 comprises a mature heavy chain variable region and a mature light chain variable region having amino acid sequences comprising SEQ ID NOs: 225 and 228, respectively. The present invention provides two exemplary humanized mature heavy chain variable regions: hu17C12VHv and hu17C12VHv2. The present invention further provides two exemplary mature light chain variable regions: hu17C12VLv1 and hu17C12VLv2. Figures 11 and 12 show the alignment of the heavy and light chain variable regions of murine 17C12 and various humanized antibodies, respectively.

[0595] The following thirteen variable region framework positions are further illustrated in the Examples for reasons such as possible effects on CDR conformation and / or binding to antigen, interactions between heavy and light chains, mediating interactions with the constant region, being sites of desirable or undesirable post-translational modifications, being rare residues at their positions in human variable region sequences and therefore potentially immunogenic, resulting in potential for aggregation, and other reasons: As detailed, two exemplified human mature light chain variable regions and two exemplified human mature light chain variable regions were considered as candidates for substitutions: 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).

[0596] Here, as elsewhere, the first mentioned residue is the residue of a humanized antibody formed by grafting a Kabat CDR, or in the case of CDR-H1, a composite Chothia-Kabat CDR, onto a human acceptor framework, and the second mentioned residue is the residue proposed to replace that residue. Thus, within the variable region frameworks the first mentioned residue is human and within the CDRs the first mentioned residue is murine.

[0597] The exemplified antibodies include any permutation or combination of the exemplified mature heavy chain variable regions and mature light chain variable regions: hu17C12VHv1 / hu17C12VLv1, hu17C12VHv1 / hu17C12VLv2, hu17C12VHv2 / hu17C12VLv1, hu17C12VHv2 / hu17C12VLv2.

[0598] The exemplified antibodies include any of the exemplified mature light chain variable regions, hu17C12VLv1 (SEQ ID NO: 234) and hu17C12VLv2 (SEQ ID NO: 235), and any of the exemplified mature heavy chain variable regions, hu17C12VHv1 (SEQ ID NO: 232) and hu17C12VHv2 (SEQ ID NO: 233).

[0599] The invention provides variants of the 17C12 humanized antibody in which the humanized mature heavy chain variable region exhibits at least 90%, 95%, 96%, 97%, 98%, or 99% identity to 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 to hu17C12VLv1 (SEQ ID NO:234) or hu17C12VLv2 (SEQ ID NO:235). In some such antibodies, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or all 13 of the back mutations or other mutations in SEQ ID NOs:232-233 and SEQ ID NOs:234-235 are retained.

[0600] In some humanized 17C12 antibodies, at least one of the following positions in the VH region is occupied by the indicated amino acid: 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, and H94 is occupied 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.

[0601] In some humanized 17C12 antibodies, at least one of the following positions in the VH region is occupied by the indicated 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.

[0602] In some humanized 17C12 antibodies, such as hu17C12VHv1, VH region positions H2, H24, H48, H67, H69, H93, and H94 are occupied by E, A, I, A, M, T, and T, respectively. In some humanized 17C12 antibodies, such as hu17C12VHv2, VH region positions H1, H2, H24, H48, H67, H69, H93, H94, H108, and H113 are occupied by E, I, A, I, A, M, T, T, L, and S, respectively.

[0603] In some humanized 17C12 antibodies, at least one of the following positions in the VL region is occupied by the indicated 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.

[0604] In some humanized 17C12 antibodies, at least one of the following positions in the VL region is occupied by the indicated amino acid: L2 is V, L36 is L, and L43 is P or S.

[0605] In some humanized 17C12 antibodies, such as hu17C12VLv1, positions L2 and 36 in the VL region are occupied by V and L, respectively. In some humanized 17C12 antibodies, such as hu17C12VLv2, positions L2, L36, and L43 in the VL region are occupied by V, L, and S, respectively.

[0606] In some humanized 17C12 antibodies, the variable heavy chain has 85% or more identity to a human sequence. In some humanized 17C12 antibodies, the variable light chain has 85% or more identity to a human sequence. In some humanized 17C12 antibodies, the variable heavy chain and the variable light chain each have 85% or more identity to a human germline sequence. In some humanized 9F5 antibodies, the three heavy chain CDRs are as defined by the Kabat / Chothia composite (SEQ ID NOs: 226, 227, and 10) and the three light chain CDRs are as defined by the Kabat / Chothia composite (SEQ ID NOs: 229-231).

[0607] An exemplary humanized antibody is a humanized form of murine 14H3, designated Hu14H3.

[0608] Murine antibody 14H3 comprises a mature heavy chain variable region and a mature light chain variable region having amino acid sequences comprising SEQ ID NO:240 and SEQ ID NO:244, respectively. The present invention provides two exemplary humanized mature heavy chain variable regions: hu14H3VHv1 and hu14H3VHv2. The present invention further provides two exemplary mature light chain variable regions: hu14H3VLv1 and hu14H3VLv2. Figures 13 and 14 show alignments of the heavy and light chain variable regions of murine 14H3 and various humanized antibodies, respectively.

[0609] For reasons such as possible effects on CDR conformation and / or binding to antigen, interactions between heavy and light chains, mediating interactions with the constant region, being sites of desirable or undesirable post-translational modifications, being rare residues at that position in human variable region sequences and therefore potentially immunogenic, gaining potential for aggregation, and other reasons, the following eight variable region framework positions were considered as candidates for substitution in the two exemplified human mature light chain variable regions and the two exemplified human mature light chain variable regions, as further detailed 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 positions were considered as candidates for substitution in two exemplified human mature heavy chain variable regions, as fur...

Claims

1. 1. An antibody or antigen-binding antibody fragment that binds to human tau, A murine antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:7 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:11, comprising three heavy chain CDRs defined by the Kabat / Chothia composite and three light chain CDRs defined by Kabat; A murine antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:219 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:11, comprising three heavy chain CDRs defined by the Kabat / Chothia composite and three light chain CDRs defined by Kabat; a murine antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:225 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:228, the three heavy chain CDRs defined by the Kabat / Chothia composite and the three light chain CDRs defined by Kabat; or A murine antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:240 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:244, comprising three heavy chain CDRs defined by the Kabat / Chothia composite and three light chain CDRs defined by Kabat; 4. An antibody or antigen-binding antibody fragment comprising:

2. 2. The antibody or antigen-binding antibody fragment of claim 1, comprising three heavy chain CDRs defined by the Kabat / Chothia composite and three light chain CDRs defined by Kabat of said murine antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:7 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:

11.

3. The three heavy chain CDRs, CDR-H1, CDR-H2, and CDR-H3, defined by the Kabat / Chothia complex have the amino acid sequences of SEQ ID NO:8, SEQ ID NO:9, and SNG, respectively, except that N or T may occupy Kabat position H28, I or V may occupy Kabat position H51, N or D may occupy Kabat position H54, and D or E may occupy Kabat position H56; and the three light chain CDRs, CDR-L1, CDR-L2, and CDR-L3, defined by Kabat have L, D, T, or Q at Kabat position L27b, and L, D, G, S, E, T, N, A, P, or I at Kabat position L27b.

3. The antibody or antigen-binding antibody fragment of claim 2, which are SEQ ID NO:12, SEQ ID NO:13, and SEQ ID NO:14, respectively, except that I, Y, E, K, G, or Q may occupy Kabat position L30, T, N, or G may occupy Kabat position L31, L, N, T, S, R, or G may occupy Kabat position L33, M, G, E, D, K, or I may occupy Kabat position L51, L, R, G, or T may occupy Kabat position L54, A or G occupies Kabat position L89, L, D, E, G, Q, T, or I occupies Kabat position L92, and E or G occupies Kabat position L93.

4. The antibody or antigen-binding antibody fragment of claim 3, wherein the Kabat / Chothia composite defined CDR-H1 has an amino acid sequence comprising SEQ ID NO:50, the Kabat defined CDR-H2 has an amino acid sequence comprising SEQ ID NO:51 or SEQ ID NO:52, the Kabat defined CDR-L1 has an amino acid sequence comprising SEQ ID NO:53, SEQ ID NO:54, or any of SEQ ID NOs:172-193, the Kabat defined CDR-L2 has an amino acid sequence comprising SEQ ID NO:55, or any of SEQ ID NOs:194-205, and the Kabat defined CDR-L3 has an amino acid sequence comprising any of SEQ ID NOs:206-213.

5. The antibody or antigen-binding antibody fragment of any one of claims 1 to 4, wherein the antibody or antigen-binding antibody fragment is a humanized antibody or an antigen-binding antibody fragment comprising a humanized mature heavy chain variable region and a humanized mature light chain variable region.

6. 6. The antibody or antigen-binding antibody fragment of claim 5, wherein the humanized mature heavy chain variable region comprises the three Kabat-defined heavy chain CDRs of the amino acid sequences of SEQ ID NO:40, SEQ ID NO:9, and SNG, and the humanized mature light chain variable region comprises the three Kabat-defined light chain CDRs of SEQ ID NO:12-14.

7. 6. The antibody or antigen-binding antibody fragment of claim 5, wherein the humanized mature heavy chain variable region has an amino acid sequence that is at least 90% identical to any one of SEQ ID NOs: 15-22 and 109-129, and the humanized mature light chain variable region has an amino acid sequence that is at least 90% identical to any one of SEQ ID NOs: 23-29, 61-108, and 130-171.

8. 8. The antibody or antigen-binding antibody fragment of claim 7, wherein at least one of the following positions in the VH region is occupied by the indicated amino acid: Kabat position H1 is occupied by Q or E, Kabat position H5 is occupied by Q or V, Kabat position H11 is occupied by L or V, Kabat position H12 is occupied by V or K, Kabat position H17 is occupied by S or T, Kabat position H20 is occupied by L or I, and t position H23 is occupied by T or K, Kabat position H28 is occupied by N or T, Kabat position H38 is occupied by K, R, or Q, Kabat position H40 is occupied by R or A, Kabat position H42 is occupied by E or G, Kabat position H43 is occupied by Q or K, Kabat position H48 is occupied by I or M, Kabat position H51 is occupied by I or V, and Kabat position H54 is occupied by N or D. Kabat position H56 is occupied by D or E, Kabat position H66 is occupied by K or R, Kabat position H69 is occupied by I or M, Kabat position H75 is occupied by S or T, Kabat position H76 is occupied by N or D, Kabat position H79 is occupied by Y, Q, D, N, or G, Kabat position H80 is occupied by L, M, P, D, G, or E, and Kabat position H81 is occupied by Q or E. Kabat position H82 is occupied by L, P, K, R, E, or N, Kabat position H82a is occupied by S or G, Kabat position H82c is occupied by L, G, D, or S, Kabat position H83 is occupied by T or R, Kabat position H93 is occupied by A or T, Kabat position H94 is occupied by S or T, Kabat position H108 is occupied by T or L, and Kabat position H109 is occupied by L or V.

9. 8. The antibody or antigen-binding antibody fragment of claim 7, wherein at least one of the following positions in the VL region is occupied by the indicated amino acid: Kabat position L3 is V or Q, Kabat position L7 is A or S, Kabat position L8 is A or P, Kabat position L9 is F or L, Kabat position L11 is N or L, Kabat position L15 is L or P, and Kabat position L17 is T or E. at Kabat position L18 is S or P, at Kabat position L27b is L, D, T, or Q, at Kabat position L27c is L, D, G, S, E, T, N, A, P, or I, at Kabat position L30 is I, Y, E, K, G, at Kabat position L31 is T, N, or G, at Kabat position L33 is L, N, T, S, R, or G, at Kabat position L37 is L, Q, G, or I, and at Kabat position L39 is at Kabat position L51 is M, G, E, D, K, or I, at Kabat position L54 is R, G, or T, at Kabat position L60 is N or D, at Kabat position L64 is G or S, at Kabat position L66 is E or G, at Kabat position L73 is L, P, or G, at Kabat position L74 is R or K, at Kabat position L75 is I, D, P, Q, or G, and at Kabat position L Kabat position L76 is S, P, or G, Kabat position L77 is R or D, Kabat position L78 is V, R, D, E, P, K, G, or Q, Kabat position L85 is V or G, Kabat position L86 is Y or T, Kabat position L89 is A or G, Kabat position L92 is L, D, E, G, Q, T, or I, Kabat position L93 is E or G, and Kabat position L100 is G or Q.

10. 8. The antibody or antigen-binding antibody fragment of claim 7, wherein the humanized mature heavy chain variable region has the amino acid sequence of any one of SEQ ID NOs: 15-22 and 109-129, and the humanized mature light chain variable region has the amino acid sequence of any one of SEQ ID NOs: 23-29, 61-108, and 130-171.

11. 3. The antibody or antigen-binding antibody fragment of claim 2, wherein the antibody or antigen-binding antibody fragment comprises a humanized mature heavy chain variable region comprising the three Kabat-defined heavy chain CDRs of the amino acid sequences of SEQ ID NO:40, SEQ ID NO:9, and SNG, and a humanized mature light chain variable region comprising the three Kabat-defined light chain CDRs of SEQ ID NO:12-14.

12. 12. The antibody or antigen-binding antibody fragment of claim 11 , comprising a humanized mature heavy chain variable region having an amino acid sequence at least 90% identical to SEQ ID NO: 214 or 215, and a humanized mature light chain variable region having an amino acid sequence at least 90% identical to SEQ ID NO: 216 or 217.

13. 13. The antibody or antigen-binding antibody fragment of claim 12, wherein at least one of the following positions in the VH region is occupied by the indicated amino acid: Kabat position H1 is occupied by Q or E, Kabat position H24 is occupied by A, Kabat position H48 is occupied by I, Kabat position H67 is occupied by A, Kabat position H69 is occupied by M, Kabat position H93 is occupied by T, and Kabat position H94 is occupied by T.

14. The antibody or antigen-binding antibody fragment of claim 12, wherein at least one of the following positions in the VL region is occupied by the indicated amino acid: Kabat position L64 is S and Kabat position L104 is V or L.

15. 13. The antibody or antigen-binding antibody fragment of claim 12, wherein the humanized mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 214 or 215, and the humanized mature light chain variable region has the amino acid sequence of SEQ ID NO: 216 or 217.

16. 2. The antibody or antigen-binding antibody fragment of claim 1, comprising three heavy chain CDRs defined by the Kabat / Chothia composite and three light chain CDRs defined by Kabat of a murine antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:219 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:

11.

17. 17. The antibody or antigen-binding antibody fragment of claim 16, wherein the antibody or antigen-binding antibody fragment is a humanized antibody or an antigen-binding antibody fragment comprising a humanized mature heavy chain variable region and a humanized mature light chain variable region.

18. 18. The antibody or antigen-binding antibody fragment of claim 17, wherein the humanized mature heavy chain variable region comprises the three Kabat-defined heavy chain CDRs of the amino acid sequences of SEQ ID NO:40, SEQ ID NO:220, and SNG, and the humanized mature light chain variable region comprises the three Kabat-defined light chain CDRs of SEQ ID NO:12-14.

19. 18. The antibody or antigen-binding antibody fragment of claim 17, wherein the humanized mature heavy chain variable region has an amino acid sequence that is at least 90% identical to SEQ ID NO: 221 or 222, and the humanized mature light chain variable region has an amino acid sequence that is at least 90% identical to SEQ ID NO: 223 or 224.

20. 20. The antibody or antigen-binding antibody fragment of claim 19, wherein at least one of the following positions in the VH region is occupied by the indicated amino acid: Kabat position H1 is occupied by Q or E, Kabat position H48 is occupied by M or I, Kabat position H93 is occupied by A or T, and Kabat position H94 is occupied by R or T.

21. The antibody or antigen-binding antibody fragment of claim 19, wherein the VL region positions are occupied by the indicated amino acids: Kabat position L64 is G or S, and Kabat position L104 is V or L.

22. 20. The antibody or antigen-binding antibody fragment of claim 19, wherein the humanized mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 221 or 222, and the humanized mature light chain variable region has the amino acid sequence of SEQ ID NO: 223 or 224.

23. 2. The antibody or antigen-binding antibody fragment of claim 1, comprising the three heavy chain CDRs defined by the Kabat / Chothia composite and the three light chain CDRs defined by Kabat of a murine antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:225 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:

228.

24. 24. The antibody or antigen-binding antibody fragment of claim 23, wherein the antibody or antigen-binding antibody fragment is a humanized antibody or an antigen-binding antibody fragment comprising a humanized mature heavy chain variable region and a humanized mature light chain variable region.

25. 25. The antibody or antigen-binding antibody fragment of claim 24, wherein the humanized mature heavy chain variable region comprises the three Kabat-defined heavy chain CDRs of the amino acid sequences of SEQ ID NO:40, SEQ ID NO:227, and SNG, and the humanized mature light chain variable region comprises the three Kabat-defined light chain CDRs of SEQ ID NO:229-231.

26. 26. The antibody or antigen-binding antibody fragment of claim 24 or 25, wherein the humanized mature heavy chain variable region has an amino acid sequence that is at least 90% identical to SEQ ID NO: 232 or 233, and the humanized mature light chain variable region has an amino acid sequence that is at least 90% identical to SEQ ID NO: 234 or 235.

27. 27. The antibody or antigen-binding antibody fragment of claim 26, wherein at least one of the following positions in the VH region is occupied by the indicated amino acid: Kabat position H1 is occupied by Q or E, Kabat position H2 is occupied by I, Kabat position H24 is occupied by A, Kabat position H48 is occupied by I, Kabat position H67 is occupied by A, Kabat position H69 is occupied by M, Kabat position H93 is occupied by T, Kabat position H94 is occupied by T, Kabat position H108 is occupied by T or L, and Kabat position H113 is occupied by R or S.

28. The antibody or antigen-binding antibody fragment of claim 26, wherein at least one of the following positions in the VL region is occupied by the indicated amino acid: Kabat position L2 is V, Kabat position L36 is L, and Kabat position L43 is P or S.

29. 27. The antibody or antigen-binding antibody fragment of claim 26, wherein the humanized mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 232 or 233, and the humanized mature light chain variable region has the amino acid sequence of SEQ ID NO: 234 or 235.

30. 2. The antibody or antigen-binding antibody fragment of claim 1, comprising three heavy chain CDRs defined by the Kabat / Chothia composite and three light chain CDRs defined by Kabat of a murine antibody characterized by a heavy chain variable region having an amino acid sequence comprising SEQ ID NO:240 and a light chain variable region having an amino acid sequence comprising SEQ ID NO:

244.

31. The antibody or antigen-binding antibody fragment of claim 30, wherein the Kabat / Chothia composite defined CDR-H1 is SEQ ID NO:241, the Kabat defined CDR-H2 is SEQ ID NO:242, and the Kabat defined CDR-H3 is SEQ ID NO:243, except that G or S can occupy Kabat position H35B, and the Kabat / Chothia composite defined CDRs CDR-L1, CDR-L2, and CDR-L3 are SEQ ID NOs:245, 246, and 247, respectively.

32. The antibody or antigen-binding antibody fragment of claim 31, wherein CDR-H1 as defined by the Kabat / Chothia complex has an amino acid sequence comprising SEQ ID NO:

275.

33. The antibody or antigen-binding antibody fragment of any one of claims 30 to 32, wherein the antibody or antigen-binding antibody fragment is a humanized antibody or an antigen-binding antibody fragment comprising a humanized mature heavy chain variable region and a humanized mature light chain variable region.

34. 34. The antibody or antigen-binding antibody fragment of claim 33, wherein the humanized mature heavy chain variable region comprises the three heavy chain CDRs defined by Kabat in SEQ ID NOs:265, 242, and 243, and the humanized mature light chain variable region comprises the three light chain CDRs defined by Kabat in SEQ ID NOs:245-247.

35. 35. The antibody or antigen-binding antibody fragment of claim 33 or 34, wherein the humanized mature heavy chain variable region has an amino acid sequence that is at least 90% identical to SEQ ID NO: 248 or 249, and the humanized mature light chain variable region has an amino acid sequence that is at least 90% identical to SEQ ID NO: 250 or 251.

36. The antibody or antigen-binding antibody fragment of claim 35, wherein at least one of the following positions in the VH region is occupied by the indicated amino acid: Kabat position H35B is occupied by S, Kabat position H108 is occupied by M or L, and Kabat position H113 is occupied by L or S.

37. The antibody or antigen-binding antibody fragment of claim 35, wherein at least one of the following positions in the VL region is occupied by the indicated amino acid: Kabat position L2 is V, Kabat position L7 is T or S, Kabat position L37 is L or Q, Kabat position L87 is F, Kabat position L100 is G or Q, and Kabat position L104 is V or L.

38. 36. The antibody or antigen-binding antibody fragment of claim 35, wherein the humanized mature heavy chain variable region has the amino acid sequence of SEQ ID NO: 248 or 249, and the humanized mature light chain variable region has the amino acid sequence of SEQ ID NO: 250 or 251.

39. 39. The antibody or antigen-binding antibody fragment of any one of claims 1 to 38 which is an antibody.

40. 39. The antibody or antigen-binding antibody fragment of any one of claims 1 to 38, which is an antigen-binding antibody fragment.

41. 40. The antibody or antigen-binding antibody fragment of claim 39, wherein the antibody has an isotype of human IgG1.

42. 42. The antibody or antigen-binding antibody fragment of any one of claims 1 to 39 and 41, wherein the mature light chain variable region is fused to a light chain constant region and the mature heavy chain variable region is fused to a heavy chain constant region.

43. 43. The antibody or antigen-binding antibody fragment of claim 42, wherein the heavy chain constant region is a mutated version of a native human heavy chain constant region that has reduced binding to Fcγ receptors compared to the native human heavy chain constant region.

44. 40. The antibody or antigen-binding antibody fragment of claim 39, wherein the antibody has a human IgG2 or IgG4 isotype.

45. A pharmaceutical composition comprising an antibody or antigen-binding antibody fragment as defined in any one of claims 1 to 44 and a pharma- ceutically acceptable carrier.

46. A nucleic acid encoding the heavy and light chains of an antibody according to any one of claims 1 to 39 and 41.

47. 1. A method for humanizing a murine antibody, comprising the steps of: (a) selecting one or more acceptor antibody sequences; (b) identifying the amino acid residues of the murine antibody that are retained; (c) synthesizing a nucleic acid encoding a humanized heavy chain comprising the Kabat / Chothia complex-defined CDRs of the heavy chain of a murine antibody and a nucleic acid encoding a humanized light chain comprising the Kabat-defined CDRs of the light chain of a murine antibody; (d) expressing the nucleic acid in a host cell to produce the humanized antibody. Including, The murine antibody 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, The murine antibody 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, the murine antibody 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; or The murine antibody 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. method.

48. 46. ​​The pharmaceutical composition of claim 45 for use in a method of inhibiting or reducing tau aggregation in a subject having or at risk of developing tau-mediated amyloidosis.

49. 46. ​​The pharmaceutical composition of claim 45 for use in a method of treating or affecting the prevention of a tau-associated disease in a subject.

50. The tau-related disease is Alzheimer's disease, Down's syndrome, mild cognitive impairment, primary age-related tauopathy, postencephalitic parkinsonism, posttraumatic dementia or dementia pugilistica, Pick's disease, Niemann-Pick type C disease, supranuclear palsy, frontotemporal dementia, frontotemporal lobar degeneration, argyrophilic grain disease, GGT (globular glial tauopathy), Guam amyotrophic lateral sclerosis / Parkinsonism dementia complex, corticobasal degeneration (CBD), Lewy body dementia, Lewy body variant of Alzheimer's disease (LBVAD), chronic traumatic encephalopathy (CTE), GGT (globular glial 50. The pharmaceutical composition of claim 49, wherein the disease is a progressive supranuclear palsy (PSP), Parkinson's disease, or progressive tauopathy.

51. 50. The pharmaceutical composition of claim 49 for use in a method for detecting tau protein deposits in a subject having or at risk of a disease associated with tau aggregation or precipitation.

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