Anti-n3pglu aβ antibody and use thereof

Abstract

An anti-N3pGlu Aβ antibody, a nucleic acid encoding same and the use thereof. The thermostability and immunogenicity in clinical use of various anti-N3pGlu Aβ antibodies have attracted widespread attention in the development of N3pGlu Aβ antibodies. Clinically, the presence of anti-drug antibodies is correlated to some extent with a shortened half-life thereof. Compared with an antibody without modification, a modified antibody exhibits higher yield and lower immunogenicity, and have better application prospects in the treatment of Alzheimer's disease.

Description

Anti-N3pGlu Aβ antibody and its uses

[0001] Cross-references to related applications

[0002] This application claims priority and benefit to Chinese patent application No. 202411514881.9 filed on October 28, 2024; Chinese patent application No. 202411868791.X filed on December 18, 2024; and Chinese patent application No. 202510482593.8 filed on April 17, 2025, all of which are incorporated herein by reference in their entirety for all purposes. Technical Field

[0003] This application relates to the field of biomedicine, specifically to anti-N3pGlu Aβ antibodies and their applications. Background Technology

[0004] Amyloid precursors can be cleaved to form Aβ polypeptides of 38-43 amino acids in length. These Aβ polypeptides then misfold to form insoluble amyloid fibrils, a process closely related to the development of Alzheimer's disease and other neurodegenerative diseases. N3pGlu Aβ is a specifically modified truncated Aβ polypeptide. N3pGlu Aβ lacks the first two amino acids at the N-terminus, and the glutamic acid at the third position undergoes pyroglutamic acid cyclization. N3pGlu Aβ exhibits a plaque-specific distribution in the brain and is more prone to aggregation, thus possessing stronger potential neurotoxicity.

[0005] Currently, there are very limited disease-modifying drugs for Alzheimer's disease, mainly focusing on monoclonal antibodies targeting Aβ, including Lecanemab developed by Biogen and Donanemab developed by Eli Lilly. Both drugs can effectively clear Aβ plaques after 18 months of treatment, but there is still a significant unmet clinical need. Donanemab (trade name Kisunla), developed by Eli Lilly, is the first antibody targeting N3pGlu Aβ and was approved by the FDA in July 2024 for the treatment of early-onset Alzheimer's disease, including mild cognitive impairment and mild Alzheimer's disease. Eli Lilly also has Remternetug, an investigational antibody targeting N3pGlu Aβ. At the 2023 AD / PD conference, Eli Lilly presented interim Phase 1 clinical data, in which 41 participants received 250, 700, 1400, or 2800 mg of Remternetug monthly for six months. Plaque reduction was dose-dependent in all cohorts, with participants receiving 2800 mg showing amyloid levels below the positive threshold within three months. Safety data remain unclear, but there were 10 cases of ARIA-E and 7 cases of ARIA-H overall, with no apparent dose-related variation. No anti-antibody antibodies or systemic infusion reactions were observed. Summary of the Invention

[0006] The thermal stability and immunogenicity of anti-N3pGlu Aβ antibodies in clinical use have attracted widespread attention. The presence of drug-resistant antibodies in clinical settings is correlated with their short half-life. Therefore, improving the stability and / or altering the immunogenicity of anti-N3pGlu Aβ antibodies is of great significance. In view of this, this disclosure is hereby made.

[0007] In a first aspect, this disclosure provides an anti-N3pGlu Aβ antibody or its antigen-binding fragment, comprising a light chain variable region (VL) and a heavy chain variable region (VH), wherein:

[0008] a) The light chain variable region comprises LCDR1 containing the amino acid sequence KSX1X2X3LLYSRGKTYLN (SEQ ID NO:91), containing X 22 LCDR2 containing the amino acid sequence VX4KX5DX6 (SEQ ID NO:165), and LCDR3 containing the amino acid sequence VQGTHYPFT (SEQ ID NO:67), wherein...

[0009] X1 is S, T, or G; X2 is Q or E; X3 is S, T, or D; X4 is S or T; X5 is L, Q, or W; X6 is S or T; and X... 22 It is either A or D; and

[0010] The heavy chain variable region contains HCDR1 with the amino acid sequence X7YDFTRYYIN (SEQ ID NO: 93) and contains WINPGX. 23 X8NX9KYX 24 EKFKX 10 HCDR2 with the amino acid sequence shown in SEQ ID NO:166, and containing EGITVX 25 The amino acid sequence shown in (SEQ ID NO:167) is HCDR3, in which...

[0011] X7 is G or A, X8 is G or S, X9 is T, A or N, X 10 Is it G or A, X 23 Is it S or L, X 24 Is it N or E, and X? 25 It is either Y or V;

[0012] or

[0013] b) The light chain variable region contains X 11 AX 12 QX 13 LCDR1, with the amino acid sequence shown in LGNWLA (SEQ ID NO:137), contains YQAX. 14 TLEX 15 LCDR2 containing the amino acid sequence shown in SEQ ID NO:138, and LCDR3 containing the amino acid sequence shown in SEQ ID NO:123, wherein,

[0014] X 11 Is it R or K, X 12 Is it S or G, X 13 Is it S or T, X 14 Is it S or T, and X? 15 It is S, A, or G; and

[0015] The heavy chain variable region contains X 16 AX 17 GFX 18 The HCDR1 shown in the FSSYPMS (SEQ ID NO: 139) amino acid sequence contains AISGSGX. 19 SX 20 YYADSVKX 21 HCDR2 containing the amino acid sequence shown in SEQ ID NO:140, and HCDR3 containing the amino acid sequence shown in SEQ ID NO:131, wherein,

[0016] X 16 Is it A, S, or D, X17 It is S, X 18 Is it T or D, X 19 Is it G or S, X 20 Is it T or A, and X? 21 Is it G or S?

[0017] Wherein, the anti-N3pGlu Aβ antibody or its antigen-binding fragment does not contain VL and VH of the following CDR combinations:

[0018] (1) LCDR1 of the amino acid sequence KSSQSLLYSRGKTYLN (SEQ ID NO:65), LCDR2 of the amino acid sequence AVSKLDS (SEQ ID NO:66), LCDR3 of the amino acid sequence VQGTHYPFT (SEQ ID NO:67), HCDR1 of the amino acid sequence GYDFTRYYIN (SEQ ID NO:79), HCDR2 of the amino acid sequence WINGPGSGNTKYNEKFKG (SEQ ID NO:80), and HCDR3 of the amino acid sequence EGITVY (SEQ ID NO:81); and

[0019] (2) LCDR1 of the amino acid sequence RASQSLGNWLA (SEQ ID NO:121), LCDR2 of the amino acid sequence YQASTLES (SEQ ID NO:122), LCDR3 of the amino acid sequence QHYKGSFWT (SEQ ID NO:123), HCDR1 of the amino acid sequence AASGFTFSSYPMS (SEQ ID NO:129), HCDR2 of the amino acid sequence AISGSGGSTYYADSVKG (SEQ ID NO:130), and HCDR3 of the amino acid sequence AREGGSGSYYNGFDY (SEQ ID NO:131).

[0020] In some embodiments, this disclosure provides an anti-N3pGlu Aβ antibody or an antigen-binding fragment thereof, comprising a light chain variable region and a heavy chain variable region, wherein:

[0021] a) The light chain variable region comprises LCDR1 containing the amino acid sequence KSX1X2X3LLYSRGKTYLN (SEQ ID NO:91), and contains AVX4KX5DX6 (SEQ ID NO:92) or X 22 LCDR2, containing the amino acid sequence shown in VX4KLDS (SEQ ID NO:162), and LCDR3, containing the amino acid sequence shown in VQGTHYPFT (SEQ ID NO:67),

[0022] Where X1 is S, T, or G, X2 is Q or E, X3 is S, T, or D, X4 is S or T, X5 is L, Q, or W, X6 is S or T, and X... 22 It is either A or D; and

[0023] The heavy chain variable region contains HCDR1 containing the amino acid X7YDFTRYYIN (SEQ ID NO: 93) and contains WINPGSX8NX9KYNEKFKX. 10 (SEQ ID NO:94) or WINPGX 23 GNX9KYX 24 HCDR2 containing the amino acid shown in EKFKG (SEQ ID NO:163), and containing EGITVY (SEQ ID NO:81) or EGITVX. 25 The amino acid HCDR3 shown in (SEQ ID NO:164) is HCDR3.

[0024] Where X7 is G or A, X8 is G or S, X9 is T, A or N, X 10 Is it G or A, X 23 Is it S or L, X 24 Is it N or E, and X? 25 It is either Y or V;

[0025] or

[0026] b) The variable region of the light chain includes, for example, X 11 AX 12 QX 13 LCDR1, as shown in the amino acid sequence LGNWLA (SEQ ID NO:137), such as YQAX 14 TLEX 15 LCDR2 with the amino acid sequence shown in SEQ ID NO:138 and LCDR3 with the amino acid sequence shown in QHYKGSFWT (SEQ ID NO:123),

[0027] Among them, X 11 Is it R or K, X 12 Is it S or G, X 13 Is it S or T, X 14 Is it S or T, X 15 It is S, A, or G; and

[0028] The heavy chain variable region contains X 16 AX 17 GFX 18 The HCDR1 of the amino acid shown in FSSYPMS (SEQ ID NO: 139) contains AISGSGX. 19 SX 20YYADSVKX 21 HCDR2 containing the amino acid shown in SEQ ID NO:140, and HCDR3 containing the amino acid shown in SEQ ID NO:131.

[0029] Among them, X 16 Is it A or S, X 17 It is S, X 18 Is it T or D, X 19 Is it G or S, X 20 Is it T or A, X 21 Is it G or S?

[0030] Wherein, the anti-N3pGlu Aβ antibody or its antigen-binding fragment does not contain VL and VH of the following CDR combinations:

[0031] (1) LCDR1 of the amino acid sequence KSSQSLLYSRGKTYLN (SEQ ID NO:65), LCDR2 of the amino acid sequence AVSKLDS (SEQ ID NO:66), LCDR3 of the amino acid sequence VQGTHYPFT (SEQ ID NO:67), HCDR1 of the amino acid sequence GYDFTRYYIN (SEQ ID NO:79), HCDR2 of the amino acid sequence WINPGSGNTKYNEKFKG (SEQ ID NO:80), and HCDR3 of the amino acid sequence EGITVY (SEQ ID NO:81); and

[0032] (2) LCDR1 of the amino acid sequence RASQSLGNWLA (SEQ ID NO:121), LCDR2 of the amino acid sequence YQASTLES (SEQ ID NO:122), LCDR3 of the amino acid sequence QHYKGSFWT (SEQ ID NO:123), HCDR1 of the amino acid sequence AASGFTFSSYPMS (SEQ ID NO:129), HCDR2 of the amino acid sequence AISGSGGSTYYADSVKG (SEQ ID NO:130), and HCDR3 of the amino acid sequence AREGGSGSYYNGFDY (SEQ ID NO:131).

[0033] In some embodiments, this disclosure provides an anti-N3pGlu Aβ antibody or an antigen-binding fragment thereof, comprising a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises LCDR1, LCDR2, and LCDR3, and the VH comprises HCDR1, HCDR2, and HCDR3 peptides, wherein LCDR1, LCDR2, LCDR3, HCDR1, HCDR2, and HCDR3 are selected from at least one group of the following combinations:

[0034] 1) LCDR1 contains the amino acid sequence shown in SEQ ID NO:68, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0035] 2) LCDR1 contains the amino acid sequence shown in SEQ ID NO:69, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0036] 3) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0037] 4) LCDR1 contains the amino acid sequence shown in SEQ ID NO:71, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0038] 5) LCDR1 contains the amino acid sequence shown in SEQ ID NO:72, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0039] 6) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0040] 7) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:74, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0041] 8) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:75, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0042] 9) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0043] 10) LCDR1 contains the amino acid sequence shown in SEQ ID NO:69, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0044] 11) LCDR1 contains the amino acid sequence shown in SEQ ID NO:71, LCDR2 contains the amino acid sequence shown in SEQ ID NO:75, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0045] 12) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0046] 13) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0047] 14) LCDR1 contains the amino acid sequence shown in SEQ ID NO:77, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0048] 15) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:75, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0049] 16) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0050] 17) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:83, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0051] 18) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:84, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0052] 19) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:85, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0053] 20) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:86, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0054] 21) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:83, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0055] 22) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:86, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0056] 23) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0057] 24) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:88, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0058] 25) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0059] 26) LCDR1 contains the amino acid sequence shown in SEQ ID NO:69, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:84, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0060] 27) LCDR1 contains the amino acid sequence shown in SEQ ID NO:69, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:86, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0061] 28) LCDR1 contains the amino acid sequence shown in SEQ ID NO:69, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:86, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0062] 29) LCDR1 contains the amino acid sequence shown in SEQ ID NO:69, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0063] 30) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:84, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0064] 31) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:86, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0065] 32) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:86, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0066] 33) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0067] 34) LCDR1 contains the amino acid sequence shown in SEQ ID NO:69, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0068] 35) LCDR1 contains the amino acid sequence shown in SEQ ID NO:69, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:88, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0069] 36) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:83, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0070] 37) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:83, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0071] 38) LCDR1 contains the amino acid sequence shown in SEQ ID NO:71, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:83, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0072] 39) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:84, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0073] 40) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:84, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0074] 41) LCDR1 contains the amino acid sequence shown in SEQ ID NO:71, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:84, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0075] 42) LCDR1 contains the amino acid sequence shown in SEQ ID NO:71, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0076] 43) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0077] 44) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:75, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0078] 45) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0079] 46) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0080] 47) LCDR1 contains the amino acid sequence shown in SEQ ID NO:71, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0081] 48) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:78, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0082] 49) LCDR1 contains the amino acid sequence shown in SEQ ID NO:71, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:89, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0083] 50) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:89, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0084] 51) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:75, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:89, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0085] 52) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:89, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0086] 53) LCDR1 contains the amino acid sequence shown in SEQ ID NO:71, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:89, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0087] 54) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:90, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0088] 55) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:78, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:90, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0089] 56) LCDR1 contains the amino acid sequence shown in SEQ ID NO:127, LCDR2 contains the amino acid sequence shown in SEQ ID NO:128, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:129, HCDR2 contains the amino acid sequence shown in SEQ ID NO:130, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:131.

[0090] 57) LCDR1 contains the amino acid sequence shown in SEQ ID NO:126, LCDR2 contains the amino acid sequence shown in SEQ ID NO:125, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:129, HCDR2 contains the amino acid sequence shown in SEQ ID NO:130, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:131.

[0091] 58) LCDR1 contains the amino acid sequence shown in SEQ ID NO:121, LCDR2 contains the amino acid sequence shown in SEQ ID NO:122, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:132, HCDR2 contains the amino acid sequence shown in SEQ ID NO:136, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:131.

[0092] 59) LCDR1 contains the amino acid sequence shown in SEQ ID NO:121, LCDR2 contains the amino acid sequence shown in SEQ ID NO:122, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:132, HCDR2 contains the amino acid sequence shown in SEQ ID NO:134, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:131.

[0093] 60) LCDR1 contains the amino acid sequence shown in SEQ ID NO:126, LCDR2 contains the amino acid sequence shown in SEQ ID NO:122, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:132, HCDR2 contains the amino acid sequence shown in SEQ ID NO:130, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:131.

[0094] 61) LCDR1 contains the amino acid sequence shown in SEQ ID NO:126, LCDR2 contains the amino acid sequence shown in SEQ ID NO:122, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:129, HCDR2 contains the amino acid sequence shown in SEQ ID NO:134, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:131.

[0095] 62) LCDR1 contains the amino acid sequence shown in SEQ ID NO:126, LCDR2 contains the amino acid sequence shown in SEQ ID NO:122, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:132, HCDR2 contains the amino acid sequence shown in SEQ ID NO:134, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:131.

[0096] 63) LCDR1 contains the amino acid sequence shown in SEQ ID NO:126, LCDR2 contains the amino acid sequence shown in SEQ ID NO:125, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:132, HCDR2 contains the amino acid sequence shown in SEQ ID NO:130, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:131.

[0097] 64) LCDR1 contains the amino acid sequence shown in SEQ ID NO:126, LCDR2 contains the amino acid sequence shown in SEQ ID NO:125, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:129, HCDR2 contains the amino acid sequence shown in SEQ ID NO:134, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:131.

[0098] 65) LCDR1 contains the amino acid sequence shown in SEQ ID NO:126, LCDR2 contains the amino acid sequence shown in SEQ ID NO:125, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:132, HCDR2 contains the amino acid sequence shown in SEQ ID NO:134, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:131.

[0099] 66) LCDR1 contains the amino acid sequence shown in SEQ ID NO:124, LCDR2 contains the amino acid sequence shown in SEQ ID NO:122, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:132, HCDR2 contains the amino acid sequence shown in SEQ ID NO:135, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:131.

[0100] 67) LCDR1 contains the amino acid sequence shown in SEQ ID NO:124, LCDR2 contains the amino acid sequence shown in SEQ ID NO:122, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:132, HCDR2 contains the amino acid sequence shown in SEQ ID NO:136, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:131.

[0101] 68) LCDR1 contains the amino acid sequence shown in SEQ ID NO:121, LCDR2 contains the amino acid sequence shown in SEQ ID NO:125, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:129, HCDR2 contains the amino acid sequence shown in SEQ ID NO:130, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:131.

[0102] 69) LCDR1 contains the amino acid sequence shown in SEQ ID NO:121, LCDR2 contains the amino acid sequence shown in SEQ ID NO:122, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:133, HCDR2 contains the amino acid sequence shown in SEQ ID NO:130, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:131.

[0103] 70) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:153, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0104] 71) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:154, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0105] 72) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:154, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:155.

[0106] 73) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:151, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:84, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0107] 74) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:152, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:84, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:81.

[0108] 75) LCDR1 contains the amino acid sequence shown in SEQ ID NO:121, LCDR2 contains the amino acid sequence shown in SEQ ID NO:160, and LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:133, HCDR2 contains the amino acid sequence shown in SEQ ID NO:161, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:131; or

[0109] 76) LCDR1 contains the amino acid sequence shown in SEQ ID NO:121, LCDR2 contains the amino acid sequence shown in SEQ ID NO:160, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:133, HCDR2 contains the amino acid sequence shown in SEQ ID NO:130, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:131.

[0110] In some embodiments, this disclosure provides an anti-N3pGlu Aβ antibody or an antigen-binding fragment thereof comprising a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL and VH are selected from at least one group of the following combinations:

[0111] 1) VL containing the amino acid sequence shown in SEQ ID NO:3 and VH containing the amino acid sequence shown in SEQ ID NO:2;

[0112] 2) VL containing the amino acid sequence shown in SEQ ID NO:4 and VH containing the amino acid sequence shown in SEQ ID NO:2;

[0113] 3) VL containing the amino acid sequence shown in SEQ ID NO:5 and VH containing the amino acid sequence shown in SEQ ID NO:2;

[0114] 4) VL containing the amino acid sequence shown in SEQ ID NO:6 and VH containing the amino acid sequence shown in SEQ ID NO:2;

[0115] 5) VL containing the amino acid sequence shown in SEQ ID NO:7 and VH containing the amino acid sequence shown in SEQ ID NO:2;

[0116] 6) VL containing the amino acid sequence shown in SEQ ID NO:8 and VH containing the amino acid sequence shown in SEQ ID NO:2;

[0117] 7) VL containing the amino acid sequence shown in SEQ ID NO:9 and VH containing the amino acid sequence shown in SEQ ID NO:2;

[0118] 8) VL containing the amino acid sequence shown in SEQ ID NO:10 and VH containing the amino acid sequence shown in SEQ ID NO:2;

[0119] 9) VL containing the amino acid sequence shown in SEQ ID NO:11 and VH containing the amino acid sequence shown in SEQ ID NO:2;

[0120] 10) VL containing the amino acid sequence shown in SEQ ID NO:12 and VH containing the amino acid sequence shown in SEQ ID NO:2;

[0121] 11) VL containing the amino acid sequence shown in SEQ ID NO:13 and VH containing the amino acid sequence shown in SEQ ID NO:2;

[0122] 12) VL containing the amino acid sequence shown in SEQ ID NO:14 and VH containing the amino acid sequence shown in SEQ ID NO:2;

[0123] 13) VL containing the amino acid sequence shown in SEQ ID NO:15 and VH containing the amino acid sequence shown in SEQ ID NO:2;

[0124] 14) VL containing the amino acid sequence shown in SEQ ID NO:16 and VH containing the amino acid sequence shown in SEQ ID NO:2;

[0125] 15) VL containing the amino acid sequence shown in SEQ ID NO:17 and VH containing the amino acid sequence shown in SEQ ID NO:2;

[0126] 16) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:20;

[0127] 17) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:21;

[0128] 18) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:22;

[0129] 19) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:23;

[0130] 20) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:24;

[0131] 21) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:25;

[0132] 22) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:26;

[0133] 23) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:27;

[0134] 24) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:28;

[0135] 25) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:29;

[0136] 26) VL containing the amino acid sequence shown in SEQ ID NO:12 and VH containing the amino acid sequence shown in SEQ ID NO:22;

[0137] 27) VL containing the amino acid sequence shown in SEQ ID NO:12 and VH containing the amino acid sequence shown in SEQ ID NO:24;

[0138] 28) VL containing the amino acid sequence shown in SEQ ID NO:12 and VH containing the amino acid sequence shown in SEQ ID NO:26;

[0139] 29) VL containing the amino acid sequence shown in SEQ ID NO:12 and VH containing the amino acid sequence shown in SEQ ID NO:29;

[0140] 30) VL containing the amino acid sequence shown in SEQ ID NO:15 and VH containing the amino acid sequence shown in SEQ ID NO:22;

[0141] 31) VL containing the amino acid sequence shown in SEQ ID NO:15 and VH containing the amino acid sequence shown in SEQ ID NO:24;

[0142] 32) VL containing the amino acid sequence shown in SEQ ID NO:15 and VH containing the amino acid sequence shown in SEQ ID NO:26;

[0143] 33) VL containing the amino acid sequence shown in SEQ ID NO:15 and VH containing the amino acid sequence shown in SEQ ID NO:29;

[0144] 34) VL containing the amino acid sequence shown in SEQ ID NO:4 and VH containing the amino acid sequence shown in SEQ ID NO:27;

[0145] 35) VL containing the amino acid sequence shown in SEQ ID NO:4 and VH containing the amino acid sequence shown in SEQ ID NO:28;

[0146] 36) VL containing the amino acid sequence shown in SEQ ID NO:11 and VH containing the amino acid sequence shown in SEQ ID NO:21;

[0147] 37) VL containing the amino acid sequence shown in SEQ ID NO:14 and VH containing the amino acid sequence shown in SEQ ID NO:21;

[0148] 38) VL containing the amino acid sequence shown in SEQ ID NO:18 and VH containing the amino acid sequence shown in SEQ ID NO:21;

[0149] 39) VL containing the amino acid sequence shown in SEQ ID NO:11 and VH containing the amino acid sequence shown in SEQ ID NO:22;

[0150] 40) VL containing the amino acid sequence shown in SEQ ID NO:14 and VH containing the amino acid sequence shown in SEQ ID NO:22;

[0151] 41) VL containing the amino acid sequence shown in SEQ ID NO:18 and VH containing the amino acid sequence shown in SEQ ID NO:22;

[0152] 42) VL containing the amino acid sequence shown in SEQ ID NO:6 and VH containing the amino acid sequence shown in SEQ ID NO:27;

[0153] 43) VL containing the amino acid sequence shown in SEQ ID NO:8 and VH containing the amino acid sequence shown in SEQ ID NO:27;

[0154] 44) VL containing the amino acid sequence shown in SEQ ID NO:10 and VH containing the amino acid sequence shown in SEQ ID NO:27;

[0155] 45) VL containing the amino acid sequence shown in SEQ ID NO:11 and VH containing the amino acid sequence shown in SEQ ID NO:27;

[0156] 46) VL containing the amino acid sequence shown in SEQ ID NO:14 and VH containing the amino acid sequence shown in SEQ ID NO:27;

[0157] 47) VL containing the amino acid sequence shown in SEQ ID NO:18 and VH containing the amino acid sequence shown in SEQ ID NO:27;

[0158] 48) VL containing the amino acid sequence shown in SEQ ID NO:19 and VH containing the amino acid sequence shown in SEQ ID NO:27;

[0159] 49) VL containing the amino acid sequence shown in SEQ ID NO:6 and VH containing the amino acid sequence shown in SEQ ID NO:30;

[0160] 50) VL containing the amino acid sequence shown in SEQ ID NO:8 and VH containing the amino acid sequence shown in SEQ ID NO:30;

[0161] 51) VL containing the amino acid sequence shown in SEQ ID NO:10 and VH containing the amino acid sequence shown in SEQ ID NO:30;

[0162] 52) VL containing the amino acid sequence shown in SEQ ID NO:14 and VH containing the amino acid sequence shown in SEQ ID NO:30;

[0163] 53) VL containing the amino acid sequence shown in SEQ ID NO:18 and VH containing the amino acid sequence shown in SEQ ID NO:30;

[0164] 54) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:31;

[0165] 55) VL containing the amino acid sequence shown in SEQ ID NO:19 and VH containing the amino acid sequence shown in SEQ ID NO:31;

[0166] 56) VL containing the amino acid sequence shown in SEQ ID NO:100 and VH containing the amino acid sequence shown in SEQ ID NO:96;

[0167] 57) VL containing the amino acid sequence shown in SEQ ID NO:101 and VH containing the amino acid sequence shown in SEQ ID NO:96;

[0168] 58) VL containing the amino acid sequence shown in SEQ ID NO:95 and VH containing the amino acid sequence shown in SEQ ID NO:106;

[0169] 59) VL containing the amino acid sequence shown in SEQ ID NO:95 and VH containing the amino acid sequence shown in SEQ ID NO:107;

[0170] 60) VL containing the amino acid sequence shown in SEQ ID NO:99 and VH containing the amino acid sequence shown in SEQ ID NO:102;

[0171] 61) VL containing the amino acid sequence shown in SEQ ID NO:99 and VH containing the amino acid sequence shown in SEQ ID NO:104;

[0172] 62) VL containing the amino acid sequence shown in SEQ ID NO:99 and VH containing the amino acid sequence shown in SEQ ID NO:107;

[0173] 63) VL containing the amino acid sequence shown in SEQ ID NO:101 and VH containing the amino acid sequence shown in SEQ ID NO:102;

[0174] 64) VL containing the amino acid sequence shown in SEQ ID NO:101 and VH containing the amino acid sequence shown in SEQ ID NO:104;

[0175] 65) VL containing the amino acid sequence shown in SEQ ID NO:101 and VH containing the amino acid sequence shown in SEQ ID NO:107;

[0176] 66) VL containing the amino acid sequence shown in SEQ ID NO:97 and VH containing the amino acid sequence shown in SEQ ID NO:105;

[0177] 67) VL containing the amino acid sequence shown in SEQ ID NO:97 and VH containing the amino acid sequence shown in SEQ ID NO:106;

[0178] 68) VL containing the amino acid sequence shown in SEQ ID NO:98 and VH containing the amino acid sequence shown in SEQ ID NO:96;

[0179] 69) VL containing the amino acid sequence shown in SEQ ID NO:95 and VH containing the amino acid sequence shown in SEQ ID NO:103;

[0180] 70) VL containing the amino acid sequence shown in SEQ ID NO:14 and VH containing the amino acid sequence shown in SEQ ID NO:143;

[0181] 71) VL containing the amino acid sequence shown in SEQ ID NO:14 and VH containing the amino acid sequence shown in SEQ ID NO:144;

[0182] 72) VL containing the amino acid sequence shown in SEQ ID NO:14 and VH containing the amino acid sequence shown in SEQ ID NO:145;

[0183] 73) VL containing the amino acid sequence shown in SEQ ID NO:141 and VH containing the amino acid sequence shown in SEQ ID NO:22;

[0184] 74) VL containing the amino acid sequence shown in SEQ ID NO:142 and VH containing the amino acid sequence shown in SEQ ID NO:22;

[0185] 75) VL containing the amino acid sequence shown in SEQ ID NO:95 and VH containing the amino acid sequence shown in SEQ ID NO:157; and

[0186] 76) VL containing the amino acid sequence shown in SEQ ID NO:156 and VH containing the amino acid sequence shown in SEQ ID NO:103.

[0187] In some embodiments, the anti-N3pGlu Aβ antibody or its antigen-binding fragment comprises a light chain (LC) and a heavy chain (HC), wherein the light chain comprises a light chain constant region of the amino acid sequence shown in SEQ ID NO: 63, and / or the heavy chain comprises a heavy chain constant region of the amino acid sequence shown in SEQ ID NO: 64.

[0188] In some embodiments, the anti-N3pGlu Aβ antibody or its antigen-binding fragment comprises a light chain (LC) and a heavy chain (HC), wherein the LC and HC are selected from at least one group of the following combinations:

[0189] 1) The light chain of the amino acid sequence shown in SEQ ID NO:34 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33;

[0190] 2) The light chain of the amino acid sequence shown in SEQ ID NO:35 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33;

[0191] 3) The light chain of the amino acid sequence shown in SEQ ID NO:36 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33;

[0192] 4) The light chain of the amino acid sequence shown in SEQ ID NO:37 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33;

[0193] 5) The light chain of the amino acid sequence shown in SEQ ID NO:38 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33;

[0194] 6) The light chain of the amino acid sequence shown in SEQ ID NO:39 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33;

[0195] 7) The light chain of the amino acid sequence shown in SEQ ID NO:40 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33;

[0196] 8) The light chain of the amino acid sequence shown in SEQ ID NO:41 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33;

[0197] 9) The light chain of the amino acid sequence shown in SEQ ID NO:42 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33;

[0198] 10) The light chain of the amino acid sequence shown in SEQ ID NO:43 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33;

[0199] 11) The light chain of the amino acid sequence shown in SEQ ID NO:44 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33;

[0200] 12) The light chain of the amino acid sequence shown in SEQ ID NO:45 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33;

[0201] 13) The light chain of the amino acid sequence shown in SEQ ID NO:46 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33;

[0202] 14) The light chain of the amino acid sequence shown in SEQ ID NO:47 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33;

[0203] 15) The light chain of the amino acid sequence shown in SEQ ID NO:48 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33;

[0204] 16) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:51;

[0205] 17) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:52;

[0206] 18) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:53;

[0207] 19) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:54;

[0208] 20) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:55;

[0209] 21) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:56;

[0210] 22) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:57;

[0211] 23) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58;

[0212] 24) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:59;

[0213] 25) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:60;

[0214] 26) The light chain of the amino acid sequence shown in SEQ ID NO:43 and the heavy chain of the amino acid sequence shown in SEQ ID NO:53;

[0215] 27) The light chain of the amino acid sequence shown in SEQ ID NO:43 and the heavy chain of the amino acid sequence shown in SEQ ID NO:55;

[0216] 28) The light chain of the amino acid sequence shown in SEQ ID NO:43 and the heavy chain of the amino acid sequence shown in SEQ ID NO:57;

[0217] 29) The light chain of the amino acid sequence shown in SEQ ID NO:43 and the heavy chain of the amino acid sequence shown in SEQ ID NO:60;

[0218] 30) The light chain of the amino acid sequence shown in SEQ ID NO:46 and the heavy chain of the amino acid sequence shown in SEQ ID NO:53;

[0219] 31) The light chain of the amino acid sequence shown in SEQ ID NO:46 and the heavy chain of the amino acid sequence shown in SEQ ID NO:55;

[0220] 32) The light chain of the amino acid sequence shown in SEQ ID NO:46 and the heavy chain of the amino acid sequence shown in SEQ ID NO:57;

[0221] 33) The light chain of the amino acid sequence shown in SEQ ID NO:46 and the heavy chain of the amino acid sequence shown in SEQ ID NO:60;

[0222] 34) The light chain of the amino acid sequence shown in SEQ ID NO:35 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58;

[0223] 35) The light chain of the amino acid sequence shown in SEQ ID NO:35 and the heavy chain of the amino acid sequence shown in SEQ ID NO:59;

[0224] 36) The light chain of the amino acid sequence shown in SEQ ID NO:42 and the heavy chain of the amino acid sequence shown in SEQ ID NO:52;

[0225] 37) The light chain of the amino acid sequence shown in SEQ ID NO:45 and the heavy chain of the amino acid sequence shown in SEQ ID NO:52;

[0226] 38) The light chain of the amino acid sequence shown in SEQ ID NO:49 and the heavy chain of the amino acid sequence shown in SEQ ID NO:52;

[0227] 39) The light chain of the amino acid sequence shown in SEQ ID NO:42 and the heavy chain of the amino acid sequence shown in SEQ ID NO:53;

[0228] 40) The light chain of the amino acid sequence shown in SEQ ID NO:45 and the heavy chain of the amino acid sequence shown in SEQ ID NO:53;

[0229] 41) The light chain of the amino acid sequence shown in SEQ ID NO:49 and the heavy chain of the amino acid sequence shown in SEQ ID NO:53;

[0230] 42) The light chain of the amino acid sequence shown in SEQ ID NO:37 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58;

[0231] 43) The light chain of the amino acid sequence shown in SEQ ID NO:39 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58;

[0232] 44) The light chain of the amino acid sequence shown in SEQ ID NO:41 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58;

[0233] 45) The light chain of the amino acid sequence shown in SEQ ID NO:42 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58;

[0234] 46) The light chain of the amino acid sequence shown in SEQ ID NO:45 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58;

[0235] 47) The light chain of the amino acid sequence shown in SEQ ID NO:49 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58;

[0236] 48) The light chain of the amino acid sequence shown in SEQ ID NO:50 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58;

[0237] 49) The light chain of the amino acid sequence shown in SEQ ID NO:37 and the heavy chain of the amino acid sequence shown in SEQ ID NO:61;

[0238] 50) The light chain of the amino acid sequence shown in SEQ ID NO:39 and the heavy chain of the amino acid sequence shown in SEQ ID NO:61;

[0239] 51) The light chain of the amino acid sequence shown in SEQ ID NO:41 and the heavy chain of the amino acid sequence shown in SEQ ID NO:61;

[0240] 52) The light chain of the amino acid sequence shown in SEQ ID NO:45 and the heavy chain of the amino acid sequence shown in SEQ ID NO:61;

[0241] 53) The light chain of the amino acid sequence shown in SEQ ID NO:49 and the heavy chain of the amino acid sequence shown in SEQ ID NO:61;

[0242] 54) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:62;

[0243] 55) The light chain of the amino acid sequence shown in SEQ ID NO:50 and the heavy chain of the amino acid sequence shown in SEQ ID NO:62;

[0244] 56) The light chain of the amino acid sequence shown in SEQ ID NO:113 and the heavy chain of the amino acid sequence shown in SEQ ID NO:109;

[0245] 57) The light chain of the amino acid sequence shown in SEQ ID NO:114 and the heavy chain of the amino acid sequence shown in SEQ ID NO:109;

[0246] 58) The light chain of the amino acid sequence shown in SEQ ID NO:108 and the heavy chain of the amino acid sequence shown in SEQ ID NO:119;

[0247] 59) The light chain of the amino acid sequence shown in SEQ ID NO:108 and the heavy chain of the amino acid sequence shown in SEQ ID NO:120;

[0248] 60) The light chain of the amino acid sequence shown in SEQ ID NO:112 and the heavy chain of the amino acid sequence shown in SEQ ID NO:115;

[0249] 61) The light chain of the amino acid sequence shown in SEQ ID NO:112 and the heavy chain of the amino acid sequence shown in SEQ ID NO:117;

[0250] 62) The light chain of the amino acid sequence shown in SEQ ID NO:112 and the heavy chain of the amino acid sequence shown in SEQ ID NO:120;

[0251] 63) The light chain of the amino acid sequence shown in SEQ ID NO:114 and the heavy chain of the amino acid sequence shown in SEQ ID NO:115;

[0252] 64) The light chain of the amino acid sequence shown in SEQ ID NO:114 and the heavy chain of the amino acid sequence shown in SEQ ID NO:117;

[0253] 65) The light chain of the amino acid sequence shown in SEQ ID NO:114 and the heavy chain of the amino acid sequence shown in SEQ ID NO:120;

[0254] 66) The light chain of the amino acid sequence shown in SEQ ID NO:110 and the heavy chain of the amino acid sequence shown in SEQ ID NO:118;

[0255] 67) The light chain of the amino acid sequence shown in SEQ ID NO:110 and the heavy chain of the amino acid sequence shown in SEQ ID NO:119;

[0256] 68) The light chain of the amino acid sequence shown in SEQ ID NO:111 and the heavy chain of the amino acid sequence shown in SEQ ID NO:109;

[0257] 69) The light chain of the amino acid sequence shown in SEQ ID NO:108 and the heavy chain of the amino acid sequence shown in SEQ ID NO:116;

[0258] 70) The light chain of the amino acid sequence shown in SEQ ID NO:45 and the heavy chain of the amino acid sequence shown in SEQ ID NO:148;

[0259] 71) The light chain of the amino acid sequence shown in SEQ ID NO:45 and the heavy chain of the amino acid sequence shown in SEQ ID NO:149;

[0260] 72) The light chain of the amino acid sequence shown in SEQ ID NO:45 and the heavy chain of the amino acid sequence shown in SEQ ID NO:150;

[0261] 73) The light chain of the amino acid sequence shown in SEQ ID NO:146 and the heavy chain of the amino acid sequence shown in SEQ ID NO:53;

[0262] 74) The light chain of the amino acid sequence shown in SEQ ID NO:147 and the heavy chain of the amino acid sequence shown in SEQ ID NO:53;

[0263] 75) The light chain of the amino acid sequence shown in SEQ ID NO:108 and the heavy chain of the amino acid sequence shown in SEQ ID NO:159;

[0264] 76) The light chain of the amino acid sequence shown in SEQ ID NO:158 and the heavy chain of the amino acid sequence shown in SEQ ID NO:116.

[0265] On the other hand, this disclosure provides a multispecific antigen-binding molecule comprising: (1) the aforementioned anti-N3pGlu Aβ antibody or its antigen-binding fragment; and (2) an antibody or its antigen-binding fragment that binds to other antigens besides N3pGlu Aβ, such other antigens including, but not limited to, TfR, Tau, APP (amyloid precursor protein) or TREM2.

[0266] In some embodiments, the multispecific antigen-binding molecule may be bispecific, trispecific, or tetraspecific; preferably, the multispecific antigen-binding molecule may be divalent, trivalent, tetravalent, pentavalent, or hexavalent.

[0267] On the other hand, this disclosure provides one or more isolated nucleic acid molecules, which may be of any length and contain nucleotides, deoxynucleotides and / or ribonucleotides, wherein the nucleic acid molecules encode the aforementioned antibody or its antigen-binding fragment, or the aforementioned multispecific antigen-binding molecule.

[0268] On the other hand, this disclosure provides a carrier comprising the aforementioned isolated nucleic acid molecules.

[0269] On the other hand, this disclosure provides a host cell containing the aforementioned nucleic acid molecules or isolated host cells of the aforementioned vectors; preferably, the host cell is a eukaryotic cell or a prokaryotic cell; more preferably, the host cell is derived from mammalian cells, yeast cells, insect cells, Escherichia coli and / or Bacillus subtilis; even more preferably, the host cell is selected from Expi293 or CHO cells.

[0270] On the other hand, this disclosure provides a method for preparing the aforementioned antibody or its antigen-binding fragment or multispecific antigen-binding molecule.

[0271] On the other hand, this disclosure provides a pharmaceutical composition comprising the aforementioned anti-N3pGlu Aβ monoclonal antibody or its antigen-binding fragment, the aforementioned multispecific antigen-binding molecule, the aforementioned nucleic acid, or the aforementioned carrier. In a preferred embodiment, the pharmaceutical composition comprises the aforementioned anti-N3pGlu Aβ monoclonal antibody or its antigen-binding fragment, the aforementioned multispecific antigen-binding molecule, the aforementioned nucleic acid, or the aforementioned carrier, as well as a pharmaceutically acceptable carrier, diluent, or excipient. In another preferred embodiment, the pharmaceutical composition further comprises one or more therapeutic ingredients.

[0272] On the other hand, this disclosure provides a method for treating Aβ-related diseases in mammals, comprising administering to a mammal in need of treatment, preferably a human, a therapeutically effective amount of the aforementioned antibody or its antigen-binding fragment, the aforementioned multispecific antigen-binding molecule or its pharmaceutical composition.

[0273] In another preferred embodiment, this disclosure provides a method for preventing human diseases associated with Aβ abnormalities, comprising administering to a patient in need an effective amount of the aforementioned antibody or antigen-binding fragment thereof, the aforementioned multispecific antigen-binding molecule or pharmaceutical composition thereof.

[0274] On the other hand, this disclosure provides the use of the aforementioned antibody or antigen-binding fragment thereof, the aforementioned multispecific antigen-binding molecule or pharmaceutical composition thereof in the preparation of a medicament for treating Aβ-related diseases.

[0275] On the other hand, this disclosure provides the use of the aforementioned antibodies or antigen-binding fragments thereof, the aforementioned multispecific antigen-binding molecules or pharmaceutical compositions thereof in the treatment of Aβ-related diseases.

[0276] On the other hand, this disclosure provides the aforementioned antibody or its antigen-binding fragment, the aforementioned multispecific antigen-binding molecule or its pharmaceutical composition for the treatment of Aβ-related diseases.

[0277] In some embodiments of the foregoing aspects, the Aβ-related disease is selected from Alzheimer's disease, pre-Alzheimer's disease, Down syndrome, and clinical or preclinical cerebral amyloid angiopathy (CAA); Alzheimer's disease is preferred.

[0278] Beneficial effects: Compared with the unmutated sequence, the mutant disclosed herein exhibits lower immunogenicity in humans, higher yield in production, and does not reduce or slightly increase the binding activity to N3pGlu Aβ3-42 fibrils. The mutant disclosed herein also shows higher phagocytic activity of Aβ plaques. Attached Figure Description

[0279] Figure 1 shows the binding activity of the mutant antibody to N3pGlu Aβ3-42 protofibrils;

[0280] Figure 2 shows the binding activity of the mutant antibody to N3pGlu Aβ3-42 protofibrils;

[0281] Figure 3 shows the binding activity of the mutant antibody to N3pGlu Aβ3-42 protofibrils;

[0282] Figure 4 shows the binding activity of the mutant antibody to N3pGlu Aβ3-42 protofibrils;

[0283] Figure 5 shows the binding activity of the mutant antibody to N3pGlu Aβ3-42 protofibrils;

[0284] Figure 6 shows the binding activity of the mutant antibody to N3pGlu Aβ3-42 protofibrils.

[0285] Figure 7 shows the binding activity of the mutant antibody to N3pGlu Aβ3-42 protofibrils;

[0286] Figure 8 shows the binding activity of the mutant antibody to N3pGlu Aβ3-42 protofibrils;

[0287] Figure 9 shows the binding activity of the mutant antibody to N3pGlu Aβ3-42 protofibrils;

[0288] Figure 10 shows the binding activity of the mutant antibody to N3pGlu Aβ3-42 protofibrils;

[0289] Figure 11 shows the binding activity of the mutant antibody to N3pGlu Aβ3-42 protofibrils;

[0290] Figure 12 shows the binding activity of the mutant antibody to N3pGlu Aβ3-42 protofibrils;

[0291] Figure 13 shows the binding activity of the mutant antibody to N3pGlu Aβ3-42 protofibrils;

[0292] Figure 14-17 shows the activity of the mutant antibody in inducing cellular phagocytosis of N3pGlu Aβ3-42 aggregates.

[0293] Terminology Definitions and Detailed Explanations

[0294] Unless otherwise defined in this application, scientific and technical terms related to this disclosure shall have the meanings understood by one of ordinary skill in the art.

[0295] Furthermore, unless otherwise stated herein, singular terms shall include plural terms, and plural terms shall include singular terms. More specifically, as used in this specification and the appended claims, unless otherwise expressly indicated, the singular forms “a” and “this” include plural indicators.

[0296] The terms “comprising,” “including,” and “having” are used interchangeably herein to indicate the inclusiveness of a solution, meaning that the solution may contain elements other than those listed. It should also be understood that the use of “comprising,” “including,” and “having” herein also provides for solutions “composed of.” For example, “a composition comprising A and B” should be understood to include compositions consisting of A and B, as well as compositions containing other components besides A and B, both falling within the scope of the aforementioned “a composition.”

[0297] When used herein, the term “and / or” includes the meaning of “and,” “or,” and “all or any other combination of elements linked by the term.”

[0298] The term "specific binding" in this article refers to the fact that antigen-binding molecules (such as antibodies) typically bind antigens with high affinity and substantially the same antigens, but do not bind unrelated antigens with high affinity.

[0299] The term "N3pGlu Aβ" refers to the amino acid sequence from Pyr to Aβ42 after the pyroglutamic acid cyclization of Glu at position 3 in the Aβ1-42 amino acid sequence. "N3pGlu Aβ" and "AβN3pE-42" refer to the same sequence and can be used interchangeably in this paper.

[0300] The term "antibody" as used in the broadest sense herein refers to a polypeptide or combination of polypeptides containing sufficient sequences from the variable regions of the immunoglobulin heavy chain and / or from the variable regions of the immunoglobulin light chain, thereby enabling specific binding to an antigen. The term "antibody" encompasses a wide range of forms and structures, provided they exhibit the desired antigen-binding activity. "Antibody" includes alternative protein scaffolds or artificial scaffolds having a transplanted complementarity-determining region (CDR) or a CDR derivative. Such scaffolds include antibody-derived scaffolds (containing mutations introduced to, for example, stabilize the three-dimensional structure of the antibody) and fully synthetic scaffolds containing, for example, biocompatible polymers. Such scaffolds may also include non-antibody-derived scaffolds, such as scaffold proteins known in the art for transplanting CDRs, including but not limited to tendinins, fibronectins, peptide aptamers, etc.

[0301] The term "antibody" includes complete antibodies and any antigen-binding fragments (i.e., "antigen-binding portions") or single chains. An "antibody" is a glycoprotein, or its antigen-binding portion, comprising at least two heavy (H) chains and two light (L) chains linked together by disulfide bonds. Each heavy chain consists of a heavy chain variable region (abbreviated VH) and a heavy chain constant region. The heavy chain constant region consists of three domains: CH1, CH2, and CH3. Each light chain consists of a light chain variable region (abbreviated VL) and a light chain constant region. The light chain constant region consists of one domain: CL. The VH and VL regions can be further subdivided into hypervariable regions called complementarity-determining regions (CDRs), which are scattered across more conserved regions called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs, arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of both heavy and light chains contain binding domains that can interact with antigens. The constant regions of antibodies mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system. Due to differences in the amino acid composition and sequence of the constant regions of immunoglobulin heavy chains, their antigenicity also differs. Accordingly, the "immunoglobulins" in this article can be divided into five classes, or isotypes of immunoglobulins: IgM, IgD, IgG, IgA, and IgE, with their corresponding heavy chains being μ, δ, γ, α, and ε chains, respectively. Within the same class of Ig, based on differences in the amino acid composition of its hinge region and the number and position of disulfide bonds in its heavy chain, it can be further divided into different subclasses; for example, IgG can be divided into IgG1, IgG2, IgG3, and IgG4, and IgA into IgA1 and IgA2. Light chains are classified as κ or λ chains based on differences in their constant regions. Each of the five classes of Ig can have either a κ or λ chain.

[0302] The term "antibody" in this article also includes antibodies that do not contain light chains, such as heavy-chain antibodies (HCAbs) produced by camels such as dromedary camels (Camelus dromedarius), Bactrian camels (Camelus bactrianus), llamas (Lama glama), guanicoes (Lama guanicoe), and alpacas (Vicugna pacos), as well as immunoglobulin new antigen receptors (IgNARs) found in cartilaginous fish such as sharks.

[0303] The term "heavy chain antibody" in this article refers to an antibody that lacks a light chain, as is commonly used in antibody therapy. This term specifically includes, but is not limited to, homodimeric antibodies that contain a VH antigen-binding domain and constant CH2 and CH3 domains in the absence of a CH1 domain.

[0304] The term "nanobody" in this article refers to a naturally occurring heavy chain antibody in animals such as camels that lacks a light chain. Cloning its variable region yields a single-domain antibody consisting only of the heavy chain variable region, also known as VHH (Variable domain of heavy chain antibody), which is the smallest functional antigen-binding fragment.

[0305] The terms "VHH domain," "nanobody," and "single domain antibody (sdAb)" in this article have the same meaning and are used interchangeably. They refer to the cloning of the variable region of a heavy chain antibody to construct a single-domain antibody consisting of only one heavy chain variable region. This is the smallest antigen-binding fragment with complete function. Typically, a naturally occurring heavy chain antibody lacking both the light chain and the heavy chain constant region 1 (CH1) is first obtained, and then the variable region of the antibody heavy chain is cloned to construct a single-domain antibody consisting of only one heavy chain variable region.

[0306] The term "scFv" (single-chain variable fragment) in this article refers to a single polypeptide chain containing VL and VH domains linked by a linker. Such scFv molecules can have a general structure: NH2-VL-linker-VH-COOH or NH2-VH-linker-VL-COOH.

[0307] The terms "flexible linker peptide," "linker peptide," or "connector" refer to a short amino acid sequence consisting of amino acids, such as glycine (G) and / or serine (S) and / or threonine residues (T) used alone or in combination, or from the hinge region of an immunoglobulin. In one embodiment, the linker peptide has a length of 5-50 amino acids, for example, 10, 15, 20, 25, or 30 amino acids. In one embodiment, the linker peptide comprises the amino acid sequence (G4S)n, where n is an integer equal to or greater than 1, for example, n is an integer of 2, 3, 4, 5, 6, or 7. In one embodiment, the linker peptide comprises the amino acid sequence TS(G4S)n, where n is an integer equal to or greater than 1, for example, n is an integer of 2, 3, 4, 5, 6, or 7. In another embodiment, the linker peptide is a hinge region derived from an immunoglobulin, such as a hinge region amino acid sequence containing “CPPC (SEQ ID NO:178)”, for example, the amino acid sequence “EPKSCDKTHTCPPCP (SEQ ID NO:179)” or “EPKSSDKTHTCPPCP (SEQ ID NO:180)”. The linker peptides that can be used to link the domains of the antibody molecules in this application may also be, for example, but not limited to, the following amino acid sequences: GGG (SEQ ID NO:168); DGGGS (SEQ ID NO:169); TGEKP (SEQ ID NO:170); GGRR (SEQ ID NO:171); EGKSSGSGSESKVD (SEQ ID NO:172); KESGSVSSEQLAQFRSLD (SEQ ID NO:173); GGRRGGGS (SEQ ID NO:174); LRQRDGERP (SEQ ID NO:175); LRQKDGGGSERP (SEQ ID NO:176); and GSTGSGSGKPGSGEGSTKG (SEQ ID NO:177).

[0308] The term "multispecific" in this document refers to the ability of an antibody or antigen-binding fragment to bind to, for example, different antigens or at least two different epitopes on the same antigen. Therefore, terms such as "bispecific," "trispecific," and "quadrispecific" refer to the number of different epitopes an antibody can bind to. For example, a conventional monospecific IgG antibody has two identical antigen-binding sites (complementary sites) and therefore can only bind to the same epitopes (rather than different epitopes). In contrast, a multispecific antibody has at least two different types of complementary sites / binding sites and therefore can bind to at least two different epitopes. As described herein, a "complementarity-determining region" refers to an antibody's antigen-binding site. Furthermore, a single "specificity" can refer to one, two, three, or more than three identical complementarity-determining regions in a single antibody (the actual number of complementarity-determining regions / binding sites in a single antibody molecule is called the "valence"). For example, a single natural IgG antibody is monospecific and bivalent because it has two identical complementary sites. Accordingly, a multispecific antibody contains at least two (different) complementarity-determining regions / binding sites. Therefore, the term "multispecific antibody" refers to an antibody having more than one complementary site and the ability to bind to two or more different epitopes. The term “multispecific antibody” specifically includes bispecific antibodies as defined above, but generally also includes proteins, such as antibodies that specifically bind to three or more different epitopes, and scaffolds, i.e. antibodies with three or more complementary sites / binding sites.

[0309] The term "valence" in this article refers to the presence of a specified number of binding sites in an antibody / antigen binding molecule. Therefore, the terms "monovalent," "divalent," "tetravalent," and "hexavalent" represent the presence of one, two, four, and six binding sites in an antibody / antigen binding molecule, respectively.

[0310] In this article, "full-length antibody," "intact antibody," and "complete antibody" are used interchangeably, referring to antibodies with structures that are substantially similar to those of natural antibodies.

[0311] The terms "antigen-binding fragment," "antigen-binding fragment," and "antibody fragment" in this article include, but are not limited to, monoclonal antibodies, polyclonal antibodies, monospecific antibodies, multispecific antibodies (e.g., bispecific antibodies), monovalent antibodies, multivalent antibodies, intact antibodies, fragments of intact antibodies, naked antibodies, conjugated antibodies, chimeric antibodies, humanized antibodies, or fully human antibodies. The terms "antigen-binding fragment," "antigen-binding fragment," and "antibody fragment" in this article can be understood as local or local variants of intact antibodies having the same antigenic determinant (CDR), monoclonal antibodies, polyclonal antibodies, monospecific antibodies, multispecific antibodies (e.g., bispecific antibodies), monovalent antibodies, multivalent antibodies, intact antibodies, fragments of intact antibodies, naked antibodies, conjugated antibodies, chimeric antibodies, humanized antibodies, or fully human antibodies, including but not limited to Fab, F(ab')2, Fab', Fab'-SH, Fd, Fv, scFv, diabody, and single-domain antibodies.

[0312] The term "humanized antibody" in this article refers to a genetically engineered non-human antibody whose amino acid sequence has been modified to increase its homology with that of a human antibody. Typically, all or part of the CDR region of a humanized antibody is derived from a non-human antibody (donor antibody), while all or part of the non-CDR region (e.g., the variable region FR and / or constant region) is derived from a human immunoglobulin (receptor antibody). Humanized antibodies generally retain or partially retain the intended properties of the donor antibody, including but not limited to antigen specificity, affinity, reactivity, ability to enhance immune cell activity, or ability to strengthen the immune response.

[0313] The term "fully human antibody" in this document refers to an antibody having variable regions in which both the FR and CDR are derived from human germline immunoglobulin sequences. Furthermore, if the antibody contains a constant region, that constant region is also derived from a human germline immunoglobulin sequence. Fully human antibodies may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced through random or site-specific mutagenesis in vitro or through somatic mutations in vivo). However, "fully human antibody" in this document does not include antibodies in which a CDR sequence derived from another mammalian species (e.g., mouse) has been grafted onto a human frame sequence.

[0314] The term "variable region" in this article refers to the region in the antibody heavy or light chain involved in enabling antibody binding to antigens. "Heavy chain variable region" is used interchangeably with "VH" and "HCVR," and "light chain variable region" is used interchangeably with "VL" and "LCVR." The variable domains of the heavy and light chains of natural antibodies generally have similar structures, with each domain containing four conserved frame regions (FRs) and three hypervariable regions (HVRs). A single VH or VL domain is sufficient to confer antigen-binding specificity.

[0315] The terms "complementarity-determining region" and "CDR" are used interchangeably in this document and generally refer to a hypervariable region (HVR) found in both the light and heavy chain variable domains. The more conserved portion of the variable domain is called the frame region (FR). As understood in the art, the amino acid position representing the hypervariable region of an antibody can vary depending on the context and various definitions known in the art. Some positions within a variable domain can be considered heterozygous hypervariable positions because these positions can be considered within a set of standards (such as IMGT, KABAT, or CCG) but outside a set of standards (such as KABAT, IMGT, or CCG). One or more of these positions can also be found in extended hypervariable regions. This application includes antibodies modified at these heterozygous hypervariable positions. The heavy chain variable region CDR may be abbreviated as HCDR, and the light chain variable region may be abbreviated as LCDR. The variable domains of the natural heavy and light chains each contain four framework regions that primarily employ a lamellar configuration, connected by three CDRs (CDR1, CDR2, and CDR3). These three CDRs form loops connecting the lamellar structures and, in some cases, form part of the lamellar structure. The CDRs in each chain are tightly held together by FR regions in the sequence FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4, and, together with CDRs from other antibody chains, contribute to the formation of antigen-binding sites on the antibody.

[0316] The term "CDR" in this paper can be labeled and defined in a manner known in the art, including but not limited to the Kabat numbering system, the CCG numbering system, or the IMGT numbering system. The tools and websites used include, but are not limited to, the AbRSA website (http: / / cao.labshare.cn / AbRSA / cdrs.php), the abysis website (www.abysis.org / abysis / sequence_input / key_annotation / key_annotation.cgi), and the IMGT website (http: / / www.imgt.org / 3Dstructure-DB / cgi / DomainGapAlign.cgi#results), as well as the MOE software. The CDR in this paper includes overlaps and subsets of amino acid residues defined in different ways.

[0317] The term "Kabat numbering system" generally refers to the immunoglobulin alignment and numbering system proposed by Elvin A. Kabat. The term "IMGT numbering system" generally refers to the numbering system based on the international ImMunoGeneTics information system (IMGT) initiated by Lefranc et al. The term "CCG numbering system" generally refers to the numbering system proposed by the Chemical Computing Group (Montreal, Quebec), which can be calculated using their MOE software. The term "North numbering" can be found in North et al., A New Clustering of Antibody CDR Loop Conformations, Journal of Molecular Biology, 406:228-256 (2011). In some embodiments, the CDR numbering system described in this disclosure preferably uses CCG numbering. In some embodiments, the CDR numbering system described in this disclosure preferably uses North numbering.

[0318] The term "heavy chain constant region" in this document refers to the carboxyl-terminal portion of the antibody heavy chain, which does not directly participate in antibody-antigen binding but exhibits effector functions, such as interaction with the Fc receptor. It has a more conserved amino acid sequence compared to the variable domains of the antibody. The "heavy chain constant region" can be selected from the CH1 domain, hinge region, CH2 domain, CH3 domain, or variants or fragments thereof. The "heavy chain constant region" includes a "full-length heavy chain constant region" and a "heavy chain constant region fragment," the former having a structure substantially similar to the natural antibody constant region, while the latter includes only a portion of the full-length heavy chain constant region. Exemplarily, a typical "full-length antibody heavy chain constant region" consists of a CH1 domain-hinge region-CH2 domain-CH3 domain; when the antibody is IgE, it also includes a CH4 domain; when the antibody is a heavy chain antibody, it does not include the CH1 domain. Exemplarily, a typical "heavy chain constant region fragment" can be selected from the Fc or CH3 domain.

[0319] The term "light chain constant region" in this article refers to the carboxyl terminus of the antibody light chain, which does not directly participate in the binding of the antibody to the antigen. The light chain constant region can be selected from the constant κ domain or the constant λ domain.

[0320] The term "Fc region" used herein is used to define a C-terminal region in an antibody heavy chain that contains at least a portion of a constant region. This term includes both native sequence Fc regions and variant Fc regions. Exemplarily, the human IgG heavy chain Fc region may extend from Cys226 or Pro230 to the C-terminus of the heavy chain. However, antibodies generated by host cells may undergo post-translational cleavage, removing one or more, particularly one or two, amino acids from the C-terminus of the heavy chain. Therefore, antibodies generated by host cells through the expression of a specific nucleic acid molecule encoding the full-length heavy chain may include the full-length heavy chain, or it may include a cleaved variant of the full-length heavy chain. This may be the case when the final two C-terminal amino acids of the heavy chain are glycine (G446) and lysine (K447, numbered according to the Kabat EU index). Therefore, the C-terminal lysine (Lys447), or the C-terminal glycine (Gly446) and lysine (Lys447) of the Fc region may or may not be present. Typically, the IgG Fc region comprises IgG CH2 and IgG CH3 domains, and optionally, may also include a complete or partial hinge region, but not the CH1 domain. The “CH2 domain” of the human IgG Fc region typically extends from an amino acid residue at approximately position 231 to an amino acid residue at approximately position 340. In one embodiment, a carbohydrate chain is attached to the CH2 domain. The CH2 domain herein can be a native sequence CH2 domain or a variant CH2 domain. The “CH3 domain” comprises the residues at the C-terminus of the CH2 domain in the Fc region (i.e., from an amino acid residue at approximately position 341 of the IgG to an amino acid residue at approximately position 447). The CH3 region herein can be a native sequence CH3 domain or a variant CH3 domain (e.g., a CH3 domain having a “knob” introduced in one chain and a corresponding “hole” introduced in the other chain; see U.S. Patent No. 5,821,333, which is expressly incorporated herein by reference). As described herein, such variant CH3 domains can be used to promote heterodimerization of two dissimilar antibody heavy chains.

[0321] Unless otherwise specified herein, the amino acid residues in the Fc region or constant region are numbered according to the EU numbering system, also known as the EU index.

[0322] The term "conservative amino acid" in this document generally refers to amino acids that belong to the same class or have similar characteristics (e.g., charge, side chain size, hydrophobicity, hydrophilicity, main chain conformation, and rigidity). For example, the amino acids in each of the following groups belong to each other's conserved amino acid residues, and substitutions of amino acid residues within a group constitute substitutions of conserved amino acids:

[0323] 1) Alanine (A), Serine (S), Threonine (T);

[0324] 2) Aspartic acid (D), glutamic acid (E);

[0325] 3) Asparagine (N), glutamine (Q);

[0326] 4) Arginine (R), Lysine (K), Histidine (H);

[0327] 5) Isoleucine (I), leucine (L), methionine (M), valine (V); and

[0328] 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W).

[0329] The term "identity" in this document can be calculated as follows: To determine the percentage of "identity" between two amino acid sequences or two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., vacancies may be introduced in one or both of the first and second amino acid sequences or nucleic acid sequences for optimal alignment, or non-homologous sequences may be discarded for comparison purposes). The amino acid residues or nucleotides at corresponding amino acid or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide at the corresponding position in the second sequence, the molecules are identical at that position. Taking into account the number of vacancies that need to be introduced for optimal alignment of the two sequences and the length of each vacancy, the percentage of identity between the two sequences varies with the common positions of the sequences.

[0330] Mathematical algorithms can be used to perform sequence comparisons and calculate the percentage of identity between two sequences. For example, using the Needlema and Wunsch algorithms in the GAP program (available at www.gcg.com) integrated into the GCG software package, the percentage of identity between two amino acid sequences can be determined using a Blossum 62 matrix or a PAM250 matrix with vacancy weights of 16, 14, 12, 10, 8, 6, or 4 and length weights of 1, 2, 3, 4, 5, or 6. As another example, using the GAP program in the GCG software package (available at www.gcg.com), the percentage of identity between two nucleotide sequences can be determined using an NWSgapdna.CMP matrix with vacancy weights of 40, 50, 60, 70, or 80 and length weights of 1, 2, 3, 4, 5, or 6. A particularly preferred set of parameters (and one set of parameters that should be used unless otherwise specified) is a Blossum 62 scoring matrix with a vacancy penalty of 12, a vacancy extension penalty of 4, and a frameshift vacancy penalty of 5. Alternatively, the PAM120 weighted remainder table, a gap length penalty of 12, and a gap penalty of 4 can be used, along with the E. Meyers and W. Miller algorithm, which has been incorporated into the ALIGN program (version 2.0), to determine the percentage of identity between two amino acid sequences or nucleotide sequences.

[0331] Additionally or alternatively, the nucleic acid and protein sequences described in this application can be further used as "query sequences" to perform searches against public databases to, for example, identify sequences of other family members or related sequences. For example, such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul et al., (1990) J. Mol. Biol. 215:403-10. BLAST nucleotide searches can be performed using the NBLAST program with a score of 100 and a word length of 12 to obtain nucleotide sequences homologous to the nucleic acid molecules of this application. BLAST protein searches can be performed using the XBLAST program with a score of 50 and a word length of 3 to obtain amino acid sequences homologous to the protein molecules of this application. For comparison purposes, alignment results with gaps can be obtained using methods such as those described by Altschul, SF, Madden, TL. AA, Zhang, J., Zhang, Z., Miller, W., & Lipman, DJ (1997). Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic acids research, 25(17), 3389-3402. Use vacancy BLAST as described in [reference needed]. When using BLAST and vacancy BLAST programs, the default parameters of the corresponding programs (e.g., XBLAST and NBLAST) can be used. See www.ncbi.nlm.nih.gov.

[0332] The term "nucleic acid" in this document includes any compound and / or substance comprising a polymer of nucleotides. Each nucleotide consists of a base, particularly a purine or pyrimidine base (i.e., cytosine (C), guanine (G), adenine (A), thymine (T), or uracil (U)), a sugar (i.e., deoxyribose or ribose), and a phosphate group. Typically, nucleic acid molecules are described by the sequence of bases, whereby the bases represent the primary structure (linear structure) of the nucleic acid molecule. The sequence of bases is typically represented as 5' to 3'. In this document, the term nucleic acid molecule encompasses deoxyribonucleic acid (DNA), including, for example, complementary DNA (cDNA) and genomic DNA, ribonucleic acid (RNA), particularly messenger RNA (mRNA), synthetic forms of DNA or RNA, and polymers containing mixtures of two or more of these molecules. Nucleic acid molecules can be linear or circular. Furthermore, the term nucleic acid molecule includes both sense and antisense strands, as well as single-stranded and double-stranded forms. Moreover, the nucleic acid molecules described herein may contain naturally occurring or non-naturally occurring nucleotides. Examples of non-naturally occurring nucleotides include modified nucleotide bases having residues with derived sugar or phosphate backbones or chemical modifications. Nucleic acid molecules also encompass DNA and RNA molecules suitable as carriers for the direct expression of the antibodies of this application in vitro and / or in vivo, such as in a host or patient. Such DNA (e.g., cDNA) or RNA (e.g., mRNA) carriers can be unmodified or modified. For example, mRNA can be chemically modified to enhance the stability of the RNA carrier and / or the expression of the encoded molecule, thereby allowing the mRNA to be injected into a subject to generate antibodies in vivo (see, for example, Stadler, CR). H., Celik, L., Hebich, B., Roth, AS, Roth, RP, ... & Sahin, U. (2017). Elimination of large tumors in mice by mRNA-encoded bispecific antibodies. Nature medicine, 23(7), 815-817. or EP2101823B1).

[0333] In this document, “isolated nucleic acid” refers to a nucleic acid molecule that has been separated from the components of its natural environment. Isolated nucleic acid includes nucleic acid molecules found in cells that typically contain such molecules, but which are located outside the chromosome or at a chromosomal location other than their natural chromosomal location.

[0334] The term "vector" in this paper refers to a nucleic acid molecule capable of amplifying another nucleic acid linked to it. This term includes vectors as self-replicating nucleic acid structures as well as vectors integrated into the genome of a host cell into which the vector has been introduced. Some vectors can direct the expression of nucleic acids operatively linked to them. Such vectors are referred to herein as "expression vectors."

[0335] The term "operably linked" or "operably linked" in this document refers to the juxtaposition of two or more target biological sequences in a manner that allows them to function in the intended way, regardless of the presence of spacers or linkers. When used with peptides, the term is intended to indicate that the peptide sequences are linked in a manner that gives the linked product the intended biological function. For example, two or more peptide sequences can be linked using self-cleaving peptides, which cause the peptide sequences to separate rather than fuse together after expression, thereby allowing each peptide sequence to perform its function. The term can also be used with polynucleotides. For instance, when a polynucleotide encoding a peptide is operably linked to a regulatory sequence (e.g., a promoter, enhancer, silencer sequence, etc.), the term is intended to indicate that the polynucleotide sequences are linked in a manner that allows the expression of the peptide to be regulated by the polynucleotide.

[0336] The term "host cell" in this article refers to a cell in which foreign nucleic acids have been introduced, including the progeny of such cells. Host cells include "transformers" and "transformed cells," which include primary transformed cells and their progeny, regardless of the number of passages. Progeny may not be identical to parental cells in their nucleic acid contents and may contain mutations. This article includes mutant progeny with the same function or biological activity as those screened or selected in the initially transformed cells.

[0337] The term "pharmaceutical composition" as used herein refers to a formulation which is present in a form that allows for the effective biological activity of the active ingredient contained therein, and which does not contain any additional ingredients that would have unacceptable toxicity to a subject administering the pharmaceutical composition.

[0338] The term "pharmaceutically acceptable carrier" as used herein includes any and all solvents, dispersion media, coating materials, surfactants, antioxidants, preservatives (e.g., antibacterial agents, antifungal agents), isotonic agents, absorption delay agents, salts, preservatives, drug stabilizers, binders, excipients, disintegrants, lubricants, sweeteners, flavoring agents, dyes, and combinations thereof, as known to those skilled in the art. Except in cases of incompatibility with the active ingredient, the use of any conventional carrier in therapeutic or pharmaceutical compositions is considered.

[0339] The term "treatment" in this article refers to surgical or therapeutic treatment aimed at preventing, mitigating (reducing) undesirable physiological changes or lesions in the treated individual, such as cancer and tumors. Beneficial or desired clinical outcomes include, but are not limited to, symptom relief, disease severity reduction, disease stability (i.e., no worsening), delay or slowing of disease progression, improvement or mitigation of disease status, and remission (whether partial or complete), whether detectable or undetectable. Individuals requiring treatment include those already suffering from the condition or disease, those susceptible to the condition or disease, or those intending to prevent the condition or disease. When the terms mitigation, relief, reduction, mitigation, or remission are used, they also include elimination, disappearance, and non-occurrence.

[0340] The term "subject" in this document refers to an organism that receives treatment for a particular disease or condition as described herein. Exemplarily, "subject" includes mammals, such as humans, primates (e.g., monkeys), or non-primate mammals, that receive treatment for a disease or condition.

[0341] The term "effective dose" in this document refers to the amount of a therapeutic agent, when administered alone or in combination with another therapeutic agent to cells, tissues, or subjects, that is effective in preventing or alleviating symptoms of a disease or the progression of that disease. "Effective dose" also refers to the amount of a compound sufficient to relieve symptoms, such as treating, curing, preventing, or alleviating an associated medical condition, or increasing the rate at which such symptoms are treated, cured, prevented, or alleviated. When an active ingredient is administered to an individual alone, the therapeutically effective dose refers to that ingredient alone. When a combination is used, the therapeutically effective dose refers to the combined amount of active ingredients that produce a therapeutic effect, regardless of whether they are administered in combination, consecutively, or simultaneously.

[0342] The term "EC50" in this document refers to the half-maximal effective concentration, which includes the antibody concentration that induces a half-range response between baseline and maximum value after a specified exposure time. EC50 essentially represents the antibody concentration in which 50% of its maximum effect is observed, and can be measured by methods known in the art.

[0343] IC 50 "(half maximal inhibitory concentration)" in this specification, unless otherwise expressly stated, refers to the concentration required to reduce the response of a target molecule or cell by approximately 50% in a specific biological assay. Such assays may include, but are not limited to, ligand binding inhibition assays, cell signaling pathway inhibition assays, cell activation inhibition assays, or other functional experiments known in the art.

[0344] The IC 50The values ​​can be obtained by measuring response curves at different concentrations and calculating them using standard statistical analysis methods (such as four-parameter logistic fitting). Unless otherwise stated, IC 50 It is expressed as molar concentration (M, μM, or nM). Those skilled in the art can reasonably select the measurement conditions based on the specific target characteristics, experimental system, and known methods.

[0345] Combination or combined administration of drugs: The term "fixed combination" refers to one or more active ingredients, such as antibodies or antigen-binding fragments thereof described in this disclosure, multispecific antibodies or immune effector cells, combined with one or more other active ingredients, and simultaneously applied to a patient or subject in a single entity (such as a mixed injectable) or in a fixed specification and dosage.

[0346] The term "non-fixed combination" refers to one or more active ingredients, such as antibodies or antigen-binding fragments thereof, multispecific antibodies or immune effector cells as described in this disclosure, and other one or more active ingredients, as separate entities, administered to a patient or subject simultaneously, jointly or sequentially (without a specific time limit), such administration of two or more active ingredients to the patient or subject to provide a therapeutically effective level.

[0347] In some embodiments, the pharmaceutical composition may be administered in different ways, such as intravenous, intraperitoneal, subcutaneous, intramuscular, local, or intradermal application.

[0348] Applying the teachings of this application, those skilled in the art can use general techniques, such as site-directed mutagenesis, to substitute amino acids within the CDR and framework sequences disclosed herein, thereby generating other variable region amino acid sequences derived from the sequences of this application. All candidate naturally occurring amino acids can be introduced at specific substitution sites. The methods disclosed herein can then be used to screen these additional variable region amino acid sequences to identify sequences having a specified in vivo function. In this manner, more sequences suitable for preparing the human-engineered antibodies of this application and their antigen-binding moieties can be identified. Preferably, the amino acid substitutions within the framework are limited to one, two, or three positions within any one or more of the four light chains and / or heavy chain framework regions disclosed herein. Preferably, the amino acid substitutions within the CDR are limited to one, two, or three positions within any one or more of the three light chains and / or heavy chain CDRs. Combinations of various changes within these framework regions and CDRs are also possible.

[0349] Immunogenicity is a crucial issue in the development of N3pGlu Aβ antibodies, and the formation of drug-resistant antibodies (ADAs) is a significant reason for the rapid decline in drug concentration in vivo. The antibody in this disclosure is designed to reduce immunogenicity and ADA formation to a certain extent, making it more suitable for drug development and clinical application. Example

[0350] Example 1. Design of CDR mutation for anti-N3pGlu Aβ antibody

[0351] Based on the control N3pGlu Aβ antibody (sequence from patent CN103068848A, named Aβ-mab, VL and VH sequences are shown in SEQ ID NO: 1 and SEQ ID NO: 2 in Table 2, respectively) and the predicted N3pGlu Aβ complex structure, the amino acid interactions at the three-dimensional structural interface were analyzed. Mutated CDR sites using various algorithms were employed to obtain mutant antibodies with improved stability. Corresponding heavy and light chain sequences were designed and synthesized. Nucleic acid fragments encoding the light or heavy chain were cloned into the pTT5 vector, and plasmids were prepared according to established standard molecular biology methods.

[0352] The antibody CDR mutation design combinations are shown in Table 1.

[0353] Table 1. Sequence combinations of the variable region of antibody CDR mutation.

[0354] The specific sequence information of the light chain variable region and heavy chain variable region of the aforementioned mutant antibody is shown in Table 2.

[0355] Table 2. Variable region sequence of mutant antibodies

[0356] Note: Mutation sites are numbered naturally. For example, the S26T mutation in the light chain means that the S at position 26 of the light chain is mutated to T, and so on.

[0357] The full-length sequence of the mutant antibody is shown in Table 3.

[0358] Table 3 Full-length sequences of the heavy and light chains of the mutant antibodies

[0359] Note: Mutation sites are numbered naturally. For example, the S26T mutation in the light chain means that the S at position 26 of the light chain is mutated to T, and so on.

[0360] The CCG numbering sequence of the mutant antibody CDR is shown in Table 4.

[0361] Table 4. CCG analysis results of the CDR region of the mutant antibody sequence.

[0362] Example 2. Production and purification of mutant antibodies

[0363] The plasmid constructed in Example 1 and the transfection reagent (Stylosin, catalog number: STF02) were added to SMM CHO-T3 (Stylosin, catalog number: MCHOT3), mixed well, and allowed to stand for 5 min. This mixture was then added to CHO cells (manufacturer: Thermo Fisher), and the cells were incubated in a shaker at 37°C with 5% CO2 at 120 rpm. On the second day after transfection, feed was added, and the shaker temperature was adjusted to 32°C for continued incubation. Cell supernatant was collected on days 4-5 after transfection.

[0364] The fusion protein in cell culture supernatant was purified using protein A packing material (purchased from Suzhou Nanomicro Technology Co., Ltd.). The protein A packing material was first equilibrated with equilibration buffer (PBS phosphate buffer, pH 7.4), and then the equilibrated packing material was added to the clear culture supernatant and incubated at 4°C for 2 hours. The sample was added to a pretreated empty column tube. After loading, the protein A column was washed with equilibration buffer, and the protein bound to the protein A column was eluted with elution buffer (0.05M acetate-sodium acetate, pH 3.5). The eluted protein was collected, neutralized with 1M Tris, pH 8.0 buffer, and concentrated using a 30kDa Amicon Ultra centrifuge ultrafiltration tube (purchased from Sigma-Aldrich). The sample was then replaced with PBS, pH 7.4. The sample was aseptically filtered through a 0.22μm filter and stored aseptically to obtain the purified Aβ-mab mutant antibody.

[0365] The purified Aβ-mab mutant antibody was analyzed for protein yield and protein concentration (A280). The purified Aβ-mab mutant antibody met the quality requirements, and the expression level did not decrease significantly. Information on protein yield and concentration after 4 days of expression in a small volume (3 mL) is shown in Table 5, and information on protein yield and concentration after 5 days of expression in a large volume (30 mL) is shown in Table 6.

[0366] Table 5. Detection and analysis of Aβ-mab mutant antibodies

[0367] Table 6. Detection and analysis of Aβ-mab mutant antibodies

[0368] Example 3. Identification of the binding activity of Aβ-mab mutant antibody

[0369] Antigen-antibody binding activity was detected using ELISA. N3pGlu Aβ3-42 peptide powder (purchased from Jier Biochemical) was dissolved in 10 mM NaOH to a final concentration of 100 μM. After vortexing, it was diluted to 50 μM with an equal volume of PBS. The solution was incubated at 37°C for 15 min, centrifuged at 4°C for 5 min, and the supernatant was transferred to a 10 kDa ultrafiltration tube and centrifuged to obtain N3pGlu Aβ3-42 fibrils. Specifically, Streptavidin antibody (purchased from Sigma, catalog number #S4762) was diluted with PBS (purchased from Meilunbio, catalog number PWL050), and then 50 μL was added to each well of a 96-well ELISA plate (purchased from Corning, catalog number #9018). The plate was incubated overnight at 4°C. The next day, the plate was washed once with PBST [PBS + 0.5% (w / w) Tween 20], and blocking buffer [PBS + 5% (w / w) skim milk] was added, and the plate was blocked at 37°C for 1 hour. Discard the blocking solution and add 50 μl of 2 μg / ml biotin-labeled N3pGlu Aβ3-42 protocellulose to each well. Incubate at 4°C for 1 hour, then wash 3 times with PBST. After incubating at 4°C for 1 hour, wash 3 times with PBST, and add 50 μl of 200 nM serially diluted mutant antibody or negative control antibody to each well. After incubating at 4°C for 1 hour, wash 3 times with PBST, add HRP (horseradish peroxidase)-labeled secondary antibody (Jackson Immuno, catalog number: 109-035-098), incubate at 4°C for 1 hour, and wash 5 times with PBST. Add 50 μl of TMB substrate (Sera Care, catalog number: 5120-0077) to each well, incubate at room temperature, and then add 50 μl of stop solution (1.0 M HCl) to each well. Read the OD450 nm value using an ELISA reader (PowerWaveHT, Biotec).

[0370] Alternatively, dilute the mutant antibody or negative control antibody to 2 μg / ml with PBS, then add 50 μl to each well of a 96-well ELISA plate and incubate overnight at 4°C. The next day, wash once with PBST, add blocking buffer, and block at 37°C for 1 hour. After incubating at 4°C for 1 hour, wash three times with PBST, and add 50 μl of biotin-labeled N3pGlu Aβ3-42 fibrils (20 μg / ml, serially diluted 5-fold) to each well. After incubating at 4°C for 1 hour, wash three times with PBST, and add HRP-labeled secondary antibody (Sigma, catalog number: S2438). After incubating at 4°C for 1 hour, wash five times with PBST. Add 50 μl of TMB substrate to each well, incubate at room temperature, and then add 50 μl of stop buffer to each well. Read the OD450 nm value using an ELISA reader.

[0371] The results are shown in Figures 1-8 and...

[0372] As shown in Tables 7, 8, and 9, the mutant antibody exhibits good binding activity to N3pGlu Aβ3-42 fibrils. hIgG1 serves as a negative control, not binding to N3pGlu Aβ3-42 fibrils.

[0373] Table 7. ELISA detection of the binding activity of mutant antibody to N3pGlu Aβ3-42 fibrils

[0374] Note: " / " indicates the poor fit.

[0375] Table 8. ELISA detection of the binding activity of mutant antibody to N3pGlu Aβ3-42 fibrils

[0376] Note: " / " indicates the poor fit.

[0377] Table 9. ELISA detection of the binding activity of mutant antibody to N3pGlu Aβ3-42 fibrils

[0378] Note: " / " indicates the poor fit.

[0379] Example 4. Identification of nonspecific binding activity of mutant antibodies

[0380] Antigen-antibody binding activity was detected using an ELISA method. Fibrinogen (Sigma, catalog number F3879) and Fibronectin (Sigma, catalog number F2006) were diluted with PBS (Meilunbio, catalog number PWL050), and then 50 μl of each 20 μg / mL solution was added to a 96-well ELISA plate (Corning, catalog number #9018) and incubated overnight at 4°C. The next day, the plate was washed once with PBST [PBS + 0.5% (w / w) Tween 20], and blocking buffer [PBS + 5% (w / w) skim milk] was added and the plate blocked at room temperature for 1 hour. The blocking buffer was discarded, and 50 μl of mutant antibody or positive control antibody diluted to 100 nM was added to each well. After incubating at room temperature for 2 hours, wash the plate three times with PBST, add HRP (horseradish peroxidase)-labeled secondary antibody (Jackson Immuno, catalog number: 109-035-098), incubate at room temperature for 1 hour, and wash the plate five times with PBST. Add 50 μl of TMB (Sera Care, catalog number: 5120-0077) substrate to each well, incubate at room temperature, and then add 50 μl of stop solution (1.0 M HCl) to each well. Read the OD450 nm value using an ELISA reader (PowerWaveHT, Biotek).

[0381] The results are shown in Table 10. The mutant antibody showed weaker nonspecific binding activity to Fibrinogen and Fibronectin than that of the control.

[0382] Table 10. ELISA detection of binding activity of mutant antibodies with fibrinogen and fibronectin.

[0383] Example 5. Design of CDR mutation for anti-N3pGlu Aβ antibody

[0384] Based on the control N3pGlu Aβ antibody (sequence from patent US10647759B2, named Aβ-mab2, VL and VH sequences are shown in Table 12 as SEQ ID NO: 95 and SEQ ID NO: 96, respectively) and the predicted N3pGlu Aβ complex structure, the amino acid interactions at the three-dimensional structure interface were analyzed. Various algorithms were used to mutate CDR sites to obtain mutant antibodies with improved stability and / or increased yield and maintained activity. Corresponding heavy and light chain sequences were designed and synthesized. Nucleic acid fragments encoding the light or heavy chain were cloned into the pTT5 vector, and plasmids were prepared according to established standard molecular biology methods.

[0385] The antibody CDR mutation design combinations are shown in Table 11.

[0386] Table 11 Sequence Combinations of Variable Regions for Antibody CDR Mutations

[0387] The specific sequence information of the light chain variable region and heavy chain variable region of the aforementioned mutant antibody is shown in Table 12.

[0388] Table 12 Variable Region Sequence of Mutant Antibodies

[0389] Note: Mutation sites are numbered naturally. For example, the R24K mutation in the light chain means that the R at position 24 of the light chain is mutated to K, and so on.

[0390] The full-length sequence of the mutant antibody is shown in Table 13.

[0391] Table 13 Full-length sequences of the heavy and light chains of the mutant antibodies

[0392] Note: Mutation sites are numbered naturally. For example, the R24K mutation in the light chain means that the R at position 24 of the light chain is mutated to K, and so on.

[0393] The CDRs of Aβ-mab2 antibodies were numbered according to the Kabat and North rules. The CDR sequences of the mutant antibodies are shown in Table 14.

[0394] Table 14 Results of CDR analysis of mutant antibody sequences

[0395] Example 6. Production and purification of mutant antibodies

[0396] The plasmid constructed in Example 4 and the transfection reagent (Stylosin, catalog number: STF02) were added to SMM CHO-T3 (Stylosin, catalog number: MCHOT3), mixed well, and allowed to stand for 5 min. This mixture was then added to CHO cells (Thermo Fisher) and cultured in a shaker at 37°C with 5% CO2 at 120 rpm. On the second day after transfection, feed was added, and the shaker temperature was adjusted to 32°C for continued culture. On the fifth day after transfection, the cell supernatant was collected.

[0397] The fusion protein in cell culture supernatant was purified using protein A packing material (purchased from Suzhou Nanomicro Technology Co., Ltd.). The protein A packing material was first equilibrated with equilibration buffer (PBS phosphate buffer, pH 7.4), and then added to the clear culture supernatant and incubated at 4°C for 2 hours. The sample was added to a pretreated empty column tube. After loading, the protein A column was washed with equilibration buffer, and the protein bound to the protein A column was eluted with elution buffer (0.05M acetate-sodium acetate, pH 3.5). The eluted protein was collected, neutralized with 1M Tris, pH 8.0 buffer, and concentrated using a 30kDa Amicon Ultra centrifuge ultrafiltration tube (purchased from Sigma-Aldrich). The sample was then replaced with PBS, pH 7.4. The sample was aseptically filtered through a 0.22μm filter and stored aseptically to obtain the purified Aβ-mab2 mutant antibody.

[0398] The purified mutant antibody was analyzed for protein yield and protein concentration (A280). The purified mutant antibody was of acceptable quality, and the expression level did not decrease significantly. Information on protein yield and concentration after 4 days of expression in a small volume (3 mL) is shown in Table 15, and information on protein yield and concentration after 5 days of expression in a large volume (30 mL) is shown in Table 16.

[0399] Table 15 Detection and Analysis of Mutant Antibodies

[0400] Table 16 Detection and Analysis of Mutant Antibodies

[0401] Example 7. Identification of the binding activity of mutant antibodies

[0402] Antigen-antibody binding activity was detected using ELISA. N3pGlu Aβ3-42 peptide powder (purchased from Jier Biochemical) was dissolved in 10 mM NaOH to a final concentration of 100 μM. After vortexing, it was diluted to 50 μM with an equal volume of PBS. The solution was incubated at 37°C for 15 min, centrifuged at 4°C for 5 min, and the supernatant was transferred to a 10 kDa ultrafiltration tube and centrifuged to obtain N3pGlu Aβ3-42 fibrils. Specifically, the mutant antibody or negative control antibody was diluted to 2 μg / ml with PBS (purchased from Meilunbio, catalog number PWL050), and then 50 μl was added to each well of a 96-well ELISA plate (purchased from Corning, catalog number #9018). The plate was incubated overnight at 4°C. The next day, the plate was washed once with PBST [PBS + 0.5% (w / w) Tween 20], and blocking buffer [PBS + 5% (w / w) skim milk] was added, blocking at 37°C for 1 hour. After incubating at 4°C for 1 hour, wash the plate three times with PBST. Add 50 μl of biotin-labeled N3pGlu Aβ3-42 fibrils (20 μg / ml, serially diluted 5-fold) to each well. After incubating at 4°C for 1 hour, wash the plate three times with PBST. Add HRP (horseradish peroxidase)-labeled secondary antibody (Sigma, catalog number: S2438). After incubating at 4°C for 1 hour, wash the plate five times with PBST. Add 50 μl of TMB (Sera Care, catalog number: 5120-0077) substrate to each well, incubate at room temperature, and then add 50 μl of stop solution (1.0 M HCl) to each well. Read the OD450 nm value using an ELISA reader (PowerWaveHT, Biotec).

[0403] Alternatively, dilute the mutant antibody or negative control antibody to 2 μg / ml with PBS, then add 50 μl to each well of a 96-well ELISA plate and incubate overnight at 4°C. The next day, wash once with PBST, add blocking buffer, and block at 37°C for 1 hour. After incubating at 4°C for 1 hour, wash three times with PBST, and add 50 μl of biotin-labeled N3pGlu Aβ3-42 fibrils (20 μg / ml, serially diluted 5-fold) to each well. After incubating at 4°C for 1 hour, wash three times with PBST, and add HRP-labeled secondary antibody (Sigma, catalog number: S2438). After incubating at 4°C for 1 hour, wash five times with PBST. Add 50 μl of TMB substrate to each well, incubate at room temperature, and then add 50 μl of stop buffer to each well. Read the OD450 nm value using an ELISA reader.

[0404] The results are shown in Figures 9-13 and Tables 17 and 18 (negative control results are not included in Table 17). The mutant antibody has good binding activity with N3pGlu Aβ3-42 protofibrils.

[0405] Table 17 ELISA detection of the binding activity of mutant antibody to N3pGlu Aβ3-42 fibrils

[0406] Table 18 ELISA detection of the binding activity of mutant antibody to N3pGlu Aβ3-42 fibrils

[0407] Example 8. Identification of nonspecific binding activity of mutant antibodies

[0408] Antigen-antibody binding activity was detected using an ELISA method. Fibrinogen (Sigma, catalog number F3879) and Fibronectin (Sigma, catalog number F2006) were diluted with PBS (Meilunbio, catalog number PWL050), and then 50 μl of each solution (1 μg / mL or 20 μg / mL) was added to each well of a 96-well ELISA plate (Corning, catalog number #9018) and incubated overnight at 4°C. The next day, the plate was washed once with PBST [PBS + 0.5% (w / w) Tween 20], and blocking buffer [PBS + 5% (w / w) skim milk] was added and the plate blocked at room temperature for 1 hour. The blocking buffer was discarded, and 50 μl of mutant antibody or positive control antibody diluted to 100 nM was added to each well. After incubating at room temperature for 2 hours, wash the plate three times with PBST, add HRP (horseradish peroxidase)-labeled secondary antibody (Jackson Immuno, catalog number: 109-035-098), incubate at room temperature for 1 hour, and wash the plate five times with PBST. Add 50 μl of TMB (Sera Care, catalog number: 5120-0077) substrate to each well, incubate at room temperature, and then add 50 μl of stop solution (1.0 M HCl) to each well. Read the OD450 nm value using an ELISA reader (PowerWaveHT, Biotek).

[0409] The results are shown in Tables 19 and 20 (different batches of experiments). The mutant antibody did not show non-specific binding activity with Fibrinogen or Fibronectin.

[0410] Table 19 ELISA detection of binding activity of mutant antibodies with fibrinogen and fibronectin

[0411] Table 20 ELISA detection of binding activity of mutant antibodies with fibrinogen and fibronectin

[0412] Example 9. Detection of the thermostability of mutant antibodies

[0413] The thermal stability of antibodies was detected using differential scanning fluorimetry (DSF). TM The dye in the Dye Kit (Applied Biosystems, catalog number 4461146) binds to exposed hydrophobic residues, and the fluorescence signal generated by the exposure of hydrophobic residues in proteins with temperature changes is monitored using a real-time quantitative PCR instrument (Thermo 7500). The dye and antibody to be tested were diluted with appropriate buffers, with a dye concentration of 2× and an antibody concentration of 0.4 mg / mL in each well, in a volume of 20 μL. Three replicates were performed per sample. The temperature was increased at a rate of 1% for 2 min, starting at 25°C and ending at 99°C. Melting curves were analyzed using Protein Thermal Shift software 1.3, and TmD represents the derivative of the melting temperature (Tm). Results for different batches are shown in Tables 21 and 22. The thermostability of the mutant antibody was comparable to that of the parental antibodies Aβ-mab and Aβ-mab2, with the mutant Aβ-mab2 showing a slightly better Tm value than Aβ-mab2.

[0414] Table 21 Melting temperatures of DSF-detected mutant antibodies

[0415] Table 22 Melting temperatures of DSF for detecting mutant antibodies

[0416] " / " indicates that it was not detected.

[0417] Example 10. Detection of phagocytic activity of mutant antibodies

[0418] The activity of antibody-dependent cellular phagocytosis (ADCP) N3pGlu Aβ3-42 aggregates was detected using the FACS method. N3pGlu Aβ3-42 and N3pGlu Aβ3-42-Cy5 peptide powders (purchased from Jier Biochemical) were dissolved separately in 10 mM NaOH to a concentration of 5 mg / ml, then diluted with PBS to 0.5 mg / ml. The peptides were mixed thoroughly at a ratio of 95% N3pGlu Aβ3-42 + 5% N3pGlu Aβ3-42-Cy5 and incubated at 37°C for 7 days. The mixture was then centrifuged at 4°C and 16000g for 5 minutes using an Eppendorf centrifuge (Eppendorf, 5424R). The supernatant was discarded, and the precipitate was resuspended in PBS to obtain 100% N3pGlu Aβ3-42 aggregates with a Cy5 fluorescent tag. To induce THP-1 cells into phagocytes for phagocytosis experiments, THP-1 cells were seeded at a rate of 2E6 / 10ml into 10cm cell culture dishes (purchased from Corning, catalog number #430167). hM-CSF was added to complete cell culture medium (RPMI 1640 + 10% FBS + 1% P / S) to a final concentration of 100 ng / ml, and the dishes were incubated at 37°C in a 5% CO2 incubator for 8 days to induce differentiation. Fresh 100 ng / ml hM-CSF-complete culture medium was replaced every 3 days to maintain cell condition. On day 8, THP-1 cells were collected from the 10cm cell culture dishes, centrifuged, and counted. The cells were then aliquoted into 96-well plates at a rate of 1E5 / well. The THP-1 cell culture medium was discarded, and each well was incubated with 80 μl of RPMI 1640 medium (containing 1% Ultra Low IgG FBS) at a concentration of 100 μg / ml fucose for 30 minutes. Antibody dilution buffer was prepared, and the antibody was diluted 10-fold starting at 100 nM using PBS. Compound activity was tested using a 6-point dose-response method. After incubation with fucose for 30 minutes, corresponding concentrations of antibody and N3pGlu Aβ3-42-Cy5 aggregates (final concentration 5 μg / ml) were added, and the cells were co-incubated with THP-1 cells for 1.5 h. After incubation, the supernatant was discarded. Cells were washed once with PBS, and after discarding the supernatant, 30 μl of 0.25% trypsin was added to each well for digestion at 4°C for 20 minutes.After digestion, 150 μl of stop solution (RPMI 1640 medium containing 10% FBS) was added to each well, and the cells were collected into 96-well U-shaped plates (Jetek, tcp002096). The cells were centrifuged at 450 rcf for 5 minutes using an Eppendorf centrifuge (5920R). After centrifugation, the supernatant was discarded, and the cells were washed twice with 200 μl of PBS per well, centrifuged at 450 rcf for 5 minutes, and the supernatant was discarded. The cells were resuspended in 300 μl of FACS buffer (PBS solution containing 1% FBS) and then analyzed by flow cytometry. The results are shown in Table 23 and Figures 14-17. The phagocytic activity induced by the mutant antibody was basically equivalent to that of Aβ-mab and Aβ-mab2, with the Aβ-mab mutant showing a higher maximum phagocytic rate than Aβ-mab (negative control results are not included in Table 23).

[0419] Table 23 Phagocytic effect of mutant antibody ADCP

[0420] All references to this application are incorporated herein by reference as if each reference were individually incorporated herein by reference. Furthermore, it should be understood that after reading the foregoing teachings of this application, those skilled in the art can make various alterations or modifications to this application, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. An anti-N3pGlu Aβ antibody or its antigen-binding fragment, comprising a light chain variable region (VL) and a heavy chain variable region (VH), wherein: a) The light chain variable region comprises LCDR1 containing the amino acid sequence KSX1X2X3LLYSRGKTYLN (SEQ ID NO: 91), containing X 22 LCDR2 containing the amino acid sequence VX4KX5DX6 (SEQ ID NO:165), and LCDR3 containing the amino acid sequence VQGTHYPFT (SEQ ID NO:67), wherein... X1 is S, T, or G. X2 is either Q or E. X3 is S, T, or D. X4 is either S or T. X5 is L, Q, or W. X6 is either S or T, and X 22 It is either A or D; and The heavy chain variable region contains HCDR1 with the amino acid sequence X7YDFTRYYIN (SEQ ID NO: 93) and contains WINPGX. 23 X8NX9KYX 24 EKFKX 10 HCDR2 with the amino acid sequence shown in SEQ ID NO:166, and containing EGITVX 25 The amino acid sequence shown in (SEQ ID NO:167) is HCDR3, in which... X7 is either G or A. X8 is either G or S. X9 is T, A, or N. X 10 Is it G or A? X 23 Is it S or L? X 24 Is it N or E, and X 25 It is either Y or V; or b) The light chain variable region contains X 11 AX 12 QX 13 LCDR1, with the amino acid sequence shown in LGNWLA (SEQ ID NO:137), contains YQAX. 14 TLEX 15 LCDR2 containing the amino acid sequence shown in SEQ ID NO:138, and LCDR3 containing the amino acid sequence shown in SEQ ID NO:123, wherein, X 11 Is it R or K? X 12 Is it S or G? X 13 Is it S or T? X 14 Is it S or T, and X 15 It is S, A, or G; and The heavy chain variable region contains X 16 AX 17 GFX 18 The HCDR1 shown in the FSSYPMS (SEQ ID NO: 139) amino acid sequence contains AISGSGX. 19 SX 20 YYADSVKX 21 HCDR2 containing the amino acid sequence shown in SEQ ID NO:140, and HCDR3 containing the amino acid sequence shown in SEQ ID NO:131, wherein, X 16 Is it A, S, or D? X 17 It's S. X 18 Is it T or D? X 19 Is it G or S? X 20 Is it T or A, and X 21 It's either G or S.

2. An anti-N3pGlu Aβ antibody or its antigen-binding fragment, comprising a light chain variable region (VL) and a heavy chain variable region (VH), wherein: a) The light chain variable region comprises LCDR1 containing the amino acid sequence KSX1X2X3LLYSRGKTYLN (SEQ ID NO:91), and contains AVX4KX5DX6 (SEQ ID NO:92) or X 22 LCDR2 containing the amino acid sequence shown in VX4KLDS (SEQ ID NO:162), and LCDR3 containing the amino acid sequence shown in VQGTHYPFT (SEQ ID NO:67), wherein... X1 is S, T, or G. X2 is either Q or E. X3 is S, T, or D. X4 is either S or T. X5 is L, Q, or W. X6 is either S or T, and X 22 It is either A or D; and The heavy chain variable region contains HCDR1 with the amino acid sequence X7YDFTRYYIN (SEQ ID NO: 93) and contains WINPGSX8NX9KYNEKFKX. 10 (SEQ ID NO:94) or WINPGX 23 GNX9KYX 24 HCDR2 with the amino acid sequence shown in EKFKG (SEQ ID NO:163), and containing EGITVY (SEQ ID NO:81) or EGITVX. 25 The amino acid sequence shown in (SEQ ID NO:164) is HCDR3, in which... X7 is either G or A. X8 is either G or S. X9 is T, A, or N. X 10 Is it G or A? X 23 Is it S or L? X 24 Is it N or E, and X 25 It is either Y or V; or b) The light chain variable region contains X 11 AX 12 QX 13 LCDR1, with the amino acid sequence shown in LGNWLA (SEQ ID NO:137), contains YQAX. 14 TLEX 15 LCDR2 containing the amino acid sequence shown in SEQ ID NO:138, and LCDR3 containing the amino acid sequence shown in SEQ ID NO:123, wherein, X 11 Is it R or K? X 12 Is it S or G? X 13 Is it S or T? X 14 Is it S or T? X 15 It is S, A, or G; and The heavy chain variable region includes, for example, X. 16 AX 17 GFX 18 HCDR1, such as AISGSGX, is an amino acid sequence shown in FSSYPMS (SEQ ID NO: 139). 19 SX 20 YYADSVKX 21 HCDR2 with the amino acid sequence shown in SEQ ID NO:140, and HCDR3 with the amino acid sequence shown in AREGGSGSYYNGFDY (SEQ ID NO:131), wherein, X 16 Is it A or S? X 17 It's S. X 18 Is it T or D? X 19 Is it G or S? X 20 Is it T or A? X 21 It's either G or S.

3. The antibody or antigen-binding fragment thereof according to claim 1 or 2, wherein VL comprises LCDR1, LCDR2, and LCDR3, and VH comprises HCDR1, HCDR2, and HCDR3, wherein LCDR1, LCDR2, LCDR3, HCDR1, HCDR2, and HCDR3 are selected from at least one group of the following combinations: 1) LCDR1 contains the amino acid sequence shown in SEQ ID NO:68, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 2) LCDR1 contains the amino acid sequence shown in SEQ ID NO:69, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 3) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 4) LCDR1 contains the amino acid sequence shown in SEQ ID NO:71, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 5) LCDR1 contains the amino acid sequence shown in SEQ ID NO:72, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 6) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 7) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:74, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 8) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:75, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 9) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 10) LCDR1 contains the amino acid sequence shown in SEQ ID NO:69, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 11) LCDR1 contains the amino acid sequence shown in SEQ ID NO:71, LCDR2 contains the amino acid sequence shown in SEQ ID NO:75, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 12) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 13) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 14) LCDR1 contains the amino acid sequence shown in SEQ ID NO:77, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 15) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:75, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 16) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:80, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 17) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:83, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 18) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:84, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 19) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:85, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 20) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:86, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 21) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:83, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 22) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:86, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 23) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 24) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:88, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 25) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 26) LCDR1 contains the amino acid sequence shown in SEQ ID NO:69, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:84, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 27) LCDR1 contains the amino acid sequence shown in SEQ ID NO:69, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:86, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 28) LCDR1 contains the amino acid sequence shown in SEQ ID NO:69, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:86, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 29) LCDR1 contains the amino acid sequence shown in SEQ ID NO:69, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 30) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:84, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 31) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:86, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 32) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:86, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 33) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 34) LCDR1 contains the amino acid sequence shown in SEQ ID NO:69, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 35) LCDR1 contains the amino acid sequence shown in SEQ ID NO:69, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:82, HCDR2 contains the amino acid sequence shown in SEQ ID NO:88, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 36) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:83, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 37) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:83, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 38) LCDR1 contains the amino acid sequence shown in SEQ ID NO:71, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:83, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 39) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:84, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 40) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:84, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 41) LCDR1 contains the amino acid sequence shown in SEQ ID NO:71, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:84, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 42) LCDR1 contains the amino acid sequence shown in SEQ ID NO:71, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 43) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 44) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:75, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 45) LCDR1 contains the amino acid sequence shown in SEQ ID NO:76, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 46) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 47) LCDR1 contains the amino acid sequence shown in SEQ ID NO:71, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 48) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:78, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:87, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 49) LCDR1 contains the amino acid sequence shown in SEQ ID NO:71, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:89, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 50) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:89, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 51) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:75, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:89, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 52) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:89, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 53) LCDR1 contains the amino acid sequence shown in SEQ ID NO:71, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:89, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 54) LCDR1 contains the amino acid sequence shown in SEQ ID NO:65, LCDR2 contains the amino acid sequence shown in SEQ ID NO:66, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:90, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 55) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:78, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:90, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 56) LCDR1 contains the amino acid sequence shown in SEQ ID NO:127, LCDR2 contains the amino acid sequence shown in SEQ ID NO:128, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:129, HCDR2 contains the amino acid sequence shown in SEQ ID NO:130, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

131. 57) LCDR1 contains the amino acid sequence shown in SEQ ID NO:126, LCDR2 contains the amino acid sequence shown in SEQ ID NO:125, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:129, HCDR2 contains the amino acid sequence shown in SEQ ID NO:130, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

131. 58) LCDR1 contains the amino acid sequence shown in SEQ ID NO:121, LCDR2 contains the amino acid sequence shown in SEQ ID NO:122, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:132, HCDR2 contains the amino acid sequence shown in SEQ ID NO:136, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

131. 59) LCDR1 contains the amino acid sequence shown in SEQ ID NO:121, LCDR2 contains the amino acid sequence shown in SEQ ID NO:122, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:132, HCDR2 contains the amino acid sequence shown in SEQ ID NO:134, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

131. 60) LCDR1 contains the amino acid sequence shown in SEQ ID NO:126, LCDR2 contains the amino acid sequence shown in SEQ ID NO:122, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:132, HCDR2 contains the amino acid sequence shown in SEQ ID NO:130, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

131. 61) LCDR1 contains the amino acid sequence shown in SEQ ID NO:126, LCDR2 contains the amino acid sequence shown in SEQ ID NO:122, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:129, HCDR2 contains the amino acid sequence shown in SEQ ID NO:134, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

131. 62) LCDR1 contains the amino acid sequence shown in SEQ ID NO:126, LCDR2 contains the amino acid sequence shown in SEQ ID NO:122, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:132, HCDR2 contains the amino acid sequence shown in SEQ ID NO:134, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

131. 63) LCDR1 contains the amino acid sequence shown in SEQ ID NO:126, LCDR2 contains the amino acid sequence shown in SEQ ID NO:125, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:132, HCDR2 contains the amino acid sequence shown in SEQ ID NO:130, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

131. 64) LCDR1 contains the amino acid sequence shown in SEQ ID NO:126, LCDR2 contains the amino acid sequence shown in SEQ ID NO:125, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:129, HCDR2 contains the amino acid sequence shown in SEQ ID NO:134, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

131. 65) LCDR1 contains the amino acid sequence shown in SEQ ID NO:126, LCDR2 contains the amino acid sequence shown in SEQ ID NO:125, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:132, HCDR2 contains the amino acid sequence shown in SEQ ID NO:134, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

131. 66) LCDR1 contains the amino acid sequence shown in SEQ ID NO:124, LCDR2 contains the amino acid sequence shown in SEQ ID NO:122, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:132, HCDR2 contains the amino acid sequence shown in SEQ ID NO:135, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

131. 67) LCDR1 contains the amino acid sequence shown in SEQ ID NO:124, LCDR2 contains the amino acid sequence shown in SEQ ID NO:122, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:132, HCDR2 contains the amino acid sequence shown in SEQ ID NO:136, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

131. 68) LCDR1 contains the amino acid sequence shown in SEQ ID NO:121, LCDR2 contains the amino acid sequence shown in SEQ ID NO:125, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:129, HCDR2 contains the amino acid sequence shown in SEQ ID NO:130, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

131. 69) LCDR1 contains the amino acid sequence shown in SEQ ID NO:121, LCDR2 contains the amino acid sequence shown in SEQ ID NO:122, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:133, HCDR2 contains the amino acid sequence shown in SEQ ID NO:130, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

131. 70) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:153, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 71) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:154, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 72) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:73, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:154, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

155. 73) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:151, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:84, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 74) LCDR1 contains the amino acid sequence shown in SEQ ID NO:70, LCDR2 contains the amino acid sequence shown in SEQ ID NO:152, LCDR3 contains the amino acid sequence shown in SEQ ID NO:67; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:79, HCDR2 contains the amino acid sequence shown in SEQ ID NO:84, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

81. 75) LCDR1 contains the amino acid sequence shown in SEQ ID NO:121, LCDR2 contains the amino acid sequence shown in SEQ ID NO:160, and LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:133, HCDR2 contains the amino acid sequence shown in SEQ ID NO:161, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:131; or 76) LCDR1 contains the amino acid sequence shown in SEQ ID NO:121, LCDR2 contains the amino acid sequence shown in SEQ ID NO:160, LCDR3 contains the amino acid sequence shown in SEQ ID NO:123; and HCDR1 contains the amino acid sequence shown in SEQ ID NO:133, HCDR2 contains the amino acid sequence shown in SEQ ID NO:130, and HCDR3 contains the amino acid sequence shown in SEQ ID NO:

131.

4. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 3, wherein, The VL and VH are selected from at least one of the following combinations: 1) VL containing the amino acid sequence shown in SEQ ID NO:3 and VH containing the amino acid sequence shown in SEQ ID NO:2; 2) VL containing the amino acid sequence shown in SEQ ID NO:4 and VH containing the amino acid sequence shown in SEQ ID NO:2; 3) VL containing the amino acid sequence shown in SEQ ID NO:5 and VH containing the amino acid sequence shown in SEQ ID NO:2; 4) VL containing the amino acid sequence shown in SEQ ID NO:6 and VH containing the amino acid sequence shown in SEQ ID NO:2; 5) VL containing the amino acid sequence shown in SEQ ID NO:7 and VH containing the amino acid sequence shown in SEQ ID NO:2; 6) VL containing the amino acid sequence shown in SEQ ID NO:8 and VH containing the amino acid sequence shown in SEQ ID NO:2; 7) VL containing the amino acid sequence shown in SEQ ID NO:9 and VH containing the amino acid sequence shown in SEQ ID NO:2; 8) VL containing the amino acid sequence shown in SEQ ID NO:10 and VH containing the amino acid sequence shown in SEQ ID NO:2; 9) VL containing the amino acid sequence shown in SEQ ID NO:11 and VH containing the amino acid sequence shown in SEQ ID NO:2; 10) VL containing the amino acid sequence shown in SEQ ID NO:12 and VH containing the amino acid sequence shown in SEQ ID NO:2; 11) VL containing the amino acid sequence shown in SEQ ID NO:13 and VH containing the amino acid sequence shown in SEQ ID NO:2; 12) VL containing the amino acid sequence shown in SEQ ID NO:14 and VH containing the amino acid sequence shown in SEQ ID NO:2; 13) VL containing the amino acid sequence shown in SEQ ID NO:15 and VH containing the amino acid sequence shown in SEQ ID NO:2; 14) VL containing the amino acid sequence shown in SEQ ID NO:16 and VH containing the amino acid sequence shown in SEQ ID NO:2; 15) VL containing the amino acid sequence shown in SEQ ID NO:17 and VH containing the amino acid sequence shown in SEQ ID NO:2; 16) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:20; 17) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:21; 18) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:22; 19) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:23; 20) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:24; 21) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:25; 22) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:26; 23) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:27; 24) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:28; 25) VL containing the amino acid sequence shown in SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:29; 26) VL containing the amino acid sequence shown in SEQ ID NO:12 and VH containing the amino acid sequence shown in SEQ ID NO:22; 27) VL containing the amino acid sequence shown in SEQ ID NO:12 and VH containing the amino acid sequence shown in SEQ ID NO:24; 28) VL containing the amino acid sequence shown in SEQ ID NO:12 and VH containing the amino acid sequence shown in SEQ ID NO:26; 29) VL containing the amino acid sequence shown in SEQ ID NO:12 and VH containing the amino acid sequence shown in SEQ ID NO:29; 30) VL containing the amino acid sequence shown in SEQ ID NO:15 and VH containing the amino acid sequence shown in SEQ ID NO:22; 31) VL containing the amino acid sequence shown in SEQ ID NO:15 and VH containing the amino acid sequence shown in SEQ ID NO:24; 32) VL containing the amino acid sequence shown in SEQ ID NO:15 and VH containing the amino acid sequence shown in SEQ ID NO:26; 33) VL containing the amino acid sequence shown in SEQ ID NO:15 and VH containing the amino acid sequence shown in SEQ ID NO:29; 34) VL containing the amino acid sequence shown in SEQ ID NO:4 and VH containing the amino acid sequence shown in SEQ ID NO:27; 35) VL containing the amino acid sequence shown in SEQ ID NO:4 and VH containing the amino acid sequence shown in SEQ ID NO:28; 36) VL containing the amino acid sequence shown in SEQ ID NO:11 and VH containing the amino acid sequence shown in SEQ ID NO:21; 37) VL containing the amino acid sequence shown in SEQ ID NO:14 and VH containing the amino acid sequence shown in SEQ ID NO:21; 38) VL containing the amino acid sequence shown in SEQ ID NO:18 and VH containing the amino acid sequence shown in SEQ ID NO:21; 39) VL containing the amino acid sequence shown in SEQ ID NO:11 and VH containing the amino acid sequence shown in SEQ ID NO:22; 40) VL containing the amino acid sequence shown in SEQ ID NO:14 and VH containing the amino acid sequence shown in SEQ ID NO:22; 41) VL containing the amino acid sequence shown in SEQ ID NO:18 and VH containing the amino acid sequence shown in SEQ ID NO:22; 42) VL containing the amino acid sequence shown in SEQ ID NO:6 and VH containing the amino acid sequence shown in SEQ ID NO:27; 43) VL containing the amino acid sequence shown in SEQ ID NO:8 and VH containing the amino acid sequence shown in SEQ ID NO:27; 44) VL containing the amino acid sequence shown in SEQ ID NO:10 and VH containing the amino acid sequence shown in SEQ ID NO:27; 45) VL containing the amino acid sequence shown in SEQ ID NO:11 and VH containing the amino acid sequence shown in SEQ ID NO:27; 46) VL containing the amino acid sequence shown in SEQ ID NO:14 and VH containing the amino acid sequence shown in SEQ ID NO:27; 47) VL containing the amino acid sequence shown in SEQ ID NO:18 and VH containing the amino acid sequence shown in SEQ ID NO:27; 48) VL containing the amino acid sequence shown in SEQ ID NO:19 and VH containing the amino acid sequence shown in SEQ ID NO:27; 49) VL containing the amino acid sequence shown in SEQ ID NO:6 and VH containing the amino acid sequence shown in SEQ ID NO:30; 50) VL containing the amino acid sequence shown in SEQ ID NO:8 and VH containing the amino acid sequence shown in SEQ ID NO:30; 51) VL containing the amino acid sequence shown in SEQ ID NO:10 and VH containing the amino acid sequence shown in SEQ ID NO:30; 52) VL containing the amino acid sequence shown in SEQ ID NO:14 and VH containing the amino acid sequence shown in SEQ ID NO:30; 53) VL containing SEQ ID NO:18 and VH containing the amino acid sequence shown in SEQ ID NO:30; 54) VL containing SEQ ID NO:1 and VH containing the amino acid sequence shown in SEQ ID NO:31; 55) VL containing the amino acid sequence shown in SEQ ID NO:19 and VH containing the amino acid sequence shown in SEQ ID NO:31; 56) VL containing the amino acid sequence shown in SEQ ID NO:100 and VH containing the amino acid sequence shown in SEQ ID NO:96; 57) VL containing the amino acid sequence shown in SEQ ID NO:101 and VH containing the amino acid sequence shown in SEQ ID NO:96; 58) VL containing the amino acid sequence shown in SEQ ID NO:95 and VH containing the amino acid sequence shown in SEQ ID NO:106; 59) VL containing the amino acid sequence shown in SEQ ID NO:95 and VH containing the amino acid sequence shown in SEQ ID NO:107; 60) VL containing the amino acid sequence shown in SEQ ID NO:99 and VH containing the amino acid sequence shown in SEQ ID NO:102; 61) VL containing the amino acid sequence shown in SEQ ID NO:99 and VH containing the amino acid sequence shown in SEQ ID NO:104; 62) VL containing the amino acid sequence shown in SEQ ID NO:99 and VH containing the amino acid sequence shown in SEQ ID NO:107; 63) VL containing the amino acid sequence shown in SEQ ID NO:101 and VH containing the amino acid sequence shown in SEQ ID NO:102; 64) VL containing the amino acid sequence shown in SEQ ID NO:101 and VH containing the amino acid sequence shown in SEQ ID NO:104; 65) VL containing the amino acid sequence shown in SEQ ID NO:101 and VH containing the amino acid sequence shown in SEQ ID NO:107; 66) VL containing the amino acid sequence shown in SEQ ID NO:97 and VH containing the amino acid sequence shown in SEQ ID NO:105; 67) VL containing the amino acid sequence shown in SEQ ID NO:97 and VH containing the amino acid sequence shown in SEQ ID NO:106; 68) VL containing the amino acid sequence shown in SEQ ID NO:98 and VH containing the amino acid sequence shown in SEQ ID NO:96; 69) VL containing the amino acid sequence shown in SEQ ID NO:95 and VH containing the amino acid sequence shown in SEQ ID NO:103; 70) VL containing the amino acid sequence shown in SEQ ID NO:14 and VH containing the amino acid sequence shown in SEQ ID NO:143; 71) VL containing the amino acid sequence shown in SEQ ID NO:14 and VH containing the amino acid sequence shown in SEQ ID NO:144; 72) VL containing the amino acid sequence shown in SEQ ID NO:14 and VH containing the amino acid sequence shown in SEQ ID NO:145; 73) VL containing the amino acid sequence shown in SEQ ID NO:141 and VH containing the amino acid sequence shown in SEQ ID NO:22; 74) VL containing the amino acid sequence shown in SEQ ID NO:142 and VH containing the amino acid sequence shown in SEQ ID NO:22; 75) VL containing the amino acid sequence shown in SEQ ID NO:95 and VH containing the amino acid sequence shown in SEQ ID NO:157; and 76) VL containing the amino acid sequence shown in SEQ ID NO:156 and VH containing the amino acid sequence shown in SEQ ID NO:

103.

5. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 4, comprising a light chain and a heavy chain, wherein, The light chain comprises a light chain constant region of the amino acid sequence shown in SEQ ID NO:63, and / or the heavy chain comprises a heavy chain constant region of the amino acid sequence shown in SEQ ID NO:

64.

6. The antibody or antigen-binding fragment thereof according to claim 5, wherein the light chain and heavy chain are selected from at least one group of the following: 1) The light chain of the amino acid sequence shown in SEQ ID NO:34 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33; 2) The light chain of the amino acid sequence shown in SEQ ID NO:35 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33; 3) The light chain of the amino acid sequence shown in SEQ ID NO:36 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33; 4) The light chain of the amino acid sequence shown in SEQ ID NO:37 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33; 5) The light chain of the amino acid sequence shown in SEQ ID NO:38 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33; 6) The light chain of the amino acid sequence shown in SEQ ID NO:39 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33; 7) The light chain of the amino acid sequence shown in SEQ ID NO:40 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33; 8) The light chain of the amino acid sequence shown in SEQ ID NO:41 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33; 9) The light chain of the amino acid sequence shown in SEQ ID NO:42 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33; 10) The light chain of the amino acid sequence shown in SEQ ID NO:43 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33; 11) The light chain of the amino acid sequence shown in SEQ ID NO:44 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33; 12) The light chain of the amino acid sequence shown in SEQ ID NO:45 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33; 13) The light chain of the amino acid sequence shown in SEQ ID NO:46 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33; 14) The light chain of the amino acid sequence shown in SEQ ID NO:47 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33; 15) The light chain of the amino acid sequence shown in SEQ ID NO:48 and the heavy chain of the amino acid sequence shown in SEQ ID NO:33; 16) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:51; 17) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:52; 18) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:53; 19) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:54; 20) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:55; 21) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:56; 22) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:57; 23) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58; 24) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:59; 25) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:60; 26) The light chain of the amino acid sequence shown in SEQ ID NO:43 and the heavy chain of the amino acid sequence shown in SEQ ID NO:53; 27) The light chain of the amino acid sequence shown in SEQ ID NO:43 and the heavy chain of the amino acid sequence shown in SEQ ID NO:55; 28) The light chain of the amino acid sequence shown in SEQ ID NO:43 and the heavy chain of the amino acid sequence shown in SEQ ID NO:57; 29) The light chain of the amino acid sequence shown in SEQ ID NO:43 and the heavy chain of the amino acid sequence shown in SEQ ID NO:60; 30) The light chain of the amino acid sequence shown in SEQ ID NO:46 and the heavy chain of the amino acid sequence shown in SEQ ID NO:53; 31) The light chain of the amino acid sequence shown in SEQ ID NO:46 and the heavy chain of the amino acid sequence shown in SEQ ID NO:55; 32) The light chain of the amino acid sequence shown in SEQ ID NO:46 and the heavy chain of the amino acid sequence shown in SEQ ID NO:57; 33) The light chain of the amino acid sequence shown in SEQ ID NO:46 and the heavy chain of the amino acid sequence shown in SEQ ID NO:60; 34) The light chain of the amino acid sequence shown in SEQ ID NO:35 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58; 35) The light chain of the amino acid sequence shown in SEQ ID NO:35 and the heavy chain of the amino acid sequence shown in SEQ ID NO:59; 36) The light chain of the amino acid sequence shown in SEQ ID NO:42 and the heavy chain of the amino acid sequence shown in SEQ ID NO:52; 37) The light chain of the amino acid sequence shown in SEQ ID NO:45 and the heavy chain of the amino acid sequence shown in SEQ ID NO:52; 38) The light chain of the amino acid sequence shown in SEQ ID NO:49 and the heavy chain of the amino acid sequence shown in SEQ ID NO:52; 39) The light chain of the amino acid sequence shown in SEQ ID NO:42 and the heavy chain of the amino acid sequence shown in SEQ ID NO:53; 40) The light chain of the amino acid sequence shown in SEQ ID NO:45 and the heavy chain of the amino acid sequence shown in SEQ ID NO:53; 41) The light chain of the amino acid sequence shown in SEQ ID NO:49 and the heavy chain of the amino acid sequence shown in SEQ ID NO:53; 42) The light chain of the amino acid sequence shown in SEQ ID NO:37 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58; 43) The light chain of the amino acid sequence shown in SEQ ID NO:39 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58; 44) The light chain of the amino acid sequence shown in SEQ ID NO:41 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58; 45) The light chain of the amino acid sequence shown in SEQ ID NO:42 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58; 46) The light chain of the amino acid sequence shown in SEQ ID NO:45 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58; 47) The light chain of the amino acid sequence shown in SEQ ID NO:49 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58; 48) The light chain of the amino acid sequence shown in SEQ ID NO:50 and the heavy chain of the amino acid sequence shown in SEQ ID NO:58; 49) The light chain of the amino acid sequence shown in SEQ ID NO:37 and the heavy chain of the amino acid sequence shown in SEQ ID NO:61; 50) The light chain of the amino acid sequence shown in SEQ ID NO:39 and the heavy chain of the amino acid sequence shown in SEQ ID NO:61; 51) The light chain of the amino acid sequence shown in SEQ ID NO:41 and the heavy chain of the amino acid sequence shown in SEQ ID NO:61; 52) The light chain of the amino acid sequence shown in SEQ ID NO:45 and the heavy chain of the amino acid sequence shown in SEQ ID NO:61; 53) The light chain of the amino acid sequence shown in SEQ ID NO:49 and the heavy chain of the amino acid sequence shown in SEQ ID NO:61; 54) The light chain of the amino acid sequence shown in SEQ ID NO:32 and the heavy chain of the amino acid sequence shown in SEQ ID NO:62; 55) The light chain of the amino acid sequence shown in SEQ ID NO:50 and the heavy chain of the amino acid sequence shown in SEQ ID NO:62; 56) The light chain of the amino acid sequence shown in SEQ ID NO:113 and the heavy chain of the amino acid sequence shown in SEQ ID NO:109; 57) The light chain of the amino acid sequence shown in SEQ ID NO:114 and the heavy chain of the amino acid sequence shown in SEQ ID NO:109; 58) The light chain of the amino acid sequence shown in SEQ ID NO:108 and the heavy chain of the amino acid sequence shown in SEQ ID NO:119; 59) The light chain of the amino acid sequence shown in SEQ ID NO:108 and the heavy chain of the amino acid sequence shown in SEQ ID NO:120; 60) The light chain of the amino acid sequence shown in SEQ ID NO:112 and the heavy chain of the amino acid sequence shown in SEQ ID NO:115; 61) The light chain of the amino acid sequence shown in SEQ ID NO:112 and the heavy chain of the amino acid sequence shown in SEQ ID NO:117; 62) The light chain of the amino acid sequence shown in SEQ ID NO:112 and the heavy chain of the amino acid sequence shown in SEQ ID NO:120; 63) The light chain of the amino acid sequence shown in SEQ ID NO:114 and the heavy chain of the amino acid sequence shown in SEQ ID NO:115; 64) The light chain of the amino acid sequence shown in SEQ ID NO:114 and the heavy chain of the amino acid sequence shown in SEQ ID NO:117; 65) The light chain of the amino acid sequence shown in SEQ ID NO:114 and the heavy chain of the amino acid sequence shown in SEQ ID NO:120; 66) The light chain of the amino acid sequence shown in SEQ ID NO:110 and the heavy chain of the amino acid sequence shown in SEQ ID NO:118; 67) The light chain of the amino acid sequence shown in SEQ ID NO:110 and the heavy chain of the amino acid sequence shown in SEQ ID NO:119; 68) The light chain of the amino acid sequence shown in SEQ ID NO:111 and the heavy chain of the amino acid sequence shown in SEQ ID NO:109; 69) The light chain of the amino acid sequence shown in SEQ ID NO:108 and the heavy chain of the amino acid sequence shown in SEQ ID NO:116; 70) The light chain of the amino acid sequence shown in SEQ ID NO:45 and the heavy chain of the amino acid sequence shown in SEQ ID NO:148; 71) The light chain of the amino acid sequence shown in SEQ ID NO:45 and the heavy chain of the amino acid sequence shown in SEQ ID NO:149; 72) The light chain of the amino acid sequence shown in SEQ ID NO:45 and the heavy chain of the amino acid sequence shown in SEQ ID NO:150; 73) The light chain of the amino acid sequence shown in SEQ ID NO:146 and the heavy chain of the amino acid sequence shown in SEQ ID NO:53; 74) The light chain of the amino acid sequence shown in SEQ ID NO:147 and the heavy chain of the amino acid sequence shown in SEQ ID NO:53; 75) The light chain of the amino acid sequence shown in SEQ ID NO:108 and the heavy chain of the amino acid sequence shown in SEQ ID NO:159; 76) The light chain of the amino acid sequence shown in SEQ ID NO:158 and the heavy chain of the amino acid sequence shown in SEQ ID NO:

116.

7. A multispecific antigen-binding molecule comprising: (1) an anti-N3pGlu Aβ antibody or an antigen-binding fragment thereof as described in any one of claims 1 to 6; and (2) an antibody or an antigen-binding fragment thereof that binds to an antigen other than N3pGlu Aβ, said other antigen including, but not limited to, TfR, Tau, APP (amyloid precursor protein) or TREM2; preferably, said multispecific antigen-binding molecule is bispecific, trispecific or tetraspecific; preferably, said multispecific antigen-binding molecule is divalent, trivalent, tetravalent, pentavalent or hexavalent.

8. An isolated nucleic acid molecule, wherein, The nucleic acid molecule is of arbitrary length and comprises nucleotides, deoxynucleotides, and / or ribonucleotides, wherein the nucleic acid molecule encodes an antibody or an antigen-binding fragment thereof as described in any one of claims 1 to 6, or a multispecific antigen-binding molecule as described in claim 7.

9. A carrier, wherein, The carrier comprises the isolated nucleic acid molecule as described in claim 8.

10. A host cell, wherein, The host cell comprises the nucleic acid molecule of claim 8 or the vector of claim 9; preferably, the host cell is a eukaryotic cell or a prokaryotic cell; more preferably, the host cell is derived from mammalian cells, yeast cells, insect cells, Escherichia coli and / or Bacillus subtilis; even more preferably, the host cell is selected from Expi293 or CHO cells.

11. A method for preparing an antibody or antigen-binding fragment thereof as described in any one of claims 1 to 6, or a multispecific antigen-binding molecule as described in claim 7.

12. A pharmaceutical composition, wherein, The pharmaceutical composition comprises an antibody or antigen-binding fragment thereof as described in any one of claims 1 to 6 or a multispecific antigen-binding molecule as described in claim 7, and a pharmaceutically acceptable carrier, diluent or excipient; preferably, the pharmaceutical composition further comprises one or more therapeutic ingredients.

13. A method for treating Aβ-related diseases in mammals, comprising administering to a mammal requiring treatment, preferably a human, a therapeutically effective amount of an antibody or antigen-binding fragment thereof as described in any one of claims 1 to 6, a multispecific antigen-binding molecule as described in claim 7, or a pharmaceutical composition as described in claim 12; preferably, the Aβ-related disease is selected from Alzheimer's disease, pre-Alzheimer's disease, Down syndrome, and clinical or preclinical CAA; preferably Alzheimer's disease.

14. A method for preventing human Aβ abnormality-related diseases, comprising administering to a patient in need an effective amount of an antibody or antigen-binding fragment thereof as described in any one of claims 1 to 6, a multispecific antigen-binding molecule as described in claim 7, or a pharmaceutical composition as described in claim 12; preferably, the Aβ abnormality-related disease is selected from Alzheimer's disease, pre-Alzheimer's disease, Down syndrome, and clinical or preclinical CAA; preferably Alzheimer's disease.

15. Use of the antibody or antigen-binding fragment thereof of any one of claims 1 to 6, the multispecific antigen-binding molecule of claim 7, or the pharmaceutical composition of claim 12 in the preparation of a medicament for treating Aβ abnormality-related diseases; preferably, the Aβ abnormality-related diseases are selected from Alzheimer's disease, pre-Alzheimer's disease, Down syndrome, and clinical or preclinical CAA (cerebral amyloid angiopathy); preferably Alzheimer's disease.

16. Use of the antibody or antigen-binding fragment thereof of any one of claims 1 to 6, the multispecific antigen-binding molecule of claim 7, or the pharmaceutical composition of claim 12 in the treatment of Aβ abnormality-related diseases; preferably, the Aβ abnormality-related diseases are selected from Alzheimer's disease, pre-Alzheimer's disease, Down syndrome, and clinical or preclinical CAA (cerebral amyloid angiopathy); preferably Alzheimer's disease.

17. The antibody or antigen-binding fragment thereof of any one of claims 1 to 6, the multispecific antigen-binding molecule of claim 7, or the pharmaceutical composition of claim 12, for treating Aβ abnormality-related diseases; preferably, the Aβ abnormality-related diseases are selected from Alzheimer's disease, pre-Alzheimer's disease, Down syndrome, and clinical or preclinical CAA (cerebral amyloid angiopathy); preferably Alzheimer's disease.

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