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Anti-Siglec-15 antibody and application thereof in preparation of medicine

An antibody, humanized antibody technology, applied in the direction of anti-animal/human immunoglobulin, antibody, application, etc., can solve the problem of lack of drugs to treat siglec-15 related diseases

Pending Publication Date: 2022-05-06
SHANGHAI JMT BIO INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] In order to overcome the lack of drugs for the treatment of siglec-15-related diseases in the prior art, the present invention provides anti-Siglec-15 antibodies and their application in the preparation of medicaments

Method used

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  • Anti-Siglec-15 antibody and application thereof in preparation of medicine
  • Anti-Siglec-15 antibody and application thereof in preparation of medicine
  • Anti-Siglec-15 antibody and application thereof in preparation of medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0118] Embodiment 1, the preparation of Siglec-15 antigen and detection protein

[0119] Use the human Siglec-15 extracellular region (F20-F263) of UniProt Sialic acid-binding Ig-like lectin 15Isoform 1 as a template to obtain the gene sequence encoding the antigen of the present invention and the protein for detection, and the antibody heavy chain Fc fragment (such as human IgG1) or His tag recombination, expressed and purified by HEK293 or Expi293 cells, to obtain the antigen and detection protein of the present invention. SG5-H52H3 (His tag), wherein the human Siglec-15 extracellular region sequence is shown in SEQ ID NO: 75, wherein 1-19 amino acid residues are signal peptide sequences, and 40-158 amino acid residues are "V- set” domain, the 168-251 amino acid residues are the “C2-set” domain; the sequence of the recombinant protein (hS15-hFc for short) obtained by linking the full-length extracellular region of human Siglec-15 with the Fc fragment of human IgG1 is shown i...

Embodiment 2

[0120] Embodiment 2, preparation of anti-human Siglec-15 hybridoma monoclonal antibody

[0121] Fifteen mice (5 C57bl / 6, 5 balb / c, and 5 SJL) were immunized with hS15-hFc emulsified with adjuvant, and then immunized with hS15-hFc or hS15-D1-hFc 2 weeks later. Serum was taken for ELISA experiment to evaluate the antiserum titer. Select mice with higher serum titers for splenocyte fusion. Three days after the last booster immunization, spleen cells were taken for fusion, and a total of 5 rounds of fusion were carried out by means of electrofusion, and 50 hybridoma cells obtained after each fusion were plated in 96-well plates. The hybridoma supernatants were screened for positive clones by binding activity assays, and further subcloned. Positive clones were screened for sequencing by combining activity assays and in vitro immune cell function assays.

[0122] Results: 5 positive hybridoma clones were screened by binding activity test and in vitro immune cell function test, re...

Embodiment 3

[0161] Example 3, the binding of anti-human Siglec-15 antibody to Siglec-15 protein from different species

[0162] The variable region of the antibody obtained by hybridoma screening was combined with the constant region of human IgG1 (N297A) to synthesize chimeric antibodies, which were named 21F6C6, 75E1F2, 124B11F6, 174C11C9 and 255C12B10 chimeric antibodies, respectively. The binding activity experiment of Siglec-15 protein from different species and chimeric antibody was carried out. Siglec-15 protein from human / mouse / cynomolgus monkey species was from Beijing Baipusaisi Biotechnology Co., Ltd. The antigen was coated with 0.5 μg / ml, and the antibody was incubated with the antigen after gradient dilution from 1 μg / ml. The secondary antibody was Anti-Human IgG antibody labeled with horseradish peroxidase, and the OD was read after adding the substrate 450 value.

[0163] Results: All five anti-Siglec-15 chimeric antibodies had binding activity to human Siglec-15 and cynom...

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Abstract

The invention relates to the field of biological pharmacy. In particular to an anti-Siglec-15 antibody or an antigen-binding fragment thereof, a nucleic acid sequence for coding the anti-Siglec-15 antibody or the antigen-binding fragment thereof, an expression vector containing the nucleic acid sequence, a host cell containing the expression vector, a composition containing the anti-Siglec-15 antibody or the antigen-binding fragment thereof, and an application of the anti-Siglec-15 antibody or the antigen-binding fragment thereof. The invention relates to a bispecific antibody or an immune linker formed by connecting the anti-Siglec-15 antibody or the antigen-binding fragment thereof with a therapeutic agent, and application of the Siglec-15 antibody or the antigen-binding fragment thereof in preparation of a medicine for treating cancer. The Siglec-15 antibody or the antigen binding fragment of the Siglec-15 antibody can be combined with Siglec-15, and is used for treating diseases related to Siglec-15.

Description

technical field [0001] The present invention relates to the field of biopharmaceuticals, in particular to an anti-Siglec-15 antibody or an antigen-binding fragment thereof, a nucleic acid sequence encoding an anti-Siglec-15 antibody or an antigen-binding fragment thereof of the present invention, an expression vector comprising the nucleic acid sequence, an expression vector comprising the expression The host cell of the carrier, the composition comprising the anti-Siglec-15 antibody or its antigen-binding fragment of the present invention, the bispecific antibody or the anti-Siglec-15 antibody of the present invention or its antigen-binding fragment and the immune linker linked to the therapeutic agent, and the present invention The invention discloses the use of Siglec-15 antibody or its antigen-binding fragment in the preparation of medicine for treating cancer. Background technique [0002] Cancer has become an important killer endangering human health. Due to populatio...

Claims

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Application Information

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IPC IPC(8): C07K16/28C12N15/13C12N5/20A61K39/395A61K45/00A61K47/68A61P35/00A61P35/02
CPCC07K16/2803A61K39/3955A61K47/6849A61K45/00A61P35/00A61P35/02C07K2317/56C07K2317/565C07K2317/24C07K2317/52C07K2317/31C07K2317/92C07K2317/73A61K2039/505A61K2300/00A61K45/06C07K2317/30C07K2317/70
Inventor 何柯廖诗骅冯旭任宝兰杜钰涵陈英娇宋利平范艺
Owner SHANGHAI JMT BIO INC
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