VEGF (Vascular Endothelial Growth Factor) antibody as well as preparation method and application thereof

An antibody, single-domain antibody technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, antibodies, etc.

Inactive Publication Date: 2014-05-07
CHENGDU INST OF BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problems existing in the existing VEGF antibody, the

Method used

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  • VEGF (Vascular Endothelial Growth Factor) antibody as well as preparation method and application thereof
  • VEGF (Vascular Endothelial Growth Factor) antibody as well as preparation method and application thereof
  • VEGF (Vascular Endothelial Growth Factor) antibody as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] Example 1 Preparation of VEGF antibody of the present invention and detection of its affinity

[0090] In this example, the single-domain antibody Vh shown in SEQ ID NO: 1 and its fusion protein Vh-double (shown in SEQ ID NO: 7), trimer Vh-trime (shown in SEQ ID NO: 8) were prepared. shown), the fusion Vh-Fc (shown in SEQ ID NO: 9), and its molecular weight, affinity, etc. were detected.

[0091] 1. Antibody preparation

[0092] 1.1 Construction of plasmid

[0093] The C-terminus of the nucleotide sequence shown in SEQ ID NO:11, SEQ ID NO:17, SEQ ID NO:18 or SEQ ID NO:20 respectively adds 6 repeated CGU or CAC nucleotide sequences, that is, in Each expressed protein is followed by a tag composed of 6 histidines, abbreviated as (His) 6 In order to facilitate the detection of expression products, each nucleotide sequence was synthesized by Jiangsu Jinweizhi Biotechnology Co., Ltd., digested with EcoRI and NcoI, and respectively ligated into pET22b vector.

[0094] 1.2...

Embodiment 2

[0126] Example 2 Preparation of VEGF antibody of the present invention and detection of affinity

[0127] The antibodies involved in this example are: the single domain antibody Vh-YDA shown in SEQ ID NO:4 and its trimer antibody Vh-YDA-trime; SEQ ID NO:2, SEQ ID NO:3, SEQ ID Single-domain antibodies Y31, Q61, 30, and 94 shown in NO:5 and SEQ ID NO:6 respectively, using Avastin single-chain antibody (scFv) as a positive control antibody (Avastin single-chain antibody is based on editor-in-chief Shen Beifen, etc., "Recombinant Antibody", Science Press, 2005, p246-p250 was constructed according to the method described).

[0128] 1. Preparation of the antibody of the present invention

[0129] 1.1 Construction of plasmid

[0130] 6 repeats are added to the C-terminus of the nucleotide sequence shown in SEQ ID NO:14, SEQ ID NO:19, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:15 or SEQ ID NO:16 The CGU or CAC nucleotide sequence, that is, a tag consisting of 6 histidines is added after...

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Abstract

The invention discloses a VEGF (Vascular Endothelial Growth Factor) single-domain antibody as well as a fusion antibody and a polymer antibody of the VEGF antibody. The invention also discloses a nucleotide sequence for encoding the antibodies, an expression vector containing the nucleotide sequence, a recombinant bacterium containing the nucleotide sequence as well as a preparation method and applications of the antibodies. The VEGF antibody is low in molecular weight, high in plasticity and wide in application prospect.

Description

technical field [0001] The present invention relates to a new VEGF antibody and its preparation method and use. Background technique [0002] Vascular endothelial growth factor (VEGF) is a homodimeric glycoprotein that promotes the division and proliferation of vascular endothelial cells and induces angiogenesis in the human body. VEGF can cause rapid tumor growth by promoting angiogenesis and is an important tumor drug target. [0003] The VEGF antibody can combine with VEGF to inhibit its physiological activity, thereby inhibiting the rapid growth of tumors. In addition, the VEGF antibody can also treat age-related macular degeneration. Commonly used antibodies include whole antibodies, Fabs, and single-chain antibodies. At present, most VEGF antibodies are whole antibodies, such as bevacizumab (Avastin), HuMV833 monoclonal antibody, VEGF-Trap antibody, and a small number of single-chain antibodies, such as the single-chain antibody of bevacizumab. These antibody drugs ...

Claims

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Application Information

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IPC IPC(8): C07K16/22C07K16/46C12N15/13C12N15/62C12N15/70C12N1/21C12P21/02A61K39/395A61P35/00A61P27/02
Inventor 何勇智丛聪代云见张勇侠李坤李春阳王保成秦娇荣赵兆王明蓉何明跃
Owner CHENGDU INST OF BIOLOGICAL PROD
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