Preparation method and application for human-derived vibrio vulnificus hemolysin (VVH) resistant antibody

A technology of Vibrio vulnificus and antibodies, which is applied in anti-bacterial immunoglobulin, botany equipment and methods, biochemical equipment and methods, etc. It can solve the problems that have not yet been seen in the research reports of humanized antibodies against Vibrio vulnificus. Achieve the effects of short circulation half-life, strong penetrating power and broad application prospects

Active Publication Date: 2015-12-30
PLA NAVY GENERAL HOSIPTAL
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  • Abstract
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Problems solved by technology

Using antibody library screening technology, there are currently available ScFv human-sourced monoclonal antibodies against various bacteria (such as Streptococcus pneumoniae, Mycobacterium tuberculosis, Yersi

Method used

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  • Preparation method and application for human-derived vibrio vulnificus hemolysin (VVH) resistant antibody
  • Preparation method and application for human-derived vibrio vulnificus hemolysin (VVH) resistant antibody
  • Preparation method and application for human-derived vibrio vulnificus hemolysin (VVH) resistant antibody

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Embodiment 1

[0036] Embodiment 1, the preparation method of anti-VVH antibody

[0037] 1. Preparation of large-capacity human phage antibody library

[0038] The method in the reference "SblatteroD, BradburyA. Exploitingrecombinationinsinglebacteriatomakelargephageantibodylibraries. NatBiotechnol, 2000; 18:74." prepares a large-capacity human phage antibody library, and the specific steps are as follows:

[0039] 1. Take out 7×10 antibodies from the original phage antibody library 12 Phage particles of cfu (cloning forming unit) superinfect BS1365 cells, and the multiplicity of infection (MOI) is 100 / 1. For specific steps, refer to the literature "Wang Yan, Wang Yuxiao, Chen Xiaosui, Hua Bing, Liu Xiaolin. For large-capacity phage antibodies Construction and identification of library expression vectors, Chinese Journal of Immunology, 2003, 19(2), 93-96.”, 37°C water bath for 1 hour, so that multiple copies of the vector can enter the same cell, mediated by cre-loxp The targeted recombina...

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Abstract

The invention discloses a preparation method and application of a human-derived VVH resistant antibody. The human-derived VVH resistant antibody is a human-derived antibody and has the capacity of neutralizing VVH. The VVH resistant antibody provided by the invention comprises a light chain variable region and a heavy chain variable region, wherein the amino acid sequence of the heavy chain variable region is 135th-260th amino acid residues from an amino terminal in the sequence 3 in a sequence table; the amino acid sequence of the light chain variable region is 1st-113th amino acid residues from the amino terminal in the sequence 3 in the sequence table. According to the antibody disclosed by the invention, immobilized VVH is used as an antigen, and a constructed human-derived natural high-capacity phage antibody library is used, so that the human-derived VVH resistant antibody is successfully screened, and through ELISA identification, the VVH resistant antibody has good specificity, and has a wide application prospect in treatment of vibrio infection.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method and application of a human anti-vibrio vulnificus hemolysin protein (VVH) antibody. Background technique [0002] Vibrio vulnificus is a common pathogenic Vibrio with prominent pathogenicity in coastal waters. It naturally exists in coastal waters and marine animals and plants. It can enter the human body through open wounds in the gastrointestinal mucosa or skin, causing cellulite Severe infection symptoms such as inflammation, septic shock, multiple organ failure, etc., even with active treatment with antibiotics, still have a high mortality rate (>50%), so there is great harm to human health and even survival and marine fisheries. . At present, the treatment of Vibrio vulnificus at home and abroad is still dominated by the traditional treatment method of antibiotics. The emergence of drug-resistant strains and drug residues have made the resear...

Claims

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Application Information

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IPC IPC(8): C07K16/12C12N15/13A61K39/40A61P31/04
Inventor 周丽君付凯飞王欲晓李军
Owner PLA NAVY GENERAL HOSIPTAL
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