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Preparation method and application of a kind of human source anti-vibrio vulnificus hemolysin protein (vvh) antibody

A technology of Vibrio vulnificus and antibodies, applied in anti-bacterial immunoglobulin, botany equipment and methods, biochemical equipment and methods, etc., can solve the problems that have not 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: 2018-07-13
PLA NAVY GENERAL HOSIPTAL
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  • Abstract
  • Description
  • Claims
  • Application Information

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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, Yersinia pestis, Klebsiella pneumoniae, etc.) and viruses (such as respiratory coronavirus, dengue virus, etc.) Chain antibody came out, but there is no research report on anti-Vibrio vulnificus humanized antibody

Method used

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  • Preparation method and application of a kind of human source anti-vibrio vulnificus hemolysin protein (vvh) antibody
  • Preparation method and application of a kind of human source anti-vibrio vulnificus hemolysin protein (vvh) antibody
  • Preparation method and application of a kind of human source anti-vibrio vulnificus hemolysin protein (vvh) 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 "Sblattero D, Bradbury A. Exploiting recombination in singlebacteria to make large phage antibody libraries. Nat Biotechnol, 2000; 18:74." The specific steps are as follows:

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

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Abstract

The invention discloses a preparation method and application of a human anti-vibrio vulnificus hemolysin protein (VVH) antibody. The human source anti-vibrio vulnificus hemolysin protein (VVH) antibody of the present invention is a human source antibody and has the ability of neutralizing the vibrio vulnificus hemolysin protein VVH. The anti-VVH antibody provided by the present invention includes a light chain variable region and a heavy chain variable region, and the amino acid sequence of the heavy chain variable region is amino acid residues 135-260 from the amino terminal of sequence 3 in the sequence listing , the amino acid sequence of the light chain variable region is amino acid residues 1-113 from the amino terminal of Sequence 3 in the sequence listing. The present invention uses solid-phase Vibrio vulnificus VVH as an antigen and uses the constructed human-derived natural large-capacity phage antibody library to successfully screen human-derived anti-Vibrio vulnificus VVH antibodies. After ELISA identification, the anti-VVH antibody of the present invention has It has good specificity and has broad application prospects in the 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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Patent Type & Authority Patents(China)
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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