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Fish Vibrio. alginolyticus anti-idiotype antibody genetic engineering vaccine and preparation method thereof

An anti-idiotype antibody, genetic engineering vaccine technology, applied in genetic engineering, botanical equipment and methods, biochemical equipment and methods, etc., can solve problems such as fish resistance, antibiotic damage, and antigen activity destruction.

Inactive Publication Date: 2013-12-25
FOURTH MILITARY MEDICAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

Long-term use of antibiotics can lead to resistance in fish, and residual antibiotics can cause harm to humans through the food chain
The safety of the attenuated vaccine is difficult to grasp; and the inactivated vaccine prepared by physical and chemical methods may destroy the antigenic activity, and the remaining impurities may cause the inflammatory reaction of the fish and cause the fish to die

Method used

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  • Fish Vibrio. alginolyticus anti-idiotype antibody genetic engineering vaccine and preparation method thereof
  • Fish Vibrio. alginolyticus anti-idiotype antibody genetic engineering vaccine and preparation method thereof
  • Fish Vibrio. alginolyticus anti-idiotype antibody genetic engineering vaccine and preparation method thereof

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Embodiment Construction

[0065] The anti-idiotypic antibody against Vibrio alginolyticus that causes bacterial fish disease has good immunogenicity and safety, and the small molecule single-chain antibody (scFv) of the variable region of the anti-idiotypic antibody constructed by genetic engineering not only has anti- The good immunogenicity and safety of the idiotype antibody also has the advantages of small molecular weight, strong penetrating power, easy mass production and low cost. To this end, we applied RT-PCR technology to amplify the heavy chain variable region gene (V H ) and the light chain variable region (V L ) gene fragment, cloned and sequenced, constructed the scFv expression vector pEr132a-AL by genetic recombination, transformed Escherichia coli BL21 and induced the expression of the fusion protein of the antibody scFv with IPTG, and immunized Ba1b / c mice with the expressed fusion protein Perform immunogenicity identification.

[0066] The present invention will be further describe...

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Abstract

The invention relates to a fish Vibrio. alginolyticus anti-idiotype antibody genetic engineering vaccine, which is characterized by: utilizing a hybridoma cell strain (2F4) of a fish Vibrio. alginolyticus anti-idiotype antibody to clone heavy chain variable region (VH) and light chain variable region (VL) genes, performing genetic recombination to construct scFv and prokaryotic expression vector PET32a-AL, transforming the product into E.coli BL21 and subjecting it to IPTG induced expression; and carrying out immunogenicity identification on expressed recombinant protein by animal immunization test. The ScFv gene sequence has an overall length of 747 base pairs, encodes 249 amino acids, is in line with the variable region gene characteristics of mouse immunoglobulin, and contains a framework region (FRs), an antigen complementary determining region (CDRs) and antibody characteristic cysteine residue. ELISA and animal immunity tests show that the scFv of the anti-idiotype antibody has the same immunogenicity with vibrio alginolyticus.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a preparation method of a fish Vibrio alginolyticus anti-idiotypic antibody genetic engineering vaccine. Background technique [0002] Vibrio alginolyticus (Vibrio.alginolyticus) is one of the bacteria that cause disease in many valuable economic fish such as flounder and eel, which seriously threatens the development of aquaculture in my country. Can bring huge economic loss to fishery and aquaculture. Traditional methods of preventing and treating Vibrio alginolyticus infection mainly use antibiotics and attenuated and inactivated vaccines. The long-term use of antibiotics can make fish resistant, and the residual antibiotics can cause harm to humans through the food chain. The safety of the attenuated vaccine is difficult to grasp; and the inactivated vaccine prepared by physical and chemical methods may destroy the antigenic activity, and the remaining im...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K39/40A61P31/04C12N15/13C12N15/70C07K16/12
Inventor 付建芳黄威权涂艳阳付菊芳
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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