Hybridoma cell strain 4A5 and vibrio splendidus flagellin FlgT monoclonal antibody

A hybridoma cell line, monoclonal antibody technology, applied in the direction of anti-bacterial immunoglobulin, immunoglobulin, biochemical equipment and methods, etc.

Inactive Publication Date: 2018-02-23
DALIAN OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, so far, there has been no report on the production of hybridoma cell lines that secrete FlgT monoclonal antibodies against Vibrio splendidus with FlgT protein

Method used

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Embodiment Construction

[0013] The hybridoma cell line 4A5 of the present invention has been preserved in the General Microorganism Center of China Microbiological Culture Collection Management Committee, and the preservation number is CGMCC No. 14311. Hybridoma cell line 4A5 secretes monoclonal antibody against flagellin FlgT of Vibrio splendidus.

[0014] The preparation method of hybridoma cell line 4A5 is as follows:

[0015] 1. Preparation of Immunizing Antigen

[0016] (1) Construction of recombinant expression vector: Design primers according to the FlgT nucleotide sequence (the FlgT nucleotide sequence is shown in SEQ ID NO.1, 762 bp) for PCR amplification, and introduce restriction endonuclease sites on both sides of the gene Point SgfI and MluI, insert expression vector pET-22b T7 promoter downstream, construct FlgT recombinant expression plasmid;

[0017] (2) Expression and purification of FlgT recombinant protein: E. coli The BL21 strain is an expression strain, and the FlgT recombina...

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Abstract

The invention discloses a hybridoma cell strain 4A5 and a vibrio splendidus flagellin FlgT monoclonal antibody. The hybridoma cell strain 4A5 is collected in China General Microbiological Culture Collection Center with the collection number of CGMCC No.14311. The hybridoma cell strain 4A5 is obtained by utilizing recombinant protein FlgT as an antigen to immunize a mouse, taking spleen cells of the mouse to be fused with SP2/0 cells, adopting a limiting dilution method to obtain the monoclonal antibody, and adopting an ELISA (Enzyme Linked Immunosorbent Assay) method to screen positive hybridoma cells; and the vibrio splendidus flagellin FlgT monoclonal antibody secreted by the hybridoma cell strain 4A5 has more specificity, and can be applied in the field of immunodetection.

Description

technical field [0001] The invention relates to a hybridoma cell strain and a monoclonal antibody, in particular to a hybridoma cell strain 4A5 and a monoclonal antibody to the flagellin FlgT of Vibrio candidiasis. Background technique [0002] Vibrio splendidus belongs to the Vibrio family ( Vibrionaceae ), is sea cucumber ( Apostichopus japonicus ) is the main pathogenic bacteria of sea cucumber allergy (rot skin syndrome) which is very common in farming. The disease is highly contagious and spreads widely, and can cause symptoms such as large-scale body surface ulcers, mouth swelling and ulceration, body atrophy, and viscera discharge in sea cucumbers. In addition, Vibrio splendidus can also cause disease and even death of various fish, bivalve molluscs, crustaceans and echinoderms, causing bleeding spots on both sides of the infected fish, especially near the tail, causing skin to rot and forming Ulcers, hyperemia at the base of rays of dorsal fin, pectoral fin, and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/12
CPCC07K16/1239
Inventor 李强黎睿君于子超王博雅叶仕根
Owner DALIAN OCEAN UNIV
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