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Staphylococcal enterotoxin micromolecule antibody and its preparation method and use

A staphylococcus enteric and small-molecule antibody technology, which is applied in the direction of analyzing materials, anti-bacterial immunoglobulin, and using carriers to introduce foreign genetic materials, can solve problems such as disappearance of secretion ability, stability defects, and hindered protein activity performance, and achieve Strong affinity and high specificity

Inactive Publication Date: 2013-07-31
TIANJIN UNIV
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  • Abstract
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Problems solved by technology

In 2009, Kalinina et al. established a prokaryotic expression system for anti-SEC1 gene remodeling antibody, but due to the lack of protein modification function of the expression system, its protein activity was blocked. The antibody was different from SEA, SEB, SED, SEE, SEG and SEI. degree of cross-reactivity, and the specific binding ability needs to be improved; in 2010, Pawan et al. prepared an anti-SEB single-chain antibody with strong specificity and high affinity through the pCANTAB5E vector system, but the study found that the secretion ability of the expression system gradually disappeared over time, and the stability Defect highlights

Method used

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  • Staphylococcal enterotoxin micromolecule antibody and its preparation method and use
  • Staphylococcal enterotoxin micromolecule antibody and its preparation method and use
  • Staphylococcal enterotoxin micromolecule antibody and its preparation method and use

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Embodiment Construction

[0030] 1. Preparation of staphylococcal enterotoxin monoclonal antibody hybridoma cell 5C12

[0031] Using SEO-GST fusion protein as the immunization antigen, immunize 6-week-old Balb / c mice 4 times, 2 weeks / time: the initial immunization adopts the method of adding the same amount of Freund's complete adjuvant to subcutaneously inject multiple points on the back of the neck, and the dose of immunization antigen is 100 μg / rat; the second immunization adopts the method of adding the same amount of Freund’s incomplete adjuvant into multiple subcutaneous injections on the back of the neck, and the dose of immune antigen is 100 μg / rat; the third and final immunization does not add adjuvant, and the tail vein is used to immunize The dose of antigen was 50 μg / monkey. The Balb / c mice that had been immunized were sacrificed, the spleen was aseptically removed, and the immune spleen cell suspension was prepared for future use. In vitro fusion of immune spleen cells and myeloma SP2 / 0 c...

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Abstract

The invention discloses a staphylococcal enterotoxin micromolecule antibody and its preparation method and use. The staphylococcal enterotoxin micromolecule antibody has an amino acid sequence shown in the formula of SEQ ID No.1 in the sequence table. The preparation method comprises the following steps of designing and constructing eukaryotic mono-promoter co-expression vectors of staphylococcal enterotoxin monoclonal antibody light chain and heavy chain variable region genes, introducing cysteine residues to C- ends of the light chain and the heavy chain to form interchain disulfide bonds by an intracellular peptide fragment self-assembling principle, and simulating antigen binding domain spatial conformation of natural antibodies to obtain high-efficiency expression stable-secretion mammal cell line and the high-singularity strong-affinity micromolecule antibody. The staphylococcal enterotoxin micromolecule antibody can be used for staphylococcal enterotoxin detection, cell indirect immunofluorescence detection and flow cytometry detection.

Description

technical field [0001] The invention relates to a genetic engineering recombinant antibody, in particular to a staphylococcus enterotoxin small molecule antibody and its preparation method and application. Background technique [0002] Staphylococcal enterotoxins (Staphylococcal enterotoxins, SEs) are a group of simple bacteria produced by plasma coagulase or thermostable nuclease-positive staphylococci, such as Staphylococcus aureus (Staphylococcus aureus, SA) and Staphylococcus epidermidis (Staphylococcus usepidermidis, SE). Peptide-chain soluble extracellular protein toxin. The toxin family is related in structure and similar in virulence, but different in antigenicity. Among them, the enterotoxin produced by Staphylococcus aureus is the most pathogenic and is a common and important pathogenic factor leading to food poisoning. So far, many countries in the world have reported outbreaks of Staphylococcus aureus one after another. Among them, the food poisoning caused by S...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/12C12N15/11C12N15/85G01N33/569
Inventor 黄金海刘鹏翀孙盈
Owner TIANJIN UNIV
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