Antibody IgG against Staphylococcus aureus serine-rich repeat protein SraP and application

A staphylococcus and serine-rich technology, applied in the direction of anti-bacterial immunoglobulin, immunoglobulin, application, etc., can solve the problems of unsatisfactory effects and achieve high protective, high affinity, and high specific effects

Active Publication Date: 2019-01-15
中国人民解放军东部战区疾病预防控制中心
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Now many companies or research institutions have carried out research on monoclonal antibodies against Staphylococcus aureus, and achieved some exciting results. Many antibodies have entered the clinical research stage, but some effects are not ideal, such as targeting bacterial pods Membrane polysaccharide serum type 5 / 8, lipoteichoic acid (LTA) and ATP-binding cassette (ABC) transporters have all failed in phase III clinical trials

Method used

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  • Antibody IgG against Staphylococcus aureus serine-rich repeat protein SraP and application
  • Antibody IgG against Staphylococcus aureus serine-rich repeat protein SraP and application
  • Antibody IgG against Staphylococcus aureus serine-rich repeat protein SraP and application

Examples

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Embodiment 1

[0020] Example 1 Preparation and screening detection of mouse monoclonal antibody IgG

[0021] 1) Using hybridoma technology to prepare anti-SraP monoclonal antibody, the specific steps are as follows:

[0022] 1. Preparation of antigen (SraP L-Lectin Recombinant protein)

[0023] Build SraP L-Lectin Expression plasmid: extract the genome of Staphylococcus aureus USA300, follow the method of bacterial DNA extraction kit, and amplify SraP by PCR with the following primers L-Lectin Modular gene fragments,

[0024] F(5'-3'): CCGGATCCTTTGCGTCAGCAGCGACG;

[0025] R(5'-3'): CACAAGCTTTATATTCGAATGTTCCAAATTGTAC;

[0026] add at both ends BamHI and HindⅢ restriction endonuclease sites. Cloning the gene fragment into the pET28a expression vector to construct pET28a-SraP L-Lectin recombinant plasmid. The recombinant plasmid was successfully constructed by double enzyme digestion and DNA sequencing.

[0027]The above plasmid was transformed into Escherichia coli BL-21 competen...

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Abstract

The invention discloses an antibody IgG against Staphylococcus aureus serine-rich repeat protein SraP and application. The antibody can specifically bind to an L-Lectin motif of the serine-rich repeatprotein, can bind to Staphylococcus aureus surface anchored protein SraP to reduce cell adhesion and invasion of Staphylococcus aureus and the number of bacteria in blood of mice, and can be used asan action target for an anti-Staphylococcus aureus infection antibody in treatment intervention.

Description

technical field [0001] The invention relates to the cloning of the SraP gene of the Staphylococcus aureus rich serine repeat protein, the preparation technology of the mouse monoclonal antibody, and the application of the monoclonal antibody in the treatment and preventive medicine of the Staphylococcus aureus infection. Background technique [0002] Serine-rich repeat proteins (SRRPs) have been in Streptococcus parasanguis since 1998 ( Streptococcus parasanguinis Since it was first discovered in FW213), SRRPs have been found in other Streptococcus, Staphylococcus aureus ( Staphylococcus aureus ), Staphylococcus epidermidis ( Staphylococcus epidermidis ) etc. were found. Currently known, SRRPs are a class of glycoproteins located on the cell surface of Gram-positive bacteria that can mediate the adhesion between bacteria and bacteria or between host cells. SRRPs can mediate not only intra-species adhesion but also inter-species adhesion, enabling bacteria to coloni...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/12C12N15/13A61K39/40A61P31/04
CPCA61K2039/505A61P31/04C07K16/1271C07K2317/24
Inventor 周婷婷岳岩朱进郑峰
Owner 中国人民解放军东部战区疾病预防控制中心
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