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Application of TRAP protein in preparing medicine for treating infection of staphylococcus aureus

A staphylococcal infection, staphylococcus technology, applied in the direction of antibacterial drugs, peptide/protein components, medical preparations containing active ingredients, etc.

Active Publication Date: 2006-12-06
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the abuse of antibiotics has led to increasing drug resistance of Staphylococcus aureus, and the drug-resistant strains that have emerged include methicillin-resistant Staphylococcus aureus (MRSA) resistant to β-lactam antibiotics, and sugar-resistant strains resistant to glycopeptide antibiotics. Peptide Staphylococcus aureus (GISA), Vancomycin-resistant Staphylococcus aureus (VRSA), etc.
Because Staphylococcus aureus is very prone to drug resistance and there is no good solution, many commonly used antibiotics are ineffective against it, so finding a drug that can effectively control Staphylococcus aureus infection and is not easy to develop drug resistance is a clinical medical waiting solved problem

Method used

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  • Application of TRAP protein in preparing medicine for treating infection of staphylococcus aureus
  • Application of TRAP protein in preparing medicine for treating infection of staphylococcus aureus

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Experimental program
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Effect test

preparation Embodiment 1

[0032] Two Staphylococcus aureus families (TRAP protein (GenBank TM Accession number: AF202641 / AY248703)) acquisition of TRAP protein. According to the method of Yang, G, et al. (2005), according to the base sequence of GenBank, primers were designed, and the TRAP gene was amplified from the genomes of RN6390B and 04018 strains by PCR, and cloned into the pET-28a vector, Transform Escherichia coli, induce expression by IPTG, use Ni 2+ Purified by affinity resin, two corresponding TRAP proteins were obtained.

preparation Embodiment 2

[0034]Preparation of natural TRAP protein Use recombinant TRAP protein to prepare polyclonal antibody, couple recombinant TRAP protein to CNBr-activated Sepherose4B to prepare affinity strain purified antibody, couple purified antibody to CNBr-activated Sepherose4B, and use affinity purification method from The TRAP protein was purified from the ultrasonic supernatant of Staphylococcus aureus RN6390B, and further purified by HPLC to obtain natural purified TRAP protein with a purity greater than 95%.

preparation Embodiment 3

[0036] Preparation of fusion protein composed of TRAP protein and 12 amino acid polypeptide: select irrelevant peptide sequence (TPSIPSLWWTTP-Shen Tong et al., Cell and Molecular Immunology 2002, 18: 623-626), design primers, and obtain expression TRAP by PCR - Gene sequence of unrelated peptide fusion protein, cloned into pET-28a vector, induced expression by IPTG, transformed into Escherichia coli, using Ni 2+ The fusion protein composed of TRAP protein and 12 amino acid polypeptide was obtained by affinity resin purification.

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Abstract

An application of the TRAP protein in preparing the medicine for treating the Staphylococcus aureus infection is disclosed. Its advantages are high suppression action to Staphylococcus aureus, generation of the antibodies in human body, and no generation of resistance for Staphylococcus aureus.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, in particular to the application of TRAP protein in pharmacy. Background technique [0002] Staphylococcus aureus (hereinafter referred to as Staphylococcus aureus, Staphylococcus aureus) belongs to the genus Staphylococcus and is an important pathogenic bacteria for humans, which can cause pneumonia, endocarditis, burn and war wound infection, sepsis, toxic shock, etc. severe infection. Staphylococcus aureus is also a type of bacteria that causes food poisoning. Staphylococcus aureus is widely distributed in air, soil, water and tableware. Humans and animals have a higher carrier rate. Staphylococcus aureus can spread on a large scale through saliva and contact in crowded areas. The number of people infected with Staphylococcus aureus in hospitals alone exceeds millions every year. It is the main pathogenic bacterium of gram-positive sepsis, and also an important pathogenic bacterium of bu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/16A61P31/04
CPCA61K38/164A61P31/04A61K38/16A61K39/085
Inventor 杨光邵宁生高亚萍刘玉董洁李少华柳川沈倍奋范明
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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