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Staphylococcus aureus Efb protein C-end antigen epitope, as well as preparation method and usage thereof

A staphylococcus, epitope technology, used in protein engineering and molecular biology and immunology

Active Publication Date: 2011-03-23
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the Efb protein has a complement inhibitory effect, it is expected to become a new type of drug for the treatment of autoimmune diseases and anti-inflammation, and the functional domain of the Efb protein as the leading molecule for the development of related drugs has not yet been reported.

Method used

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  • Staphylococcus aureus Efb protein C-end antigen epitope, as well as preparation method and usage thereof
  • Staphylococcus aureus Efb protein C-end antigen epitope, as well as preparation method and usage thereof
  • Staphylococcus aureus Efb protein C-end antigen epitope, as well as preparation method and usage thereof

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Experimental program
Comparison scheme
Effect test

Embodiment

[0043] 1. Phage display peptide library screening for anti-rEfb antibody-binding peptides

[0044] Ph.D.-12 peptide library kit was purchased from NEB Company, with a library capacity of 2.7×10 9 , with a titer of 1.5×10 13 / μl.. Escherichia coli ER2537 is the host strain of the peptide library. For the screening process, refer to the instruction manual of the phage display peptide library. Each round of screening was coated with anti-rEfb antibody (10 μg per well), and 1 × 10 11 Phage. The degree of phage enrichment was calculated by "input / output ratio". After three rounds of screening, positive clones were selected for sequencing. The specific binding of positive phage clones to anti-rEfb antibody was determined by ELISA. Anti-rEfb antibody (10 μg per well) was coated on a 96-well plate (Nunc Company) overnight at 4°C. Unbound antibodies were poured off and blocked with 3% BSA at 37°C for 1 hour. Put 1×10 in each hole 9 Phage were incubated at 37°C for 2 hours. T...

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Abstract

The invention belongs to the field of protein engineering, molecular biology and immunology, in particular to a protein Efb antigen epitope as the source of staphylococcus aureus, as well as a preparation method and a usage thereof. In the invention, a strip of antigen epitope peptide EC1 is obtained by recombining and expressing the Efb protein in vitro and combining a bacteriophage technique; and the enzyme-linked immuno sorbent assay (ELISA) is captured through a CH50 test and C3 / Fibrinogen and the tests of the corresponding mutant protein are established to determine that the EC1 peptide has the function of inhabiting the complement, and thus, the EC1 peptide has good application prospects in the research and the development of medicines for treating autoimmune diseases and medicines for resisting inflammation.

Description

Technical field: [0001] The present invention relates to the fields of protein engineering and molecular biology and immunology, specifically, the present invention relates to the protein Efb ( E xcelluar f ibrinogen- b inding protein) antigenic epitope and its preparation method and use. Background technique: [0002] Staphylococcus aureus (Staphylococcus aureus) is a widespread Gram-positive pathogen that can cause a variety of purulent inflammations, and even life-threatening sepsis, endocarditis, pneumonia, and meningitis et al [Foster T J (2005) Nat Rev Microbiol 1289(3): 948-958]. The persistent infection of Staphylococcus aureus is related to its ability to produce a variety of immune regulatory molecules that interact with the host's immune system. Among them, superantigen and Protein A protein are relatively well-studied immune regulatory proteins, which can respectively affect T cell-mediated Cellular immunity and antibody-mediated humoral immune responses [Fan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C12N15/31A61K38/10A61K31/7088A61P31/00A61P37/02
Inventor 杨光高亚萍张鑫董洁刘玉
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA