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Application of novel polypeptide in diagnosis of rheumatoid arthritis

A technology for arthritis and rheumatoids, applied in the field of immunological diagnosis, which can solve problems such as unclear pathogenesis and self-antigen detection antibodies

Inactive Publication Date: 2012-05-30
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

[0004] The appearance of a large number of autoantibodies is one of the important characteristics of autoimmune diseases, but because its pathogenesis is still unclear, it is difficult to directly use autoantigens to detect antibodies

Method used

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  • Application of novel polypeptide in diagnosis of rheumatoid arthritis
  • Application of novel polypeptide in diagnosis of rheumatoid arthritis
  • Application of novel polypeptide in diagnosis of rheumatoid arthritis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0020] 1. Phage display peptide library screening for RA-specific antibody-binding peptides

[0021] 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 total IgG in RA patient serum (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 total IgG antibodies was determined by ELISA. Total IgG 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 ...

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Abstract

The invention provides novel polypeptide used for diagnosis of rheumatoid arthritis. An IgG (immunoglobulin G) type antibody of antigen of the polypeptide has high sensitivity and specificity in diagnosis of rheumatoid arthritis.

Description

Technical field: [0001] The invention relates to the field of immunological diagnosis, in particular, the invention relates to a preparation method of a novel polypeptide (RAP) and its application in the diagnosis of rheumatoid arthritis. Background technique: [0002] Rheumatoid arthritis (RA) is a systemic autoimmune disease based on joint synovial inflammation and bone destruction. The prevalence rate in my country is 0.34%. There are more than 5 million RA patients in China. Most of them are young and middle-aged laborers, and it is a major disabling disease that seriously endangers human health. So far, the pathogenesis of this disease is still unclear, and the treatment is mainly non-specific symptomatic treatment, which temporarily relieves and improves the condition, but cannot fundamentally control the occurrence of synovial inflammation and the progress of bone destruction. [0003] In recent years, a citrulline-related autoantibody system with high specificity for...

Claims

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Application Information

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IPC IPC(8): C07K7/08C12N15/12A61K38/10G01N33/53C12Q1/68
Inventor 杨光高亚萍柳川董洁刘玉
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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