Beta-fodrin antigen epitope polypeptide, and its screening method and use

An antigenic epitope, lining protein technology, applied in the direction of peptide sources, animal/human peptides, material testing products, etc., can solve the problem of lack of antigen sources and specific diagnostic methods, etc.

Inactive Publication Date: 2007-03-28
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among the currently known diagnostic antigens (proteins or polypeptides) for Sjogren's syndrome, there is still a lack of a marker with strong specificity and high positive rate for human Sjögren's syndrome; the lack of stable antigen sources and specific diagnostic methods is also a problem core

Method used

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  • Beta-fodrin antigen epitope polypeptide, and its screening method and use
  • Beta-fodrin antigen epitope polypeptide, and its screening method and use
  • Beta-fodrin antigen epitope polypeptide, and its screening method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Screening and identification of β-fodrin epitope polypeptide

[0041] The aforementioned research work has proved that the β-fodrin fusion protein purified by genetic engineering can specifically bind to the anti-β-fodrin antibody in the serum of patients with Sjogren's syndrome, and then it can be purified by specific methods. Detected (Masataka K, Tetsuroh O, Yoko O, Junichi K, and Yutaka K, Autoantibodies to theAmino-Terminal Fragment of β-Fodrin Expressed in Glandular Epithelial Cells in Patientswith sjogren's Syndrome. The Journal of Immunology, 2001, 167: 5449- 5456.). In the experiment of the present invention, the inventor selected the amino acid sequence corresponding to the ORF nucleic acid sequence of the full-length β-fodrin gene as the object, and adopted five international standard methods for predicting antigenic epitopes, including Standard, Karplus, Emini, Amphiphi, and Pellequer. The secondary structure (hydrophilicity, hydrophobicity, etc.)...

Embodiment 2

[0066]Example 2 Detection of β-fodrin epitope polypeptide on anti-β-fodrin epitope polypeptide IgG antibody in patient serum and its value evaluation

[0067] In the experiment of the present invention, for the amino acid characteristics of the synthetic polypeptide and its reaction with the antibody, after a series of adjustments of the pH value and the concentration of the reactant ion, we determined that the self-prepared PBS with pH 9.6 was used as the coating buffer; Corresponding experiments were carried out on the concentration of the coating antigen, and the β-fodrin epitope polypeptide SEQ ID NO.1 was serially diluted and then reacted. The results showed that the self-prepared PBS coating buffer with pH 9.6 above will The peptide was diluted to 0.5 μg / ml, 100 μl was added to each well, and after incubation at 37°C for 4 hours, the serum antibody detection was performed, and the effect was the best; in addition, with regard to the prevention of non-specific reactions, b...

Embodiment 3

[0114] Example 3 Application of β-fodrin epitope polypeptide as an antigen in the preparation of clinical Sjogren's syndrome diagnostic kit

[0115] The present inventors used the sandwich method solid-phase enzyme-linked immunosorbent assay (ELISA), coated the β-fodrin epitope polypeptide antigen on a microwell plate, and prepared β-fodrin epitope polypeptide IgG The kit for clinical diagnosis is used for the quantitative detection of anti-β-fodrin epitope polypeptide IgG antibody, which can complete the detection of 96 people in total. The specific content is as follows:

[0116] 1. Kit composition materials

[0117] One detachable microwell plate (1×8) with 12 strips coated with highly purified β-fodrin epitope polypeptide;

[0118] β-fodrin epitope polypeptide IgG standard: 0 / 6.3 / 12.5 / 25 / 50 / 100U / ml, 1 bottle each, 1.5ml per bottle;

[0119] 1 bottle of quality control serum (including IgG and negative quality control), 1.5ml;

[0120] 1 bottle of concentrated sample buff...

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Abstract

The invention relates beta-fordin antigen epitope polypeptide screening for gaining amino acid sequence corresponded with the optimum beta-fordin polypeptide. The beta-fordin antigent epitope polypeptide includes the following two that one is formed by the amino acid sequence showed in SEQ ID NO.1; another is derived from the above amino acid sequence by replacing, missing or adding one or many amino acid with antigen epitope function. It can used to specially test beta-fordin polypeptide IgG antibody for Sjogren syndrome patient. The invention also supplies the beta-fordin polypeptide IgG antibody immunology method, and its application used as antigen in Sjogren syndrome medicine preparation.

Description

technical field [0001] The invention belongs to the field of immunology, and relates to the screening of antigenic epitope polypeptides, the establishment of immunological methods and the application of clinical diagnostic reagents, in particular to the screening of β-fodrin antigenic epitope polypeptides to obtain the best β-fodrin antigenic polypeptides with diagnostic significance. - The amino acid sequence corresponding to the fodrin epitope polypeptide, and the establishment of an immunological method for detecting IgG antibodies to the β-fodrin epitope polypeptide in serum of patients, and the application of using it as an antigen in the preparation of drugs for diagnosing Sjögren's syndrome. technical background [0002] Sjogren's syndrome (SS) is a systemic autoimmune disease mainly characterized by mouth and eye dryness and involving multiple systems and organs throughout the body. It can be divided into primary and secondary Sjögren's syndrome. The disease mainly m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435G01N33/53
Inventor 陈巧林
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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