Alpha-fodrin antigen epi-position polypeptide mixture and its use

A technology of fodrin and antigenic epitope, which is applied in the direction of peptide, hybrid peptide, specific peptide, etc., can solve the problems of lack of in-depth research on antigenic epitope polypeptide, lack of methods for simultaneous detection of α-fodrin antibody, etc.

Inactive Publication Date: 2011-09-21
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

Exploratory research including serum antibody detection and epitope peptide screening has been carried out in the field of Sjogren's syndrome-related antigen-α-fodrin, but there are still many problems to be clarified and resolved, such as: high specificity, There is still a lack of in-depth research on the epitope polypeptides for the detection of highly sensitive α-fodrin IgG and IgA antibodies; there is still a lack of methods for the simultaneous detection of α-fodrin IgG / IgA antibodies

Method used

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  • Alpha-fodrin antigen epi-position polypeptide mixture and its use
  • Alpha-fodrin antigen epi-position polypeptide mixture and its use
  • Alpha-fodrin antigen epi-position polypeptide mixture and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Screening and identification of α-fodrin epitope polypeptide

[0048] The aforementioned research work has proved that the α-fodrin fusion protein purified by genetic engineering can specifically bind to the anti-α-fodrin antibody (IgA or IgG) in the serum of patients with Sjögren's syndrome, and then pass specific It can be detected by the method of (Cheney R, Hirokawa N, Levine J, Willard M, Intracellular movementof Fodrin.Cell Motil, 1983, 3(5-6): 649-55.Haneji, N., Nakamura T, Takio K , et al, Identification of α-Fodrin as a candidate autoantigen in primary Sjogren's Syndrome.Science, 1997, 276:604-7.Watanabc T, Tsuchida T, Kanda N, et al, Anti-α-Fodrin antibodies in Sjogren's Syndrome and lupus erythematosus. Arch Dermatol, 1999, 135: 535-539. Witte T, Matthias T, Arnett C F, et al, IgA and IgG autoantibodies against α-Fodrin as marker for Sjogren's Syndrome. J Rheumatol, 2000, 27: 2617-20.) . In the experiment of the present invention, the inventor sel...

Embodiment 2

[0083] Example 2 Mixing concentration design of α-fodrin antigen epitope polypeptide mixture

[0084] In order to meet the needs of "mixed ELISA" combined with "antibody spectrum ELISA" method detection, in the present invention, according to the α-fodrin epitope polypeptide (SEQ ID NO.1 and SEQ ID NO.2) as a separate detection antigen, using The monospecific ELISA method was used to detect the serum antibody results of 10 cases of α-fodrin IgG antibody positive and 10 cases of α-fodrin IgA antibody positive in patients diagnosed with Sjogren's syndrome. Using these two polypeptides as mixed antigens, adjusting the different ratio concentrations of the two polypeptides, the serum IgG / IgA total antibodies of 20 patients with clinically confirmed Sjogren's syndrome were detected by "mixed ELISA" combined with "antibody profile ELISA". Through preliminary exploration and comparison results, further select the different proportioning concentrations of the synthetic polypeptide of ...

Embodiment 3

[0091] Example 3 Detection of Anti-α-fodrin Antigen Epitope Polypeptide Mixture on Anti-α-fodrin Polypeptide IgG / IgA Total Antibody in Serum of Patients and Evaluation of Its Value

[0092] In the experiment of the present invention, aiming at the amino acid characteristics of the synthetic polypeptide and the reaction of the mixed antigen and the antibody, after a series of pH value and reaction ion concentration adjustments, we determined that the self-prepared 1×PBS with pH 8.0 was used as the coating buffer At the same time, corresponding experiments were carried out on the mixing ratio and optimal concentration of the coated polypeptide mixture, and the α-fodrin polypeptide SEQ ID NO.1 and SEQ ID NO.2 were mixed in different proportions and then reacted. The results showed that the above pH 8.0 self-prepared 1×PBS coating buffer was used to mix and dilute the two antigen polypeptides at a ratio of 11:14 to 0.7 μg / ml, add 100 μl to each well, and incubate at 37°C for 4 hour...

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Abstract

The invention relates to screen for antigen epitope polypeptide, and building up of immunology detection method and the manufacturing and application for reagent for clinical diagnosis, especially for alpha-cyst albumen antigen epitope polypeptide.

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 detection methods and the preparation and application of clinical diagnostic reagents, in particular to the screening of α-fodrin antigenic epitope polypeptides and α-fodrin Design and composition of antigen epitope polypeptide mixture, obtain the amino acid sequences corresponding to two optimal α-fodrin epitope polypeptides with diagnostic significance, and establish a method for detecting the total antibody of α-fodrin polypeptide IgG / IgA in serum of patients An immunological method, and the application of two synthetic optimal α-fodrin epitope polypeptides as mixed antigens in the preparation of drugs for diagnosing Sjögren's syndrome. [0002] Specifically, the present invention relates to a novel epitope polypeptide and its corresponding amino acid sequence of human Sjögren's syndrome antigenic protein...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00G01N33/53G01N33/68
Inventor 陈巧林曾令文
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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