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

A technology for lining proteins and antigenic epitopes, which can be applied to peptides, hybrid peptides, specific peptides, etc., can solve the problems of lack of in-depth research on antigenic epitope peptides, and the lack of methods for simultaneous detection of α-lined protein antibodies.

Inactive Publication Date: 2007-06-13
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

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Embodiment 1

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

[0048] The aforementioned research work has proved that the α-cell protein fusion protein purified by genetic engineering can specifically bind to the anti-α-cell protein antibody (IgA or IgG) in the serum of patients with Sjogren’s syndrome, and then through specific It can be detected by the method (Cheney R, Hirokawa N, Levine J, Willard M, Intracellular movement of Fodrin. Cell Motil, 1983, 3(5-6): 649-55. Haneji, N., Nakamura T, Takio K , Et al, Identification of a-Fodrin as a candidate autoantigen in primary Sjogren's Syndrome. Science, 1997, 276: 604-7. Watanabc T, Tsuchida T, Kanda N, et al, Anti-a-Fodrin antibodies in Sjogren's Syndrome and lupus erythematosus.Arch Dermatol, 1999, 135: 535-539. Witte T, Matthias T, Amett CF, 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, t...

Embodiment 2

[0082] Example 2 Mixing concentration design of α-cell protein epitope polypeptide mixture

[0083] In order to meet the detection requirements of "mixed ELISA" combined with "antibody profile ELISA" method, in the present invention, the α-consin epitope polypeptides (SEQ ID NO. 1 and SEQ ID NO. 2) are used as separate detection antigens respectively. The single-specific ELISA method was used to detect the results of serum antibodies in 10 clinical patients with positive α-consin IgG antibody and 10 patients with positive α-consin IgA antibody. Using these two peptides as mixed antigens, adjusting the different proportions of the two peptides, the total serum IgG / IgA antibodies of these 20 patients with clinically diagnosed Sjogren’s syndrome were tested by "mixed ELISA" combined with "antibody profile ELISA". Through preliminary exploration and comparison results, the different ratios (1:2, 6:7, 9:14, 5:7, 11:14, 6) of the synthetic polypeptide of SEQ ID NO. 1 and the synthetic p...

Embodiment 3

[0090] Example 3 Detection and evaluation of the anti-α-consin polypeptide IgG / IgA total antibody in the serum of patients with the α-consin epitope polypeptide mixture

[0091] In the experiment of the present invention, aiming at the amino acid characteristics of synthetic peptides and the reaction of mixed antigens and antibodies, after a series of pH and reactive ion concentration adjustments, we determined to use self-prepared 1×PBS with pH 8.0 as the coating buffer ; At the same time, corresponding experiments were carried out for the mixing ratio and optimal concentration of the coated polypeptide mixture, and the α-cell protein polypeptide SEQ ID NO.1 and SEQ ID NO.2 were mixed in different proportions and reacted. The results showed that the two antigen peptides were mixed and diluted to 0.7μg / ml in the ratio of 11:14 with the above self-made 1×PBS coating buffer with pH 8.0, 100μl was added to each well, and incubated at 37°C for 4 hours. Serum antibody detection has the...

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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 epitope polypeptides, the establishment of immunological detection methods, and the preparation and application of clinical diagnostic reagents, in particular to the screening of α-cell protein epitope polypeptides and α-cell protein The design and composition of the epitope peptide mixture to obtain the two best diagnostically meaningful amino acid sequences corresponding to the epitope peptides of the α-cell protein, and establish a method for detecting the total antibody of the α-cell protein peptide IgG / IgA in the patient’s serum Immunological methods, and the application of the two best synthesized α-cell protein epitope polypeptides as mixed antigens in the preparation of drugs for the diagnosis of Sjogren’s syndrome. [0002] Specifically, the present invention relates to a new epitope polypeptide of human Sjogren’s syndrome antigen protein and its corresponding amino ac...

Claims

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Application Information

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