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Colloidal gold testing card for testing sjogren's syndrome alpha-fodrin antibody

A lining protein and detection card technology is applied in the field of rapid detection products for Sjögren's syndrome alpha- lining protein antibody, which can solve the problems of unsuitable on-site rapid detection and complicated operation, and achieve rapid and rapid detection and clear and easy results. The effect of recognition

Inactive Publication Date: 2011-11-09
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In recent years, the commonly used antibody detection method is mostly ELISA method, but the detection requires special equipment, and requires a long time of pre-hybridization and hybridization washing process, and requires professional and technical personnel to carry out, the operation is complicated, and it is not suitable for rapid on-site detection

Method used

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  • Colloidal gold testing card for testing sjogren's syndrome alpha-fodrin antibody
  • Colloidal gold testing card for testing sjogren's syndrome alpha-fodrin antibody
  • Colloidal gold testing card for testing sjogren's syndrome alpha-fodrin antibody

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Embodiment 1

[0018] Example 1: Preparation of colloidal gold test card for detecting Sjogren's syndrome α-fodrin antibody

[0019] 1. Preparation of macromolecule-coupled α-fodrin epitope polypeptide:

[0020] According to the research results of the α-fodrin epitope region, combined with computer software to predict the trend of the α-fodrin epitope, the peptides of the 46-59 amino acid region of α-fodrin were designed and synthesized according to the antigenic trend index. The solid-phase method is artificially synthesized, and the HPLC purification purity is >95%. The polypeptide is coupled with BSA, OVA, KLH and other macromolecules by the EDC method, and the HPLC purification purity is >98%.

[0021] 2. Preparation of nano gold (colloidal gold):

[0022] Add 100ml of 0.01% HAuCL to a 500ML round bottom flask 4 solution, magnetic stirring and heating to boiling; then add 2ml 1% sodium citrate to the above solution, after the solution turns wine red, continue to boil for 10 minutes, s...

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Abstract

The invention relates to a colloidal gold testing card for testing sjogren's syndrome alpha-fodrin autoantibody. The testing card comprises orderly arranged components of, from left to right: a sample pad, glass fiber, a nitrocellulose membrane, and absorbent paper. Recombinant protein G labeled by nano-sized colloidal gold is applied on the glass fiber. Macromolecular-protein-coupled alpha-fodrin epitope polypeptide and recombinant protein G resistant antibody are fixed on the nitrocellulose membrane, wherein the sequence of alpha-fodrin epitope polypeptide is represented by SEQ ID NO: 1. Recombinant protein G resistant antibody fixed on the end close to the absorbent paper forms a control line, and macromolecule-protein-coupled alpha-fodrin epitope polypeptide fixed on the end close to the glass fiber forms a test line. The colloidal gold testing card provided by the present invention has advantages of easy preparation and quick testing. With the testing card, no professional technicist or expensive equipment is required. The testing card is suitable for large batch testing, and is suitable to be used in primary areas. The testing card has auxiliary diagnosis functions on early-stage sjogren's syndrome, and intermediate and advanced stage sjogren's syndromes.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a rapid detection product of Sjogren's syndrome α-fodrin (α-fordin) protein antibody. Background technique [0002] Primary sjögren's syndrome (pSS) is an autoimmune disease that causes abnormal cellular and humoral immune responses under the influence of multiple factors such as genetics, viral infection, and abnormal sex hormones. Foreign and domestic studies by He Jing have confirmed that α-fodrin is closely related to the pathogenesis of primary Sjögren's syndrome, which has important reference significance for the diagnosis of this disease. [0003] Chen Qiaolin et al. further used bioinformatics methods to predict and analyze the α-fodrin epitope, and recombinantly expressed the α-fodrin fragment containing overlapping sequences in vitro as a fusion protein, and screened and analyzed the α-fodrin epitope. Identification, and it was found that amino acids 1 to 59 in its sequence ...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/552
Inventor 方志远曾令文
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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