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Colloidal gold chromatography anti-Sp100 antibody detection test paper and preparation method thereof

An antibody detection and colloidal gold technology, which is applied in the field of medical immunology, can solve the problem that the colloidal gold immunoassay test paper has not come out.

Active Publication Date: 2012-08-01
SHANGHAI KEXIN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0020] In terms of autoimmune diseases, there are currently some test strip products for detecting autoimmune diseases in foreign markets, but colloidal gold immune test strips for anti-Sp100 antibodies have not yet come out in domestic and foreign markets

Method used

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  • Colloidal gold chromatography anti-Sp100 antibody detection test paper and preparation method thereof
  • Colloidal gold chromatography anti-Sp100 antibody detection test paper and preparation method thereof
  • Colloidal gold chromatography anti-Sp100 antibody detection test paper and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] Example 1 Sp100 Antigen Protein Preparation

[0080] The Sp100 antigenic protein used in this test paper is a recombinant gene constructed by gene cloning technology, and then the all-human Sp100 antigenic protein is successfully expressed by prokaryotic expression technology. Wherein, the Sp100 antigen protein is derived from purified genetic engineering antigen protein.

Embodiment 2

[0081] Example 2 Antibody Preparation

[0082] Antibody A, Antibody C, and Antibody B were prepared by the following method. Anti-human IgG in Antibody A, Antibody C and Antibody B can generally be produced by multiple subcutaneous (sc) or intraperitoneal (ip) injections of purified immunogens and adjuvants on animals.

[0083] The injection solution was obtained by mixing 0.05mg~1mg of immune preparations (respectively for goats or mice) with 3 times the volume of Freund's complete adjuvant, and injected the injection solution into multiple parts of the animal skin. 5 to 1 / 10 of the original amount of human IgG in Freund's complete adjuvant mixture was injected subcutaneously into multiple sites to boost the immunization. After 7-14 days, the animals were bled, and the serum anti-human IgG titer was measured. Animals were boosted until titers plateaued. Methods for producing polyclonal antibodies are described in many immunology textbooks, for example, "Common Experimental...

Embodiment 3

[0086] Colloidal gold solution preparation

[0087] 0.01% HAuCl 4 Heat the solution to boiling, quickly add every 100mL HAuCl 4 Add an appropriate amount of reducing agent solution to the solution, and the color changes from blue, then light blue, to blue, then red after heating, and transparent orange-red after boiling for 7-10 minutes. Then filter with ultrafiltration or microporous membrane (0.45μm) to remove the polymer and other impurities that may be mixed therein. The prepared colloidal gold should be pure, translucent, free of sediment and floating matter, and discard when oily matter and a large amount of black granular precipitate appear on the liquid surface.

[0088] Wherein the reducing agent used can be trisodium citrate (Frens 1973), tannic acid-trisodium citrate (Slot and Gueeze 1985), white phosphorus, preferably use trisodium citrate, more preferably use 1% trisodium citrate sodium.

[0089] The glass containers used should be absolutely clean, and must b...

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Abstract

The invention discloses a colloidal gold chromatography anti-Sp100 antibody detection test paper and a preparation method thereof. The colloidal gold chromatography anti-Sp100 antibody detection test paper comprises a sample pad, a combination pad, a cellulose nitrate coated film and a water absorption pad, and the sample pad, the combination pad, the cellulose nitrate coated film and the water absorption pad are pasted on a base plate sequentially from one side of the base plate to the other side of the base plate, wherein the combination pad is coated with a gold-labeled antibody a and a gold-labeled antibody b; and the cellulose nitrate coated film is provided with a detection line and a quality control line, the detection line is coated with an Sp100 antigen protein, and the quality control line is coated with a gold-labeled antibody c. By adopting indirect immunoassay to introduce the Sp100 antigen protein in the invention to carry out process optimization on the combination pad and the sample pad, so high sensitivity, high specificity and high accuracy detection performances of the anti-Sp100 antibody are realized, and a reference basis is provided for the auxiliary diagnosis of primary biliary cirrhosis.

Description

technical field [0001] The invention belongs to the field of medical immunization applications, and in particular relates to a test paper for detecting anti-Sp100 antibodies by colloidal gold immunochromatography technology and a preparation method thereof. Background technique [0002] A variety of autoantibodies can be detected in PBC patients, such as anti-nuclear antibody (ANA), anti-mitochondrial antibody (AMA), anti-smooth muscle antibody (SMA), anti-liver-kidney microsomal antibody (LKM-1) and anti-soluble acidic nucleoprotein (Sp100) antibody, anti-nuclear membrane glycoprotein (Gp210) antibody, anti-promyelocytic leukemia protein (PML), anti-soluble liver antigen / hepatopancreatic antigen (SLA / LP) antibody, etc., the most important antibody is AMA, PBC is often accompanied by high-titre AMA, and AMA appears early in the course of the disease is a characteristic of this disease. [0003] AMA was first discovered in 1965 by WalRer et al. in the serum of PBC patients u...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558G01N33/532
Inventor 韩永俊高成秀张玥葛文斌孙宏彬钱杰
Owner SHANGHAI KEXIN BIOTECH
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