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Colloidal gold chromatography anti-Ro52 antibody detection test paper and preparation method thereof

A technology of antibody detection and colloidal gold, which is applied in the field of medical immunization and can solve the problems that colloidal gold immunization 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

[0011] 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-Ro52 antibodies have not yet come out in domestic and foreign markets

Method used

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  • Colloidal gold chromatography anti-Ro52 antibody detection test paper and preparation method thereof
  • Colloidal gold chromatography anti-Ro52 antibody detection test paper and preparation method thereof
  • Colloidal gold chromatography anti-Ro52 antibody detection test paper and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Example 1 Preparation of RO52 Antigen Protein

[0071] The RO52 antigen protein used in this test paper is constructed by gene cloning technology to construct recombinant gene, and then the RO52 antigen protein which is all human origin is successfully expressed by prokaryotic expression technology. Wherein, the RO52 antigen protein is derived from purified genetic engineering antigen protein.

Embodiment 2

[0072] Example 2 Antibody Preparation

[0073] 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.

[0074] 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

[0077] Colloidal gold solution preparation

[0078] 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.

[0079] 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.

[0080] The glass containers used should be absolutely clean, and must b...

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Abstract

The invention discloses a colloidal gold chromatography anti-Ro52 antibody detection test paper and a preparation method thereof. The colloidal gold chromatography anti-Ro52 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 Ro52 antigen protein, and the quality control line is coated with a gold-labeled antibody c. By adopting indirect immunoassay to introduce the Ro52 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-Ro52 antibody are realized, and a reference basis is provided for the auxiliary diagnosis of Sjogren's syndrome.

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-Ro52 antibodies by colloidal gold immunochromatography technology and a preparation method thereof. Background technique [0002] SS-A antigen is a small ribonucleoprotein consisting of an RNA molecule and two different proteins (molecular weights of 52 and 60kDa, respectively). The specificity of anti-SS-A 52kDa is higher, while the antibody against SS-A 52kDa alone has no disease specificity, and it is easy to cross-react with other antibodies. [0003] In the differential diagnosis, anti-SSA / Ro antibodies are highly specific for Sjogren's syndrome and SLE. In particular, the immunoreactivity of anti-Ro antibodies with 60KD Ro protein (Ro60) and 52KD Ro protein (Ro52) was used to distinguish SLE from primary Sjogren's syndrome, respectively. SS-A / Ro antibodies are found in 85%-95% of Sjögren's syndrome patients. The...

Claims

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Application Information

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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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