Preparation method of rheumatoid factor antigen, detection kit and preparation method of detection kit

Abstract

The invention relates to a preparation method of a rheumatoid factor antigen, a detection kit and a preparation method of the detection kit. The preparation method of rheumatoid factor antigen comprises the following steps: uniformly mixing an IgG solution with a denaturing buffer at 4 DEG C; after IgG molecules and the denaturant are fully reacted, carrying out heat polymerization reaction in a constant-temperature water bath at 55-65 DEG C for 30-60 min to obtain multimeric modified IgG; filtering to remove impurities by using a dialysis buffer; diluting to the proper concentration with a stock buffer solution and preserving, and applying the prepared rheumatoid factor antigen to the detection kit. Compared with the prior art, the preparation method of rheumatoid factor antigen, detection kit and preparation method of detection kit can greatly improve the reagent performance of such products, and has a simple preparation method, the difference between the preparation batches is controllable, and large-batch industrial production can be performed.

Description

technical field [0001] The present invention relates to biological reagents, in particular to detection kits, in particular to the preparation of a highly specific polymeric IgG (rheumatoid factor antigen) and a rheumatoid factor latex with wide linear range, high sensitivity and low preparation cost Immunoturbidimetric detection kit and preparation method thereof. Background technique [0002] Rheumatoid factor (Rheumatoid Factor, RF) is an autoimmune antibody targeting denatured IgG, which was first discovered in the serum of patients with rheumatoid arthritis. Its essence is a kind of immunoglobulin, mainly 19S IgM rheumatoid factor, and 7S IgG and IgA, while IgE and IgD rheumatoid factor are relatively rare. Rheumatoid factor can recognize the CH2 and CH3 parts of denatured IgG Fc fragments, and has no species specificity. When the two form immune complexes, they can activate complement and cause tissue damage. RF mainly exists in the serum and joint fluid of patients ...

AI Technical Summary

Problems solved by technology

[0004] At present, the domestic RF latex immunoturbidimetric kits in the market generally have poor reaction specificity

The main raw material denatured IgG (rheumatoid factor antigen) is usually prepared by heat denaturation, which cannot guarantee to maintain the exposed state of the binding site, and the stability and batch-to-batch difference are poor; which further leads to the linear range and sensitivity of the kit. , specificity, stability and other indicators are not ideal, these technical issues need further in-depth research

[0005] Chinese patent CN105911285A discloses a kit for measuring rheumatoid factor, aiming to solve the problem of complex operation and low measurement accuracy in the detection process of rheumatoid factor in the prior art. The kit used is composed of reagent R1 and reagent R2. Grouped into kits, wherein the R1 reagent contains Tris buffer 30-230mmol / L, Tween-80 2.0-6.0mL / L, polyethylene glycol-8000 5-15g / L, EDTA 30-50mmol / L, azide Sodium 0.4~1.0g / L, the solvent is purified water; R2 reagent contains Tris buffer 50~150mmol / L, glycerol 5~25mmol / L, sodium azide 0.4~1.0g / L, bovine serum albumin 20~60g / L L latex-coated anti-human rheumatism factor antibody 0.5-4.0%, and its solvent is purified water; wherein, the latex-coated anti-human rheumatism factor antibody is a monoclonal complete monoclonal containing the Fab functional site that can specifically bind to the anti-human rheumatism factor antibody Antibodies or antibody fragments, so there are fewer rheumatoid factor binding sites on the latex microspheres per unit surface area, resulting in a narrow detection linear range, which cannot meet the clinical detection requirements

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