Preparation method of allergen-specific IgE antibody composite quality control product and allergen-specific IgE antibody composite quality control product

Abstract

The invention discloses a preparation method of an allergen-specific IgE antibody composite quality control product and the allergen-specific IgE antibody composite quality control product. The preparation method specifically comprises the following steps: preparing an allergen-specific IgE antibody; obtaining a target gene, and obtaining a constant region gene sequence and a variable region gene sequence of human IgE; the method comprises the following steps: designing a primer containing an XhoI restriction enzyme cutting site according to a heavy chain constant region gene sequence of human IgE, designing a primer containing an Hind III restriction enzyme cutting site according to a light chain constant region gene sequence of human IgE, and constructing an expression empty vector containing a light chain signal peptide, a heavy chain signal peptide, a human IgE heavy chain and a light chain C region gene fragment; constructing recombinant expression vectors of different allergens; transfecting the cells; cloning, recognizing and screening; the quality control product has the advantages of high titer, higher purity, favorable dilution linearity and favorable stability, and has higher use value in the aspects of quality control, conventional quality control, performance verification and product development of large-scale production.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a preparation method of an allergen-specific IgE antibody composite quality control product and the allergen-specific IgE antibody composite quality control product prepared by the method. Background technique [0002] Type I allergic diseases are quite common, and the total incidence rate of the population is as high as 10% to 30%. It is a major worldwide health problem and is listed by the World Health Organization (WHO) as one of the three major diseases that should be prevented and controlled in the 21st century. one. The occurrence of allergic diseases, IgE antibodies play a key role. The detection of allergen-specific IgE antibodies is of great significance in the auxiliary diagnosis, treatment and prevention of allergic diseases. The detection of allergen IgE generally adopts immunodiagnostic methods, such as enzyme-linked immunosorbent assay, flu...

AI Technical Summary

Problems solved by technology

At present, the conventional method of preparing IgE antibody quality control products is mainly to collect allergy-positive serum, which has huge defects. First, positive serum is not easy to collect, the cost is very high, and the supply is often insufficient; The amount is small, and it is difficult to produce large-scale and industrialized products. Third, the stability of natural blood products is poor, and they are easy to deteriorate or degrade during processing, packaging, transportation and use. It is difficult to be effective as a routine quality control product. Ensure consistency; Fourth, the composite quality control product used for the detection of multiple allergen-specific IgE antibodies will use multiple positive sera to mix, and the mixing and debugging of multiple specific IgE antibodies will be very cumbersome and difficult to adjust the value. The mixing of sera from different donor sources can also cause serum agglutination due to immune reactions, resulting in huge waste

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