Antigen protein and kit for detecting diphtheria antibody and preparation method

Abstract

The invention relates to a method for preparing an antigen protein for detecting a diphtheria antibody. The method is characterized by comprising the steps as follows: S1: infecting a baculovirus expression vector carrying a CRM197 protein expression cassette into insect cells so as to obtain infected insect cells; S2: carrying out cultivation and proliferation on the infected insect cells; and S3: carrying out crushing on the proliferated insect cells, and extracting the antigen protein in the insect cells. The invention further relates to an antigen protein prepared by the method and furtherrelates to a kit comprising the antigen protein. An expressed CRM197 protein is safe, non-toxic, high in yield and easy to separate and purify, meanwhile, has very high affinity with the diphtheria antibody, and is good in specificity.

Description

technical field [0001] The field of diphtheria prevention and treatment of the present invention, more particularly, relates to a method for preparing an antigenic protein for detecting diphtheria antibody, the antigenic protein prepared by the method, and a kit including the antigenic protein. Background technique [0002] Diphtheria is an acute upper respiratory infectious disease caused by Gram-positive Corynebacterium diphtheriae transmitted by air droplets. Diphtheria exotoxin is the main pathogenic factor. Its clinical features are mucous membrane congestion and swelling in the pharynx, larynx, nose, etc., with gray-white pseudomembrane formation, and systemic poisoning symptoms caused by bacterial exotoxins. In severe cases, toxic myocarditis and peripheral nerve paralysis may occur. Before the introduction of the diphtheria vaccine, diphtheria was a leading cause of death in children. Diphtheria is still prevalent in many developing countries. Diphtheria poses gre...

AI Technical Summary

Problems solved by technology

However, the expression product recombinant protein CRM197 protein obtained by this method is mainly expressed in the form of inclusion bodies, which needs to undergo multiple purification operations such as denaturation, renaturation, anion column and molecular sieve, which is not conducive to large-scale industrial production

Moreover, the E. coli expression system is a prokaryotic expression system, which lacks some necessary protein post-translational modifications, and often cannot form a soluble protein in a natural conformation. The epitope, specificity, and affinity used to detect antibodies will directly affect the detection results. accuracy

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