Preparation method of novel coronavirus nucleocapsid protein monoclonal antibody

Abstract

The invention discloses a preparation method of a novel coronavirus nucleocapsid protein monoclonal antibody. The preparation method comprises the following steps: S1, synthesizing polypeptides; S2, preparing an immunogen; S3, preparing a screening antigen; S4, immunizing animals; S5, fusing cells; S6, screening and detecting; S7, establishing a stable cell strain; S8, producing the monoclonal antibody; S9, coupling the antibody; and S10, screening paired antibodies. The polypeptides are synthesized by selecting different antigen epitopes at two ends of N protein respectively and are used as antigens; the prepared monoclonal antibody can realize direct pairing; in a polypeptide synthesis process, a connecting bridge is added and steric hindrance is reduced, so that the antigen epitopes aresufficiently exposed on the surface of carrier protein and the antigens can be rapidly identified; recombinant protein is selected as a detection antigen, so that the risk that the screened antibodyonly identifies polypeptide antigens and does not identify the N protein is avoided; and whether a sample contains novel coronaviruses or not can be directly detected by detecting the content of the Nprotein.

Description

technical field [0001] The invention belongs to the technical field of monoclonal antibody preparation, and in particular relates to a preparation method of a monoclonal antibody to a novel coronavirus nucleocapsid protein. Background technique [0002] Since December 2019, a number of cases of pneumonia of unknown cause have been discovered one after another, and it has been confirmed as an acute respiratory infectious disease caused by a new type of coronavirus infection. Based on the current epidemiological investigation, the incubation period is generally 3-7 days, and the longest does not exceed 14 days. The clinical manifestations of this epidemic are mainly fever, fatigue, and dry cough. A small number of patients have symptoms such as nasal congestion, runny nose, and diarrhea. Some patients only showed low-grade fever, mild fatigue, etc., without pneumonia, and mostly recovered after 1 week. The World Health Organization named the pathogen that caused this case of...

AI Technical Summary

Problems solved by technology

[0003] The detection antibodies used in the existing novel coronavirus detection kits cannot be directly paired, and during the preparation of the detection antibodies, due to the insufficient exposure of the antigenic epitopes, the speed of antigen recognition is reduced. At the same time, when preparing the detection antibodies, Both use the coronavirus as the antigen, and the coronavirus is easy to combine with the N protein to form a helical nucleocapsid protein, which reduces the accuracy of the test results

Images