Monoclone antibody of swine fever virus resistant wild strain E2 protein, preparation method and application thereof

Abstract

The invention discloses a monoclonal antibody against virulent strain E2 protein of classical swine fever virus and a hybridoma cell strain secreting the monoclonal antibody. The hybridoma cell strain is obtained by using hog cholera lapinized virus vaccine strain E2 protein expressed by Baculovirus as tolerogen, selecting Shimen strain E2 protein as immunogen, immunizing mouse by cyclophosphamide immunosuppression method, carrying out cell fusion, and sieving hybridoma cell strain capable of stably secreting monoclonal antibody against E2 protein. The monoclonal antibody can react with Shimen strain and can produce specific reaction with virulent strain of classical swine fever viruses of 1.1, 2.1, 2.2 and 2.3 gene sub-groups. The monoclonal antibody has neutralization activity and does not react with hog cholera lapinized virus vaccine strain, so that the monoclonal antibody can be used for differentiating virulent strain of classical swine fever virus and hog cholera lapinized virus vaccine strain, which establishes the foundation for establishing a method for differentiating wild virus infection of classical swine fever and vaccine immunity and for researching the molecular difference between CSFV virulent strain and mild strain.

Description

technical field [0001] The present invention relates to a monoclonal antibody, in particular to a monoclonal antibody against the E2 protein of the wild strain of classical swine fever virus, and the present invention also relates to a hybridoma cell strain secreting the anti-E2 protein monoclonal antibody and the anti-E2 protein monoclonal antibody and the monoclonal antibody in classical swine fever The application in clinical diagnosis belongs to the field of animal disease inspection and quarantine. Background technique [0002] Classical swine fever (CSF) is a highly contagious infectious disease characterized by high fever and hemorrhage caused by classical swine fever virus (CSFV), and is one of the major infectious diseases that endanger the pig industry , the World Organization for Animal Health (OIE) included it in the OIE List of Diseases (OIE Listeddiseases), and my country also listed it as a first-class infectious disease. CSFV belongs to the genus Pestivirus o...

AI Technical Summary

Problems solved by technology

The biggest disadvantage of conventional attenuated vaccine immunization is that it is difficult to distinguish it from natural infection by serological methods

Scholars at home and abroad have prepared a variety of CSFV monoclonal antibodies (Wensvoort G, Terpstra C, Boonstra J, et al. Production of monoclonal antibodies against swine fever virus and their use in laboratory diagnosis. Vet Microbiol, 1986, 12 (2): 101 -108; Qiu Huishen, Wang Zaishi, Fang Guoan. Research on Diagnostic Reagents of Monoclonal Antibody of Swine Fever. China Livestock and Poultry Infectious Diseases, 1991, 6: 20-26; 1991, 6: 20-26; Ma Gang, Li Zuosheng, Yu Xing Long et al. Preparation of monoclonal antibody against classical swine fever virus E2 protein and preliminary analysis of its epitope. Chinese Journal of Veterinary Medicine, 2002, 22(2): 121-124; Yang Yanyan, Zhang Gaiping, Wang Xuannian. List of classical swine fever virus Preparation of cloned antibody and its immunohistochemical study. Chinese Veterinary Science, 2006, 36(10): 782-786; Hou Qiang, Peng Wuping, Sun Yuan, etc. Prokaryotic expression of the gene encoding the main antigenic region of the E2 protein of classical swine fever virus and the preparation of its monoclonal antibody. Chinese Veterinary Science, 2008, 38 (1): 1-5.), however, due to the high homology of swine fever wild strain and attenuated vaccine strain, most monoclonal antibodies and pig Both wild and attenuated pestivirus strains can react, and cannot effectively distinguish wild virus infection from vaccine virus, which largely limits the application of CSFV monoclonal antibodies in the clinical diagnosis of classical swine fever

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