Preparation Of Monoclonal Antibody Against G Protein Of Rabies Virus

Rabies is a deadly zoonotic infectious disease caused by rabies virus infection.Rabies continues to threaten the lives of warm-blooded animals,especially in Asia and Africa,where there is a high incidence of human and animal rabies.Therefore,in areas with high incidence of rabies,rabies virus infection not only causes economic losses,but also causes serious public health safety problems.The hosts of rabies virus are mainly domestic animals(dogs,pigs,cattle,horses,sheep,etc.)and wild animals.In recent years,there have been many reports of rabies virus infection caused by biting humans by domestic animals and pets.Once the disease,because the virus invasion is the central nervous system,resulting in a case fatality rate of 100%,is the highest human case fatality of acute infectious diseases.The rabies virus has no specific animal host.There are five structural proteins of rabies virus: nucleoprotein,phosphoprotein,matrix protein,glycoprotein and RNA polymerase large protein.When rabies virus invades the central nervous system of the brain,it can lead to fatal brain neuritis.While the rabies virus glycoprotein(G)can induce the host to produce Neutralizing antibody,which can effectively stimulate the mass production of T lymphocytes,and the rabies virus glycoprotein Neutralizing antibody can effectively neutralize the free virus,and then inhibit the invasion and replication of the virus.Glycoprotein can protect the receptor and prevent the rabies virus from attacking the receptor in the brain.Because of the important role of the G protein of rabies virus in virus invasion and immune protection,it has become one of the most thoroughly studied proteins.In recent years,researchers have focused on the establishment and screening of Mc Ab with neutralizing activity.In this study,an effective G protein monoclonal antibody was screened,which is essential for the study of the function of G protein of rabies virus,the diagnosis of rabies virus infection,the development of rabies virus vaccine,and the screening of rabies antiviral drugs.In this study,in order to prepare an anti-rabies virus G protein monoclonal antibody,attenuated vaccine ERA strain whole virus was purified by density gradient centrifugation method,and BALB/c mice were immunized by intrapermuscle injection.After three times of routine immunization and one time of enhanced immunization,spleen cells of mice were collected and cultured with myeloma cells for fusion.The anti-rabies virus G protein monoclonal antibody was screened by IFA method in the supernatant of culture.After 3 to 4subclones,the monoclonal antibody against rabies virus G protein was secreted stably,and ascites was prepared.Finally,two monoclonal antibodies were screened and named as G3E8 and G4G12.The subclass of monoclonal antibodies were all mouse Ig G3.Western Bolt identification showed that the anti-rabies virus G protein monoclonal antibody could react with the whole virus,and the reaction band was 67 k Da.Virus neutralization experiments showed that the two monoclonal antibodies had neutralization activity of rabies virus.The two monoclonal antibodies had two different G protein epitopes.In this experiment,the monoclonal antibody against G protein of rabies virus was successfully prepared and the biological information of the monoclonal antibody was comprehensively identified.The purpose of this study was to prepare the monoclonal antibody against the G protein of rabies virus with high school and activity,which laid the foundation for the study of the mechanism of rabies virus invasion,the development of the quantitative detection kit of the neutralizing antibody against rabies virus and the antiviral treatment of rabies virus after exposure.