Preparation And Characterization Of Monoclonal Antibody Against HBV S Protein

Objectives: Hepatitis B virus is a very dangerous pathogen for human health. Till now, there is no effective therapeutic methods and solutions for chronic Hepatitis B virus infection. HBsAg is a sign of infecting HBV, and the first signal appearing in the serum while people infect HBV. It has inportant significance in clinically type B hepatitis early diagnosis and HBV scientific research. The specific monoclonal antibodies (McAbs) against HBV S protein were prepared by hybridoma cell techniques, which served as the basement for further study.Methods:The recombinant Hepatitis B vaccine was confirmed by SDS-PAGE analysis. Immunogenicity of the recombinant Hepatitis B vaccine was confirmed by indirect ELISA.Balb/c mice were immunized with the recombinant Hepatitis B vaccine. Splenocytes of the immunized mice were collected and fused with the mouse myeloma cell line SP2/0 cells. The hybridoma cells that secreted anti-HBV S protein McAb antibodies were cloned with limited dilution method. The immunoglobulin (Ig) subtype, the ascites titers, and the affinity of the obtained McAbs were determined by indirect ELISA. The specificity of McAbs was tested by indirect ELISA and western blot analysis.Results:The recombinant Hepatitis B vaccine contains HBV S protein in high purity. The S protein hasn't been degraded, and is in good specificity and immunogenicity.One of hybridomas was screened out, designated 4D2. Chromosome analysis revealed that the obtained hybridomas were with the universal characteristics of the monoclonal hybridoma cells which secreted McAb, and the Ig subtype of 4D2 McAb was IgG1. The ascites titers of 4D2 McAb was 1:108. The relative affinity of 4D2 McAb was determined as 105. Western blot identification suggested that the McAbs reacted specifically with HBV S protein.Conclusions: One of high titer, specific McAbs against HBV S protein has been successfully prepared and primarily identified, which may be useful in the development of a rapid and convenient diagnostic kit for detection of viral hepatitis B.