Preparation And Identification Of The Monoclonal Antibodies Against GBD Of Glucan Binding Protein A From Mutans Streptococci

Dental caries is one of the three chief non-contagious diseases which threaten human health. Mutans streptococci (MS) are the primary cariogenic agents, and Streptococcus mutans and Streptococcus sobrinus are believed to have the closest relationship with human caries. Adhesion and proliferation of MS are the basis of cariogensis. At present, the study of anticaries mainly focuses on a series of events involving the adherence, accumulation and proliferation of MS on tooth surfaces. Researchers have done many studies on cloning, functions and anticaries immunization of the genes related to adherence and accumulation. These genes includ surface protein antigen I / II (Ag I / II), glucosyltransferases (GTFs) and glucan binding proteins (Gbps). Ag I / II, GTFs and GbpB canprovide protection against dental caries. GbpA from S. mutans contains a C-terminal GBD homologous to the GBDs of the GTFs, but the effects of this GBD of GbpA on anticaries immunization has not yet been reported.METHODS: Balb/c mice were administered with the expressed and purified fusion protein GBD of GbpA. Then the spleen of immunized mice were collected and hybridized with SP2/0 cells. Monoclonal antibodies were prepared with the routine methods. The monoclonal antibodies were identificated by ELISA and Western blot. RESULTS: Six strains of hybridomas were obtained. Titre oi ascites reached dilution of 10-6. Isotype of antibodies were determined. CONCLUSION: Monoclonal antibodies against the GBD of GbpA frorr S.mutans can be obtained by routine hybridoma technic, which will make the sequential preparation of gene technological realized.