Study on antibody against AST made by immunogen with different carrier proteins

Objective: To compare the immunogenity of the AST artificial antigen with two different carrier proteins,bovine serum albumin( BSA) and keyhole limpet hemocyanin( KLH). Methods: The hapten Astragaloside Ⅳ( AST) was linked to carrier protein BSA or KLH as immunogen,and linked ovalbumin( OVA) as coating antigen by the sodium periodate method. Then the 6-8 weeks BALB /c mice were immunized with AST-BSA or AST-KLH. The titer and specificity of the antiserum were detected by indirect enzymelinked immunosorbent assay( iELISA) and indirect competitive enzyme-linked immunosorbent assay( icELISA) respectively. Results:The antiserum of the mice immunized with AST-KLH had better specificity than which immunized with AST-BSA. The IC50 of ASTKLH antiserum was about 5 μg /ml in the detection by icELISA. The high titer of both antiserums was about 1∶ 51 200 after immunizing mouse. Conclusion: The conjugate of AST-KLH was testified to have a better immunogenity in comparison with AST-BSA,this investigation would be the foundation of the further research of AST immunoassay.