RESUMO
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 enzyme-linked 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 AST-KLH 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.
