Preparation of hybridoma cell strains secreting monoclonal antibodies against domic acid

To obtain stable and efficient hybridoma cell strains secreting monoclonal antibody(McAb) against domic acid(DA),completed antigen(DA-KLH) and coating antigen(DA-BSA) were synthesized by active ester method.BALB/c mice were immunized with DA-KLH and hybridoma cell strains secreting McAb against DA were screened by cell fusion,and the immunological titers of DA-McAb were determined.Capture ELISA method was used to determine isotypes of the mouse monoclonal antibody and the indirect ELISA method was performed to determine the titers of McAbs.The protein G affinity chromatography protein purification method was used to purify ascites.Results showed that: two strains of hybridomas,2B1and 4C2,which could secret McAb against DA steadily were successfully prepared and the isotypes were IgG1 and IgG2a.The titer of purified McAb was 1∶64 000 and the protein concentration of purified ascites were 1.27 and 0.675 mg/mL respectively.It could be demonstrated that the successful establishment of hybridoma cell strains secreting McAb against DA supply a firm foundation for developing a gold immunochromatography assay that can detect DA as quickly,economically and simplely as possible in future research.