Abstract:
Objective To prepare monoclonal antibodies(McAb)against okadaic acid(OA)using B lymphocytes hybrid technique.
Methods To prepare immunogen of OA-BSA, hapten of OA is coupled to bovine serum albumin(BSA)according to carbodiimide method.Splenic cells of mice immunized by OA-BSA produce antibodies against OA.T he cell fusion is carried out by mixing splenic cells with the myeloma cells using polyethylene glycol(PEG).After the cell fusion, hybridoma cells are screened in a HAT medium.Hybridoma cells strains secreting monoclonal antibodies against OA are selected out by using enzyme-linked immunosorbent assay(ELISA).
Results According to carbodiimide method, antigens of OA-BSA and OA-OVA are successfully prepared from minute amount hapten of OA.Antibodies against OA are positive in sera of mice, which shows splenic cells can secrete antibodies against OA.After three times detection of antibodies, three hybridoma cells strains secreting monoclonal antibodies against OA are obtained.
Conclusion The successful preparation of monoclonal antibodies against OA supply a firm substantial foundation for developing an ELISA method that can determine OA as quickly and economically as possible in our country and furnish a experience that can be used as a reference on developing a method by which other shellfish toxins can be determined.
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