Preparation of Monoclonal Antibody against Okadaic Acid and Development of a Sensitive ELISA for Detection Diarrhetic Shellfish Poisoning in Shellfish
Liu Renyan, Daoyan Xu, Liang Yubo
Abstract
Liu Renyan, Daoyan Xu, Liang Yubo
Abstract
A rapid method for detection diarrhetic shellfish poisoning (DSP), a marine biotoxin, were developed. Monoclonal antibody against OA (okadaic acid) was prepared by using the technique of cell fusion. This antibody was applied in development the idc-ELISA (indirect competitive enzyme-linked immunosorbent assay) for detection of OA. Using OA-KLH as the immunogen to immunize BALB/c mice, splenocytes from the immunized mice were fused with murine myeloma cells. Three positive hybridoma cells strains that could secrete steadily anti-OA monoclonal antibody were obtained after three times of cloning. The idc-ELISA method with detection limit of 0.781 ng/ml was developed using 1# monoclonal antibody showing highest specificity against OA. Okadaic acid added was detected with average recoveries of 92.5% for scallop.
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A rapid method for detection diarrhetic shellfish poisoning (DSP), a marine biotoxin, were developed. Monoclonal antibody against OA (okadaic acid) was prepared by using the technique of cell fusion. This antibody was applied in development the idc-ELISA (indirect competitive enzyme-linked immunosorbent assay) for detection of OA. Using OA-KLH as the immunogen to immunize BALB/c mice, splenocytes from the immunized mice were fused with murine myeloma cells. Three positive hybridoma cells strains that could secrete steadily anti-OA monoclonal antibody were obtained after three times of cloning. The idc-ELISA method with detection limit of 0.781 ng/ml was developed using 1# monoclonal antibody showing highest specificity against OA. Okadaic acid added was detected with average recoveries of 92.5% for scallop.
Key concepts: Diarrhetic shellfish poisoning, Okadaic acid, Monoclonal antibody, Immunogen, Antibody, Chemistry, Molecular biology, Biology