Title :
Preparation of Monoclonal Antibody against Okadaic Acid and Development of a Sensitive ELISA for Detection Diarrhetic Shellfish Poisoning in Shellfish
Author :
Renyan, Liu ; Daoyan, Xu ; Yubo, Liang
Author_Institution :
Nat. Marine Environ. Monitoring Center, Dalian, China
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.
Keywords :
biological techniques; cellular biophysics; enzymes; molecular biophysics; organic compounds; cell fusion; cloning; detection diarrhetic shellfish poisoning; hybridoma cell strains; immunized mice; immunogen; indirect competitive enzyme-linked immunosorbent assay; marine biotoxin; monoclonal antibody; murine myeloma cells; okadaic acid; splenocytes; Buffer storage; Digital signal processing; Immune system; Information analysis; Instruments; Marine technology; Mice; Performance analysis; Phase measurement; Proteins; ELISA; diarrhetic shellfish poisoning; monoclonal antibody; okadaic acid;
Conference_Titel :
Environmental Science and Information Application Technology, 2009. ESIAT 2009. International Conference on
Conference_Location :
Wuhan
Print_ISBN :
978-0-7695-3682-8
DOI :
10.1109/ESIAT.2009.101