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À propos de : Development of a Monoclonal Antibody against Domoic Acidand Its Application in Enzyme-Linked Immunosorbent Assayand Colloidal Gold Immunostrip        

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  • Development of a Monoclonal Antibody against Domoic Acidand Its Application in Enzyme-Linked Immunosorbent Assayand Colloidal Gold Immunostrip
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  • A monoclonal antibody (mAb) specific to domoic acid was produced from a stable hybridoma cellline, 9F1F11, generated by the fusion of P3/NS1/1-AG4-1 myeloma cells with spleen cells isolatedfrom a Balb/c mouse immunized with domoic acid−keyhole limpet hemocyanin. The 9F1F11 mAbbelongs to the immunoglobulin G1 (κ-chain) isotype. A competitive direct enzyme-linked immunosorbent assay (cdELISA) and a competitive indirect ELISA were established for antibody characterization.In the cdELISA, the concentration causing 50% inhibition (IC50) of binding of domoic acid−horseradishperoxidase to the antibody by domoic acid was found to be 0.58 ng/mL. A sensitive and rapid mAb-based colloidal gold immunostrip was also developed. The immunostrip assay, which has a detectionlimit of 5 ng/mL for domoic acid, can be completed in 10 min. Analysis of domoic acid in blue musselsamples revealed that data obtained from immunostrip were in a good agreement with those obtainedfrom cdELISA. The mAb-based cdELISA and immunostrip assay established in this study weresensitive and accurate for rapid screening of domoic acid in shellfish samples. Keywords: Domoic acid; monoclonal antibodies; ELISA; colloidal gold immunostrip
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