期刊
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
卷 62, 期 6, 页码 1254-1260出版社
AMER CHEMICAL SOC
DOI: 10.1021/jf404827s
关键词
enzyme-linked immunosorbent assay; gold nanoparticle immunochromatographic strip; monoclonal antibody; okadaic acid
资金
- National Science Council of the Taiwan [NSC 100-2923-B-040-001-MY2, 101-2313-B-040-005]
Okadaic acid (OA) is a common marine biotoxin that accumulates in bivalves and causes diarrhetic shellfish poisoning (DSP). This study generated a monoclonal antibody (mAb) specific to OA from a hybridoma cell line, 6BIA3, which was obtained by fusion of myeloma cells (P3/NS1/1-AG4-1) with spleen cells isolated from a BALB/c mouse immunized with OA-gamma-globulin. The 6BIA3 mAb belongs to the immunoglobulin GI (kappa chain) isotype. Both competitive direct and indirect enzyme-linked immunosorbent assays (ELISAs) were established for characterization of the antibody. The concentrations causing 50% inhibition of binding of OA-horseradish peroxidase to the antibody by OA were calculated to be 0.077 ng/mL in the cdELISA. A rapid and sensitive mAb-based gold nanoparticle immunochromatographic strip was also established. This proposed strip has a detection limit of 5 ng/mL for OA and can be finished in 10 min. Extensive analyses of 20 seafood samples with ELISA revealed that 10 were slightly contaminated with OA, with a mean concentration of 0.892 ng/g. Analysis of OA in shellfish samples showed that data acquired by the immunochromatographic strip agreed well with those acquired by the ELISA. The mAb-based ELISA and immunochromatographic strip assay developed in this study have adequate sensitivity and accuracy for rapid screening of OA in shellfish samples.
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