Production of monoclonal antibody for okadaic acid and its utilization in an ultrasensitive enzyme-linked immunosorbent assay and one-step immunochromatographic strip
Journal
Journal of Agricultural and Food Chemistry
Journal Volume
62
Journal Issue
6
Pages
1254-1260
Date Issued
2014
Author(s)
Abstract
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, 6B1A3, 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-γ-globulin. The 6B1A3 mAb belongs to the immunoglobulin G1 (κ 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. © 2014 American Chemical Society.
Subjects
enzyme-linked immunosorbent assay; gold nanoparticle immunochromatographic strip; monoclonal antibody; okadaic acid
SDGs
Other Subjects
monoclonal antibody; okadaic acid; animal; article; Bagg albino mouse; biosynthesis; cell line; enzyme linked immunosorbent assay; female; hybridoma; immunoaffinity chromatography; immunology; methodology; mouse; sensitivity and specificity; shellfish; shellfish poisoning; test strip; Animals; Antibodies, Monoclonal; Cell Line; Enzyme-Linked Immunosorbent Assay; Female; Hybridomas; Immunochromatography; Mice; Mice, Inbred BALB C; Okadaic Acid; Reagent Strips; Sensitivity and Specificity; Shellfish; Shellfish Poisoning; Assays; Bioassay; Cell culture; Enzymes; Gold; Metal nanoparticles; Monoclonal antibodies; Nanoparticles; Screening; Shellfish; Diarrhetic shellfish poisoning; Enzyme linked immunosorbent assay; Gold Nanoparticles; Horse-radish peroxidase; Immunochromatographic strips; Mean concentrations; Monoclonal antibodies (mAb); Okadaic acid; Antibodies; Armoracia rusticana; Bivalvia
Type
journal article