Production of Monoclonal Antibody for Okadaic Acid and Its Utilization in an Ultrasensitive Enzyme-Linked Immunosorbent Assay and One-Step Immunochromatographic Strip

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.

Development of novel monoclonal antibodies-based ultrasensitive enzyme-linked immunosorbent assay and rapid immunochromatographic strip for aflatoxin B1 detection

“…For conjugation, AFB1 mAbs were dialyzed in boric acid-borax buffer (pH 8) for 24 h at 4 C and the gold nanoparticle solution was adjusted to pH 8.0 with 0.1 M K 2 CO 3 (pH 11.5) (Liu et al, 2013;Liu et al, 2014). Five mL of anti-AFB1A mAb (1 mg/ml) was added dropwise to 2 ml of gold nanoparticle solution with gentle stirring.…”

“…Gold nanoparticles are conjugated with antibody as a tracer that provides a rapid visual evidence of the result (Liu, Hsu, Lu, & Yu, 2013;Liu, Hung, Lu, Chou, & Yu, 2014;Liu, Tsao, Wang, & Yu, 2008;Shim et al, 2007;Xiulan, Xiaolian, Jian, Zhou, & Chu, 2005). During the immunostrip assay, interaction occurs when test samples co-migrate with the antibody-gold nanoparticles that are conjugated along the strip.…”

“…During the immunostrip assay, interaction occurs when test samples co-migrate with the antibody-gold nanoparticles that are conjugated along the strip. The whole process takes less than 10 min and the results can be read without the aid of skilled personnel and instruments, making the method highly feasible for the on-site detection of toxin (Liu et al, 2013;Liu et al, 2014).…”

Periconceptional undernutrition modifies endocrine profiles and hepatic gene expression in sheep

“…() and Liu et al. () found that levels of ADIPOR2 mRNA are increased in the liver after 48 h of fasting. Liu et al.…”

Sensitive enzyme‐linked immunosorbent assay and gold nanoparticle immunochromatocgraphic strip for rapid detecting chloramphenicol in food

“…Gold nanoparticles were applied in immunostrip which is a promising immunoassay method to detect small molecular weight compound (Liu, Hung, Lu, Chou, & Yu, ; Shiu, Wang, & Yu, ; Xie et al, ). The principle of immunostrip relies on the movement of gold nanoparticle‐antibody conjugates and test sample along membrane strips on which the binding interactions occur.…”