Key Laboratory of Pathogenic Fungi and Mycotoxins of Fujian Province, Key Laboratory of Biopesticide and Chemical Biology of Education Ministry, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, China.
Key Laboratory of Pathogenic Fungi and Mycotoxins of Fujian Province, Key Laboratory of Biopesticide and Chemical Biology of Education Ministry, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, China.
J Hazard Mater. 2017 Oct 5;339:154-160. doi: 10.1016/j.jhazmat.2017.06.030. Epub 2017 Jun 16.
Okadaic Acid (OA), a small seafood-borne toxin secreted by Dinophysis and Prorocentrum dinoflagellates, is generally distributed in various species of shellfish and has caused diarrhetic shellfish poisoning (DSP). In view of OA toxin threat to humans and animals, it is essential to develop a rapid, accurate and sensitive method for the detection and quantification of OA in real samples. In this study, a monoclonal antibody named 10E8 was screened by cells fusion of Sp2/0 with spleen cells isolated from immunized mouse, and the isotype of McAb 10E8 was belonged to IgG1. The resulted McAb 10E8 displayed higher specificity to OA antigen, with the highest affinity of 2.66×10L/moL until now. Indirect competitive ELISA (ic-ELISA) indicated that the linear range to detect OA was 20-750ng/mL. The limit of detection (LOD) was 12pg/mL, and the recovery average was (84.04±5.08)%. The LOD of colloidal gold immunoassay by naked eye and strip reader was 1ng/mL and 100pg/mL, respectively, with an average recovery of (88.0275±4.4225)%. Therefore, the developed ELISA and colloidal gold immunoassay based on this McAb can be used for OA detection in real samples.
冈田酸（OA）是一种由腰鞭毛藻和原甲藻分泌的小型海鲜源性毒素，广泛分布于各种贝类中，并已导致腹泻性贝类中毒（DSP）。鉴于 OA 毒素对人类和动物的威胁，开发一种快速、准确和敏感的方法来检测和定量真实样本中的 OA 至关重要。在这项研究中，通过 Sp2/0 细胞与免疫小鼠分离的脾细胞融合筛选出一种名为 10E8 的单克隆抗体，McAb 10E8 的同种型属于 IgG1。得到的 McAb 10E8 对 OA 抗原表现出更高的特异性，亲和力最高可达 2.66×10L/mol，这是迄今为止的最高记录。间接竞争 ELISA（ic-ELISA）表明，检测 OA 的线性范围为 20-750ng/mL。检测限（LOD）为 12pg/mL，平均回收率为（84.04±5.08）%。肉眼和条带阅读器的胶体金免疫测定的 LOD 分别为 1ng/mL 和 100pg/mL，平均回收率分别为（88.0275±4.4225）%。因此，基于该 McAb 开发的 ELISA 和胶体金免疫测定可用于真实样本中的 OA 检测。
