基于高灵敏度和特异性单克隆抗体的免疫分析法检测牡蛎和绿贻贝中的冈田酸

IF 1.7 3区农林科学 Q3 CHEMISTRY, APPLIED

Food and Agricultural Immunology Pub Date : 2022-05-25 DOI:10.1080/09540105.2022.2076812

Cheng Liang, Yuxiang Ji, Jiyong Ma, Chundong Zhang, Hongwei Zhao

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引用次数: 8

摘要

冈田酸(OA)是几种鞭毛藻产生的海洋生物毒素之一，可在海洋动物体内积累。食用受oa污染的海鲜可导致腹泻性贝类中毒。许多国家已经建立了监管限制，以限制海鲜中OA的水平。在本研究中，我们报道了一种新的免疫原产生的针对OA的高敏感单克隆抗体(mAb)。获得了两株单克隆杂交瘤细胞株(M8和M12)产生单抗的最佳敏感性和特异性。利用获得的单抗- m12建立间接竞争ELISA (icELISA)， 50%抑制浓度为0.15 ng mL−1，工作范围为0.02 ~ 2.74 ng mL−1。开发的icELISA比以前的报道更敏感。HPLC-MS验证了icELISA的准确性和工作范围，表明所建立的icELISA适用于牡蛎和青贻贝中OA的快速检测。

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Development of a highly sensitive and specific monoclonal antibody-based immunoassay for detection of okadaic acid in oysters and green mussels

ABSTRACT Okadaic acid (OA), one of marine biotoxins produced by several species of dinoflagellates, can accumulate in marine animals. The consumption of OA-contaminated seafood can cause diarrhoeic shellfish poisoning. Many countries have established regulatory restriction to limit the level of OA in seafood. In the present study, we report a highly sensitive monoclonal antibody (mAb) against OA produced by a new immunogen. Two monoclonal hybridoma cell lines (M8 and M12) that producing mAb with the best sensitivity and specificity were obtained. The obtained mAb-M12 was used to develop an indirect competitive ELISA (icELISA) with 50% inhibition concentrations of 0.15 ng mL−1 and a working range of 0.02–2.74 ng mL−1. The developed icELISA was more sensitive than the previous reports. HPLC-MS confirmed the accuracy and the working range of icELISA, suggesting that the developed icELISA is suitable for the rapid detection of OA in oysters and green mussels.

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来源期刊

Food and Agricultural Immunology 农林科学-毒理学

CiteScore

5.30

自引率

6.70%

发文量

审稿时长

2 months

期刊介绍： Food and Agricultural Immunology is an international open access journal publishing original immunological research with applications in food, agricultural, environmental and veterinary science. Submissions describing the use of immunological techniques and methods are particularly welcomed. The journal aims to expand our understanding of the interactions at the interface of food and immune systems including studies on: -Development of diagnostic systems – all types of ligand-based assays, e.g. antibody, aptamer -Application of ligand-based assays for the detection or identification of molecules of interest in food science, agricultural research, veterinary investigations and clinical systems relating to food allergy or sensitivity to agricultural chemicals -Effects of food on the immune system -Studies on allergy and allergic reactions -Investigations into food allergies -Development of allergen-free food systems -Development of novel assay formats -Applications of assay systems to the monitoring of food items in relation to safety and labelling -Food quality issues, e.g. speciation, adulteration and contamination -Comparisons between different analytical techniques The journal publishes research and review articles and is essential reading for food scientists, immunologists and all those concerned with the interaction between food and immune systems.