An Enzyme Immunoassay Developed for the Determination of Methylene Blue in Aquatic Products Based on a Novel Hapten

文献类型： 外文期刊

第一作者： Huang, Xuanyun

作者： Huang, Xuanyun;Wang, Liqing;Huang, Xuanyun;Yang, Guangxin;Tang, Yunyu

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关键词： methylene blue; hapten design; icELISA; polyclonal antibody

期刊名称：CHEMOSENSORS （ 影响因子：4.2； 五年影响因子：4.2 ）

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年卷期： 2024 年 12 卷 4 期

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收录情况： SCI

摘要： Methylene blue (MB) is a chemical dye which is used as an alternative drug for malachite green. In this study, based on computational chemistry, a novel hapten (TM) of MB was designed by comparing the conformational and electronic properties of MB. TM was synthesized and further conjugated with bovine serum albumin (BSA) and Ovalbumin (OVA). Then, the polyclonal antibodies were obtained by immunization with the immunogen of TM-BSA. Under optimal conditions, a sensitive indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed for the determination of MB, with an IC50 value of 41.5 mu g L-1 and displaying a cross reaction of 78.2 similar to 88.9% with three different MB metabolins. The limit of detection (LOD) and the limit of quantitation (LOQ) for MB were validated to be 4.8 mu g/kg and 6.0 mu g/kg, respectively. Spiking experiments showed recoveries between 82.3% and 84.3%, with a relative standard deviation (RSD) of <= 6%. The results showed the generated polyclonal antibodies from the new hapten TM were reliable and could be used for detecting MB in fishery productions.

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