Preparation monoclonal -type anti-idiotype antibody of zearalenone and development of green ELISA quantitative detecting technique
PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY(2020)
摘要
Immunoassay has been widely used in the screening of mycotoxins, which may be hazardous to the operator or the environment. This study was to develop a green way to measure zearalenone (ZEN) with a monoclonal beta-type anti-idiotype antibody (Ab2 beta) against ZEN in place of ZEN standard. Six monoclonal beta-type anti-idiotype antibodies were prepared. The 50% inhibitory concentration (IC50) value to ZEN of the six antibodies was between 34.45 +/- 1.12-182.12 +/- 15.40 nM. A green ELISA was then developed and validated. The quantitative conversion formula between ZEN and the monoclonal Ab2 beta against ZEN was y = 0.092x(0.722), R-2 = 0.990. The working range was 2.63-100.64 ng ml(-1). The recovery rate in spiked feed samples was from 82.15% to 102.79%, and the within-assay and between-assay coefficient variation (CV) level were less than 10.00%. A good correlation was obtained by high-performance liquid chromatography method (HPLC) to validate the developed method.
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关键词
Zearalenone,monoclonal antibody,beta-type anti-idiotype antibody,indirect competitive enzyme-linked immunosorbent assay
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