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基于酶联免疫分析方法检测畜禽饲料中恩拉霉素

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本文建立了一种应用于检测畜禽饲料中恩拉霉素的间接竞争酶联免疫分析法.采用羰基二咪唑法将生物活性高的恩拉霉素A组分合成的半抗原偶联卵清蛋白,构建检测原;同时将杂交瘤细胞株2H1F91免疫BALB/c小鼠,制备特异性识别恩拉霉素的单克隆抗体.在此基础上,构建了恩拉霉素的间接竞争酶联免疫分析法的标准曲线,该曲线IC50为91.61 ng/mL,线性范围为25.73～471.39 ng/mL,检出限为13.11 ng/mL.猪饲料样品的添加回收率为91.60％～95.75％,鸡饲料的添加回收率为89.33％～94.80％,并且二者的变异系数均低于12％,这表明建立的方法结果可靠,适用于畜禽饲料中恩拉霉素含量的快速检测.
Establishment of an Enzyme-Linked Immunosorbent Assay for the Detection of Enramycin in Livestock and Poultry Feed
An indirect competitive enzyme-linked immunosorbent assay(ic-ELISA)was established for the detection of enramycin in livestock and poultry feed.The carbonyl diimidazole method was used to couple OVA with a semi-antigen synthesized from the biologically active enramycin A component.The hybridoma cell line 2H1F91 was immunized to BALB/c mice,and a monoclonal antibody specifically recognizing enramycin was prepared.On this basis,a standard curve was constructed for the indirect competitive enzyme immunoassay of enramycin,which had an IC50 of 91.61 ng/mL,a linear range of 25.73-471.39 ng/mL,and a limit of detection of 13.11 ng/mL.The recoveries of pork feed samples ranged from 91.60％ to 95.75％ and those of chicken feed samples ranged from 89.33％ to 94.80％,and the coefficients of variation were lower than 12％ in both cases,which indicated that the developed ic-ELISA method was reliable and suitable for the rapid determination of enramycin in livestock and poultry feeds.

EnramycinHybridoma cells resistant to enramycin monocloning antibodyIndirect competitive enzyme-linked immunosorbent assay(ic-ELISA)Pork feedChicken feed

蔡永辉、王康、陈敏、王冰清

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浙江工商大学食品与生物工程学院,浙江 杭州 310018

恩拉霉素 单克隆抗体 间接竞争性酶联免疫吸附分析 猪饲料 鸡饲料

浙江省重中之重一级学科项目校企合作研发项目浙江省"十四五"省级大学生校外实践基地建设项目国家一流专业平台项目浙江工商大学校级精品在线开放课程建设项目

2017SIAR2012018330101002573浙教办函[2023]41号1110XJ05201201110XJ2922015

2024

10.13526/j.issn.1006-6144.2024.05.013

分析科学学报
武汉大学,北京大学,南京大学

分析科学学报

CSTPCD北大核心
影响因子：0.717
ISSN：1006-6144
年,卷(期)：2024.40(5)