Development of a Sensitive Monoclonal Antibody-Based Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Determination of Monensin in Edible Chicken Tissues
Development of a Sensitive Monoclonal Antibody-Based Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Determination of Monensin in Edible Chicken Tissues
复制标题
开发基于单克隆抗体的灵敏间接竞争酶联免疫吸附测定法，用于测定可食用鸡组织中的莫能菌素
DOI：
10.1007/s12161-019-01461-3
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发表时间：
2019
影响因子：
2.9
通讯作者：
Yuan Zonghui
中科院分区：
文献类型：
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作者：
Li Langhong;Pan Yuanhu;Tao Yanfei;Chen Donmei;Wang Yulian;Wang Xu;Liu Zhenli;Peng Dapeng;Yuan Zonghui
为了检测鸡组织中莫能菌素（Monensin，MON）的残留，建立了基于单克隆抗体（mAb）的间接竞争酶联免疫吸附试验（ic-ELISA）。通过向MON盐中加入盐酸获得半抗原MON，然后与载体蛋白偶联。在接种雌性Balb/c小鼠和细胞融合后，获得一个细胞系MON/4C 9。MON/4C 9单克隆抗体对MON的IC 50为1.28 μg L-1，在此基础上，建立了优化的免疫吸附试验（ic-ELISA）方法。各种样品基质中MON的检测限和定量限从0.38 μg kg-1到1.01 μg kg-1不等。鸡组织中的回收率为71.9%~ 116.9%，批内和批间变异系数均小于18%。此外，所开发的方法也表现出与样品进行的HPLC-MS的结果呈正相关。结果表明，所制备的单克隆抗体和所建立的间接ELISA方法可用于鸡组织中MON的检测。
To monitor monensin (MON) residues in chicken tissues, a monoclonal antibody (mAb)–based indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed in this study. The haptenMON was obtained by adding hydrochloric acid to a MON salt, and then was coupled with carrier protein. After the inoculation of female Balb/c mice and cell fusions, one cell line, MON/4C9, was obtained. The MON/4C9 antibody exhibited the ability to specifically recognize MON with IC501.28 μg L−1.Based on this mAb, an optimized ic-ELISA protocol was performed using only methanol-water (8:2,v/v) in chicken tissue samples. The limits of detection and the limits of quantification of MON in various sample matrices varied from 0.38 to 1.01 μg kg−1. The recoveries ranged from 71.9 to 116.9% in chicken tissues, and the intra- and inter-assay CVs were all less than 18%. Moreover, the developed method also exhibited a positive correlation with the results of HPLC-MS conducted on the samples. These results suggest that the prepared mAb and the developed ic-ELISA method will be a useful tool for detecting MON in chicken tissues.