Jiang Hongtao, Xu Xinxin, Song Shanshan, Wu Aihong, Liu Liqiang, Kuang Hua, Xu Chuanlai
State Key Lab of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, People's Republic of China.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2022 Jun;39(6):1053-1064. doi: 10.1080/19440049.2022.2048089. Epub 2022 Apr 29.
In this study, a monoclonal antibody (mAb) 1G5 against novobiocin with high sensitivity and specificity was prepared from a newly-designed hapten. According to the results of an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA), the 50%-inhibitory concentration of the anti-novobiocin mAb was 6.9 ng/mL and the cross-reactivity was less than 0.1% to its analogues. Furthermore, a rapid colloidal gold immunochromatographic assay (ICA) was successfully developed for the determination of novobiocin in spiked samples. Two calibration curves were established respectively, for beef and chicken samples. The ICA results showed a visual colorimetric value of 50 ng/mL and a cut-off value of 300 ng/mL in beef samples. The ICA results of chicken samples were almost the same as that of beef. When quantitative detection was performed using a strip reader, the detection ranges for quantitative analysis in beef and chicken were 23.7-287.5 and 19.7-263.8 µg/kg respectively. Recoveries were between 82.7 and 95.3% for beef samples with the coefficient of variation (CV) ranging from 2.5 to 5.1%. Recoveries were in the range of 89.6-105.5% with the CV ranging from 2.9% to 6.3% for chicken samples. Importantly, these results from the ICA were highly consistent with the results obtained by LC-MS/MS. Therefore, this ICA could be used as an alternative means for the rapid determination of NOV in a large number of beef and chicken samples.
在本研究中，从新设计的半抗原制备了一种对新生霉素具有高灵敏度和特异性的单克隆抗体（mAb）1G5。根据间接竞争酶联免疫吸附测定（ic-ELISA）的结果，抗新生霉素单克隆抗体的50%抑制浓度为6.9 ng/mL，与其类似物的交叉反应率小于0.1%。此外，成功开发了一种快速胶体金免疫层析测定法（ICA），用于测定加标样品中的新生霉素。分别为牛肉和鸡肉样品建立了两条校准曲线。ICA结果显示，牛肉样品的目视比色值为50 ng/mL，临界值为300 ng/mL。鸡肉样品的ICA结果与牛肉样品几乎相同。使用试纸读数器进行定量检测时，牛肉和鸡肉定量分析的检测范围分别为23.7-287.5和19.7-263.8 μg/kg。牛肉样品的回收率在82.7%至95.3%之间，变异系数（CV）为2.5%至5.1%。鸡肉样品的回收率在89.6%-105.5%之间，CV为2.9%至6.3%。重要的是，ICA的这些结果与LC-MS/MS获得的结果高度一致。因此，这种ICA可作为快速测定大量牛肉和鸡肉样品中新生霉素的替代方法。
