State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China.
Jiangxi Province Center for Disease Control and Prevention, Nanchang 330047, China.
J Dairy Sci. 2019 Jul;102(7):6037-6046. doi: 10.3168/jds.2018-16048. Epub 2019 May 2.
The use of the heterologous competitive strategy has become a vital method to improve the sensitivity of ELISA. In this work, we prepared an anti-enrofloxacin (ENR) mAb with ENR-bovine serum albumin (BSA) as immunogen. The molecular descriptors of quinolones were then used to screen heterologous coating antigens for the detection of ENR based on an ensemble learning method to improve the sensitivity of the ELISA. Results indicated that 6 of the 7 selected heterologous competitive antigens could enhance the sensitivity of ELISA. The ELISA sensitivity for the detection of ENR with sarafloxacin-BSA as heterologous coating antigen was improved 10-fold (in PBS) and 6-fold (in milk) compared with that with ENR-BSA as homologous antigen. The strategy can effectively screen suitable heterologous competitive antigens to improve the sensitivity of ELISA, followed by preparation of mAb with no additional modification to the corresponding immunogen.
利用异源竞争策略已成为提高 ELISA 灵敏度的重要方法。本工作以恩诺沙星-牛血清白蛋白（BSA）为免疫原，制备了抗恩诺沙星（ENR）单克隆抗体。然后，使用喹诺酮类药物的分子描述符，基于集成学习方法筛选用于检测 ENR 的异源包被抗原，以提高 ELISA 的灵敏度。结果表明，7 种选择的异源竞争抗原中的 6 种可以提高 ELISA 的灵敏度。与以 ENR-BSA 作为同源抗原相比，以沙拉沙星-BSA 作为异源包被抗原的 ELISA 检测 ENR 的灵敏度在 PBS 中提高了 10 倍，在牛奶中提高了 6 倍。该策略可以有效地筛选合适的异源竞争抗原，提高 ELISA 的灵敏度，而无需对相应的免疫原进行额外修饰。
