半抗原
化学
抗体
单克隆抗体
检出限
IC50型
色谱法
生物化学
生物
免疫学
体外
作者
Rui Li,Xingya Sun,Yingjie Zhang,Peipei Li,Nan Li,Qing Shen,Kai Wen,Xuezhi Yu,Jianzhong Shen,Yantong Pan,Zhanhui Wang
标识
DOI:10.1016/j.foodchem.2024.139198
摘要
The preparation of high specificity and affinity antibodies is challenging due to limited information on characteristic groups of haptens in traditional design strategy. In this study, we first predicted characteristic groups of flurogestone acetate (FGA) using quantitative analysis of molecular surface combined with atomic charge distribution. Subsequently, FGA haptens were rationally designed to expose these identified characteristic groups fully. As a result, seven monoclonal antibodies were obtained with satisfactory performance, exhibiting IC50 values from 0.17 to 0.45 μg/L and negligible cross-reactivities below 1% to other 18 hormones. The antibody recognition mechanism further confirmed hydrogen bonds and hydrophobic interactions involving predicted FGA characteristic groups and specific amino acids in the antibodies contributed to their high specificity and affinity. Finally, one selective and sensitive ic-ELISA was developed for FGA determination with a detection limit as low as 0.12 μg/L, providing an efficient tool for timely monitoring of FGA in goat milk samples.
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