Li Chenglong, Yang Huijuan, Yu Wenbo, Yu Xuezhi, Wen Kai, Shen Jianzhong, Wang Zhanhui
College of Veterinary Medicine, China Agricultural University, 100193 Beijing, People's Republic of China.
Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing Laboratory for Food Quality and Safety, 100193 Beijing, People's Republic of China.
Anal Chem. 2021 Apr 20;93(15):6008-6012. doi: 10.1021/acs.analchem.1c00633. Epub 2021 Mar 17.
The use of organic solvents to extract chemical contaminants for an immunoassay is mostly inevitable. On this occasion, the intolerance of natural antibodies against organic solvent is detrimental to the performance of immunoassays in terms of sensitivity, assay time, accuracy, and precision. Few studies have focused on improving the low tolerance of natural antibodies to organic solvents for analytical purposes. In this study, we engineered the monoclonal antibody (mAb) 4D11 to sulfonamides through CDR grafting by using one proven highly stable humanized antibody (hAb) 4D5 for the first time. The engineered antibody hAb 4D11 showed significantly improved tolerance abilities to acetonitrile (2% to 20%) and methanol (10% to 20%), and retained the highly affinity and class-specificity to sulfonamides. This study provided a general strategy to improve antibody tolerance to organic solvents and was greatly beneficial to the robust development of immunoassays.
使用有机溶剂提取用于免疫分析的化学污染物大多是不可避免的。在这种情况下，天然抗体对有机溶剂的不耐受性在灵敏度、分析时间、准确性和精密度方面对免疫分析的性能有害。很少有研究专注于提高天然抗体对有机溶剂的低耐受性以用于分析目的。在本研究中，我们首次通过使用一种经证实高度稳定的人源化抗体（hAb）4D5，通过互补决定区（CDR）移植对磺胺类药物工程化改造单克隆抗体（mAb）4D11。工程化抗体hAb 4D11对乙腈（2%至20%）和甲醇（10%至20%）的耐受能力显著提高，并保留了对磺胺类药物的高亲和力和类特异性。本研究提供了一种提高抗体对有机溶剂耐受性的通用策略，对免疫分析的稳健发展非常有益。
