College of Veterinary Medicine, China Agricultural University, Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing Laboratory of Food Quality and Safety, Beijing 100193, China.
College of Veterinary Medicine, China Agricultural University, Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing Laboratory of Food Quality and Safety, Beijing 100193, China; Beijing Vocational College of Agriculture, Beijing 102442, China.
J Hazard Mater. 2022 Feb 15;424(Pt D):127689. doi: 10.1016/j.jhazmat.2021.127689. Epub 2021 Nov 11.
Monoclonal antibodies (mAbs) are useful biological tools for research, diagnostics, and pharmaceuticals. Here, we proposed a new mAb discovery platform named the two-step cell selection method (TCSM) for mAbs production of some small molecule haptens as antibiotic, toxins, and pesticides. The first step was performed by a fluorescence-activated cell sorter to enrich the hapten-specific B cells, the second step was an image-based precise pick of single hapten-specific hybridoma cells by confocal laser scanning microscopy. In this study, we used tetramethylenedisulfotetramine (TETS) as a model analyte, which is a highly lethal neurotoxic rodenticide. The TETS-specific hybridoma cells selection was completed within 10 days by the TCSM, compared with at least 40 days in the traditional hybridoma method (THM). The half maximal inhibitory concentration (IC) of the best mAb 1G6 for TETS in the TCSM was 1.98 ng mL, and that of mAb 2B6 in the THM was 11.49 ng mL. Antibody-TETS recognition also showed more interactions in mAb 1G6 than in mAb 2B6. Then, the mAb 1G6 was then successfully applied to develop an icELISA for TETS in biological samples with satisfactory sensitivity, accuracy and precision. The results demonstrated that the TCSM was a feasible and efficient method for mAb discovering of poisonous hapten molecules.
单克隆抗体（mAbs）是用于研究、诊断和制药的有用的生物工具。在这里，我们提出了一种新的 mAb 发现平台，称为两步细胞选择法（TCSM），用于生产某些小分子半抗原作为抗生素、毒素和杀虫剂的 mAbs。第一步通过荧光激活细胞分选仪对半抗原特异性 B 细胞进行富集，第二步通过共聚焦激光扫描显微镜对单个半抗原特异性杂交瘤细胞进行基于图像的精确挑选。在这项研究中，我们使用四亚甲基二砜四胺（TETS）作为模型分析物，TETS 是一种高度致命的神经毒鼠剂。使用 TCSM 在 10 天内完成了 TETS 特异性杂交瘤细胞的选择，而传统杂交瘤方法（THM）至少需要 40 天。在 TCSM 中，针对 TETS 的最佳 mAb 1G6 的半最大抑制浓度（IC）为 1.98 ng mL，而在 THM 中，mAb 2B6 的 IC 为 11.49 ng mL。mAb 1G6 与 TETS 的抗体识别也显示出比 mAb 2B6 更多的相互作用。然后，mAb 1G6 成功地应用于开发用于生物样品中 TETS 的 icELISA，具有令人满意的灵敏度、准确性和精密度。结果表明，TCSM 是一种可行且有效的方法，用于发现有毒半抗原分子的 mAb。
