Highly efficient and precise two-step cell selection method for tetramethylenedisulfotetramine-specific monoclonal...

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 (IC₅₀) of the best mAb 1G6 for TETS in the TCSM was 1.98 ng mL^-1, and that of mAb 2B6 in the THM was 11.49 ng mL^-1. 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.