ABSTRACT
Acquisition of the genes encoding variable regions of paired heavy and light chains (VH:VL) is crucial, but it is a labor and cost-intensive process in traditional methods. This study presents a novel method in which all processing steps for acquiring natively paired VH:VL genes from single cells are finished in a single microfluidic chip. The microfluidic chip performs single-cell trap/in situ fluorescent examination of antibody specificity/cell lysis/gene amplification all at single-cell level. By a proof-of-concept validation of efficiently acquiring paired VH:VL genes of anti-RBD (which is a key protein of SARS-CoV-2 virus) mAbs from single hybridomas, the microfluidic chip has been proved capable of remarkably improving cell loss/human labor/time cost, and more importantly, determinacy of native VH:VL genes pairing which is one of the most decisive factors of effectiveness for antibody discovery. © 2021 SPIE.