Title: High-throughput single-cell activity-based screening and sequencing of antibodies using droplet microfluidics
Author: Annabelle Grard, Adam Woolfe, Guillaume Mottet, Marcel Reichen, Carlos Castrillon, Vera Menrath, Sami Ellouze, Adeline Poitou, Raphal Doineau, Luis Briseno-Roa, Pablo Canales-Herrerias, Pascaline Mary, Gregory Rose, Charina Ortega, Matthieu Delinc, Sosthene Essono, Bin Jia, Bruno Iannascoli, Odile Richard-Le Goff, Roshan Kumar, Samantha N. Stewart, Yannick Pousse, Bingqing Shen, Kevin Grosselin, Baptiste Saudemont, Antoine Sautel-Caill, Alexei Godina, Scott McNamara, Klaus Eyer, Gal A. Millot, Jean Baudry, Patrick England, Clment Nizak, Allan Jensen, Andrew D. Griffiths, Pierre Bruhns, Colin Brenan
Abstract: Mining the antibody repertoire of plasma cells and plasmablasts could enable the discovery of useful antibodies for therapeutic or research purposes1. We present a method for high-throughput, single-cell screening of IgG-secreting primary cells to characterize antibody binding to soluble and membrane-bound antigens. CelliGO is a droplet microfluidics system that combines high-throughput screening for IgG activity, using fluorescence-based in-droplet single-cell bioassays2, with sequencing of paired antibody V genes, using in-droplet single-cell barcoded reverse transcription. We analyzed IgG repertoire diversity, clonal expansion and somatic hypermutation in cells from mice immunized with a vaccine target, a multifunctional enzyme or a membrane-bound cancer target. Immunization with these antigens yielded 100–1,000 IgG sequences per mouse. We generated 77 recombinant antibodies from the identified sequences and found that 93% recognized the soluble antigen and 14% the membrane antigen. The platform also allowed recovery of ~450–900 IgG sequences from ~2,200 IgG-secreting activated human memory B cells, activated ex vivo, demonstrating its versatility.