美国1CellBiO公司Colin Brenan、法国巴斯德研究所Pierre Bruhns、巴黎第九大学Andrew D. Griffiths等研究人员合作利用液滴微流控技术实现单细胞水平的抗体筛选与测序。该项研究成果于2020年3月30日在线发表在《自然—生物技术》杂志上。

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

Issue&Volume: 2020-03-30

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.

DOI: 10.1038/s41587-020-0466-7

Source: https://www.nature.com/articles/s41587-020-0466-7