Journal Article

High-throughput single-cell activity-based screening and sequencing of antibodies using droplet microfluidics

Doineau, Raphaël, Stewart, Samantha N., Gérard, Annabelle, Canales-Herrerias, Pablo, Delincé, Matthieu, Castrillon, Carlos, Richard-Le Goff, Odile, Mottet, Guillaume, Ortega, Charina, McNamara, Scott, Ellouze, Sami, Essono, Sosthene, Jia, Bin, Sautel-Caillé, Antoine, Bruhns, Pierre, Iannascoli, Bruno, Kumar, Roshan, Pousse, Yannick, Godina, Alexei, Baudry, Jean, Poitou, Adeline, Rose, Gregory, Brenan, Colin, Eyer, Klaus, Briseno-Roa, Luis, Mary, Pascaline, Reichen, Marcel, Nizak, Clément, Woolfe, Adam, Saudemont, Baptiste, Griffiths, Andrew D., Millot, Gaël A., Grosselin, Kevin, Menrath, Vera, England, Patrick, Jensen, Allan, Shen, Bingqing

2020

Overview

Mining the antibody repertoire of plasma cells and plasmablasts could enable the discovery of useful antibodies for therapeutic or research purposes 1 . 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. Celli GO is a droplet microfluidics system that combines high-throughput screening for IgG activity, using fluorescence-based in-droplet single-cell bioassays 2 , 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. Millions of primary IgG-secreting cells from mouse and human are characterized for activity and antibody sequence at the single-cell level.

• Publisher: Nature Publishing Group US,Nature Publishing Group
• Subject: 631/250/2152/2153/1291 / 631/61/24/590 / Agriculture / Animals / Antibodies / Antibodies - genetics / Antigens / Antigens - immunology / B-Lymphocytes - immunology / Bioinformatics / Biomedical and Life Sciences / Biomedical Engineering/Biotechnology / Biomedicine / Biotechnology / Cancer Vaccines - immunology / DNA - analysis / DNA - genetics / DNA sequencing / Droplets / Fluorescence / Genetic aspects / High-Throughput Nucleotide Sequencing - instrumentation / High-Throughput Nucleotide Sequencing - methods / High-throughput screening / Humans / Immunization / Immunoglobulin G / Immunoglobulin G - genetics / Immunological memory / Immunology / Letter / Life Sciences / Lymphocytes B / Membranes / Memory cells / Methods / Mice / Microfluidic Analytical Techniques - instrumentation / Microfluidics / Nucleotide sequencing / Plasma cells / Reverse transcription / Screening / Single-Cell Analysis - instrumentation / Single-Cell Analysis - methods / Somatic hypermutation / Viral antibodies