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

Annabelle Gérard (1) , Adam Woolfe (1) , Guillaume Mottet (2) , Marcel Reichen (1) , Carlos Castrillón (2, 3, 4) , Vera Menrath (1) , Sami Ellouze (1) , Adeline Poitou (1) , Raphaël Doineau (1, 3, 4) , Luis Briseño-Roa (1) , Pablo Canales-Herrerias (2, 3, 5) , Pascaline Mary (6) , Gregory Rose (6) , Charina Ortega (6) , Matthieu Delincé (6) , Sosthène Essono (6) , Bin Jia (7) , Bruno Iannascoli (2) , Odile Richard-Le Goff (2) , Roshan Kumar (6) , Samantha N Stewart (6) , Yannick Pousse (1) , Bingqing Shen (1) , Kevin Grosselin (1, 4) , Baptiste Saudemont (2, 4) , Antoine Sautel-Caillé (4) , Alexei Godina (4) , Scott Mcnamara (1) , Klaus Eyer (5) , Gaël A Millot (8) , Jean Baudry (5) , Patrick England (9, 10) , Clément Nizak (4) , Allan Jensen (1) , Andrew D Griffiths (4) , Pierre Bruhns (2) , Colin Brenan (1, 6)

1 HiFiBiO Therapeutics SAS [Paris]
2 Anticorps en thérapie et pathologie - Antibodies in Therapy and Pathology
3 FdV - Ecole Doctorale Frontières du Vivant - Programme Bettencourt
4 CBI - Chimie-Biologie-Innovation (UMR 8231)
5 LCMD - Laboratoire Colloïdes et Matériaux Divisés
6 HiFiBiO Therapeutics Inc. [Cambridge]
7 Abbvie Bioresearch Center [Worcester]
8 Hub Bioinformatique et Biostatistique - Bioinformatics and Biostatistics HUB
9 Biophysique Moléculaire (Plate-forme)
10 Biophysique des macromolécules et leurs interactions

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.

Domains: Life Sciences [q-bio]; Immunology

Annabelle Gérard, Adam Woolfe, Guillaume Mottet, Marcel Reichen, Carlos Castrillón, et al.. High-throughput single-cell activity-based screening and sequencing of antibodies using droplet microfluidics. Nature Biotechnology, 2020, 38, pp.715-721. ⟨10.1038/s41587-020-0466-7⟩. ⟨pasteur-02505776⟩