High-throughput single-cell activity-based screening and sequencing of antibodies using droplet micr
Author: ["Annabelle Gérard","Adam Woolfe","Guillaume Mottet","Marcel Reichen","Carlos Castrillon","Vera Menrath","Sami Ellouze","Adeline Poitou","Raphaël Doineau","Luis Briseno-Roa","Pablo Canales-Herrerias","Pascaline Mary","Gregory Rose","Charina Ortega","Matth
Publication: Nature Biotechnology
CITE.CC academic search helps you expand the influence of your papers.
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. Millions of primary IgG-secreting cells from mouse and human are characterized for activity and antibody sequence at the single-cell level.
Cite this article
Gérard, A., Woolfe, A., Mottet, G. et al. High-throughput single-cell activity-based screening and sequencing of antibodies using droplet microfluidics. Nat Biotechnol 38, 715–721 (2020). https://doi.org/10.1038/s41587-020-0466-7