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© Research
Publication : Nature biotechnology

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

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Nature biotechnology - 30 Mar 2020

Gérard A, Woolfe A, Mottet G, Reichen M, Castrillon C, Menrath V, Ellouze S, Poitou A, Doineau R, Briseno-Roa L, Canales-Herrerias P, Mary P, Rose G, Ortega C, Delincé M, Essono S, Jia B, Iannascoli B, Richard-Le Goff O, Kumar R, Stewart SN, Pousse Y, Shen B, Grosselin K, Saudemont B, Sautel-Caillé A, Godina A, McNamara S, Eyer K, Millot GA, Baudry J, England P, Nizak C, Jensen A, Griffiths AD, Bruhns P, Brenan C

Link to Pubmed [PMID] – 32231335

Nat. Biotechnol. 2020 Mar;

Mining the antibody repertoire of plasma cells and plasmablasts could enable the discovery of useful antibodies for therapeutic or research purposes. 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 bioassays, 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.

https://www.ncbi.nlm.nih.gov/pubmed/32231335