Details

Autor(en) / Beteiligte

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

Titel

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

Ist Teil von

• Nature biotechnology, 2020-06, Vol.38 (6), p.715-721

Ort / Verlag

New York: Nature Publishing Group US

Erscheinungsjahr

2020

Quelle

MEDLINE

Beschreibungen/Notizen

• 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.