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  • 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

    Nature biotechnology, 06/2020, Letnik: 38, Številka: 6
    Journal Article

    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.