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Journal articles
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 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 Millot
8
Jean Baudry
5
Patrick England
9, 10
Clément Nizak
4
Allan Jensen
1
Andrew Griffiths
4
Pierre Bruhns
2, *
Colin Brenan
1, 6
*
Corresponding author
Carlos Castrillón 2, 3, 4
Author
Vera Menrath 1
Author
Sami Ellouze 1
Author
Adeline Poitou 1
Author
Raphaël Doineau 1, 3, 4
Author
Luis Briseño-Roa 1
Author
Pablo Canales-Herrerias 2, 3, 5
Author
Pascaline Mary 6
Author
Samantha N Stewart 6
Author
Yannick Pousse 1
Author
Bingqing Shen 1
Author
Kevin Grosselin 1, 4
Author
Baptiste Saudemont 2, 4
Author IdHAL : baptiste-saudemont ORCID : https://orcid.org/0000-0002-8360-3565
Antoine Sautel-Caillé 4
Author
Alexei Godina 4
Author
Scott Mcnamara 1
Author
Klaus Eyer 5
Author
Gaël A Millot 8
Author IdHAL : gael-millot ORCID : https://orcid.org/0000-0002-0591-3509
Jean Baudry 5
Author
Patrick England 9, 10
Author IdHAL : patrick-england ORCID : https://orcid.org/0000-0001-6410-5918
Clément Nizak 4
Author
Allan Jensen 1
Author
Andrew D Griffiths 4
Author
Pierre Bruhns 2
Correspondent author IdHAL : pierre-bruhns ResearcherId : http://www.researcherid.com/rid/F-5567-2013 ORCID : https://orcid.org/0000-0002-4709-8936
Colin Brenan 1, 6
Author
1
HiFiBiO Therapeutics SAS [Paris] (Paris - France)
2
Anticorps en thérapie et pathologie - Antibodies in Therapy and Pathology (Département d'Immunologie, 25-28, rue du Docteur Roux, 75724 Paris cedex 15 - France)
- Institut Pasteur [Paris] (25-28, rue du docteur Roux, 75724 Paris cedex 15 - France)
- INSERM - Institut National de la Santé et de la Recherche Médicale : U1222 (101, rue de Tolbiac, 75013 Paris - France)
3
FdV - Ecole Doctorale Frontières du Vivant - Programme Bettencourt (France)
- UPD5 - Université Paris Descartes - Paris 5 (12, rue de l'École de Médecine - 75270 Paris cedex 06 - France)
- PRES Sorbonne Paris Cité (France)
4
CBI - Chimie-Biologie-Innovation (10 rue Vauquelin 75231 Paris Cedex 05 - France)
- ESPCI Paris - Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (10 rue Vauquelin, 75231 Paris cedex 05 - France)
- PSL - Université Paris sciences et lettres (60 rue Mazarine 75006 Paris - France)
- SU - Sorbonne Université : UMR8231 (21 rue de l’École de médecine - 75006 Paris - France)
- CNRS - Centre National de la Recherche Scientifique : UMR8231 (France)
5
LCMD - Laboratoire Colloïdes et Matériaux Divisés (10 rue Vauquelin 75005 Paris - France)
- CNRS - Centre National de la Recherche Scientifique : UMR7195 (France)
- ESPCI Paris - Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (10 rue Vauquelin, 75231 Paris cedex 05 - France)
- PSL - Université Paris sciences et lettres (60 rue Mazarine 75006 Paris - France)
6
HiFiBiO Therapeutics Inc. [Cambridge] (Cambridge, MA, USA - United States)
7
Abbvie Bioresearch Center [Worcester] (Worcester, MA - United States)
8
Hub Bioinformatique et Biostatistique - Bioinformatics and Biostatistics HUB (C3BI, 25-28 rue du Docteur Roux, 75724 Paris cedex 15 - France)
- Institut Pasteur [Paris] (25-28, rue du docteur Roux, 75724 Paris cedex 15 - France)
- CNRS - Centre National de la Recherche Scientifique : USR3756 (France)
9
Biophysique Moléculaire (Plate-forme) (Citech
25-28, rue du Docteur Roux F-75724 Paris Cedex 15 - France)
- Institut Pasteur [Paris] (25-28, rue du docteur Roux, 75724 Paris cedex 15 - France)
- CNRS - Centre National de la Recherche Scientifique : UMR3528 (France)
10
Biophysique des macromolécules et leurs interactions (25-28 rue du Docteur Roux F-75724 Paris Cedex 15 - France)
- Institut Pasteur [Paris] (25-28, rue du docteur Roux, 75724 Paris cedex 15 - France)
- CNRS - Centre National de la Recherche Scientifique : UMR3528 (France)
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.
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Journal articles
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https://hal-pasteur.archives-ouvertes.fr/pasteur-02505776
Contributor : Pierre Bruhns <>
Submitted on : Wednesday, March 11, 2020 - 5:11:54 PM
Last modification on : Wednesday, October 14, 2020 - 3:45:10 AM
Long-term archiving on: : Friday, June 12, 2020 - 5:15:40 PM
Contributor : Pierre Bruhns <>
Submitted on : Wednesday, March 11, 2020 - 5:11:54 PM
Last modification on : Wednesday, October 14, 2020 - 3:45:10 AM
Long-term archiving on: : Friday, June 12, 2020 - 5:15:40 PM
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Bruhns_Nature_Biotech_2020.pdf
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Distributed under a Creative Commons Attribution - NonCommercial - NoDerivatives 4.0 International License
Identifiers
- HAL Id : pasteur-02505776, version 1
- DOI : 10.1038/s41587-020-0466-7
- PUBMED : 32231335
Collections
PASTEUR | INC-CNRS | SORBONNE-UNIVERSITE | PARISTECH | CNRS | UNIV-PARIS | PSL | ESPCI-PSL | ESPCI | LCMD
Citation
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, Nature Publishing Group, 2020, pp.715-721. ⟨10.1038/s41587-020-0466-7⟩. ⟨pasteur-02505776⟩
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