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 N 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 A Millot
(8)
,
Jean Baudry
(5)
,
Patrick England
(9, 10)
,
Clément Nizak
(4)
,
Allan Jensen
(1)
,
Andrew D Griffiths
(4)
,
Pierre Bruhns
(2)
,
Colin Brenan
(1, 6)
1
HiFiBiO Therapeutics SAS [Paris]
2 Anticorps en thérapie et pathologie - Antibodies in Therapy and Pathology
3 FdV - Ecole Doctorale Frontières du Vivant - Programme Bettencourt
4 CBI - Chimie-Biologie-Innovation (UMR 8231)
5 LCMD - Laboratoire Colloïdes et Matériaux Divisés
6 HiFiBiO Therapeutics Inc. [Cambridge]
7 Abbvie Bioresearch Center [Worcester]
8 Hub Bioinformatique et Biostatistique - Bioinformatics and Biostatistics HUB
9 Biophysique Moléculaire (Plate-forme)
10 Biophysique des macromolécules et leurs interactions
2 Anticorps en thérapie et pathologie - Antibodies in Therapy and Pathology
3 FdV - Ecole Doctorale Frontières du Vivant - Programme Bettencourt
4 CBI - Chimie-Biologie-Innovation (UMR 8231)
5 LCMD - Laboratoire Colloïdes et Matériaux Divisés
6 HiFiBiO Therapeutics Inc. [Cambridge]
7 Abbvie Bioresearch Center [Worcester]
8 Hub Bioinformatique et Biostatistique - Bioinformatics and Biostatistics HUB
9 Biophysique Moléculaire (Plate-forme)
10 Biophysique des macromolécules et leurs interactions
Guillaume Mottet
- Function : Author
- PersonId : 961563
Raphaël Doineau
- Function : Author
- PersonId : 1232080
- IdHAL : raphael-doineau
- ORCID : 0000-0003-0817-7778
- IdRef : 236676474
Odile Richard-Le Goff
- Function : Author
- PersonId : 750176
- IdHAL : odile-richard-le-goff
- ORCID : 0000-0002-6263-2375
Kevin Grosselin
- Function : Author
- PersonId : 799439
- ORCID : 0000-0001-6189-3197
Baptiste Saudemont
- Function : Author
- PersonId : 734931
- IdHAL : baptiste-saudemont
- ORCID : 0000-0002-8360-3565
Klaus Eyer
- Function : Author
- PersonId : 784887
- ORCID : 0000-0001-9344-5110
Gaël A Millot
- Function : Author
- PersonId : 185072
- IdHAL : gael-millot
- ORCID : 0000-0002-0591-3509
Jean Baudry
- Function : Author
- PersonId : 741526
- IdHAL : jean-baudry
- ORCID : 0000-0002-5526-499X
- IdRef : 075107228
Patrick England
- Function : Author
- PersonId : 735030
- IdHAL : patrick-england
- ORCID : 0000-0001-6410-5918
Pierre Bruhns
Connectez-vous pour contacter l'auteur
- Function : Correspondent author
- PersonId : 183222
- IdHAL : pierre-bruhns
- ORCID : 0000-0002-4709-8936
- IdRef : 168916401
Connectez-vous pour contacter l'auteur
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.
Domains
Immunology
Origin : Files produced by the author(s)
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