TRAPnSeq allows high-throughput profiling of antigen-specific antibody-secreting cells.
IgE
IgG
antibody-secreting cells
plasma cells
plasmablasts
repertoire analysis
secretion trap
Journal
Cell reports methods
ISSN: 2667-2375
Titre abrégé: Cell Rep Methods
Pays: United States
ID NLM: 9918227360606676
Informations de publication
Date de publication:
24 07 2023
24 07 2023
Historique:
received:
10
02
2023
revised:
06
05
2023
accepted:
15
06
2023
medline:
4
8
2023
pubmed:
3
8
2023
entrez:
3
8
2023
Statut:
epublish
Résumé
Following activation by cognate antigen, B cells undergo fine-tuning of their antigen receptors and may ultimately differentiate into antibody-secreting cells (ASCs). While antigen-specific B cells that express surface receptors (B cell receptors [BCRs]) can be readily cloned and sequenced following flow sorting, antigen-specific ASCs that lack surface BCRs cannot be easily profiled. Here, we report an approach, TRAPnSeq (antigen specificity mapping through immunoglobulin [Ig] secretion TRAP and Sequencing), that allows capture of secreted antibodies on the surface of ASCs, which in turn enables high-throughput screening of single ASCs against large antigen panels. This approach incorporates flow cytometry, standard microfluidic platforms, and DNA-barcoding technologies to characterize antigen-specific ASCs through single-cell V(D)J, RNA, and antigen barcode sequencing. We show the utility of TRAPnSeq by profiling antigen-specific IgG and IgE ASCs from both mice and humans and highlight its capacity to accelerate therapeutic antibody discovery from ASCs.
Identifiants
pubmed: 37533642
doi: 10.1016/j.crmeth.2023.100522
pii: S2667-2375(23)00151-0
pmc: PMC10391570
doi:
Substances chimiques
Antigens
0
Antibodies
0
Receptors, Antigen, B-Cell
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
100522Informations de copyright
© 2023 The Authors.
Déclaration de conflit d'intérêts
This study was sponsored by Regeneron Pharmaceuticals, Inc. All authors are current or former employees of Regeneron and may hold stock options in the company. S.A., J.C.D., A.V., B.K., I.S., G.S.A., M.A.S., W.K.L., and J.M.O. are inventors on a pending US patent application on TRAPnSeq (“Methods of Mapping Antigen Specificity to Antibody-Secreting Cells”). A.J.M. is an inventor on a pending US patent application (#16/363,774; “Humanized Rodents for Testing Therapeutic Agents”).
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