TRAPnSeq allows high-throughput profiling of antigen-specific antibody-secreting cells.

Autor:	Asrat S; Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA., Devlin JC; Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA., Vecchione A; Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA., Klotz B; Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA., Setliff I; Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA., Srivastava D; Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA., Limnander A; Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA., Rafique A; Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA., Adler C; Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA., Porter S; Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA., Murphy AJ; Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA., Atwal GS; Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA., Sleeman MA; Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA., Lim WK; Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA., Orengo JM; Regeneron Pharmaceuticals, Tarrytown, NY 10591, USA.
Jazyk:	angličtina
Zdroj:	Cell reports methods [Cell Rep Methods] 2023 Jul 07; Vol. 3 (7), pp. 100522. Date of Electronic Publication: 2023 Jul 07 (Print Publication: 2023).
DOI:	10.1016/j.crmeth.2023.100522
Abstrakt:	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. Competing Interests: 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”). (© 2023 The Authors.)
Databáze:	MEDLINE
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