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STAR Protoc DOI:10.1016/j.xpro.2022.101910

HLA-I immunopeptidome profiling of human cells infected with high-containment enveloped viruses.

Publication TypeJournal Article
Year of Publication2022
AuthorsWeingarten-Gabbay, S, Pearlman, LR, Chen, D-Y, Klaeger, S, Taylor, HB, Welch, NL, Keskin, DB, Carr, SA, Abelin, JG, Saeed, M, Sabeti, PC
JournalSTAR Protoc
Volume3
Issue4
Pages101910
Date Published2022 Dec 16
ISSN2666-1667
KeywordsHistocompatibility Antigens Class I, Humans, Mass Spectrometry, Peptides, Viruses
Abstract

Immunopeptidome profiling of infected cells is a powerful technique for detecting viral peptides that are naturally processed and loaded onto class I human leukocyte antigens (HLAs-I). Here, we provide a protocol for preparing samples for immunopeptidome profiling that can inactivate enveloped viruses while still preserving the integrity of the HLA-I complex. We detail steps for lysate preparation of infected cells followed by HLA-I immunoprecipitation and virus inactivation. We further describe peptide purification for mass spectrometry outside a high-containment facility. For complete details on the use and execution of this protocol, please refer to Weingarten-Gabbay et al. (2021)..

DOI10.1016/j.xpro.2022.101910
Pubmed

https://www.ncbi.nlm.nih.gov/pubmed/36595954?dopt=Abstract

Alternate JournalSTAR Protoc
PubMed ID36595954
PubMed Central IDPMC9731565
Grant ListU19 AI110818 / AI / NIAID NIH HHS / United States