ARTEMIS: A Novel Mass-Spec Platform for HLA-Restricted Self and Disease-Associated Peptide Discovery

Frontiers in Immunology

卷:12

Mi-Youn Brusniak¹  Kathryn A. K. Finton²  Chance Brock²  Roland K. Strong²  Lisa A. Jones³  Chenwei Lin³  Philip R. Gafken³  Andrew J. Fioré-Gartland⁴ 

[1] Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA, United States;

[2] Division of Basic Science, Fred Hutchinson Cancer Research Center, Seattle, WA, United States;

[3] Proteomics Shared Resource, Fred Hutchinson Cancer Research Center, Seattle, WA, United States;

[4] Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA, United States;

关键词: MHC class I;    peptide-HLA complex;    mass spectrometry;    immunotherapy;    ligandome analysis;   

DOI  :  10.3389/fimmu.2021.658372

来源: DOAJ

【 摘 要 】

Conventional immunoprecipitation/mass spectroscopy identification of HLA-restricted peptides remains the purview of specializing laboratories, due to the complexity of the methodology, and requires computational post-analysis to assign peptides to individual alleles when using pan-HLA antibodies. We have addressed these limitations with ARTEMIS: a simple, robust, and flexible platform for peptide discovery across ligandomes, optionally including specific proteins-of-interest, that combines novel, secreted HLA-I discovery reagents spanning multiple alleles, optimized lentiviral transduction, and streamlined affinity-tag purification to improve upon conventional methods. This platform fills a middle ground between existing techniques: sensitive and adaptable, but easy and affordable enough to be widely employed by general laboratories. We used ARTEMIS to catalog allele-specific ligandomes from HEK293 cells for seven classical HLA alleles and compared results across replicates, against computational predictions, and against high-quality conventional datasets. We also applied ARTEMIS to identify potentially useful, novel HLA-restricted peptide targets from oncovirus oncoproteins and tumor-associated antigens.

【 授权许可】

Unknown