Purification of HLA Immunopeptidomes from Human Thymus

Producción científica: Capítulo del libroCapítuloInvestigaciónrevisión exhaustiva

Resumen

Mass spectrometry has become an essential technique for the analysis of peptide repertoires presented by MHC molecules to T lymphocytes. Years ago, analyses of MHC peptidomes were performed using a great number of cells, and cell lines were chosen as the main peptide source. Mass spectrometry devices have been improved in terms of sensitivity and resolution, making feasible the analysis of samples with relatively small amounts of cells. Thus, analyses of MHC peptide repertoires from different tissue samples are now available. Here, I describe a protocol to process human thymus samples to purify HLA class I- or HLA-DR-associated peptidomes. For that, cells are lysed using a nonionic detergent together with a mechanical cell rupture. Immunopeptidomes are purified by immunoaffinity chromatography. The peptide pool is fractionated by ionic chromatography. Finally, peptide fragmentation and identification are conducted by LC-MS/MS and the use of MASCOT search engine.

Idioma originalInglés
Título de la publicación alojadaMethods in Molecular Biology
Páginas127-136
Número de páginas10
Volumen2420
DOI
EstadoPublicada - 15 dic 2021

Serie de la publicación

NombreMethods in molecular biology (Clifton, N.J.)
EditorialHumana Press
ISSN (versión impresa)1064-3745

Huella

Profundice en los temas de investigación de 'Purification of HLA Immunopeptidomes from Human Thymus'. En conjunto forman una huella única.

Citar esto