Purification of HLA Immunopeptidomes from Human Thymus

Producció científica: Capítol de llibreCapítolRecercaAvaluat per experts

Resum

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 originalEnglish
Títol de la publicacióMethods in Molecular Biology
Pàgines127-136
Nombre de pàgines10
Volum2420
DOIs
Estat de la publicacióPublicada - 15 de des. 2021

Sèrie de publicacions

NomMethods in molecular biology (Clifton, N.J.)
EditorHumana Press
ISSN (imprès)1064-3745

Fingerprint

Navegar pels temes de recerca de 'Purification of HLA Immunopeptidomes from Human Thymus'. Junts formen un fingerprint únic.

Com citar-ho