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Abstract
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.
Original language | English |
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Title of host publication | Methods in Molecular Biology |
Pages | 127-136 |
Number of pages | 10 |
Volume | 2420 |
DOIs | |
Publication status | Published - 15 Dec 2021 |
Publication series
Name | Methods in molecular biology (Clifton, N.J.) |
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Publisher | Humana Press |
ISSN (Print) | 1064-3745 |
Keywords
- Antigen presentation
- Human leukocyte antigen (HLA)
- Immunopeptidome
- Mass spectrometry
- Thymus
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Dive into the research topics of 'Purification of HLA Immunopeptidomes from Human Thymus'. Together they form a unique fingerprint.Projects
- 1 Finished
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MECANISMOS DE TOLERANCIA EN LA DIABETES DE TIPO 1: PROCESAMIENTO DE ANTIGENOS PEPTIDICOS Y LIPIDICOS Y MECANISMOS DE REGULACION EN TIMO Y EN PERIFERIA
Jaraquemada Perez de Guzman, M. D. (Principal Investigator), Alvarez, I. (Co-Investigador/a Principal), Pizarro Lozano, E. (Investigator), Roura Mir, C. (Investigator) & Pastor Moreno, A. (Collaborator)
Spanish Ministry of Economy and Competitiveness (MINECO), European Regional Development Fund (FEDER)
1/01/19 → 31/07/22
Project: Research Projects and Other Grants