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High-Throughput, Fast, and Sensitive Immunopeptidomics Sample Processing for Mass Spectrometry.
Journal article

High-Throughput, Fast, and Sensitive Immunopeptidomics Sample Processing for Mass Spectrometry.

  • Marino F Ludwig Centre for Cancer Research, University of Lausanne, Epalinges, Switzerland.
  • Chong C Ludwig Centre for Cancer Research, University of Lausanne, Epalinges, Switzerland.
  • Michaux J Ludwig Centre for Cancer Research, University of Lausanne, Epalinges, Switzerland.
  • Bassani-Sternberg M Ludwig Centre for Cancer Research, University of Lausanne, Epalinges, Switzerland. Michal.Bassani@chuv.ch.
  • 2019-01-23
Published in:
  • Methods in molecular biology (Clifton, N.J.). - 2019
Affinity purification
Antigen processing and presentation
HLA class I and HLA class II
Human leukocyte antigen
Immunopeptidomics
Mass spectrometry
Cell Line
Chromatography, High Pressure Liquid
High-Throughput Screening Assays
Histocompatibility Antigens Class I
Histocompatibility Antigens Class II
Humans
Hybridomas
Neoplasms
Peptides
Tandem Mass Spectrometry
Time Factors
English Comprehensive knowledge of the HLA class I and class II peptides presented to T cells is crucial for designing innovative therapeutics against cancer and other diseases. So far, methodologies for recovery of HLA class I and II peptides for subsequent mass spectrometry-based analysis have been a major limitation. In this chapter we describe a detailed protocol for a high-throughput, reproducible, and sensitive immunoaffinity-purification of HLA-I and HLA-II peptides from up to 96 samples in a plate format, suitable for tissue samples and cell lines. Our methodology reduces sample handling, has a competitive peptide yield, and can be completed within 5 h. This simplified pipeline is applicable for basic and clinical applications.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://susi.usi.ch/global/documents/106580
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