Protocol for purification and identification of MHC class I immunopeptidome from cancer cell lines

Sonali V Mohan; Keshava K Datta; Rebekah Ziegman; Corey Smith; Harsha Gowda

doi:10.1016/j.xpro.2021.100385

Protocol for purification and identification of MHC class I immunopeptidome from cancer cell lines

STAR Protoc. 2021 Mar 18;2(1):100385. doi: 10.1016/j.xpro.2021.100385. eCollection 2021 Mar 19.

Authors

Sonali V Mohan^{1

2}, Keshava K Datta¹, Rebekah Ziegman¹, Corey Smith^{3

4

5}, Harsha Gowda^{1

2

3}

Affiliations

¹ Cancer Precision Medicine Group, QIMR Berghofer Medical Research Institute, Herston, QLD, Australia.
² Faculty of Medicine, The University of Queensland, Herston, QLD, Australia.
³ School of Biomedical Sciences, Faculty of Health, Queensland University of Technology, Brisbane, QLD, Australia.
⁴ QIMR Centre for Immunotherapy and Vaccine Development, QIMR Berghofer Medical Research Institute, Brisbane, QLD, Australia.
⁵ Translational and Human Immunology Laboratory, QIMR Berghofer Medical Research Institute, Brisbane, QLD, Australia.

Abstract

Major histocompatibility complexes (MHC) play a critical role in immunity by presenting peptides on the cell surface for T cell recognition. Identification of these peptides can be valuable to develop vaccines or immunotherapeutic strategies for infectious diseases and cancers. Mass spectrometry is the only tool available for unbiased identification of the immunopeptidome. Here, we describe a protocol for purification and identification of MHC class I peptides, including in-house purification of anti-MHC-antibody from hybridoma cells and the LC-MS/MS analysis of MHC-I bound peptides.

Keywords: Cancer; Immunology; Mass spectrometry; Proteomics.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line
Cell Membrane / immunology
Chromatography, Liquid / methods
HLA Antigens / immunology
HLA Antigens / isolation & purification
Histocompatibility Antigens Class I / chemistry
Histocompatibility Antigens Class I / immunology
Histocompatibility Antigens Class I / isolation & purification*
Humans
Major Histocompatibility Complex / immunology
Peptides / chemistry
Peptides / immunology*
Peptides / isolation & purification*
Proteins / isolation & purification
Proteomics / methods
Tandem Mass Spectrometry / methods

Substances

HLA Antigens
Histocompatibility Antigens Class I
Peptides
Proteins