Purification and HPLC-MS analysis of a naturally processed HCMV-derived peptide isolated from the HEK-293T/HLA-E+/Ul40+ cell transfectants and presented at the cell surface in the context of HLA-E

J Immunol Methods. 2007 Apr 30;322(1-2):128-36. doi: 10.1016/j.jim.2007.01.018. Epub 2007 Feb 21.

Abstract

A new method for isolation and characterization of peptides presented in the context of the nonclassical human leukocytes antigen (HLA) class I molecule HLA-E was developed. A combination of different chromatographic steps coupled with electrospray mass spectrometry allowed us to detect the presence of small amounts of a naturally processed human Cytomegalovirus (HCMV)-derived peptide isolated from the HEK-293T/HLA-E+/UL40+ transfected cells of from HELA cell line. The peptide sequence was confirmed by tandem mass spectrometry (MS/MS). This approach provides a versatile and sensitive method for direct identification of MHC class I-binding peptides that might be derive from different pathogen or tumor-associated proteins.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Cell Membrane / chemistry
  • Cells, Cultured
  • Chromatography, High Pressure Liquid / methods*
  • Cytomegalovirus
  • HLA Antigens / chemistry
  • HLA Antigens / immunology*
  • HLA-E Antigens
  • HeLa Cells
  • Histocompatibility Antigens Class I / chemistry
  • Histocompatibility Antigens Class I / immunology*
  • Humans
  • Mass Spectrometry / methods*
  • Molecular Sequence Data
  • Peptides / chemistry
  • Peptides / immunology*
  • Peptides / isolation & purification*
  • Sensitivity and Specificity
  • Tandem Mass Spectrometry
  • Transfection

Substances

  • HLA Antigens
  • Histocompatibility Antigens Class I
  • Peptides