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

E. Millo, G. Pietra, A. Armirotti, P. Vacca, M. C. Mingari, L. Moretta, G. Damonte

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)128-136
Number of pages9
JournalJournal of Immunological Methods
Volume322
Issue number1-2
DOIs
Publication statusPublished - Apr 30 2007

Keywords

  • Cytomegalovirus
  • High-performance liquid chromatography
  • HLA-E
  • mass-spectrometry
  • Peptides

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

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