Analysis of the degradation mechanisms of MHC class I-presented tumor antigenic peptides by high performance liquid chromatography/electrospray ionization mass spectrometry: application to the design of peptidase-resistant analogs

Rapid Commun Mass Spectrom. 1998;12(9):557-64. doi: 10.1002/(SICI)1097-0231(19980515)12:9<557::AID-RCM199>3.0.CO;2-D.

Abstract

Peptide vaccines based on the use of MHC class I restricted epitopes are currently assayed for anti-tumor and anti-viral immunotherapy. With the aim of designing minimally modified, peptidase-resistant analogs, we developed a rational approach based on a detailed understanding of the degradation mechanism of peptides in serum. Degradation of murine tumor antigen P198 and human tumor antigen MAGE-3.A1 was followed by on line high performance liquid chromatography/electrospray ionization mass spectrometry (HPLC/ESI-MS). This method provided high precision and sensitivity for rapid and direct analysis of degradation fragments in a complex mixture and, very importantly, precise identification of transient degradation fragments present at low concentrations. The design of structurally modified analogs, and the analysis of their degradation by on-line HPLC/ESI-MS, allowed us to to demonstrate the efficiency of local modifications in the protection of a given peptide bond towards a specific peptidase activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid
  • Epitopes
  • Genes, MHC Class I / immunology*
  • Humans
  • Mass Spectrometry
  • Peptide Hydrolases / metabolism*
  • Peptides / analysis
  • Peptides / metabolism*
  • Spectrophotometry, Ultraviolet

Substances

  • Epitopes
  • Peptides
  • Peptide Hydrolases