ANALYSIS OF THE DEGRADATION MECHANISMS OF MHC CLASS I-PRESENTED TUMORANTIGENIC PEPTIDES BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY ELECTROSPRAY-IONIZATION MASS-SPECTROMETRY - APPLICATION TO THE DESIGN OF PEPTIDASE-RESISTANT ANALOGS
M. Ayyoub et al., ANALYSIS OF THE DEGRADATION MECHANISMS OF MHC CLASS I-PRESENTED TUMORANTIGENIC PEPTIDES BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY ELECTROSPRAY-IONIZATION MASS-SPECTROMETRY - APPLICATION TO THE DESIGN OF PEPTIDASE-RESISTANT ANALOGS, Rapid communications in mass spectrometry, 12(9), 1998, pp. 557-564
Peptide vaccines based on the use of MHC class I restricted epitopes a
re 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 de
sign of structurally modified analogs, and the analysis of their degra
dation by on-line HPLC/ESI-MS, allowed us to demonstrate the efficienc
y of local modifications in the protection of a given peptide bond tow
ards a specific peptidase activity. (C) 1998 John Wiley & Sons, Ltd.