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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
Author(s) -
Ayyoub Maha,
Monsarrat Bernard,
Mazarguil Honoré,
Gairin Jean Edouard
Publication year - 1998
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/(sici)1097-0231(19980515)12:9<557::aid-rcm199>3.0.co;2-d
Subject(s) - chemistry , electrospray ionization , chromatography , mass spectrometry , high performance liquid chromatography , peptide , degradation (telecommunications) , electrospray , epitope , liquid chromatography–mass spectrometry , antigen , tandem mass spectrometry , biochemistry , telecommunications , biology , computer science , genetics
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 demonstrate the efficiency of local modifications in the protection of a given peptide bond towards a specific peptidase activity. © 1998 John Wiley & Sons, Ltd.