Pertussis toxoid structure: a collaboration and comparison of two-dimensional liquid chromatography-tandem mass spectrometry, ultraperformance liquid chromatography-mass spectrometryE, and capillary liquid chromatography-matrix-assisted laser desorption ionization-tandem mass spectrometry.

ABSTRACT

The overall structure of pertussis toxoid has been established by analysis of its tryptic digest using two-dimensional liquid chromatography-tandem mass spectrometry (2D-LC-MS/MS), capillary liquid chromatography-matrix-assisted laser desorption ionization-tandem mass spectrometry (CapLC-MALDI-MS/MS), and ultraperformance liquid chromatography-mass spectrometry(E) (UPLC-MS(E)). In addition to oxidation and hydrolysis of amino acids losses of terminal peptides are observed. On-line UPLC-MS(E) generated a similar sequence coverage as the other two methods that involved off-line fraction collection. In light of recent favorable comparisons to data-dependent acquisition, UPLC-MS(E) should be the initial method of choice for analysis of a peptide mixture of moderate complexity.