The use of post-source decay in matrix-assisted laser desorption/ionisation mass spectrometry to delineate T cell determinants.
The identification of naturally processed peptides presented by molecules of the major histocompatibility complex (MHC) has progressed significantly over the past decade. The elution of peptides from immunoaffinity purified complexes of MHC class I or class II molecules has provided highly specific biochemical information regarding the nature of endogenous peptides capable of binding to and being presented by particular MHC alleles. Whilst Edman chemistry is sufficient for the identification of abundant or homogeneous immunodominant peptides contained in samples of fractionated peptides, mass spectrometry has proved more powerful for sequencing less abundant species present in the typically heterogeneous fractions of eluted peptides. This review focuses on the characterisation of T cell determinants by matrix-assisted laser desorption/ionisation (MALDI)-time-of-flight (TOF) mass spectrometry (MS). We demonstrate, with specific examples, the utility of post-source decay in MALDI-TOF MS for the characterisation of the amino acid sequences of both native and modified T cell determinants. The potential advantages and pitfalls of this technique relative to the more commonly used forms of tandem mass spectrometry in electrospray and ion spray modes of ionisation as well as hybrid quadrupole-quadrupole-TOF instruments are discussed. We highlight the complementarity between these techniques and discuss the advantages in the combined use of both MALDI- and electrospray-based instrumentation in epitope identification strategies.
The Department of Microbiology and Immunology, University of Melbourne, 3052, Victoria, Parkville, Australia. a.purcell@microbiology.unimelb.edu.au
A W
AW
Purcell
J J
JJ
Gorman
D000818:Animals; D006801:Humans; D008285:Major Histocompatibility Complex / Q000276:immunology*; D013058:Mass Spectrometry / Q000379:methods*; D013601:T-Lymphocytes / Q000276:immunology*