1. Schematic representation of our multi-dimensional chromatography off-line MALDI-TOF analysis of the complex immunoproteome of class I molecules (A), and a comparison of class I-bound peptides isolated from the surface of untransfected parental APC (positive spectra) and HLA B*2705 transfected APC (negative spectra) (B).
Schematic representation of our multi-dimensional chromatography off-line MALDI-TOF analysis of the complex immunoproteome of class I molecules (A), and a comparison of class I-bound peptides isolated from the surface of untransfected parental APC (positive spectra) and HLA B*2705 transfected APC (negative spectra) (B). This analysis was performed using a Bruker Reflex mass spectrometer (Bruker-Franzen Analytik, GMBH, Bremen, Germany) operated exclusively in the reflectron mode as described elsewhere (4, 48, 66, 187). Aliquots of each fraction (1–2 μl or ∼1% of the fraction) were mixed with an equal volume of matrix solution (α-cyano-4-hydroxycinnamic acid (10 mg/ml) in acetonitrile-ethanol 1:1 v/v), spotted uniformly onto a target, and dried for analysis. Replicate analysis and care with sample preparation can ensure high reproducibility and confidence in the differential analysis of class I ligands.
A. W. Purcell, and J. J. Gorman Mol Cell Proteomics 2004;3:
© 2004 The American Society for Biochemistry and Molecular Biology