APPLICATION OF MULTIDIMENSIONAL AFFINITY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY AND ELECTROSPRAY-IONIZATION LIQUID-CHROMATOGRAPHY MASS-SPECTROMETRY TO THE CHARACTERIZATION OF GLYCOSYLATION IN SINGLE-CHAIN PLASMINOGEN-ACTIVATOR INITIAL RESULTS

Preliminary results are presented using a combination of affinity chro matography, reversed-phase HPLC and electrospray ionization mass spect rometry to produced peptide maps for N-linked, O-linked and non-glycos ylated peptides from an endoproteinase LysC digest of DSPA alpha 1, a recombinant DNA derived glycoprotein. Although the system was used to identify a number of major N-linked structures, notably complex biante nnary structures attached to asparagine 362, no O-linked glycopeptides from the possible 4 attachment sites were identified. The system did, however, demonstrate the feasibility of the approach and the applicab ility of the instrumental system.