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

J Chromatogr A. 1996 Oct 25;750(1-2):35-42. doi: 10.1016/0021-9673(96)00528-6.

Abstract

Preliminary results are presented using a combination of affinity chromatography, reversed-phase HPLC and electrospray ionization mass spectrometry to produced peptide maps for N-linked, O-linked and non-glycosylated 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 biantennary 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 applicability of the instrumental system.

Publication types

  • Comparative Study

MeSH terms

  • Carbohydrate Sequence
  • Chromatography, High Pressure Liquid / instrumentation
  • Chromatography, High Pressure Liquid / methods*
  • Concanavalin A / chemistry
  • Glycosylation
  • Haptens
  • Lectins / chemistry
  • Mass Spectrometry / methods*
  • Molecular Sequence Data
  • Peptide Mapping
  • Plasminogen Activators / analysis*
  • Plasminogen Activators / chemistry
  • Sensitivity and Specificity

Substances

  • Haptens
  • Lectins
  • Concanavalin A
  • Plasminogen Activators