Semiautomated high-sensitivity profiling of human blood serum glycoproteins through lectin preconcentration and multidimensional chromatography/tandem mass spectrometry

Milan Madera, Yehia Mechref, Iveta Klouckova, Milos V. Novotny

Research output: Contribution to journalArticlepeer-review

67 Scopus citations

Abstract

We describe an effective analytical approach to identify trace glycoproteins in a small volume of human serum. The system is based on automatable affinity enrichment through silica-based lectin microcolumns and a further separation of the retained glycoproteins on a reversed-phase liquid chromatography with superficially porous packing, operating at high temperature. The fractionated sample is further directed into a 96-well plate for trypsinization and LC-MS/MS analysis. Using a major-component depleted blood serum (16 μg total protein), we were able to identify 271 glycoproteins through this analytical system.

Original languageEnglish
Pages (from-to)2348-2363
Number of pages16
JournalJournal of Proteome Research
Volume5
Issue number9
DOIs
StatePublished - Sep 2006

Keywords

  • Glycoprotein Enrichment
  • Human Blood Serum Glycoproteins
  • LC-MS/MS
  • Microcolumn Lectin Affinity Chromatography
  • On-line Coupling of Microcolumn Lectin Affinity Chromatography and Poroshell LC-Columns
  • Protein Separation

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