Journal
JOURNAL OF PROTEOME RESEARCH
Volume 5, Issue 9, Pages 2348-2363Publisher
AMER CHEMICAL SOC
DOI: 10.1021/pr060169x
Keywords
microcolumn lectin affinity chromatography; glycoprotein enrichment; protein separation; human blood serum glycoproteins; on-line coupling of microcolumn lectin affinity; chromatography and poroshell LC-Columns; LC-MS/MS
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Funding
- NCRR NIH HHS [RR018942] Funding Source: Medline
- NIGMS NIH HHS [GM24349] Funding Source: Medline
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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 mu g total protein), we were able to identify 271 glycoproteins through this analytical system.
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