Title: Microchip capillary gel electrophoresis combined with lectin affinity enrichment employing magnetic beads for glycoprotein analysis
Language: English
Authors: Engel, Nicole Y. 
Weiss, Victor U.
Wenz, Christian 
Glück, Susanne 
Rüfer, Andreas 
Kratzmeier, Martin 
Marchetti-Deschmann, Martina 
Allmaier, Günter
Category: Research Article
Forschungsartikel
Issue Date: 2017
Journal: Analytical and Bioanalytical Chemistry
ISSN: 1618-2650
Abstract: 
Due to the constant search for reliable methods to investigate glycoproteins in complex biological samples, an alternative approach combining affinity enrichment with rapid and sensitive analysis on-a-chip is presented. Glycoproteins were specifically captured by lectin-coated magnetic beads, eluted by competitive sugars, and investigated with microchip capillary gel electrophoresis (MCGE), i.e., CGE-on-a-chip. We compared our results to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) data, which turned out to be in very good agreement. While SDS-PAGE offers the possibility of subsequent mass spectrometric analysis of captured and separated analytes, MCGE scores with time savings, higher throughput, and lower sample consumption as well as quality control (QC) and process analytical technology (PAT) applicability. Due to these advantages, a lectin-based glycoprotein capture protocol can easily be optimized. In our case, two different types of magnetic beads were tested and compared regarding lectin binding. The selectivity of our strategy was demonstrated with a set of model glycoproteins, as well as with human serum and serum depleted from high-abundance proteins. The specificity of the capturing method was investigated revealing to a certain degree an unspecific binding between each sample and the beads themselves, which has to be considered for any specific enrichment and data interpretation. In addition, two glycoproteins from Trichoderma atroviride, a fungus with mycoparasitic activity and only barely studied glycoproteome, were enriched by means of a lectin and so identified for the first time.
Keywords: Affinity enrichment; Glycoprotein; Lab-on-a-chip; Lectin; Magnetic beads
DOI: 10.1007/s00216-017-0615-0
Library ID: AC15189807
URN: urn:nbn:at:at-ubtuw:3-4389
Organisation: E164 - Institut für Chemische Technologien und Analytik 
Publication Type: Article
Artikel
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