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Title
Microchip capillary gel electrophoresis combined with lectin affinity enrichment employing magnetic beads for glycoprotein analysis
AuthorEngel, Nicole Y. ; Weiss, Victor U. ; Wenz, Christian ; Glück, Susanne ; Rüfer, Andreas ; Kratzmeier, Martin ; Marchetti-Deschmann, Martina ; Allmaier, Günter
Published in
Analytical and Bioanalytical Chemistry, 2017, Vol. 409, Issue 28, page 6625-6634
PublishedSpringer Berlin Heidelberg, 2017
LanguageEnglish
Document typeJournal Article
Keywords (EN)Affinity enrichment / Glycoprotein / Lab-on-a-chip / Lectin / Magnetic beads
ISSN1618-2650
URNurn:nbn:at:at-ubtuw:3-4389 Persistent Identifier (URN)
DOI10.1007/s00216-017-0615-0 
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 The work is publicly available
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Microchip capillary gel electrophoresis combined with lectin affinity enrichment employing magnetic beads for glycoprotein analysis [2.22 mb]Supplementary material [0.78 mb]
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Abstract (English)

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.

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CC-BY-License (4.0)Creative Commons Attribution 4.0 International License