High-throughput lectin magnetic bead array-coupled tandem mass spectrometry for glycoprotein biomarker discovery

Choi, Eunju, Loo, Dorothy, Dennis, James W., O'Leary, Caroline A. and Hill, Michelle M. (2011) High-throughput lectin magnetic bead array-coupled tandem mass spectrometry for glycoprotein biomarker discovery. Electrophoresis, 32 24: 3564-3575. doi:10.1002/elps.201100341


Author Choi, Eunju
Loo, Dorothy
Dennis, James W.
O'Leary, Caroline A.
Hill, Michelle M.
Title High-throughput lectin magnetic bead array-coupled tandem mass spectrometry for glycoprotein biomarker discovery
Journal name Electrophoresis   Check publisher's open access policy
ISSN 0173-0835
1522-2683
Publication date 2011-12
Sub-type Article (original research)
DOI 10.1002/elps.201100341
Volume 32
Issue 24
Start page 3564
End page 3575
Total pages 12
Editor Yehia Mechref
Place of publication Weinheim, Germany
Publisher Wiley - VCH Verlag
Collection year 2012
Language eng
Abstract Alterations in protein glycosylation occur during development and progression of many diseases, hence glycomics and glycoproteomics have emerged as important tools in glycobiomarker discovery. High-throughput glycan profiling can now be achieved with the recent developments in MS-based techniques. To enable identification and rapid monitoring of glycosylation changes in serum proteins, we developed a semi-automated high-throughput glycoprotein biomarker discovery platform termed lectin magnetic bead array-coupled tandem mass spectrometry (LeMBA-MS) which includes (i) effective single-step serum glycoprotein isolation using a panel of 20 individual lectin-coated magnetic beads in microplate format, (ii) on-bead trypsin digestion, and (iii) nanoLC-MS/MS with lectin exclusion list. With use of appropriate sequence databases, LeMBA-MS can detect glycosylation changes regardless of the species. By spiking known amounts of titrated ovalbumin to a serum sample, we report nanomolar sensitivity, and linearity of response of LeMBA-MS using concanavalin A-coupled beads. Neuraminidase treatment led to reduction of binding to sialic acid-binding lectins. Interestingly, we found that desialylation caused increased binding of haptoglobin and hemopexin to mannose-specific lectins, pointing to the importance of identifying a signature of lectin-binding. High-throughput LeMBA-MS to generate glycosylation signatures will facilitate glycobiomarker discovery. LeMBA can be coupled to down-stream detection platforms for validation, making it a truly versatile platform.
Keyword Biomarker discovery
Glycoproteome fractionation
Glycosylation
Magnetic beads
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Special Issue: Focus On Glycomics & Glycoproteomics

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2012 Collection
School of Veterinary Science Publications
UQ Diamantina Institute Publications
 
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