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

Eun ju Choi, Dorothy Loo, James W. Dennis, Caroline A. O'Leary, Michelle M. Hill

Research output: Contribution to journalArticle

28 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)3564-3575
Number of pages12
JournalElectrophoresis
Volume32
Issue number24
DOIs
StatePublished - Dec 1 2011
Externally publishedYes

Fingerprint

Biomarkers
Tandem Mass Spectrometry
Lectins
Mass spectrometry
Glycoproteins
Throughput
Glycosylation
Mannose-Binding Lectins
Hemopexin
Glycomics
Haptoglobins
Ovalbumin
Neuraminidase
N-Acetylneuraminic Acid
Concanavalin A
Serum
Trypsin
Polysaccharides
Disease Progression
Blood Proteins

Keywords

  • Biomarker discovery
  • Glycoproteome fractionation
  • Glycosylation
  • Magnetic beads
  • Multilectin affinity

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

Cite this

High-throughput lectin magnetic bead array-coupled tandem mass spectrometry for glycoprotein biomarker discovery. / Choi, Eun ju; Loo, Dorothy; Dennis, James W.; O'Leary, Caroline A.; Hill, Michelle M.

In: Electrophoresis, Vol. 32, No. 24, 01.12.2011, p. 3564-3575.

Research output: Contribution to journalArticle

Choi, Eun ju ; Loo, Dorothy ; Dennis, James W. ; O'Leary, Caroline A. ; Hill, Michelle M. / High-throughput lectin magnetic bead array-coupled tandem mass spectrometry for glycoprotein biomarker discovery. In: Electrophoresis. 2011 ; Vol. 32, No. 24. pp. 3564-3575.
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