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

Electrophoresis. 2011 Dec;32(24):3564-75. doi: 10.1002/elps.201100341.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers / analysis
  • Biomarkers / blood
  • Biomarkers / metabolism
  • Biomarkers, Tumor / blood
  • Biomarkers, Tumor / isolation & purification
  • Biomarkers, Tumor / metabolism
  • Blood Proteins / chemistry
  • Concanavalin A / chemistry
  • Dogs
  • Electrophoresis, Polyacrylamide Gel
  • Glycoproteins / blood
  • Glycoproteins / isolation & purification*
  • Glycoproteins / metabolism
  • Glycosylation
  • High-Throughput Screening Assays
  • Humans
  • Lectins / chemistry
  • Lectins / metabolism*
  • Magnets*
  • Microspheres
  • Neuraminidase / chemistry
  • Proteomics
  • Sensitivity and Specificity
  • Tandem Mass Spectrometry / instrumentation*
  • Tandem Mass Spectrometry / methods


  • Biomarkers
  • Biomarkers, Tumor
  • Blood Proteins
  • Glycoproteins
  • Lectins
  • Concanavalin A
  • Neuraminidase