Enrichment of glycoproteins using nanoscale chelating concanavalin A monolithic capillary chromatography

Shun Feng, Na Yang, Subramaniam Pennathur, Steve Goodison, David M. Lubman

Research output: Contribution to journalArticle

60 Scopus citations

Abstract

Immobilized lectin chromatography can be employed for glycoprotein enrichment, but commonly used columns have limitations of yield and resolution. To improve efficiency and to make the technique applicable to minimal sample material, we have developed a nanoscale chelating Concanavalin A (Con A) monolithic capillary prepared using GMA-EDMA (glycidyl methacrylate-co-ethylene dimethacrylate) as polymeric support. Con A was immobilized on Cu(II)-charged iminodiacetic acid (IDA) regenerable sorbents by forming a IDA:Cu(II):Con A sandwich affinity structure that has high column capacity, as well as stability. When compared with conventional Con A lectin chromatography, the monolithic capillary enabled the better reproducible detection of over double the number of unique N-glycoproteins in human urine samples. Utility for analysis of minimal biological samples was confirmed by the successful elucidation of glycoprotein profiles in mouse urine samples at the microliter scale. The improved efficiency of the nanoscale monolithic capillary will impact the analysis of glycoproteins in complex biological samples, especially where only limited material may be available.

Original languageEnglish (US)
Pages (from-to)3776-3783
Number of pages8
JournalAnalytical Chemistry
Volume81
Issue number10
DOIs
StatePublished - May 15 2009

ASJC Scopus subject areas

  • Analytical Chemistry

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