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Issue 9, 2012
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Use of magnetic hydrazide-modified polymer microspheres for enrichment of Francisella tularensisglycoproteins

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Abstract

The field of microbial proteomics has currently experienced a boom in the discovery of glycosylated proteins of various pathogenic bacteria as potential mediators of host–pathogen interactions. The presence of glycoproteins has recently been discovered in a Gram-negative pathogenic bacterium Francisella tularensis, utilizing glycoprotein detection and isolation techniques in combination with mass spectrometry. The isolation of glycoproteins is a prerequisite for their subsequent mass-spectrometric identification. Current glycoprotein isolation/enrichment methods comprise lectin affinity chromatography, aminophenylboronic acid and hydrazide-based enrichment. The use of magnetic microspheres containing functional groups is nowadays among state-of-art separation methodologies owing to an ease of manipulation, a speed of separation, and a minimum of non-specific protein adsorption. In the present study, novel magnetic hydrazide-modified poly(2-hydroxyethyl methacrylate) (PHEMA) microspheres were developed using a multi-step swelling and polymerization method with subsequent precipitation of magnetic iron oxides within the pores of the particles. The microspheres had a regular shape, size of 4 μm and contained 0.18 mmol hydrazide groups per g; the magnetic microspheres were employed for specific enrichment of Francisella tularensis glycoproteins. Effectiveness of the newly prepared magnetic microspheres for glycoprotein enrichment was proved by comparison with commercial hydrazide-functionalized microparticles.

Graphical abstract: Use of magnetic hydrazide-modified polymer microspheres for enrichment of Francisella tularensis glycoproteins

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Article information


Submitted
25 Oct 2011
Accepted
21 Dec 2011
First published
30 Jan 2012

Soft Matter, 2012,8, 2775-2786
Article type
Paper

Use of magnetic hydrazide-modified polymer microspheres for enrichment of Francisella tularensis glycoproteins

D. Horák, L. Balonová, B. F. Mann, Z. Plichta, L. Hernychová, M. V. Novotny and J. Stulík, Soft Matter, 2012, 8, 2775
DOI: 10.1039/C2SM07036G

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