Application of a pH responsive multimodal hydrophobic interaction chromatography medium for the analysis of glycosylated proteins

Protein glycosylation has significant effects on the structure and function of proteins. The efficient separation and enrichment of glycoproteins from complex biological samples is one key aspect and represents a major bottleneck of glycoproteome research. In this paper, we have explored pH multimod...

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Bibliographic Details
Published in:Journal of Chromatography A 2011-02, Vol.1218 (5), p.678-683
Main Authors: Kallberg, K., Becker, K., Bülow, L.
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Deglycosylation
Fundamental and applied biological sciences. Psychology
Glycoproteins
HIC
Immunoglobulin
Invertase
Multimodal
pH responsive polymers
Proteins
Ribonuclease
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Summary:Protein glycosylation has significant effects on the structure and function of proteins. The efficient separation and enrichment of glycoproteins from complex biological samples is one key aspect and represents a major bottleneck of glycoproteome research. In this paper, we have explored pH multimodal hydrophobic interaction chromatography to separate glycosylated from non-glycosylated forms of proteins. Three different proteins, ribonuclease, invertase and IgG, have been examined and different glycoforms have been identified. The media itself shows strong responsiveness to small variations in pH, which makes it possible to fine-tune the chromatographic conditions according to the properties of the protein isolated. Optimal glycoprotein separation has been obtained at pH 4. The pH responsive multimodal HIC medium in contrast to conventional HIC media is able to resolve contaminating DNA.
ISSN:0021-9673
DOI:10.1016/j.chroma.2010.11.080