Abstract
Capillary electrophoresis (CE) is a versatile analytical method used to characterize glycoproteins. We have used several modes of CE separation such as CE-SDS gel, imaged capillary isoelectric focusing (icIEF), and capillary zone electrophoresis (CZE) to study therapeutic glycoprotein products. CE-SDS gel is applied to characterize the glycan occupancy and number of glycosylation sites, and icIEF is used to study the charge heterogeneities due to sialic acids in glycoproteins. To further characterize the glycoprotein, removal of N-linked glycans is necessary and a CZE technique is employed to analyze each glycan moiety. Examples from a monoclonal antibody, erythropoietin, and granulocyte colony-stimulating factor are presented here to demonstrate the utility of these CE modes. The details of sample preparation and separation conditions for each CE mode are described in this chapter.
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Acknowledgments
The authors would like to thank our colleagues in Bioprocess and GlycoFi for providing us with material and technical support. We gratefully thank Anna Mach for CE-SDS gel, Brian Peklansky for icIEF, and Catherine Lancaster for CZE N-glycan experimental support. Finally, we thank our management support Drs. Yang Wang and Michael W. Washabaugh.
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Rustandi, R.R., Anderson, C., Hamm, M. (2013). Application of Capillary Electrophoresis in Glycoprotein Analysis. In: Beck, A. (eds) Glycosylation Engineering of Biopharmaceuticals. Methods in Molecular Biology, vol 988. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-327-5_11
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DOI: https://doi.org/10.1007/978-1-62703-327-5_11
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Publisher Name: Humana Press, Totowa, NJ
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