Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
9748962 | Journal of Chromatography A | 2005 | 12 Pages |
Abstract
A simple high-resolution capillary zone electrophoresis (CZE) method capable of rapidly assessing the micro-heterogeneity of a 24Â kDa molecular weight glycoprotein, has been developed. Separation is carried out using a bare silica capillary at a pH of 2.5 in a commercially available electrophoresis buffer system composed of triethanolamine and phosphoric acid. Over 30 peaks were detected within a run time of 15Â min using a 27Â cm capillary and approximately 60 peaks were detected using a 77Â cm capillary. Although most of the peaks arise from differences in the oligosaccharide structures present on the one glycosylation site on this molecule, other forms of micro-heterogeneity due to the presence of the nonglycosylated form of this glycoprotein and various types of chemical degradation, e.g., deamidation, are also responsible for the multitude of peaks observed. Although the exact chemical identity of each peak in the resulting electropherogram of this glycoprotein is not known, useful information can be obtained for assessing comparability, stability, and batch consistency. Factors impacting the resolution, precision, accuracy, and robustness of the assay are also discussed along with inherent advantages and limitations associated with measuring the micro-heterogeneity of intact glycoproteins.
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Steven A. Berkowitz, Huijuan Zhong, Martha Berardino, Zoran Sosic, Joseph Siemiatkoski, Ira S. Krull, Rohin Mhatre,