Glycosylation is a key product quality attribute for many biotherapeutic proteins expressed in CHO cells. N-linked glycans may display macro- and micro-heterogeneity; the degree of this variation can depend on several factors, including cell line, media/feeds, and process. As a consequence, it has often been challenging to achieve and maintain preferred glycosylation profiles from cell culture development through bioreactor scale-up. In order to address these challenges, we have developed a new feed technology in conjunction with a unique fed-batch process…
Author Archives: Baburaj Kunnummal
Rapid and Simple Sample Preparation for High Throughput, High Resolution and Sensitive Glycan Analysis by Capillary Electrophoresis
Here, we report the simple and rapid GlycanAssureâ„¢ workflow that combines high throughput and high resolution glycan analysis of 96 samples in 7-9 hours using Applied Biosystemsâ„¢ 3500xL 24-capillary electrophoresis system. The process eliminates vacuum drying and highly toxic cyanoborohydride in the labeling reaction. Use of Dynabeadsâ„¢ magnetic beads for glycan purification post deglycosylation and removal of free dyes after labeling streamlines the process for automation. Capillary electrophoresis can detect less than 0.2 fmol/μL of labeled glycans. Two proprietary fluorescent…