Targeting Desired N-Linked Glycosylation Profiles Through The Use Of Glycosylation Enhancing Feeds And High Throughput And High Resolution N-Glycan Analysis By Multi Capillary Electrophoresis

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thermofisher-upstream-n-glycan-PH_2017Glycosylation 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 that together has been shown not only to maximize protein titers but also to modulate glycan profiles.

Precise targeting of desired N-glycan profiles necessitated the need for an analysis method that is rapid, simple, high throughput and preserves the integrity of N-glycan structure. Current N-glycan analysis methods consist of sample prep steps that are laborious and tedious. The labeling reaction typically requires vacuum drying of purified glycans and the use of toxic reducing chemicals such as sodium cyanoborohydride.

Here we report development of an integrated N-glycan analysis platform that can analyze 96 samples in 8hrs, consist of simple magnetic bead based sample prep, multi capillary CE instrument that can analyze 8 or 24 samples in parallel and glycan specific software with novel features. Our glycan analysis workflow in conjunction with unique fed-batch process can address the needs of biopharmaceutical companies interested in precise targeting of desired glycan profiles in their biomolecules

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