Chinese hamster ovary (CHO) cells are commonly used in biomanufacturing of recombinant protein therapeutics (including monoclonal antibodies) due to the human-like post translational modifications, such as glycosylation. Glycosylation of monoclonal antibodies (mAbs) is of particular interest due to the effect various glycan identities have on the effector function, structure and stability of the mAb, thus making it an important product quality attribute. To achieve a desired therapeutic effect, it could be necessary to produce an antibody with a specific glycosylation profile.  In order to produce this antibody with specific glycoforms we must understand how changes to culturing conditions impact the glycosylation profile of the mAb. In this work we detect changes in glycosylation of CHO produced VRC01- a broadly neutralizing antibody currently undergoing clinical trials to treat HIV, following changes to the culturing conditions.  More specifically we studied the impact of ammonia stress on both young and aged CHO cells, with different gassing and feeding protocols. We found that increased ammonia concentrations measured in the culture for all conditions tested led to a decrease in total and terminal galactosylation, as well as a decrease in sialylation of the associated glycans. Future work will look into how the conserved Asn<sup>297</sup> Fc glycan is impacted compared to Fab glycans under various culturing conditions.

Impact of ammonia stress on glycosylation of the VRC01 monoclonal antibody produced in CHO cells

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.