Abstract
Monoclonal antibodies (mAbs) are commonly glycosylated and show varying levels of galactose attachment. A set of experiments in our work showed that the galactosylation level of mAbs was altered by the culture conditions of hybridoma cells. The uridine diphosphate galactose (UDP-Gal) is one of the substrates of galactosylation. Based on that, we proposed a two-step model to predict N-linked glycoform profiles by solely using extracellular metabolites from cell culture. At the first step, the flux level of UDP-Gal in each culture was estimated based on a computational flux balance analysis (FBA); its level was found to be linear with the galactosylation degree on mAbs. At the second step, the glycoform profiles especially for G0F (agalactosylated), G1F (monogalactosylated) and G2F (digalactosylated) were predicted by a kinetic model. The model outputs well matched with the experimental data. Our study demonstrated that the integrated mathematical approach combining FBA and kinetic model is a promising strategy to predict glycoform profiles for mAbs during cell culture processes.
Highlights
Monoclonal antibodies have achieved remarkable success in treating inflammatory disease, infectious diseases, autoimmune diseases and cancer [1]
The flux balance analysis (FBA) and kinetic model were applied to this phase
Due to the lack of glycoform profile data at earlier days, we used glycoform profiles of Monoclonal antibodies (mAbs) at day 5 (Figure 5), assuming that the cross-batch glycoform profiles variation at day 5 in cultures could still represent the early point variation, even though the glycan for the mAbs produced at late culture would have been affected
Summary
Monoclonal antibodies (mAbs) have achieved remarkable success in treating inflammatory disease, infectious diseases, autoimmune diseases and cancer [1]. N-linked glycoform profiles are product critical quality attributes (CQAs) which must be controlled in the production of mAbs [2,3]. Poor control of glycosylation can lead to immunogenicity and product efficacy issues, resulting in product rejections [2,4]. The variation of galactosylation (the level of galactose attachment) could raise some batch consistency concerns for mAb pharmaceuticals, oncology products, for which galactosylation can significantly alter effector functions [4,5]. Glycosylation is often analyzed in products at the end of processing. The glycoform profiles on mAbs should be monitored throughout the duration of a given process and likely be fine-tuned. A box of “tools” to modulate N-linked glycan in process has been
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
