Dr. Jonathan Bones, Principal Investigator, Characterization and Comparability Laboratory, National Institute for Bioprocessing Research and Training (NIBRT), and a speaker at the Optimising Cell Culture Technology conference shares insight on his upcoming presentation.
Investigating Alterations in the CHO N-Glycome Following Exposure to Leachables from Single-Use Bioreactors
The cellular N-glycome of CHO cells, characterized using two dimensional liquid chromatography with high resolution mass spectrometry, is highly complex, consisting of a dominant high mannose series and a broad range of complex N-glycans bearing varying degrees of sialylation and poly-lactosamine motifs. The dynamics of the CHO N-glycome is described following exposure to leachable compounds from single use bioreactors. Correlation with product quality attributes, including mAb glycosylation, were also investigated.
Q1. What has your research uncovered regarding the dynamics of the CHO N-glycome following exposure to leachable compounds from single-use bioreactors?
We've been studying the effect of extractable and leachable compounds and their potential effect of CHO cell culture and the quality attributes of the expressed product. We've seen that exposure to certain leachable compounds can cause the cellular glycosylation to change. We've looked at the potential effect on the expressed monoclonal antibody and have seen slight, but not statistically significant changes in the glycans present on the mAb.
Q2. Are there specific components of single-use bioreactors that appear to be causing N-glycome problems for CHO cells? (Do you suggest manufacturers change the fabrication used in single-use bioreactors?)
We've taken two approaches, whole bag extraction and also targeted spike in of previously reported problematic leachables. We've also seen from our extractable and leachable profiling that the manufacturers are responding to this issue as the E&L profile of new single use bioreactors is significantly cleaner.
Q3. What is your ‘take-home’ advice for scientists cultivating CHO cells?
Evaluate your single-use options before selection to find the one that has no detrimental effect on your cell line. We're seeing that certain lines are more sensitive than others, so the time used in selecting the right single-use bioreactor option is time well spent.
Q4. Do you have any advice or suggestions for those visiting Lisbon for the first time?
Enjoy themselves! It's a nice city with plenty to do and good weather, a great location for this meeting.
To learn more about Dr. Jonathan Bones’ presentation at the 2018 Bioprocessing Summit Europe, visit bioprocessingeurope.com/cell-culture