Supplementary Figure 8: Production of IFNα-2b on the InSCyT system.

Dose size was 12ug¹⁸. (a) Yields for production of IFNα-2b using the InSCyT system during the Process Qualification Run. Wet cell weight (WCW) (black) and cumulative unpurified (orange) and formulated (blue) doses of IFNα-2b are shown. Grey circles show individual data points. Complete process yield is approximately 11%. Additional product quality data from the process qualification run for IFNα-2b alongside a reference drug substance. Analysis of purity by isoelectric focusing (IEF). M, IEF protein marker. Example RPLC and SEC chromatograms used to quantify purity of IFNα-2b (representative of all four time points). Differences in chromatographic behavior of InSCyT IFNα-2b and the reference standard were confirmed by MALDI as a naturally-occurring C-terminal truncation. (b) Fermentograms and UV profiles from chromatography operations on the InSCyT system during the process qualification run for IFNα-2b. DO, dissolved oxygen; SLPM, standard liters per minute. (c) Product quality data for InSCyT-produced IFNα-2b analyzed by LCMS. Peptide mapping used to confirm identity (96.3% sequence coverage). Example LCMS chromatograms used to quantify purity, including base peak chromatograms and extracted-ion chromatograms of N-terminal variant.