Effect of N-Acetylcystein on Butyrate-Treated Chinese Hamster Ovary Cells To Improve the Production of Recombinant Human Interferon-β-1a

Abstract

Sodium butyrate (NaBu) is used as a productivity enhancer for the production of therapeutic recombinant proteins in Chinese hamster ovary (CHO) cells. However, NaBu is well-known for having a cytotoxic effect, thereby inducing apoptosis. As an endeavor to reduce this defect, we studied 11 antioxidants known for inhibiting apoptosis, according to a Plackett-Burman statistical design on CHO cells producing recombinant interferon-beta-1a (IFN-beta). None of the antioxidants that we tested were as effective as N-acetylcystein (NAC) from the point of view of maintaining long-term survival of CHO cells and increasing the production of IFN-beta. In 7.5-L perfusion bioreactor cultures, the addition of NaBu and NAC elongated the culture period to almost 200 h throughout production phase and increased the production yield by 2-fold compared to control cultures containing only NaBu. Glycosylation patterns of produced IFN-beta at each run were also compared in IEF analysis. IEF profiles of where NaBu and NAC were added showed to be more isoforms with a lower pI than those of the control run. The sialic acid content was also increased by 17.7% according to HPLC analysis. Taken together, the data obtained demonstrate that the addition of NAC has positive effects on the elongation of the culture period, improving the production and increasing the sialylation of IFN-beta in NaBu-treated CHO cells.

Perfusion Processes

“…Cell arrest obtained by hypothermia or addition of a chemical, e.g. butyrate, has been reported for perfusion; given that the cell specific productivity is comparable or increased and that the protein quality is correct or improved (Ducommun et al 2002a;Chotteau et al 2001;Chen et al 2004;Kim et al 2004;Angepat et al 2005;Oh et al 2005;Meuwly et al 2006;Ahn et al 2008;Zhang et al 2014a). Another advantage of lowering the temperature under 35.5 C is that the proteolytic activity is lower compared to 37 C (Chuppa et al 1997).…”

Process parameter shifting: Part I. Effect of DOT, pH, and temperature on the performance of Epo‐Fc expressing CHO cells cultivated in controlled batch bioreactors

“…Process parameters such as pH, temperature and dissolved oxygen tension (DOT) have been shown to exert a significant effect on cell line performance. In contrast to reports where improvements in process performance have been achieved by the addition of nutrient supplements (Oh et al, 2005;Sung et al, 2004) or by means of metabolic or genetic engineering (Fussenegger and Betenbaugh, 2002), the improvement of process performance by simple parameter optimization offers a cheap and valuable strategy for process optimization. In this context, the pH is a parameter known to significantly influence cell growth, recombinant protein production, cell metabolism and protein glycosylation (Borys et al, 1993;Kurano et al, 1990;Link et al, 2004;Miller et al, 2000;Yoon et al, 2005;Zanghi et al, 1999b).…”

Quercetin treatment changes fluxes in the primary metabolism and increases culture longevity and recombinant α1-antitrypsin production in human AGE1.HN cells

“…It was reported that the addition of, e.g., DMSO, glycerol, or sodium butyrate can enhance the production of biopharmaceuticals in different mammalian cells (Liu and Chen 2007;Oh et al 2005;Rodriguez et al 2005;Cherlet and Marc 2000). Despite an increase in specific productivity in many producers, treatment with these chemicals results usually in a reduced growth rate that can reduce the final product titer compared to untreated cells (Rodriguez et al 2005).…”