The bioactivity and fractionation of peptide hydrolysates in cultures of CHO cells

Abstract

Peptide hydrolysate supplements in mammalian cell cultures provide enhanced growth and productivity. The objective of this study was to compare the bioactivity of ten different commercially available hydrolysates from plant, microbial, and animal sources. The peptide hydrolysates were tested as supplements to cultures of Chinese hamster ovary (CHO) cells that produce human beta interferon (β-IFN). A soy hydrolysate was shown to support high cell growth but not protein productivity compared to an animal component hydrolysate (Primatone RL). On the other hand, a yeast hydrolysate showed lower cell growth, but comparable productivity of the recombinant protein. Glycosylation analysis showed that the glycan profile of β-IFN produced in yeast hydrolysate supplemented cultures was equivalent to that from Primatone RL-supplemented cultures. Fractionation of the yeast hydrolysate and Primatone RL produced a similar protein-assayed pattern except for one extra peak at around 1 kDa in the Primatone RL profile. A fraction taken at a molecular weight range of 1.5-1.7 kDa showed the highest growth promoting activity in both samples. However, four other fractions in yeast hydrolysate and two in Primatone RL at lower molecular weights showed some growth promoting activity. In conclusion, the yeast hydrolysates provided a good alternative to the animal sourced Primatone RL for high productivity of β-IFN from CHO cells with equivalent glycosylation.

Autophagy‐inducing peptide increases CHO cell monoclonal antibody production in batch and fed‐batch cultures

“…The use of an alternative, more specific agent that could be more readily incorporated into an industrial bioprocess and at lower concentrations is preferable. Indeed, current media formulations often contain peptides as part of hydrolysates added to the medium to increase growth and productivity (Spearman et al, 2014). This made the use of a unique and specific cell‐permeable, autophagy‐inducing peptide (AIP), derived from the autophagy protein Beclin 1 (Shoji‐Kawata et al, 2013), suitable for this purpose.…”

Model‐based identification of cell‐cycle‐dependent metabolism and putative autocrine effects in antibody producing CHO cell culture

“…In the past, model-based approaches were used to identify a putative autocrine factor that influences the growth and death rates of various hybridoma cell lines in continuous and perfusion culture (Zeng, 1996;Zeng & Deckwer, 1999). Recent studies mainly focused on the effects of lowmolecular weight peptides (1-10 kDa) from hydrolysates on the growth and productivity of mammalian cell culture (Chabanon et al, 2008;Franěk & Fussenegger, 2005;Michiels et al, 2011;Spearman, Lodewyks, Richmond, & Butler, 2014). To the authors' knowledge, the chemical structure, its formation, mechanism of action, and degradation of putative autocrine factors are still unknown.…”

Applications and analysis of hydrolysates in animal cell culture

“…In CHO cells, Spearman et al . ( 2014 ) showed that soy hydrolysate promotes cell growth but did not increase protein productivity; while yeast hydrolysate reduces cell growth, but achieved a comparable level of protein productivity and glycosylation profile as an animal-based hydrolysate. This suggests that yeast hydrolysate would be a good alternative to animal-based hydrolysate if the quality of the protein produced is the most important attribute.…”

“…In a more recent study, cottonseed-derived hydrolysate enhances the galactosylation of CHO-S-RTX and CHO-EG2 cells, which in turn improves the product quality (Obaidi et al 2021 ). Furthermore, the glycosylation profile of the protein produced by CHO cells is equivalent between yeast and animal-based hydrolysate (Spearman et al 2014 ). This study demonstrated that biopharmaceutical manufacturers should consider the use of yeast hydrolysates to improve protein productivity and glycosylation of the product.…”