Abstract
Microcarriers are widely used for the large-scale culture of attachment-dependent cells with increased cell densities and, ultimately, higher product yield. In these processes, the specific culture conditions can affect the quality of the product, which is closely related to its glycosylation pattern. Furthermore, the lack of studies in the area reinforces the need to better understand the effects of microcarrier culture in product glycosylation. Consequently, in this work, the glycosylation profile of a monoclonal antibody (mAb) produced by adherent CHO-K1 cells grown in Cytodex 3 was evaluated under different conditions, and compared to that obtained of typical adherent cultures. It was found that microcarrier cultures result in a glycosylation profile with different characteristics from T-flask cultures, with a general increase in galactosylation and decrease in fucosylation levels, both with a potentially positive impact on mAb activity. Sialylation also varied but without a general tendency. This study then showed that the specific culture conditions used in microcarrier culture influence the mAb glycan profile, and each functional element (galactose, core fucose, sialic acid) is independently affected by these conditions. In particular, great reductions of fucosylation (from 79 to 55%) were obtained when using half volume at inoculation, and notable decreases in sialylation (from 23 to 2%) and glycoform heterogeneity (from 20 to 11 glycoforms) were observed for shake flask culture, potentially associated with the improved cell densities achieved in these culture vessels.
Highlights
Microcarrier systems are the most well-known technology for the large-scale culture of adherent cells in biotechnology (Chu and Robinson 2001; Kong et al 1999; Ziao et al 2002)
MAb-producing Chinese hamster ovary (CHO)-K1 cells were cultured in microporous Cytodex 3 carriers, under different culture conditions to evaluate their impact on the glycosylation profile of the monoclonal antibody (mAb)
The effect of microcarrier culture and different culture conditions on the glycosylation pattern of a mAb produced by CHO-K1 cells was evaluated
Summary
Microcarrier systems are the most well-known technology for the large-scale culture of adherent cells in biotechnology (Chu and Robinson 2001; Kong et al 1999; Ziao et al 2002). The characteristics of the microcarriers are important for the success of the culture (Butler 1996; Ozturk and Hu 2006), but other factors must be considered, such as the size and type of culture vessel (Wang et al 2002; Wu et al 1998), and the culture environment (Nam et al 2008; Ng et al 1996). It is important to recognize that the specific conditions of the culture can influence product quality, through effects on protein glycosylation. It has become evident that glycosylation plays critical roles in protein effectiveness in vivo (Jenkins and Curling 1994), and the ability to perform this posttranslational modification is a major reason for the current choice of mammalian cells as hosts for recombinant therapeutic protein production (Nam et al 2008)
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