Abstract
At present, nearly 70% of recombinant therapeutic proteins (RTPs) are produced by Chinese hamster ovary (CHO) cells, and serum-free medium (SFM) is necessary for their culture to produce RTPs. In this review, the history and key components of SFM are first summarized, and its preparation and experimental design are described. Some small molecule compound additives can improve the yield and quality of RTP. The function and possible mechanisms of these additives are also reviewed here. Finally, the future perspectives of SFM use with CHO cells for RTP production are discussed.
Highlights
Chinese hamster ovary (CHO) cells are the preferred expression system for recombinant therapeutic protein (RTP) production, and nearly 70% of recombinant therapeutic proteins (RTPs) are currently generated by CHO cells (Pereira et al, 2018; Stolfa et al, 2018)
Plackett Burman method (PBM) has been used to design the conduction of 24 groups of experiments, and results showed that 20 types of components can promote cell growth and increase the recombinant protein yield by 45% (Kato et al, 2018)
Hong et al (2011) observed that with 3 mM of NaB, cell viability fell below 80% after day four in GFcontaining medium, but it remained over 80% until day 18 in growth factor (GF)-deficient medium
Summary
Chinese hamster ovary (CHO) cells are the preferred expression system for recombinant therapeutic protein (RTP) production, and nearly 70% of RTPs are currently generated by CHO cells (Pereira et al, 2018; Stolfa et al, 2018). Optimizing the ratio of various amino acid concentrations is an effective way to increase the density of living cells and the expression level of recombinant proteins during the culture process (Zhang et al, 2015). The expression rate of the tissue type plasminogen activator can be increased by adding antioxidants (ascorbic acid and reduced glutathione) to the CHO cell culture process (Allen et al, 2008). Inorganic salt components in the medium mainly include sodium, potassium, magnesium, calcium, and phosphorus Their main function is to help maintain a stable osmotic pressure during the cell culture process because each cell line has its tolerable osmotic pressure range (Qin et al, 2019). Through real-time monitoring of glucose, amino acids, and vitamins, fatty acids and trace elements in the cell culture process can appropriately adjust each medium component (Zhang et al, 2013). Quantitative PCR and protein electrophoresis are used to analyze changes in the mRNA and protein levels of each enzyme involved in the metabolism process in the cell, and some additional components are specially added to the culture medium to regulate the physiological metabolism of the cells (Torkashvand et al, 2015). Ganne and Mignot (1991) effectively controlled cell proliferation, apoptosis, and transgene expression by regulating the cell physiological metabolism through adding small molecules to the medium
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