Expression systems for therapeutic glycoprotein production

A simplified polyethylenimine-mediated transfection process for large-scale and high-throughput applications

“…The need for recombinant proteins with post-translational modifications has fostered the development of mammalian cell-based expression systems rather than prokaryotes and lower eukaryotes [6]. Transient gene expression in mammalian cells is widely used for the fast production of recombinant proteins, mainly for pre-clinical in vitro and in vivo applications.…”

Production of recombinant coagulation factors: Are humans the best host cells?

“…34 This fact can be attributed to some key advantages: i) robust growth in chemically-defined and serum-free suspension culture, (ii) a reasonable safety profile regarding human pathogenic virus replication, (iii) ease generation of cell clones which are able to stably express the recombinant protein in sufficient yields and acceptable quality for human use, and (iv) ability to express recombinant proteins with human-like post-translational modifications. 35,36 However, recombinant therapeutic proteins produced in non-human cells may contain glycan epitopes, mainly Gala1,3-gal (a-Gal) residues and N-glycolylneuraminic acid (Neu5Gc), that are antigenic to humans, 34,37 and can potentially affect the efficacy of the recombinant product, since all humans tested have circulating antibodies against these epitopes. 38,39 Although few adverse immunogenic reactions have been ascribed to CHOderived recombinant proteins, this cell line possesses the enzymes (a1,3 galactosyltransferase and CMPNeu5Ac hydroxylase) responsible for the formation of this non-human epitope.…”

Oxidative stress in fungal fermentation processes: the roles of alternative respiration

“…For example, short doubling times and readily manipulated genomes, but also have the additional benefits of eukaryotes such as improved protein folding and many post-translational modifications (Durocher & Butler 2009;Yurimoto & Sakai 2009). Furthermore, filamentous fungi and yeasts are more robust than mammalian cells and can produce some enzymes in excess of 100g/l in fermenters (Cherry 2003) and are easily adapted to fermentation processes in simple and cost-effective culture media (Gasser and Mattanovich 2007;Durocher & Butler 2009).…”

“…For example, short doubling times and readily manipulated genomes, but also have the additional benefits of eukaryotes such as improved protein folding and many post-translational modifications (Durocher & Butler 2009;Yurimoto & Sakai 2009). Furthermore, filamentous fungi and yeasts are more robust than mammalian cells and can produce some enzymes in excess of 100g/l in fermenters (Cherry 2003) and are easily adapted to fermentation processes in simple and cost-effective culture media (Gasser and Mattanovich 2007;Durocher & Butler 2009). Even though filamentous fungi and yeasts have now been used industrially for decades, and are now used for production of some of our most complex and potent drugs, they are still not adequately characterized at the physiological and genetic level, particularly in terms of protein modification and secretion metabolism (Durocher & Butler 2009) and especially in relation to their physiological responses to the stressful growing environment in bioreactors (Li et al 2009 ;Bai et al 2003a).…”