Considerations for Glycoprotein Production

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 2762; p. 329
• Main Author: Clarke, Elizabeth C
• Format: Journal Article
• Language: English
• Published: United States 2024
• Subjects: Animals; CHO Cells; Cricetinae; Cricetulus; Glycoproteins - chemistry; Glycosylation; Humans; Recombinant Proteins - metabolism; Expression system; Escherichia coli; Glycoprotein; Human endothelial kidney; O-linked glycosylation; Glycans; Glycosylation; Complex glycans; Pichia pastoris; Baculovirus; Chinese hamster ovary cells; Recombinant protein production; N-linked glycosylation
• DOI: 10.1007/978-1-0716-3666-4_20

This chapter is intended to provide insights for researchers aiming to choose an appropriate expression system for the production of recombinant glycoproteins. Producing glycoproteins is complex, as glycosylation patterns are determined by the availability and abundance of specific enzymes rather than a direct genetic blueprint. Furthermore, the cell systems often employed for protein production are evolutionarily distinct, leading to significantly different glycosylation when utilized for glycoprotein production. The selection of an appropriate production system depends on the intended applications and desired characteristics of the protein. Whether the goal is to produce glycoproteins mimicking native conditions or to intentionally alter glycan structures for specific purposes, such as enhancing immunogenicity in vaccines, understanding glycosylation present in the different systems and in different growth conditions is essential. This chapter will cover Escherichia coli, baculovirus/insect cell systems, Pichia pastoris, as well as different mammalian cell culture systems including Chinese hamster ovary (CHO) cells, human endothelial kidney (HEK) cell lines, and baby hamster kidney (BHK) cells.