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. 2005 Oct 28;50(1-3):57–76. doi: 10.1007/s10616-005-4537-x

Optimisation of the Cellular Metabolism of Glycosylation for Recombinant Proteins Produced by Mammalian Cell Systems

M Butler 1,✉
PMCID: PMC3476007  PMID: 19003071

Abstract

Many biopharmaceuticals are now produced as secreted glycoproteins from mammalian cell culture. The glycosylation profile of these proteins is essential to ensure structural stability and biological and clinical activity. However, the ability to control the glycosylation is limited by our understanding of the parameters that affect the heterogeneity of added glycan structures. It is clear that the glycosylation process is affected by a number of factors including the 3-dimensional structure of the protein, the enzyme repertoire of the host cell, the transit time in the Golgi and the availability of intracellular sugar-nucleotide donors. From a process development perspective there are many culture parameters that can be controlled to enable a consistent glycosylation profile to emerge from each batch culture. A further, but more difficult goal is to control the culture conditions to enable the enrichment of specific glycoforms identified with desirable biological activities. The purpose of this paper is to discuss the cellular metabolism associated with protein glycosylation and review the attempts to manipulate, control or engineer this metabolism to allow the expression of human glycosylation profiles in producer lines such as genetically engineered Chinese hamster ovary (CHO) cells.

Keywords: Antennarity, CHO cells, Fucosylation, Galactosylation, Glycoprotein, Glycosylation, Golgi, N-glycan, O-glycan, Sequon, Sialylation

Glossary

ADCC

antibody-mediated cytotoxicity

BHK

baby hamster kidney (cells)

C2GnT

core 2 GlcNAC transferase (UDP-GlcNAc: Galβ1,3GalNAc-R β1,6-N-acetyl glucosaminyl transferase)

CHO

Chinese hamster ovary (cells)

CMP

cytidine monophosphate

DO

dissolved oxygen

EPO

erythropoietin

ER

endoplasmic reticulum

FT

fucosyl transferase

G0

agalactosylated glycans

G1

monogalactosylated glycans

G2

digalactosylated glycans

GalNAc

N-acetyl galactosamine

GDM

GDP mannose 4,6 dehydratase

GDP

guanosine diphosphate

GFAT

glutamine: fructose 6-phosphate amidotransferase

GlcNAc

N-acetyl glucosamine

GnT

N-acetyl glucosaminyl transferase

HIV

human immunodeficiency virus

IFN

interferon

IgG

immunoglobulin

LAMP

lysosomal membrane glycoprotein

ManNAc

N-acetyl mannosamine

mPL-I

mouse placental lactogen I

NANA

N-acetyl-neuraminic acid

NGNA

N-glycolyl-neuraminic acid

OST

oligosaccharyltransferase

ST

sialyl transferase

ST3Gal1

sialyl transferase 3 (CMP-sialic acid: Galβ1,3GalNAc2,3 sialyl transferase)

TIMP

tissue inhibitors of metalloproteinases

t-PA

tissue plasminogen activator

UDP

uridine diphosphate

UTP

uridine triphosphate

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