title: A theoretical estimate for nucleotide sugar demand towards Chinese Hamster Ovary cellular glycosylation.
abstract: Glycosylation greatly influences the safety and efficacy of many of the highest-selling recombinant therapeutic proteins (rTPs). In order to define optimal cell culture feeding strategies that control rTP glycosylation, it is necessary to know how nucleotide sugars (NSs) are consumed towards host cell and rTP glycosylation. Here, we present a theoretical framework that integrates the reported glycoproteome of CHO cells, the number of N-linked and O-GalNAc glycosylation sites on individual host cell proteins (HCPs), and the carbohydrate content of CHO glycosphingolipids to estimate the demand of NSs towards CHO cell glycosylation. We have identified the most abundant N-linked and O-GalNAc CHO glycoproteins, obtained the weighted frequency of N-linked and O-GalNAc glycosites across the CHO cell proteome, and have derived stoichiometric coefficients for NS consumption towards CHO cell glycosylation. By combining the obtained stoichiometric coefficients with previously reported data for specific growth and productivity of CHO cells, we observe that the demand of NSs towards glycosylation is significant and, thus, is required to better understand the burden of glycosylation on cellular metabolism. The estimated demand of NSs towards CHO cell glycosylation can be used to rationally design feeding strategies that ensure optimal and consistent rTP glycosylation.
authors.affiliation: School of Chemical &Bioprocess Engineering, University College Dublin, Belfield campus, Dublin 4, Ireland.|Centre for Process Systems Engineering, Department of Chemical Engineering, Imperial College London, South Kensington Campus, London SW7 2AZ, U.K.
authors.forename: Ioscani Jimenez
authors.initials: IJ
authors.lastname: Del Val
authors.affiliation: Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, U.K.|Centre for Synthetic Biology and Innovation, Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK.
authors.forename: Karen M
authors.initials: KM
authors.lastname: Polizzi
authors.affiliation: Centre for Process Systems Engineering, Department of Chemical Engineering, Imperial College London, South Kensington Campus, London SW7 2AZ, U.K.
authors.forename: Cleo
authors.initials: C
authors.lastname: Kontoravdi
mesh_terms: D000818:Animals; D016466:CHO Cells; D002241:Carbohydrates / Q000502:physiology; D018929:Cell Culture Techniques; D006224:Cricetinae; D003412:Cricetulus; D006023:Glycoproteins / Q000378:metabolism; D006031:Glycosylation; D009711:Nucleotides / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D000073893:Sugars / Q000378:metabolism*
chemical_list: D002241:Carbohydrates; D006023:Glycoproteins; D009711:Nucleotides; D020543:Proteome; D011994:Recombinant Proteins; D000073893:Sugars