Low glucose depletes glycan precursors, reduces site occupancy and galactosylation of a monoclonal antibody in CHO cell culture

VILLACRES, Carina, Venkata S. TAYI, Erika LATTOVÁ, Helene PERREAULT and Michael BUTLER. Low glucose depletes glycan precursors, reduces site occupancy and galactosylation of a monoclonal antibody in CHO cell culture. BIOTECHNOLOGY JOURNAL. WEINHEIM: WILEY-VCH Verlag GmbH, 2015, vol. 10, No 7, p. 1051-1066. ISSN 1860-6768. Available from: https://dx.doi.org/10.1002/biot.201400662.

Basic information

• Original name: Low glucose depletes glycan precursors, reduces site occupancy and galactosylation of a monoclonal antibody in CHO cell culture
• Authors: VILLACRES, Carina (124 Canada), Venkata S. TAYI (124 Canada), Erika LATTOVÁ (703 Slovakia, guarantor, belonging to the institution), Helene PERREAULT (124 Canada) and Michael BUTLER (124 Canada).
• Edition: BIOTECHNOLOGY JOURNAL, WEINHEIM, WILEY-VCH Verlag GmbH, 2015, 1860-6768.

Other information

• Original language: English
• Type of outcome: Article in a journal
• Field of Study: 30102 Immunology
• Country of publisher: Germany
• Confidentiality degree: is not subject to a state or trade secret
• WWW: URL
• Impact factor: Impact factor: 3.781
• RIV identification code: RIV/00216224:14740/15:00084232
• Organization unit: Central European Institute of Technology
• Doi: http://dx.doi.org/10.1002/biot.201400662
• UT WoS: 000357738900012
• Keywords in English: Adenylate energy charge; Chinese hamster ovary (CHO) cells; Glucose depletion; N-glycosylation; Nucleotide sugars
• Tags: rivok
• Tags: International impact, Reviewed

Abstract

Controlled feeding of glucose has been employed previously to enhance the productivity of recombinant glycoproteins but there is a concern that low concentrations of glucose could limit the synthesis of precursors of glycosylation. Here we investigate the effect of glucose depletion on the metabolism, productivity and glycosylation of a chimeric human-llama monoclonal antibody secreted by CHO cells. The cells were inoculated into media containing varying concentrations of glucose. Glucose depletion occurred in cultures with an initial glucose 5.5 mM and seeded at low density (2.5 x 10(5) cells/mL) or at high cell inoculum (2.5 x 10(6) cells/mL) at higher glucose concentration (up to 25 mM). Glucose-depleted cultures produced non-glycosylated Mabs (up to 51%), lower galactosylation index (GI <0.43) and decreased sialylation (by 85%) as measured by mass spectrometry and HPLC. At low glucose a reduced intracellular pool of nucleotides (0.03-0.23 fmoles/cell) was measured as well as a low adenylate energy charge (<0.57). Low glucose also reduced GDP-sugars (by 77%) and UDP-hexosamines (by 90%). The data indicate that under glucose deprivation, low levels of intracellular nucleotides and nucleotide sugars reduced the availability of the immediate precursors of glycosylation. These results are important when applied to the design of fed-batch cultures.