J 2015

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, Michael BUTLER et. al.

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

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

30102 Immunology

Country of publisher

Germany

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

Impact factor

Impact factor: 3.781

RIV identification code

RIV/00216224:14740/15:00084232

Organization unit

Central European Institute of Technology

DOI

UT WoS

000357738900012

Keywords in English

Adenylate energy charge; Chinese hamster ovary (CHO) cells; Glucose depletion; N-glycosylation; Nucleotide sugars

Tags

Tags

International impact, Reviewed
Změněno: 31/3/2016 10:06, Mgr. Eva Špillingová

Abstract

V originále

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.