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. |
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@article{1313797, author = {Villacres, Carina and Tayi, Venkata S. and Lattová, Erika and Perreault, Helene and Butler, Michael}, article_location = {WEINHEIM}, article_number = {7}, doi = {http://dx.doi.org/10.1002/biot.201400662}, keywords = {Adenylate energy charge; Chinese hamster ovary (CHO) cells; Glucose depletion; N-glycosylation; Nucleotide sugars}, language = {eng}, issn = {1860-6768}, journal = {BIOTECHNOLOGY JOURNAL}, title = {Low glucose depletes glycan precursors, reduces site occupancy and galactosylation of a monoclonal antibody in CHO cell culture}, url = {http://onlinelibrary.wiley.com/doi/10.1002/biot.201400662/abstract}, volume = {10}, year = {2015} }
TY - JOUR ID - 1313797 AU - Villacres, Carina - Tayi, Venkata S. - Lattová, Erika - Perreault, Helene - Butler, Michael PY - 2015 TI - Low glucose depletes glycan precursors, reduces site occupancy and galactosylation of a monoclonal antibody in CHO cell culture JF - BIOTECHNOLOGY JOURNAL VL - 10 IS - 7 SP - 1051-1066 EP - 1051-1066 PB - WILEY-VCH Verlag GmbH SN - 18606768 KW - Adenylate energy charge KW - Chinese hamster ovary (CHO) cells KW - Glucose depletion KW - N-glycosylation KW - Nucleotide sugars UR - http://onlinelibrary.wiley.com/doi/10.1002/biot.201400662/abstract L2 - http://onlinelibrary.wiley.com/doi/10.1002/biot.201400662/abstract N2 - 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. ER -
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. \textit{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.
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