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

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.

Základní údaje

Originální název

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

Autoři

VILLACRES, Carina; Venkata S. TAYI; Erika LATTOVÁ; Helene PERREAULT a Michael BUTLER

Vydání

BIOTECHNOLOGY JOURNAL, WEINHEIM, WILEY-VCH Verlag GmbH, 2015, 1860-6768

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

30102 Immunology

Stát vydavatele

Německo

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 3.781

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14740/15:00084232

Organizační jednotka

Středoevropský technologický institut

Klíčová slova anglicky

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

Štítky

rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 31. 3. 2016 10:06, Mgr. Eva Špillingová

Anotace

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.

Citovat

VILLACRES, Carina; Venkata S. TAYI; Erika LATTOVÁ; Helene PERREAULT a 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, roč. 10, č. 7, s. 1051-1066. ISSN 1860-6768. Dostupné z: https://doi.org/10.1002/biot.201400662.

@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 = {https://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 a 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, roč.~10, č.~7, s.~1051-1066. ISSN~1860-6768. Dostupné z: https://doi.org/10.1002/biot.201400662.

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