High-throughput profiling of nucleotides and nucleotide sugars to evaluate their impact on antibody N-glycosylation
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- 作者：Thomas K. Villigera ; 1 ; Robert F. Steinhoffb ; 1 ; Marija Ivarssona ; Thomas Solacroupc ; Matthieu Stettlerc ; Hervé ; Brolyc ; Jasmin Krismerb ; Martin Pabstb ; Renato Zenobib ; Massimo Morbidellia ; Miroslav Soosa ; d ; miroslav.soos@chem.ethz.ch" class="auth_mail" title="E-mail the corresponding author
- 关键词：9-AA ; 9-aminoacridine ; ADP ; adenosine diphosphate ; AMP ; adenosine monophosphate ; ATP ; adenosine triphosphate ; CDP ; cytidine diphosphate ; CDP ; cytidine diphosphate ; CMP ; cytidine monophosphate ; CMP-Neu5Ac ; cytidine diphosphate N-acetylneuraminic acid ; CTP ; cytidine triphosphate ; DWP ; deep well plate ; GDP ; guanosine diphosphate ; GDP-Fuc ; guanosine diphosphate fucose ; GDP-Hex ; guanosine diphosphate hexose ; GDP-Man ; guanosine diphosphate mannose ; GMP ; guanosine monophosphate ; GTP ; guanosine triphosp
- 刊名：Journal of Biotechnology
- 出版年：2016
- 出版时间：10 July 2016
- 年：2016
- 卷：229
- 期：Complete
- 页码：3-12
- 全文大小：1043 K
文摘
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Modulation of mAb N-linked glycosylation in micro-bioreactors using different feed additives.
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A high-throughput MALDI-TOF-MS technique reduced the analysis time for intracellular nucleotides and nucleotide sugars more than one order of magnitude.
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UDP-Gal synthesis appeared to be limiting, however a further increase of UDP-GlcNAc or GDP-Fuc left the glycosylation process mostly unchanged.
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Manganese and asparagine supplementation significantly altered the glycosylation profiles without changing intracellular components.
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Scalability was demonstrated by a pilot-scale bioreactor.