ArticleAnalytical and bioanalytical chemistry2020
A high-resolution measurement of nucleotide sugars by using ion-pair reverse chromatography and tandem columns.
Article in Analytical and bioanalytical chemistry, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed, 11 citations in OpenAlex.
- Temporal Galactose-Manganese Feeding in Fed-Batch and Perfusion Bioreactors Modulates UDP-Galactose Pools for Enhanced mAb Glycosylation Homogeneity.Biotechnology and bioengineering · 2025Article
- Mixed-phase weak anion-exchange/reversed-phase LC-MS/MS for analysis of nucleotide sugars in human fibroblasts.Analytical and bioanalytical chemistry · 2024Article
- Determination of UDP-Glucose and UDP-Galactose in Maize by Hydrophilic Interaction Liquid Chromatography and Tandem Mass Spectrometry.Journal of analytical methods in chemistry · 2022Article
- Desalting Paper Spay Mass Spectrometry (DPS-MS) for Rapid Detection of Glycans and Glycoconjugates.International journal of mass spectrometry · 2021Article
- An Ion Chromatography-Ultrahigh-Resolution-MSAnalytical chemistry · 2021Article
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
Abstract
N-Linked glycosylation is a cellular process transferring sugars from glycosyl donors to proteins or lipids. Biopharmaceutical products widely produced by culturing mammalian cells such as Chinese hamster ovary (CHO) cells are typically glycosylated during biosynthesis. For some biologics, the N-linked glycan is a critical quality attribute of the drugs. Nucleotide sugars are the glycan donors and impact the intracellular glycosylation process. In current analytical methods, robust separation of nucleotide sugar isomers such as UDP glucose and UDP galactose remains a challenge because of their structural similarity. In this study, we developed a strategy to resolve the separation of major nucleotide sugars including challenging isomers based on the use of ion-pair reverse phase (IP-RP) chromatography. The strategy applies core-shell columns and connects multiple columns in tandem to increase separation power and ultimately enables high-resolution detection of nucleotide sugars from cell extracts. The key parameters in the IP-RP method, including temperature, mobile phase, and flow rates, have been systematically evaluated in this work and the theoretical mechanisms of the chromatographic behavior were proposed. Graphical abstract.
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