Intracellular nucleotide and nucleotide sugar contents of cultured CHO cells determined by a fast, sensitive, and high-resolution ion-pair RP-HPLC
type: Dataset
description: Analysis of intracellular nucleotide and nucleotide sugar contents is essential in studying protein glycosylation of mammalian cells. Nucleotides and nucleotide sugars are the donor substrates of glycosyltransferases, and nucleotides are involved in cellular energy metabolism and its regulation. A sensitive and reproducible ion-pair reverse-phase high-performance liquid chromatography (RP-HPLC) method has been developed, allowing the direct and simultaneous detection and quantification of some essential nucleotides and nucleotide sugars. After a perchloric acid extraction, 13 molecules (8 nucleotides and 5 nucleotide sugars) were separated, including activated sugars such as UDP-glucose, UDP-galactose, GDP-mannose, UDP-N-acetylglucosamine, and UDP-N-acetylgalactosamine. To validate the analytical parameters, the reproducibility, linearity of calibration curves, detection limits, and recovery were evaluated for standard mixtures and cell extracts. The developed method is capable of resolving picomolar quantities of nucleotides and nucleotide sugars in a single chromatographic run. The HPLC method was then applied to quantify intracellular levels of nucleotides and nucleotide sugars of Chinese hamster ovary (CHO) cells cultivated in a bioreactor batch process. Evolutions of the titers of nucleotides and nucleotide sugars during the batch process are discussed.
issued: 2026-05-15T09:35:31.290021
language: en
modified: 2026-05-15T09:35:31.290027
contact point: Organization
  fn: CCSD
keyword: cho-cells
keyword: energy-charge
keyword: glycosylation
keyword: hplc
keyword: infoeu-reposemanticsarticle
keyword: journal-articles
keyword: nucleotide-sugars
keyword: nucleotides
keyword: spigprocengineering-sciences-physicschemical-and-process-engineering
landing page: 0003-2697

Intracellular nucleotide and nucleotide sugar contents of cultured CHO cells determined by a fast, sensitive, and high-resolution ion-pair RP-HPLC
type: Distribution
format: HTML
issued: 2026-05-15T09:35:31.295155
modified: 2026-05-15T09:35:31.264694
access URL: https://hal.science/hal-00077315

test_moissonnage_selune
type: Agent