A high-resolution measurement of nucleotide sugars by using ion-pair reverse chromatography and tandem columns.
Chinese hamster ovary
Columns in tandem
Core-shell
Ion-pair reverse chromatography
Nucleotide sugars
UDP sugars
Journal
Analytical and bioanalytical chemistry
ISSN: 1618-2650
Titre abrégé: Anal Bioanal Chem
Pays: Germany
ID NLM: 101134327
Informations de publication
Date de publication:
Jun 2020
Jun 2020
Historique:
received:
21
01
2020
accepted:
18
03
2020
revised:
08
03
2020
pubmed:
18
4
2020
medline:
28
1
2021
entrez:
18
4
2020
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 32300845
doi: 10.1007/s00216-020-02608-6
pii: 10.1007/s00216-020-02608-6
doi:
Substances chimiques
Nucleoside Diphosphate Sugars
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
3683-3693Subventions
Organisme : National Science Foundation
ID : 1706731
Références
Sha S, Agarabi C, Brorson K, Lee DY, Yoon S. N-Glycosylation design and control of therapeutic monoclonal antibodies. Trends Biotechnol. 2016;34(10):835–46.
pubmed: 27016033
Lalonde ME, Durocher Y. Therapeutic glycoprotein production in mammalian cells. J Biotechnol. 2017;251:128–40.
pubmed: 28465209
Higel F, Seidl A, Sorgel F, Friess W. N-glycosylation heterogeneity and the influence on structure, function and pharmacokinetics of monoclonal antibodies and Fc fusion proteins. Eur J Pharm Biopharm. 2016;100:94–100.
pubmed: 26775146
Wang Q, Chung CY, Yang W, Yang G, Chough S, Chen Y, et al. Combining butyrated ManNAc with glycoengineered CHO cells improves EPO glycan quality and production. Biotechnol J. 2019;14(4):e1800186.
pubmed: 30221828
Wang Q, Yang G, Wang T, Yang W, Betenbaugh MJ, Zhang H. Characterization of intact glycopeptides reveals the impact of culture media on site-specific glycosylation of EPO-Fc fusion protein generated by CHO-GS cells. Biotechnol Bioeng. 2019;116(9):2303–15.
pubmed: 31062865
Villiger TK, Steinhoff RF, Ivarsson M, Solacroup T, Stettler M, Broly H, et al. High-throughput profiling of nucleotides and nucleotide sugars to evaluate their impact on antibody N-glycosylation. J Biotechnol. 2016;229:3–12.
pubmed: 27131894
Blondeel EJ, Braasch K, McGill T, Chang D, Engel C, Spearman M, et al. Tuning a MAb glycan profile in cell culture: supplementing N-acetylglucosamine to favour G0 glycans without compromising productivity and cell growth. J Biotechnol. 2015;214:105–12.
pubmed: 26387447
Jedrzejewski PM, del Val IJ, Constantinou A, Dell A, Haslam SM, Polizzi KM, et al. Towards controlling the glycoform: a model framework linking extracellular metabolites to antibody glycosylation. Int J Mol Sci. 2014;15(3):4492–522.
pubmed: 24637934
pmcid: 3975410
Kochanowski N, Blanchard F, Cacan R, Chirat F, Guedon E, Marc A, et al. Influence of intracellular nucleotide and nucleotide sugar contents on recombinant interferon-gamma glycosylation during batch and fed-batch cultures of CHO cells. Biotechnol Bioeng. 2008;100(4):721–33.
pubmed: 18496872
Wong NS, Wati L, Nissom PM, Feng HT, Lee MM, Yap MG. An investigation of intracellular glycosylation activities in CHO cells: effects of nucleotide sugar precursor feeding. Biotechnol Bioeng. 2010;107(2):321–36.
pubmed: 20506284
Sumit M, Dolatshahi S, Chu AA, Cote K, Scarcelli JJ, Marshall JK, et al. Dissecting N-glycosylation dynamics in Chinese hamster ovary cells fed-batch cultures using time course omics analyses. iScience. 2019;12:102–20.
pubmed: 30682623
pmcid: 6352710
Naik HM, Majewska NI, Betenbaugh MJ. Impact of nucleotide sugar metabolism on protein N-glycosylation in Chinese hamster ovary (CHO) cell culture. Curr Opin Chem Eng. 2018;22:167–76.
Sha S, Yoon S. An investigation of nucleotide sugar dynamics under the galactose supplementation in CHO cell culture. Process Biochem. 2019;81:165–74.
Jimenez del Val I, Nagy JM, Kontoravdi C. A dynamic mathematical model for monoclonal antibody N-linked glycosylation and nucleotide sugar donor transport within a maturing Golgi apparatus. Biotechnol Prog. 2011;27(6):1730–43.
pubmed: 21956887
Del Val IJ, Polizzi KM, Kontoravdi C. A theoretical estimate for nucleotide sugar demand towards Chinese hamster ovary cellular glycosylation. Sci Rep. 2016;6:28547.
pubmed: 27345611
pmcid: 4921913
Sha S, Huang Z, Agarabi C, Lute S, Brorson K, Yoon S. Prediction of N-linked glycoform profiles of monoclonal antibody with extracellular metabolites and two-step intracellular models. Processes. 2019;7(4):227.
Burleigh SC, van de Laar T, Stroop CJM, van Grunsven WMJ, O'Donoghue N, Rudd PM, et al. Synergizing metabolic flux analysis and nucleotide sugar metabolism to understand the control of glycosylation of recombinant protein in CHO cells. BMC Biotechnol. 2011;11(1):95.
pubmed: 22008152
pmcid: 3219575
Sha S, Huang Z, Wang Z, Yoon S. Mechanistic modeling and applications for CHO cell culture development and production. Curr Opin Chem Eng. 2018;22:54–61.
Räbinä J, Mäki M, Savilahti EM, Järvinen N, Penttilä L, Renkonen R. Analysis of nucleotide sugars from cell lysates by ion-pair solid-phase extraction and reversed-phase high-performance liquid chromatography. Glycoconj J. 2001;18(10):799–805.
pubmed: 12441669
Nakajima K, Taniguchi N. Simultaneous quantification of nucleotide sugar metabolism by LC and LC-MS. In: Taniguchi N, et al., editors. Glycoscience: biology and medicine. Tokyo: Springer Japan; 2015. p. 103–10.
Kochanowski N, Blanchard F, Cacan R, Chirat F, Guedon E, Marc A, et al. Intracellular nucleotide and nucleotide sugar contents of cultured CHO cells determined by a fast, sensitive, and high-resolution ion-pair RP-HPLC. Anal Biochem. 2006;348(2):243–51.
pubmed: 16325757
del Val IJ, Kyriakopoulos S, Polizzi KM, Kontoravdi C. An optimized method for extraction and quantification of nucleotides and nucleotide sugars from mammalian cells. Anal Biochem. 2013;443(2):172–80.
pubmed: 24036437
Jorge TF, Florêncio MH, Ribeiro-Barros AI, António C. Quantification and structural characterization of raffinose family oligosaccharides in Casuarina glauca plant tissues by porous graphitic carbon electrospray quadrupole ion trap mass spectrometry. Int J Mass Spectrom. 2017;413:127–34.
Warth B, Siegwart G, Lemmens M, Krska R, Adam G, Schuhmacher R. Hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry for the quantification of uridine diphosphate-glucose, uridine diphosphate-glucuronic acid, deoxynivalenol and its glucoside: in-house validation and application to wheat. J Chromatogr A. 2015;1423:183–9.
pubmed: 26554298
Eastwood H, Xia F, Lo MC, Zhou J, Jordan JB, McCarter J, et al. Development of a nucleotide sugar purification method using a mixed mode column & mass spectrometry detection. J Pharm Biomed Anal. 2015;115:402–9.
pubmed: 26279371
Ito J, Herter T, Baidoo EE, Lao J, Vega-Sanchez ME, Michelle Smith-Moritz A, et al. Analysis of plant nucleotide sugars by hydrophilic interaction liquid chromatography and tandem mass spectrometry. Anal Biochem. 2014;448:14–22.
pubmed: 24299991
Pabst M, Grass J, Fischl R, Léonard R, Jin C, Hinterkörner G, et al. Nucleotide and nucleotide sugar analysis by liquid chromatography-electrospray ionization-mass spectrometry on surface-conditioned porous graphitic carbon. Anal Chem. 2010;82(23):9782–8.
pubmed: 21043458
pmcid: 2995335
Wang T, Chen X, Li L, Cao Y, Zhao L, Chai Y, et al. Characterization of nucleotides and nucleotide sugars in Candida albicans by high performance liquid chromatography–mass spectrometry with a porous graphite carbon column. Anal Lett. 2014;47(2):234–49.
Feng HT, Wong N, Wee S, Lee MM. Simultaneous determination of 19 intracellular nucleotides and nucleotide sugars in Chinese hamster ovary cells by capillary electrophoresis. J Chromatogr B Analyt Technol Biomed Life Sci. 2008;870(1):131–4.
pubmed: 18541463
Ryll T, Wagner R. Improved ion-pair high-performance liquid chromatographic method for the quantification of a wide variety of nucleotides and sugar-nucleotides in animal cells. J Chromatogr B Biomed Sci Appl. 1991;570(1):77–88.
St Amand MM, Radhakrishnan D, Robinson AS, Ogunnaike BA. Identification of manipulated variables for a glycosylation control strategy. Biotechnol Bioeng. 2014;111(10):1957–70.
pubmed: 24728980
Nakajima K, Kitazume S, Angata T, Fujinawa R, Ohtsubo K, Miyoshi E, et al. Simultaneous determination of nucleotide sugars with ion-pair reversed-phase HPLC. Glycobiology. 2010;20(7):865–71.
pubmed: 20371511
Fekete S, Ganzler K, Fekete J. Efficiency of the new sub-2 mum core-shell (Kinetex) column in practice, applied for small and large molecule separation. J Pharm Biomed Anal. 2011;54(3):482–90.
pubmed: 20940092
Zhang J, Raglione T, Wang Q, Kleintop B, Tomasella F, Liang X. Regeneration of tetrabutylammonium ion-pairing reagent distribution in a gradient elution of reversed phase ion-pair chromatography. J Chromatogr Sci. 2011;49:825–31.
pubmed: 22080812
Lazarowski ER, Shea DA, Boucher RC, Harden TK. Release of cellular UDP-glucose as a potential extracellular signaling molecule. Mol Pharmacol. 2003;63(5):1190–7.
pubmed: 12695547