Molecules 2014, 19(7), 9070-9088; doi:10.3390/molecules19079070

Characterization of Intact Neo-Glycoproteins by Hydrophilic Interaction Liquid Chromatography

1email, 1email, 1email, 1email, 2email, 1email, 1email and 1,* email
Received: 12 May 2014; in revised form: 18 June 2014 / Accepted: 19 June 2014 / Published: 30 June 2014
(This article belongs to the Special Issue Oligosaccharides and Glyco-Conjugates)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract: In this study, an HPLC HILIC-UV method was developed for the analysis of intact neo-glycoproteins. During method development the experimental conditions evaluated involved different HILIC columns (TSKgel Amide-80 and ZIC-pHILIC), and water-acetonitrile mixtures containing various types of acids and salts. The final selected method was based on a TSKgel Amide-80 column and a mobile phase composed of acetonitrile and water both containing 10 mM HClO4. The influence of temperature and sample preparation on the chromatographic performances of the HILIC method was also investigated. The method was applied to the separation of neo-glycoproteins prepared starting from the model protein RNase A by chemical conjugation of different glycans. Using the method here reported it was possible to monitor by UV detection the glycosylation reaction and assess the distribution of neo-glycoprotein isoforms without laborious sample workup prior to analysis.
Keywords: hydrophilic interaction liquid chromatography; intact glycoproteins; semi-synthetic glycosylation monitoring; neo-glycoproteins
PDF Full-text Download PDF Full-Text [2269 KB, Updated Version, uploaded 4 July 2014 10:59 CEST]
The original version is still available [803 KB, uploaded 30 June 2014 11:19 CEST]

Export to BibTeX |

MDPI and ACS Style

Pedrali, A.; Tengattini, S.; Marrubini, G.; Bavaro, T.; Hemström, P.; Massolini, G.; Terreni, M.; Temporini, C. Characterization of Intact Neo-Glycoproteins by Hydrophilic Interaction Liquid Chromatography. Molecules 2014, 19, 9070-9088.

AMA Style

Pedrali A, Tengattini S, Marrubini G, Bavaro T, Hemström P, Massolini G, Terreni M, Temporini C. Characterization of Intact Neo-Glycoproteins by Hydrophilic Interaction Liquid Chromatography. Molecules. 2014; 19(7):9070-9088.

Chicago/Turabian Style

Pedrali, Alice; Tengattini, Sara; Marrubini, Giorgio; Bavaro, Teodora; Hemström, Petrus; Massolini, Gabriella; Terreni, Marco; Temporini, Caterina. 2014. "Characterization of Intact Neo-Glycoproteins by Hydrophilic Interaction Liquid Chromatography." Molecules 19, no. 7: 9070-9088.

Molecules EISSN 1420-3049 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert