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How to separate you, glycopeptides?

Members of Laboratory of Biomolecular Structure and Function participated in a research published in the Analytical and Bioanalytical Chemistry Journal. The manuscript focuses on separation of glycopeptides using reverse-phase chromatography (RPLC), hydrophilic interaction chromatography (HILIC) and its combination with ion-exchange chromatography (HILIC/IEC).

Glycosylation is a frequent modification necessary for proper function of a number of proteins, and its alteration can indicate inflammatory or cancer diseases. A glycoprotein is composed of a set of biomolecules with the same primary structure but with various glycans attached to different glycosylation sites, It is therefore difficult to analyze unless you have an effective analyte separation method and a sensitive mass spectrometric detection. HILIC is an advantageous alternative to conventional RPLC in glycopeptide separation, as it achieves better resolution of individual peptide glycoforms.

A wide range of functionalized HILIC stationary phases is available, however for the new ones – especially those combining several distinct retention mechanisms – the possible benefit for glycoproteomics is not fully explored. Therefore, the authors of the manuscript [LINK https://rdcu.be/ckMUy] compared the separation of IgG glycopeptides on phases containing 1/ polyhydroxylated silica (pure HILIC), 2/ unfunctionalized silica (HILIC/acidic IEC), and 3/ aminopropylated silica (HILIC/basic IEC). Among other findings, they showed groups of analytes that are better separated on pure HILIC, or on one of HILIC/IEC columns.

Published: Jun 22, 2021 04:35 PM

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