Separation and Identification of Glycan Anomers Using Ultrahigh-Resolution Ion-Mobility Spectrometry and Cryogenic Ion S
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J. Am. Soc. Mass Spectrom. (2019) DOI: 10.1007/s13361-019-02333-0
RESEARCH ARTICLE
Separation and Identification of Glycan Anomers Using Ultrahigh-Resolution Ion-Mobility Spectrometry and Cryogenic Ion Spectroscopy Stephan Warnke,1 Ahmed Ben Faleh,1 Valeriu Scutelnic,1,2 Thomas R. Rizzo1 1
Laboratoire de Chimie Physique Moléculaire, EPFL SB ISIC LCPM, École Polytechnique Fédérale de Lausanne, Station 6, CH1015, Lausanne, Switzerland 2 Present Address: Department of Chemistry, University of California, Berkeley, CA 94720, USA
Abstract. The analysis of carbohydrates, or glycans, is challenging for established structuresensitive gas-phase methods. The multitude of possible stereo-, regio-, and structural isomers makes them substantially more complex to analyze than DNA or proteins, and no one method is currently able to fully resolve them. While the combination of tandem mass spectrometry (MS) and ion-mobility spectrometry (IMS) have made important inroads in glycan analysis, in many cases, this approach is still not able to identify the precise isomeric form. To advance the techniques available for glycan analysis, we employ two important innovations. First, we perform ultrahigh-resolution mobility separation using structures for lossless ion manipulations (SLIM) for isomer separation and pre-selection. We then complement this IMS-MS stage with a cryogenic IR spectroscopic dimension since a glycan’s vibrational spectrum provides a fingerprint that is extremely sensitive to the precise isomeric form. Using this unique approach in conjunction with oxygen-18 isotopic labeling, we show on a range of disaccharides how the two α and β anomers that every reducing glycan adopts in solution can be readily separated by mobility and identified based on their IR spectra. In addition to highlighting the power of our technique to detect minute differences in the structure of isomeric carbohydrates, these results provide the means to determine if and when anomericity is retained during collision-induced dissociation (CID) of larger glycans. Keywords: Glycans, Carbohydrates, Anomers, Glucose, Ion-mobility spectrometry, Ion spectroscopy, Mass spectrometry, SLIM Received: 27 June 2019/Revised: 29 August 2019/Accepted: 29 August 2019
Introduction
T
he diversity in both the structure and function of glycans is perhaps unparalleled in biological systems. As individual molecules or as decoration on proteins and lipids, they play a vital role in a myriad of physiological processes and take an active part in the molecular choreography that represents life
Electronic supplementary material The online version of this article (https:// doi.org/10.1007/s13361-019-02333-0) contains supplementary material, which is available to authorized users. Correspondence to: Thomas Rizzo; e-mail: [email protected]
[1–4]. The paramount importance of glycans is not, however, reflected in the current state of the analytical tools available for their analysis. Techniques combining liquid chromatography (LC) and mass spectrometry (MS) [5, 6], which a
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