Advancing the characterization of glycan dynamics with in-electrospray ionization hydrogen/deuterium exchange-mass spectrometry.

Glycans are biomolecules that have various essential roles in human health and disease. Carbohydrates are composed of isomeric features ranging from differences in composition, linkage, and configuration, resulting in complex and branched molecules. The structural diversity of carbohydrates influences their biological functions. Although various analytical techniques exist to study carbohydrates, mass spectrometry (MS) remains one of the most widely used methods because it provides structural information. Hydrogen/deuterium exchange (HDX-MS) is a labeling technique that monitors the exchanging labile hydrogen atoms for molecules across different timepoints. Traditionally, HDX-MS has been used to study protein structure and dynamics; however, this deuterium labeling method has become an emerging technique to study carbohydrates. The hydroxyl groups in carbohydrate structures exchange rapidly, resulting in limitations for traditional HDX when analyzing carbohydrates. In-electrospray ionization (in-ESI HDX-MS) enables rapid labeling and can be applied to study the solvated states of metal-adducted carbohydrates. The goal of this work is to develop in-ESI HDX-MS methods to study carbohydrate dynamics and structures. In Chapter 2, ESI droplet sizes and lifetimes are altered which changes the HDX reaction time, and thereby produces different magnitudes of HDX. The dynamics of solvated carbohydrates are monitored and are demonstrated to influence HDX. In Chapter 3, HX-Express, an HDX data analysis software, is used to interpret rapid in-ESI HDX data for several analytes, including a peptide, carbohydrates, and a glycopeptide. This work demonstrates a way to analyze complex spectral data resulting from different types of protonated and/or sodiated and potassiated adducts. In Chapter 4, the repeatability of in-ESI HDX using different-sized tips is validated to improve in-ESI HDX methods for carbohydrate studies. Collectively, this work facilitates the development of in-ESI HDX-MS methods to characterize the dynamics and structures of carbohydrate isomers.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Doctoral

Grantor

Baylor University.

Year dc:date.issued

2025

Author dc:creator

• Quintero, Ana Victoria, 1997-

Advisor dc:contributor.advisor

• Gallagher, Elyssia S.

• Mass spectrometry.
• In-electrospray ionization hydrogen-deuterium exchange (in-ESI HDX)
• Carbohydrates.
• Hydrogen-deuterium exchange mass spectrometry (in-ESI HDX-MS)

Statement dc:rights

• Baylor University works are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. Contact libraryquestions@baylor.edu for inquiries about permission.

Language dc:language.iso

en

Handle dc:identifier.uri

https://hdl.handle.net/2104/13962

OAI identifier oai:identifier

oai:baylor-ir.tdl.org:2104/13962