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Utilizing liquid chromatography, ion mobility spectrometry, and mass spectrometry to assess INLIGHT™ derivatized N-linked glycans in biological samples.
| Content Provider | Europe PMC |
|---|---|
| Author | Butler, Karen E. Kalmar, Jaclyn Gowen Muddiman, David C. Baker, Erin S. |
| Abstract | Glycosylation is a ubiquitous co- and post-translational modification involved in the sorting, folding, and trafficking of proteins in biological systems; in humans, >50% of gene products are glycosylated with the cellular machinery of glycosylation compromising ~2% of the genome. Perturbations in glycosylation have been implicated in a variety of diseases including neurodegenerative diseases and certain types of cancer. However, understanding the relationship between a glycan and its biological role is often difficult due to the numerous glycan isomers that exist. To address this challenge, nanoflow liquid chromatography, ion mobility spectrometry, and mass spectrometry (nLC-IMS-MS) were combined with the Individuality Normalization when Labeling with the Isotopic Glycan Hydrazide Tags (INLIGHT™) strategy to study a series of glycan standards and those enzymatically released from the glycoproteins horseradish peroxidase, fetuin, and pooled human plasma. The combination of IMS and the natural (NAT) and stable-isotope label (SIL) in the INLIGHT™ strategy provided additional confidence for each glycan identification due to the mobility aligned NAT- and SIL-labeled glycans and further capabilities for isomer examinations. Additionally, molecular trend lines based on the IMS and MS dimensions were investigated for the INLIGHT™ derivatized glycans, facilitating rapid identification of putative glycans in complex biological samples.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00216-021-03570-7. |
| ISSN | 16182642 |
| Journal | Analytical and Bioanalytical Chemistry |
| Volume Number | 414 |
| PubMed Central reference number | PMC8336533 |
| Issue Number | 1 |
| PubMed reference number | 34347113 |
| e-ISSN | 16182650 |
| DOI | 10.1007/s00216-021-03570-7 |
| Language | English |
| Publisher | Springer Berlin Heidelberg |
| Publisher Date | 2021-08-04 |
| Publisher Place | Berlin/Heidelberg |
| Access Restriction | Open |
| Rights License | This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic. © Springer-Verlag GmbH Germany, part of Springer Nature 2021 |
| Subject Keyword | Glycomics Ion mobility spectrometry INLIGHT™ N-Linked glycans LC-IMS-MS |
| Content Type | Text |
| Resource Type | Article |
| Subject | Analytical Chemistry Biochemistry |
