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Probing Blood Plasma Protein Glycosylation with Infrared Spectroscopy.

Authors :
Voronina L
Fleischmann F
Šimunović J
Ludwig C
Novokmet M
Žigman M
Source :
Analytical chemistry [Anal Chem] 2024 Feb 07. Date of Electronic Publication: 2024 Feb 07.
Publication Year :
2024
Publisher :
Ahead of Print

Abstract

The health state of an individual is closely linked to the glycosylation patterns of his or her blood plasma proteins. However, obtaining this information requires cost- and time-efficient analytical methods. We put forward infrared spectroscopy, which allows label-free analysis of protein glycosylation but so far has only been applied to analysis of individual proteins. Although spectral information does not directly provide the molecular structure of the glycans, it is sensitive to changes therein and covers all types of glycosidic linkages. Combining single-step ion exchange chromatography with infrared spectroscopy, we developed a workflow that enables the separation and analysis of major protein classes in blood plasma. Our results demonstrate that infrared spectroscopy can identify different patterns and global levels of glycosylation of intact plasma proteins. To showcase the strengths and limitations of the proposed approach, we compare the glycoforms of human and bovine alpha-1-acid glycoproteins, which exhibit highly variable global levels of glycosylation. To independently evaluate our conclusions, the glycan moieties of human alpha-1-acid glycoprotein were further analyzed using an established glycomics workflow. Importantly, the chromatographic separation of blood plasma improves the detection of aberrant glycoforms of a given protein as compared to infrared spectroscopy of bulk plasma. The presented approach allows a time-efficient comparison of glycosylation patterns of multiple plasma proteins, opening new avenues for biomedical probing.

Details

Language :
English
ISSN :
1520-6882
Database :
MEDLINE
Journal :
Analytical chemistry
Publication Type :
Academic Journal
Accession number :
38324652
Full Text :
https://doi.org/10.1021/acs.analchem.3c03589