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Issue 13, 2020
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A precise and versatile platform for rapid glycosylation analysis of brain tissue

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Abstract

Glycans influence a variety of aspects of tissue development and pathophysiology, harboring a wealth of biochemical information exploitable for the discovery of new biomarkers. The development of versatile and precise glycoanalytical platforms is crucial to bring to the scientific community reliable tools to unveil the glycan-encoded biochemical information. In this work, a novel platform for brain glycan analysis, named Lysate in-Solution Deglycosylation (LSD), is presented, validated and compared to similar methodologies reported in the literature. Briefly, within the LSD workflow, brain tissue is lysed, proteins are purified via methanol/chloroform extraction, and directly deglycosylated with PNGase F. Released N-glycans are purified using a second round of methanol/chloroform extraction, labelled with fluorescent tags, and cleaned-up prior to LC-FLR/MS analysis. LSD is a quick, sensitive, robust, precise, specific and versatile glycomic workflow, compatible with multiple glyco(proteo)mics analyses of the same sample and eventually applicable to other solid tissues. LSD was devised as a plug-and-play platform for quick glycosylation analysis, and was designed to be compatible with a variety of analytical set-ups (i.e., diverse detection systems), thus enabling the extraction of an additional layer of information from different studies (i.e., proteomics) without the need to collect fresh biological materials. In comparison to analogous N-glycomic workflows, LSD performs reliably on the smallest amount of starting brain tissue and, to the best of the authors' knowledge, is currently the most thoroughly validated neuro-N-glycomic method in the field-related literature, thus representing a significant step forward on the road towards glycan-based biomedical applications.

Graphical abstract: A precise and versatile platform for rapid glycosylation analysis of brain tissue

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Supplementary files

Article information


Submitted
02 Dec 2019
Accepted
28 Feb 2020
First published
28 Feb 2020

Anal. Methods, 2020,12, 1786-1797
Article type
Technical Note

A precise and versatile platform for rapid glycosylation analysis of brain tissue

M. Vacchini, L. Cipolla, O. Gornik, G. Lauc and T. Klarić, Anal. Methods, 2020, 12, 1786
DOI: 10.1039/C9AY02596K

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