CSF N-Glycomics Using MALDI MS Techniques

Angela Messina; Donata Agata Romeo; Rita Barone; Luisa Sturiale; Angelo Palmigiano; Mario Zappia; Domenico Garozzo

doi:10.1007/978-1-0716-3774-6_4

CSF N-Glycomics Using MALDI MS Techniques

Methods Mol Biol. 2024:2785:49-65. doi: 10.1007/978-1-0716-3774-6_4.

Authors

Angela Messina¹, Donata Agata Romeo¹, Rita Barone^{1

2}, Luisa Sturiale¹, Angelo Palmigiano¹, Mario Zappia³, Domenico Garozzo⁴

Affiliations

¹ CNR, Istituto per i Polimeri, Compositi e i Biomateriali Catania, Catania, Italy.
² Pediatric Neurology Unit, Department of Pediatrics, University of Catania, Catania, Italy.
³ Section of Neurosciences-Department GF Ingrassia, University of Catania, Catania, Italy.
⁴ CNR, Istituto per i Polimeri, Compositi e i Biomateriali Catania, Catania, Italy. domenico.garozzo@cnr.it.

PMID: 38427187
DOI: 10.1007/978-1-0716-3774-6_4

Abstract

In this chapter, we will present the methodology currently applied in our laboratory for the structural elucidation of the cerebrospinal fluid (CSF) N-glycome. N-glycans are released from denatured carboxymethylated glycoproteins by digestion with peptide-N-glycosidase F (PNGase F) and purified using both C18 Sep-Pak^® and porous graphitized carbon (PGC) HyperSep™ Hypercarb™ solid phase extraction (SPE) cartridges. The glycan pool is subsequently permethylated to increase mass spectrometry sensitivity. Molecular assignments are performed through matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF MS) analysis considering either the protein N-linked glycosylation pathway or MALDI TOF MS/MS data. Each stage has been optimized to obtain high-quality mass spectra in reflector mode with an optimal signal-to-noise ratio up to m/z 4800. This method has been successfully adopted to associate specific N-glycome profiles to the early and the advanced phases of Alzheimer's disease (AD).

Keywords: AD; CSF; MALDI MS; MCI; N-glycans.

MeSH terms

Glycomics* / methods
Glycoproteins / chemistry
Glycosylation
Polysaccharides / chemistry
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods
Tandem Mass Spectrometry*

Substances

Glycoproteins
Polysaccharides