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Biomarkers for Alzheimer’s Disease Drug Development pp 75–91Cite as

CSF N-Glycomics Using MALDI MS Techniques in Alzheimer’s Disease

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1750))

Abstract

In this chapter, we 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.

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Acknowledgements

Generous donation from Stellalucente Trust is gratefully acknowledged. This chapter is dedicated to our dear friend and colleague Francesco Le Pira, University of Catania, who too early passed away.

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Authors and Affiliations

  1. CNR, Istituto per i Polimeri, Compositi e i Biomateriali Catania, Catania, Italy

    Angelo Palmigiano, Angela Messina, Rosaria Ornella Bua, Rita Barone, Luisa Sturiale & Domenico Garozzo

  2. Pediatric Neurology Unit, Department of Pediatrics, University of Catania, Catania, Italy

    Rita Barone

  3. Section of Neurosciences, Department of GF Ingrassia, University of Catania, Catania, Italy

    Mario Zappia

Authors
  1. Angelo Palmigiano
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  2. Angela Messina
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  3. Rosaria Ornella Bua
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  4. Rita Barone
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  5. Luisa Sturiale
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  6. Mario Zappia
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  7. Domenico Garozzo
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Corresponding author

Correspondence to Domenico Garozzo .

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Editors and Affiliations

  1. Department of Psychiatry and Psychotherapy, Ludwig-Maximilians-Universität München, Munich, Germany

    Robert Perneczky

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Palmigiano, A. et al. (2018). CSF N-Glycomics Using MALDI MS Techniques in Alzheimer’s Disease. In: Perneczky, R. (eds) Biomarkers for Alzheimer’s Disease Drug Development. Methods in Molecular Biology, vol 1750. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7704-8_5

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  • DOI: https://doi.org/10.1007/978-1-4939-7704-8_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7703-1

  • Online ISBN: 978-1-4939-7704-8

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