Glycoprotein enrichment method using a selective magnetic nano-probe platform (MNP) functionalized with lectins.
Protein post-translational modifications (PTMs) have increasingly become a research field of incredible importance to fully understand the regulation of biological processes in health and disease. Among PTMs, glycosylation is one of the most studied for which contributed the development and improvement of enrichment techniques. Nowadays, glycoprotein enrichment methods are based on lectin affinity, covalent interactions, and hydrophilic interaction liquid chromatography (HILIC). Nonetheless, the nanotechnology era has fetched new methods to enrich glycoproteins from complex samples as human biological fluids. For instance, magnetic nanoparticles (MNPs) are being used as an interesting enrichment approach allowing a better characterization of glycoproteins and glycopeptides.In this chapter, we describe an enrichment method based on MNPs functionalized with lectins (Concavalin A, wheat germ agglutinin, and Maackia amurensis lectin) to enrich specific sets of glycoproteins from biological fluids. Moreover, it is proposed a bioinformatic strategy to deal with data retrieved from mass spectrometry analysis of enriched samples aiming the identification of relevant biological processes modulated by a given stimuli and, ultimately, of new biomarkers for disease screening/management.
Department of Chemistry, Mass Spectrometry Centre, QOPNA, University of Aveiro, 3810-193, Aveiro, Portugal.
Marta
M
Cova
Rui
R
Oliveira-Silva
José Alexandre
JA
Ferreira
Rita
R
Ferreira
Francisco
F
Amado
Ana Luísa
AL
Daniel-da-Silva
Rui
R
Vitorino
D002853:Chromatography, Liquid; D019295:Computational Biology; D006023:Glycoproteins / Q000097:blood / Q000737:chemistry* / Q000378:metabolism* / Q000652:urine; D006031:Glycosylation; D006801:Humans; D037102:Lectins / Q000378:metabolism*; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D059748:Proteolysis; D040901:Proteomics; D013048:Specimen Handling; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D006023:Glycoproteins; D037102:Lectins; D058185:Magnetite Nanoparticles