title: Enrichment and identification of glycoproteins in human saliva using lectin magnetic bead arrays.
abstract: Aberrant glycosylation of proteins is a hallmark of tumorigenesis and could provide diagnostic value in cancer detection. Human saliva is an ideal source of glycoproteins due to the relatively high proportion of glycosylated proteins in the salivary proteome. Moreover, saliva collection is noninvasive and technically straightforward, and the sample collection and storage is relatively easy. Although differential glycosylation of proteins can be indicative of disease states, identification of differential glycosylation from clinical samples is not trivial. To facilitate salivary glycoprotein biomarker discovery, we optimized a method for differential glycoprotein enrichment from human saliva based on lectin magnetic bead arrays (saLeMBA). Selected lectins from distinct reactivity groups were used in the saLeMBA platform to enrich salivary glycoproteins from healthy volunteer saliva. The technical reproducibility of saLeMBA was analyzed with liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify the glycosylated proteins enriched by each lectin. Our saLeMBA platform enabled robust glycoprotein enrichment in a glycoprotein- and lectin-specific manner consistent with known protein-specific glycan profiles. We demonstrated that saLeMBA is a reliable method to enrich and detect glycoproteins present in human saliva.
authors.affiliation: The University of Queensland Diamantina Institute, The University of Queensland, Translational Research Institute, Woolloongabba, Queensland, 4102, Australia.
authors.forename: Michael
authors.initials: M
authors.lastname: Caragata
authors.affiliation: The University of Queensland Diamantina Institute, The University of Queensland, Translational Research Institute, Woolloongabba, Queensland, 4102, Australia.
authors.forename: Alok K
authors.initials: AK
authors.lastname: Shah
authors.affiliation: School of Chemistry and Molecular Biosciences, The University of Queensland, St. Lucia, Queensland, 4072, Australia.
authors.forename: Benjamin L
authors.initials: BL
authors.lastname: Schulz
authors.affiliation: The University of Queensland Diamantina Institute, The University of Queensland, Translational Research Institute, Woolloongabba, Queensland, 4102, Australia. Electronic address: m.hill2@uq.edu.au.
authors.forename: Michelle M
authors.initials: MM
authors.lastname: Hill
authors.affiliation: School of Biomedical Sciences, Institute of Biomedical Innovations, Queensland University of Technology, Kelvin Grove, and Translational Research Institute, Woolloongabba, Queensland, 4102, Australia. Electronic address: chamindie.punyadeera@qut.edu.au.
authors.forename: Chamindie
authors.initials: C
authors.lastname: Punyadeera
mesh_terms: D014408:Biomarkers, Tumor / Q000032:analysis / Q000302:isolation & purification; D002853:Chromatography, Liquid / Q000379:methods; D006023:Glycoproteins / Q000032:analysis* / Q000302:isolation & purification; D006031:Glycosylation; D006801:Humans; D037102:Lectins / Q000737:chemistry*; D059346:Magnets / Q000737:chemistry*; D009369:Neoplasms / Q000175:diagnosis; D040901:Proteomics / Q000379:methods; D015203:Reproducibility of Results; D012463:Saliva / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D014408:Biomarkers, Tumor; D006023:Glycoproteins; D037102:Lectins
keywords: Biomarker discovery; Glycosylation; Lectins; Magnetic beads; Saliva proteomics