title: Three-Dimensionally Functionalized Reverse Phase Glycoprotein Array for Cancer Biomarker Discovery and Validation.
abstract: Glycoproteins have vast structural diversity that plays an important role in many biological processes and have great potential as disease biomarkers. Here, we report a novel functionalized reverse phase protein array (RPPA), termed polymer-based reverse phase glycoprotein array (polyGPA), to capture and profile glycoproteomes specifically, and validate glycoproteins. Nitrocellulose membrane functionalized with globular hydroxyaminodendrimers was used to covalently capture preoxidized glycans on glycoproteins from complex protein samples such as biofluids. The captured glycoproteins were subsequently detected using the same validated antibodies as in RPPA. We demonstrated the outstanding specificity, sensitivity, and quantitative capabilities of polyGPA by capturing and detecting purified as well as endogenous α-1-acid glycoprotein (AGP) in human plasma. We further applied quantitative N-glycoproteomics and the strategy to validate a panel of glycoproteins identified as potential biomarkers for bladder cancer by analyzing urine glycoproteins from bladder cancer patients or matched healthy individuals.
authors.forename: Li
authors.initials: L
authors.lastname: Pan
authors.forename: Hillary Andaluz
authors.initials: HA
authors.lastname: Aguilar
authors.forename: Linna
authors.initials: L
authors.lastname: Wang
authors.affiliation: Tymora Analytical Operations , West Lafayette, Indiana 47906, United States.
authors.forename: Anton
authors.initials: A
authors.lastname: Iliuk
authors.affiliation: Tymora Analytical Operations , West Lafayette, Indiana 47906, United States.|Center for Cancer Research, Purdue University , West Lafayette, Indiana 47907, United States.
authors.forename: W Andy
authors.initials: WA
authors.lastname: Tao
mesh_terms: D014408:Biomarkers, Tumor / Q000032:analysis*; D006023:Glycoproteins / Q000737:chemistry*; D006801:Humans; D040081:Protein Array Analysis / Q000379:methods*; D001749:Urinary Bladder Neoplasms / Q000175:diagnosis* / Q000378:metabolism
chemical_list: D014408:Biomarkers, Tumor; D006023:Glycoproteins