title: Bladder cancer associated glycoprotein signatures revealed by urinary proteomic profiling.
abstract: Current methods in the noninvasive detection and surveillance of bladder cancer via urine analysis include voided urine cytology (VUC) and some diagnostic urinary protein biomarkers; however, due to the poor sensitivity of VUC and high false-positive rates of currently available protein assays, detection of bladder cancer via urinalysis remains a challenge. In the study presented here, a rapid, high-sensitivity technique was developed to profile the N-linked glycoprotein component in naturally micturated human urine specimens. Concanavalin A (Con A) affinity chromatography coupled to nanoflow liquid chromatography was utilized to separate the complex peptide mixture prior to a linear ion trap MS analysis. Of 186 proteins identified with high confidence by multiple analyses, 40% were secreted proteins, 18% membrane proteins, and 14% extracellular proteins. In this study, the presence of several proteins appeared to be associated with the presence of bladder cancer, including alpha-1B-glycoprotein that was detected in all tumor-bearing patient samples but in none of the samples obtained from non-tumor-bearing individuals. The combination of Con A affinity chromatography and nano-LC/MS/MS provides an initial investigation of N-glycoproteins in complex biological samples and facilitates the identification of potential biomarkers of bladder cancer in noninvasively obtained human urine.
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, Michigan, USA.
authors.forename: Paweena
authors.initials: P
authors.lastname: Kreunin
authors.forename: Jia
authors.initials: J
authors.lastname: Zhao
authors.forename: Charles
authors.initials: C
authors.lastname: Rosser
authors.forename: Virginia
authors.initials: V
authors.lastname: Urquidi
authors.forename: David M
authors.initials: DM
authors.lastname: Lubman
authors.forename: Steve
authors.initials: S
authors.lastname: Goodison
mesh_terms: D000595:Amino Acid Sequence; D014408:Biomarkers, Tumor / Q000652:urine*; D001798:Blood Proteins / Q000652:urine*; D002846:Chromatography, Affinity / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D003208:Concanavalin A / Q000737:chemistry; D006023:Glycoproteins / Q000652:urine*; D006801:Humans; D007136:Immunoglobulins / Q000652:urine*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D036103:Nanotechnology / Q000379:methods; D040081:Protein Array Analysis / Q000379:methods; D040901:Proteomics / Q000379:methods*; D016482:Urinalysis / Q000379:methods*; D001749:Urinary Bladder Neoplasms / Q000652:urine*
chemical_list: C048566:A1BG protein, human; D014408:Biomarkers, Tumor; D001798:Blood Proteins; D006023:Glycoproteins; D007136:Immunoglobulins; D003208:Concanavalin A