Bladder Cancer Associated Glycoprotein Signatures Revealed by Urinary Proteomic Profiling

J Proteome Res. 2007 Jul;6(7):2631-9. doi: 10.1021/pr0700807. Epub 2007 May 23.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Biomarkers, Tumor / urine*
  • Blood Proteins / urine*
  • Chromatography, Affinity / methods
  • Chromatography, Liquid / methods
  • Concanavalin A / chemistry
  • Glycoproteins / urine*
  • Humans
  • Immunoglobulins / urine*
  • Mass Spectrometry
  • Molecular Sequence Data
  • Nanotechnology / methods
  • Protein Array Analysis / methods
  • Proteomics / methods*
  • Urinalysis / methods*
  • Urinary Bladder Neoplasms / urine*

Substances

  • A1BG protein, human
  • Biomarkers, Tumor
  • Blood Proteins
  • Glycoproteins
  • Immunoglobulins
  • Concanavalin A