title: Capillary electrophoresis coupled to mass spectrometer for automated and robust polypeptide determination in body fluids for clinical use.
abstract: We describe the application of capillary electrophoresis (CE) coupled on-line to an electrospray ionization-time of flight-mass spectrometer (ESI-TOF-MS) to the analysis of human urine and serum for the identification of biomarkers for clinical diagnostics. CE-MS led to display > 1000 polypeptides present in complex biological samples within 45-60 min in a single analysis run. To extract the information of the CE-MS spectra in a timely fashion, a software was designed to automatically deconvolute and normalize the spectra. Both urine and serum contain several hundred polypeptides in samples from healthy individuals. Hence, it is possible to establish typical "normal urine" or "normal serum" polypeptide patterns. Samples from patients with different diseases display polypeptide patterns that differ significantly from those obtained from healthy individuals. Examining series of patients with the same disease allowed the establishment of polypeptide patterns typical for specific diseases. This permits the search for marker peptides specific for diseases. The data indicate that a single polypeptide present in all patients with the same disease, but absent in all healthy control individuals does not exist. The combination of several polypeptides found in either urine or serum or both are forming a specific pattern, which is indicative not only for the particular disease, but also for the stage of disease. CE-MS detects many polypeptides in single samples and the application of the software to the search of identical polypeptides excreted in urine allows the unbiased diagnosis based on a pattern and does not rely on single disease markers.
authors.affiliation: Mosaiques Diagnostics & Therapeutics AG.
authors.forename: Thorsten
authors.initials: T
authors.lastname: Kaiser
authors.affiliation: Mosaiques Diagnostics & Therapeutics AG.
authors.forename: Stefan
authors.initials: S
authors.lastname: Wittke
authors.affiliation: Hannover Medical School, Department of Toxicology, Hannover, Germany.
authors.forename: Ingo
authors.initials: I
authors.lastname: Just
authors.affiliation: Mosaiques Diagnostics & Therapeutics AG.
authors.forename: Ronald
authors.initials: R
authors.lastname: Krebs
authors.affiliation: Mosaiques Diagnostics & Therapeutics AG.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Bartel
authors.affiliation: Hannover Medical School, Department of Nephrology.
authors.forename: Danilo
authors.initials: D
authors.lastname: Fliser
authors.affiliation: Mosaiques Diagnostics & Therapeutics AG.|Hannover Medical School, Department of Nephrology.
authors.forename: Harald
authors.initials: H
authors.lastname: Mischak
authors.affiliation: Mosaiques Diagnostics & Therapeutics AG.
authors.forename: Eva M
authors.initials: EM
authors.lastname: Weissinger
mesh_terms: D001331:Automation; D015415:Biomarkers / Q000032:analysis / Q000737:chemistry; D001826:Body Fluids / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D010455:Peptides / Q000097:blood* / Q000652:urine*; D012016:Reference Values; D012984:Software; D013997:Time Factors
chemical_list: D015415:Biomarkers; D010455:Peptides