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Anal Chem. 2005 Nov 15;77(22):7163-71.

Combined top-down and bottom-up mass spectrometric approach to characterization of biomarkers for renal disease.

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  • 1Ion Cyclotron Resonance Program, National High Magnetic Field Laboratory, Florida State University, Tallahassee, 32310-4005, USA.

Abstract

Here we describe a mass spectrometry (MS) approach for biomarker discovery and structural characterization, based on both top-down and bottom-up analyses. Capillary electrophoresis (CE) coupled to electrospray ionization (ESI) time-of-flight (TOF) MS serves to separate and mass-measure the thousands of polypeptides contained in human urine. Statistical analysis of the differences between healthy control samples and patients with focal-segmental glomerulosclerosis, membranous glomerulonephritis, minimal change disease, IgA nephropathy, and diabetic nephropathy validates multiple biomarkers for the control and each of the diseases. To identify those biomarkers, we employ preparative CE, enabling direct infusion ESI MS analysis, followed by sample manipulation and reanalysis where necessary. We show how tandem Fourier transform ion cyclotron resonance (FT-ICR) MS identifies these sometimes large (>8 kDa) biomarkers. Critically, we maintain connectivity between the CE TOF MS data and the ICR data used for biomarker identification.

PMID:
16285662
[PubMed - indexed for MEDLINE]
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