Miniaturized monolithic disks for immunoadsorption of cardiac biomarkers from serum |
| |
Authors: | N. Delmotte U. Kobold T. Meier A. Gallusser A. Strancar C. G. Huber |
| |
Affiliation: | (1) Department of Chemistry, Instrumental Analysis and Bioanalysis, Saarland University, P.O. Box 151150, 66041 Saarbrücken, Germany;(2) Roche Diagnostics GmbH, Nonnenwald 2, 82377 Penzberg, Germany;(3) Bia Separations, Ljubljana, Slovenia |
| |
Abstract: | Immunoadsorbers based on 2.0 × 6.0 mm i.d., epoxy-bearing, methacrylate-based monolithic disks were developed in order to target myoglobin and N-terminal pro-natriuretic peptide (NT-proBNP), two biomarkers involved in cardiovascular disease. In both cases, antibodies were successfully coupled to the polymeric disk material. The developed immunoadsorbers permitted the selective isolation of myoglobin and NT-proBNP from human serum. Myoglobin was successfully isolated and detected from serum samples at concentrations down to 250 fmol μL−1. However, the affinity of the antibodies was not sufficient for the analysis of low-concentration clinical samples. Frontal analysis of anti-NT-proBNP disks revealed the ability of the immunoadsorber to bind up to 250 pmol NT-proBNP, which is more than sufficient for the analysis of clinical samples. Anti-NT-proBNP disks showed good stability over more than 18 months and excellent batch-to-batch reproducibility. Moreover, anti-NT-proBNP disks permitted the isolation of NT-proBNP at concentrations down to 750 amol μL−1 in serum, corresponding to concentrations of strongly diseased patients. Using reversed-phase trapping columns, the detection of NT-proBNP eluted from immunoadsorbers by mass spectrometry was achieved for concentrations down to 7.8 fmol μL−1. |
| |
Keywords: | Immunoadsorber Monolithic disk Affinity chromatography Liquid chromatography Mass spectrometry Myoglobin N-terminal natriuretic peptide |
本文献已被 PubMed SpringerLink 等数据库收录! |
|