Miniaturized monolithic disks for immunoadsorption of cardiac biomarkers from serum.
Immunoadsorbers based on 2.0 x 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 microL(-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 microL(-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 microL(-1).
Delmotte
N
N
Department of Chemistry, Instrumental Analysis and Bioanalysis, Saarland University, 66041, Saarbrücken, Germany.
Kobold
U
U
Meier
T
T
Gallusser
A
A
Strancar
A
A
Huber
C G
CG
D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D015415:Biomarkers / Q000097:blood; D002138:Calibration; D002318:Cardiovascular Diseases / Q000097:blood* / Q000175:diagnosis; D002846:Chromatography, Affinity / Q000379:methods; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007118:Immunoassay / Q000379:methods; D007163:Immunosorbent Techniques; D007164:Immunosorbents / Q000737:chemistry*; D013058:Mass Spectrometry; D008689:Methacrylates / Q000737:chemistry; D008904:Miniaturization; D009211:Myoglobin / Q000097:blood* / Q000276:immunology; D020097:Natriuretic Peptide, Brain / Q000097:blood* / Q000276:immunology; D010446:Peptide Fragments / Q000097:blood* / Q000276:immunology; D015203:Reproducibility of Results
D000906:Antibodies; D000911:Antibodies, Monoclonal; D015415:Biomarkers; D007164:Immunosorbents; D008689:Methacrylates; D009211:Myoglobin; D010446:Peptide Fragments; C109794:pro-brain natriuretic peptide (1-76); D020097:Natriuretic Peptide, Brain