Enzyme Immunoassays for Fragments (Epitopes) of Human Proatrial Natriuretic Peptides
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Engelbert Hartter
Abstract
Several peptides derived from the N-terminal sequence of pro-atrial natriuretic peptide (proANP) have been tested successfully as markers of heart disease. We have developed specific and sensitive competitive enzyme immunoassays for fragments [1–30] and [31–67] of proANP. Antisera were raised in sheep against synthetic peptides predicted to be highly immunogenic. Binding specificity was determined by epitope mapping. Microtiter plates were coated with antibody specific for the Fc region of sheep IgG to capture the affinity-purified specific anti-proANP antibodies in an oriented and reproducible form. Synthetic proANP calibrators or diluted samples were incubated simultaneously with biotinylated peptide and binding was quantitated using streptavidin-peroxidase and TMB. Immunoreactive proANP could be measured in diluted plasma, serum and urine. The detection limits of the proANP[1–30] and proANP[31–67] assays were 2.5 and 10 pmol/l respectively. The linearity of samples diluted beyond the recommended assay conditions was good. Recoveries of added standard peptides ranged from 102 to 112%. Circulating concentrations of immunoreactive proANP in 115 healthy subjects ranged from 0.11 to 0.47 nmol/l proANP[1–30] and 0.18 to 0.79 nmol/l proANP[31–67]. In patients with cardiac disease, proANP levels were increased significantly. The reference interval of proANP[31–67] in urine was 0.09 to 1.7 nmol/l, several-fold higher than proANP[1–30] (<0.03 to 1.1 nmol/l). After storage for 6 months at −20°C there was no detectable decrease in immunoreactivity.
Copyright © 2000 by Walter de Gruyter GmbH & Co. KG
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- The Millenium 2000; Laboratory Medicine – from Where? to Where?
- Regulation of Lipid and Lipoprotein Metabolism by PPAR Activators
- Laboratory Screening for Erythropoietin Abuse in Sport: an Emerging Challenge
- Antioxidant Enzyme Activities and Total Nitrite/ Nitrate Levels in the Collar Model. Effect of Nicardipine
- Enzyme Immunoassays for Fragments (Epitopes) of Human Proatrial Natriuretic Peptides
- Transient Nature of Interference Effects from Heterophile Antibodies: Examples of Interference with Cardiac Marker Measurements
- Comparison of Procalcitonin, sCD14 and Interleukin-6 Values in Septic Patients
- Changes in Bone Turnover and Deoxypyridinoline Levels in Epileptic Patients
- Analytical Performance of the Fully Automated AxSYM(r) Estradiol Assay
- Impacts of Laboratory Methodology on Medical Thinking in the 19th Century
- Reference Intervals for Erythropoietin Measurement by the JCL EIA Assay Need to Be Determined Locally
- The Physical Basis of Biochemistry. By Peter R. Bergethon
- European Science. Science funding: the European dimension. A discussion meeting held at the Royal Society, Tuesday 17 March, 1998
- IFCC News
