A solid-phase immobilized epitope immunoassay (SPIE-IA) permitting very sensitive and specific measurement of angiotensin II in plasma

We have developed a new enzyme immunometric assay for angiotensin II (AII) based on SPIE-IA technology (solid-phase immobilized epitope-immunoassay). A monoclonal antibody with optimal properties (mAb3 131) was selected from a series of 19 anti-AII mAbs. The mAb had to be purified from ascitic fluid...

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Bibliographic Details
Published in:Journal of immunological methods 1999-08, Vol.228 (1), p.37-47
Main Authors: Volland, Hervé, Pradelles, Philippe, Ronco, Pierre, Azizi, Michel, Simon, Dominique, Créminon, Christophe, Grassi, Jacques
Format: Article
Language:English
Subjects:
Acetylcholinesterase
Amino Acid Sequence
Angiotensin II
Angiotensin II - analogs & derivatives
Angiotensin II - blood
Angiotensin II - immunology
Animals
Antibodies, Monoclonal
Biological and medical sciences
Biotechnology
Chromatography, High Pressure Liquid
Enzyme immunometric assay
Epitopes
Evaluation Studies as Topic
Fundamental and applied biological sciences. Psychology
Humans
Immunoenzyme Techniques - methods
Immunoenzyme Techniques - statistics & numerical data
Methods. Procedures. Technologies
Others
Plasma
Radioimmunoassay
Reproducibility of Results
Sensitivity and Specificity
Various methods and equipments
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Summary:We have developed a new enzyme immunometric assay for angiotensin II (AII) based on SPIE-IA technology (solid-phase immobilized epitope-immunoassay). A monoclonal antibody with optimal properties (mAb3 131) was selected from a series of 19 anti-AII mAbs. The mAb had to be purified from ascitic fluid in a specific manner in order to remove endogenous AII from the antibody-binding sites. We established a sensitive (minimum detectable concentration 0.5 pg/ml) and precise (CV below 15% in the 2–100 pg/ml range) SPIE-IA. Using different AII-related peptides, we observed that this new assay has a specificity profile that compares favourably with the corresponding competitive immunoassay. We have used the assay to measure AII in 42 plasma samples, and demonstrated a good correlation with values obtained using a commercial radioimmunoassay. Assay specificity was supported by HPLC fractionation experiments, confirming the absence of interference induced by endogenous AII-related products.
ISSN:0022-1759
1872-7905
DOI:10.1016/S0022-1759(99)00097-6