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Spontaneously beating neonatal rat heart myocyte culture - A model to characterize angiotensin II at1 receptor autoantibodies in patients with preeclampsia

Item Type:Article
Title:Spontaneously beating neonatal rat heart myocyte culture - A model to characterize angiotensin II at1 receptor autoantibodies in patients with preeclampsia
Creators Name:Wallukat, G. and Nissen, E. and Neichel, D. and Harris, J.
Abstract:This report focuses on angiotensin II AT1 receptor autoantibodies (anti-AT1-AABs) in preeclamptic women. An enzyme-linked immunosorbent assay was described. Biotinylated peptide was incubated with anti-AT1-AABs. Streptavidin-coated magnetic particles bind the protein-autoantibody complex. Detection of anti-AT1-AABs was performed using anti-human IgG3 peroxidase-coupled antibody. The color reaction of tetramethylbenzidine solution was stopped by adding 0.5 M H2SO4. Optical density was measured at 450 nm (620 nm reference filter). Seventy-nine percent of anti-AT1-AAB-positive patients (measured by bioassay) showed an increase in optical density (>145%). The same biotinylated peptide was successfully used for purification of 6/6 anti-AT1-AABs. Chronotropic effects of purified antibodies were registered on primary cultured neonatal rat cardiomyocytes with the computer imaging system IMAGOQUANT. Western blot of coimmunoprecipitation of angiotensin II AT1 receptor shows one band (molecular weight >40.0 kDa) in potassium thiocyanate eluate.
Keywords:Angiotensin II AT1 Receptor, Autoantibodies, ELISA, Neonatal Rat Cardiomyocytes, Animals, Rats
Source:In Vitro Cellular & Developmental Biology Animal
ISSN:1071-2690
Publisher:Soc In Vitro Biology (U.S.A.)
Volume:38
Number:7
Page Range:376-377
Date:July 2002
Official Publication:https://doi.org/10.1290/1071-2690(2002)038<0376:SBNRHM>2.0.CO;2
PubMed:View item in PubMed

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