Development of an enzyme‐linked immunosorbent assay and a beta‐1 adrenergic receptor–based assay for monitoring the drug atenolol

Abstract

Abstract-Two approaches for monitoring atenolol (ATL) were applied: an immunochemical assay and a competitive-binding assay, based on the interaction between ATL and its target receptor, b1 adrenergic receptor (b1AR). Polyclonal antibodies (Abs) for ATL were generated, and a highly specific microplate immunochemical assay, that is, an enzyme-linked immunosorbent assay (ELISA), for its detection was developed. The ATL ELISA exhibited I50 and limit of detection (I20) values of 0.15 AE 0.048 and 0.032 AE 0.016 ng/ml, respectively, and the Abs did not cross-react with any of the tested beta-blocker drugs. Furthermore, a human b1AR (h-b1AR) was stably expressed in Spodoptera frugiperda cells (Sf9). The receptor was employed to develop a competitive-binding assay that monitored binding of ATL in the presence of isoproteranol by quantification of secondary messenger, cyclic adenosine monophosphate (cAMP), levels in the transfected cells. The assay showed that the recombinant h-b1AR was functional, could bind the agonistic ligand isoproterenol as well as the antagonist ATL, as indicated by a dose-dependent elevation of cAMP in the presence of isoproteranol, and decrease after ATL addition. The highly efficient and sensitive ELISA and the receptor assay represent two methods suitable for efficient and cost-effective large-scale, high-throughput monitoring of ATL in environmental, agricultural, and biological samples. Environ.

Экскреция тестостерона и эстрадиола культивируемой мидией Mytilus galloprovincialis Lam. (Чёрное море)

“…Гонады и отобранную дозатором суспензию половых продуктов от каждой особи гомогенизировали в фарфоровой ступке с добавлением минимального количества этанола (не более 10 % от объема гонад или суспензии половых продуктов). Низкая концентрация этанола в пробе (5-10 %) существенно не влияет на результат иммуноферментного анализа (Sapir et al, 2013). Пробы хранили в течение суток при 4-8 °С, так как эстрадиол стабилен именно при такой температуре (Меньшиков, 2003).…”

Molecularly-imprinted silica nanoparticles for rapid and selective detection of atenolol in artificial urine samples

“…There have been several analytical methods such as high performance liquid chromatography (HPLC) [10], liquid chromatography-mass spectrometry (LC-MS) [11], gas chromatography-mass spectrometry (GC-MS) [12], potentiometry [13], spectrofluorimetry [14], and enzymelinked immunoassay (ELISA) [15] reported for determination of atenolol in different biological samples. The aforementioned chromatographic methods are generally suffer from pure selectivity due to the presence of potential interferences which limits sensitivity of the methods.…”