4.5 Article

Creation of a quick and sensitive fluorescent immunosensor for detecting the mineralocorticoid steroid hormone aldosterone

出版社

PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.jsbmb.2022.106118

关键词

Aldosterone; Essential hypertension; Monoclonal antibody; Phage display; Fluorescence quenching

资金

  1. National Natural Science Foundation of China [21775064]
  2. Science and Tech-nology Planning Project of Weifang City, Shandong Province of China [2020YQFK014]
  3. JSPS KAKENHI [JP18H03851]
  4. Japan Society for the Promotion of Science, Japan
  5. World Research Hub (WRH) Program of International Research Fron-tiers Initiative, Tokyo Institute of Technology

向作者/读者索取更多资源

Aldosterone is a hormone that regulates sodium ion and water reabsorption in the kidney. In this study, the gene encoding the anti-aldosterone antibody A2E11 was cloned and analyzed, and a sensor was developed to detect aldosterone. The sensor showed a detection limit of 24.1 pg/mL with a wide detection range, and it could accurately detect aldosterone within 2 minutes. This sensor has potential applications in the diagnosis of diseases such as hypertension.
Aldosterone (ALD) is a steroid hormone secreted by the zona glomerulosa of the adrenal cortex that mainly acts on the kidney to regulate sodium ion and water reabsorption. Detection of ALD plays an important role in the diagnosis of primary aldosteronism in patients with hypertension. For the first time, the gene encoding the antiALD antibody, A2E11, was successfully cloned and analyzed using phage display technology. The antibody had an affinity of 2.5 nM against ALD, and after binding to ALD, it reached saturation within 5 s. Using this antibody, a Quenchbody (Q-body) was constructed by labeling the N-termini of heavy and light chains of the antigenbinding fragment of A2E11 with the fluorescent dye ATTO520 to detect ALD based on the principle of photoinduced electron transfer. The sensor detected ALD in 2 min, and the limit of detection was 24.1 pg/mL with a wide detection range from 24.1 pg/mL to 10 mu g/mL and a half-maximal effective concentration of 42.3 ng/mL. At the highest concentration of ALD in the assay, the fluorescence intensity increased by 5.0-fold compared to the original fluorescence intensity of the Q-body solution. The Q-body could be applied to analyze 50% of human serum without a significant influence of the matrix. The recoveries of ALD in spiked serum samples with the Qbody assay were confirmed to range from 90.3% to 98.2%, suggesting their potential applications in the diagnosis of diseases, such as essential hypertension.

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