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通过 G-四链体适体检测人血管紧张素原蛋白

Fluorescence detection of the human angiotensinogen protein by the G-quadruplex aptamer.

机构信息

College of Science, Harbin Institute of Technology (Shenzhen), Shenzhen, Guangdong, 518055, China.

Medical and Molecular Microbiology Unit, Department of Medicine, Faculty of Science and Medicine, University of Fribourg, Fribourg, 1700, Switzerland.

出版信息

Analyst. 2022 Sep 12;147(18):4040-4048. doi: 10.1039/d2an01057g.

Abstract

Noncanonical G-quadruplex nucleic acid structures can be used as probes in biosensors for the detection of metal ions, proteins and nucleic acids. Angiotensinogen (AGT) is a glycosylated globulin found in serum, which can regulate blood pressure and body fluid homeostasis. AGT is an important part of the renin-angiotensin system (RAS) and can be potentially used as a biomarker of diseases with RAS alterations. G-quadruplex based biosensors can detect targets with high accuracy and speed and low cost. Employing the magnetic bead enrichment method we constructed a reliable and efficient fluorescent biosensor platform for G-quadruplex based detection of the human AGT protein. The primary antibody in our biosensor is recognized by fluorescently labeled secondary antibodies, which leads to the detection of AGT captured by the G-quadruplex aptamer coupled magnetic beads. This G-quadruplex based fluorescent biosensor designed with a detection limit of 5 × 10 mg mL was used for the successful detection of AGT at the cellular level. Our G-quadruplex based fluorescent biosensor will contribute to the more reliable and efficient detection of AGT.

摘要

非经典 G-四链体核酸结构可用作生物传感器中的探针,用于检测金属离子、蛋白质和核酸。血管紧张素原 (AGT) 是血清中发现的一种糖基化球蛋白,可调节血压和体液平衡。AGT 是肾素-血管紧张素系统 (RAS) 的重要组成部分,可作为 RAS 改变相关疾病的潜在生物标志物。基于 G-四链体的生物传感器可以高精度、高速和低成本地检测目标物。我们采用磁珠富集法构建了一种可靠且高效的基于 G-四链体的荧光生物传感器平台,用于检测人 AGT 蛋白。我们的生物传感器中的一抗被荧光标记的二抗识别,从而检测与 G-四链体适体偶联磁珠结合的 AGT。该基于 G-四链体的荧光生物传感器的检测限为 5×10mg·mL,可成功检测细胞水平的 AGT。我们基于 G-四链体的荧光生物传感器将有助于更可靠、高效地检测 AGT。

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