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钙调节发光蛋白 Obelin 作为 SELEX 监测和 DNA 适体亲和力评估的工具。

The Ca -Regulated Photoprotein Obelin as a Tool for SELEX Monitoring and DNA Aptamer Affinity Evaluation.

机构信息

Institute of Biophysics SB RAS, Federal Research Center KSC SB RAS, Krasnoyarsk, Russia.

Siberian Federal University, Krasnoyarsk, Russia.

出版信息

Photochem Photobiol. 2020 Sep;96(5):1041-1046. doi: 10.1111/php.13274. Epub 2020 May 28.

Abstract

Bioluminescent solid-phase analysis was proposed to monitor the selection process and to determine binding characteristics of the aptamer-target complexes during design and development of the specific aptamers. The assay involves Ca -regulated photoprotein obelin as a simple, sensitive and fast reporter. Applicability and the prospects of the approach were exemplified by identification of DNA aptamers to cardiac troponin I, a highly specific early biomarker for acute myocardial infarction. Two structurally different aptamers specific to various epitopes of troponin I were obtained and then tested in a model bioluminescent assay.

摘要

生物发光固相分析被提议用于监测选择过程,并在特定适体的设计和开发过程中确定适体-靶复合物的结合特性。该测定法涉及钙调节发光蛋白 obelin 作为一种简单、灵敏和快速的报告蛋白。该方法的适用性和前景通过鉴定心肌肌钙蛋白 I 的 DNA 适体得到了例证,心肌肌钙蛋白 I 是急性心肌梗死的高度特异性早期生物标志物。获得了针对肌钙蛋白 I 的不同表位的两种结构不同的特异性适体,然后在模型生物发光测定中进行了测试。

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