[논문]Aptamer-linked in vitro expression assay for ultrasensitive detection of biomarkers

Baek, Min-Seok; Lee, Kyung-Ho; Byun, Ju-Young; Shin, Yong-Beom; Kim, Dong-Myung

doi:10.1016/j.aca.2020.12.036

[해외논문] Aptamer-linked in vitro expression assay for ultrasensitive detection of biomarkers

Analytica chimica acta : an international journal devoted to all branches of analytical chemistry, v.1146, 2021년, pp.118 - 123

Baek, Min-Seok (Department of Chemical Engineering and Applied Chemistry, Chungnam National University) , Lee, Kyung-Ho (Department of Chemical Engineering and Applied Chemistry, Chungnam National University) , Byun, Ju-Young (BioNano Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) , Shin, Yong-Beom (BioNano Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) , Kim, Dong-Myung (Department of Chemical Engineering and Applied Chemistry, Chungnam National University)

Abstract ▼ AI-Helper

Abstract Signal amplification is a key step that determines the sensitivity of molecular assays. Although studies on aptamers have mostly focused on their target-binding ability, taking advantage of the gene-coding function of nucleic acids, we demonstrate here that aptamers can be engineered into diagnostic reagents that can both recognize a target and generate highly amplified detection signals. We developed a strategy that employs a ‘readable’ aptamer that consists of a single-stranded aptamer and a double-stranded reporter gene. After binding to its target via the aptamer region, the reporter gene of the readable aptamer produces amplified number of signal-generating enzymes through a subsequent in vitro expression reaction. In contrast to conventional enzyme-conjugation methods, this method allows the generation of far more amplified detection signals, thereby markedly increasing the sensitivity of detection enough to analyze a target present in aM concentrations. Highlights Efficiency of signal amplification determines the sensitivity of a bioassay method. Compared to antibodies, aptamers carry substantial advantages, including lower production cost and higher stability. Readable aptamers can both recognize and produce amplified number of signal-generating enzymes. The ALIVE assay that employs the readable aptamers could detect thrombin present in attomolar concentrations. Graphical abstract [DISPLAY OMISSION]

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