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Chem. Eng. J., Volume 477, 1 December 2023, 147002 | https://doi.org/10.1016/j.cej.2023.147002 A highly sensitive sandwich-type electrochemical aptasensor using a pair of novel truncated aptamers for the detection of vaspin

Eun Ryung Kim ^a, Thi Thanh-Thao Dang ^a, Su Jin Lee ^b, Thi Thanh-Qui Nguyen ^a, Jee-Woong Park ^c, Man Bock Gu ^a

^aDepartment of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Republic of Korea

^bDrug Manufacturing Center, Daegu-Gyeongbuk Medical Innovation Foundation (K-MEDI Hub), Daegu 41061, Republic of Korea

^cMedical Device Development Center, Daegu-Gyeongbuk Medical Innovation Foundation (K-MEDI Hub), Daegu 41061, Republic of Korea

Corresponding author : Man Bock Gu

Abstract

Visceral adipose tissue-derived serpin (vaspin) is a biomarker associated with obesity and type 2 diabetes, both of which are prevalent and increasing in numbers worldwide. However, the sensitivity of existing aptasensors for vaspin detection does not yet match the reported serum levels. Aptamer sequences could contain nonessential nucleotides, which can be truncated out to potentially enhance their functionality and applicability. In this study, we developed a novel truncation method, a nonessentials-guided in situ truncation strategy to generate high-affinity aptamers by directly removing the nonessential portions of the aptamer sequences using exonucleases from the aptamer-protein complex kept stably. We applied this novel strategy to the original vaspin-binding aptamers, V1 and V49, resulting in two truncated aptamers, V1(3′-5′)–26 and V49(3′-5′)–30. These truncated aptamers showed enhanced binding affinity and specificity compared to their original aptamers. Confirmed to have different binding sites, this pair of truncated aptamers was successfully incorporated into a sandwich-type electrochemical aptasensor. This aptasensor achieved 16-fold higher sensitivity than the one using untruncated aptamers and enabled specific detection of vaspin in a buffer and even sensitive detection in serum within the physiological range. The reduced size of the aptamers could potentially cover the surface of the sensing platform, blocking nonspecific binding, thereby lowering the blank value, and improving the signal-to-noise ratio and limit of detection. This approach highlights the potential of the nonessentials-guided in situ truncation strategy for developing improved truncated aptamers, examining aptamer binding sites, and enhancing the analytical performance of biosensing platforms.

논문정보

형식Research article
게재일2023년 12월 (BRIC 등록일 2023-11-07)
연구진국내 연구진
분야 바이오·의료융합 > 바이오센싱 및 나노바이오물질

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