适体
肌氨酸
指数富集配体系统进化
检出限
DNA
化学
荧光
SELEX适体技术
寡核苷酸
线性范围
分子生物学
色谱法
生物化学
生物
氨基酸
核糖核酸
基因
物理
量子力学
甘氨酸
作者
Canan Özyurt,Zeynep Çelik Canbay,Erhan Dinçkaya,Serap Evran
标识
DOI:10.1016/j.ijbiomac.2019.02.030
摘要
Sarcosine is an amino acid derivative, which is considered as a key metabolite in various metabolic processes. Therefore, simple and sensitive detection methods are needed for further understanding its metabolic role and diagnostic value. In this study, we developed a novel method that meets the need for practical and sensitive detection in a complex medium mimicking urine conditions. For this aim, we selected sarcosine-specific DNA aptamers using graphene oxide-assisted systemic evolution of ligands by exponential enrichment (GO-SELEX). The candidate aptamers were labeled with 6-carboxyfluorescein (6-FAM) at their 5' ends. Two aptamers, namely 9S and 13S produced a significant fluorescence signal upon sarcosine binding. Both aptamers enabled a sensitive analysis with a detection limit of 0.5 pM. The linear detection ranged between 5 pM and 50 μM for 9S aptamer, while 13S aptamer enabled a wider linear detection range between 5 pM and 500 μM. The aptamer-based assay allowed rapid detection with no need for chemical derivatization of sarcosine and sophisticated instruments. Moreover, the aptamer-based assay was free of interference from urea and human serum albumin.
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