A new lateral flow strip assay (LFSA) using a pair of aptamers for the detection of Vaspin
A new lateral flow strip assay (LFSA) using a pair of aptamers for the detection of Vaspin
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DOI：
10.1016/j.bios.2016.11.061
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发表时间：
2017-07-15
影响因子：
12.6
通讯作者：
Gu, Man Bock
中科院分区：
文献类型：
--
作者：
Raston, Nurul Hanun Ahmad;Van-Thuan Nguyen;Gu, Man Bock
使用一对用金纳米粒子 (AuNP) 功能化的适体设计并成功开发了一种使用一对适体的新型侧流试纸条测定 (LFSA)。这种新型 LFSA 生物传感器系统利用同源适体二重体在两个不同的结合位点与靶蛋白 vaspin 结合，并对 vaspin 表现出灵敏且高度选择性的响应。 LFSA 中的夹心型格式是基于固定在链霉亲和素包被膜上的生物素标记的初级 V1 适体作为捕获探针和与 AuNP 缀合的次级 V49 适体作为信号探针而开发的。使用该 LFSA，在缓冲液和血清条件下，可在高达 5 nM 的 vaspin 可检测浓度范围内清晰地观察到 vaspin。本研究中开发的 LFSA 的灵敏度在缓冲液中为 0.137 至 25 nM，在加标人血清中为 0.105 至 25 nM。该 LFSA 在缓冲液和加标人血清条件下的检测限 (LOD) 分别为 0.137 nM 和 0.105 nM。因此，本研究成功开发了一种简单而有效的用于 vaspin 检测的 LFSA，并且这种开发不仅限于该靶标，还包括具有一对适体的其他靶标，无需任何特殊且费力的仪器。该系统作为筛选工具特别有用，可通过生成的一对适体快速现场检测任何目标。
A new lateral flow strip assay (LFSA) using a pair of aptamers has been designed and successfully developed using a pair of aptamers-functionalized with gold nanoparticles (AuNPs). This new LFSA biosensor system utilizes a cognate aptamer duo binding to vaspin, a target protein, at the two different binding sites, and exhibited a sensitive and highly selective response to vaspin. A sandwich-type format in LFSA was developed based on biotin-labeled primary V1 aptamer immobilized on streptavidin coated membrane as a capturing probe and secondary V49 aptamer conjugated with AuNPs as a signaling probe. Using this LFSA, vaspin could be visibly observed within the detectable concentrations of vaspin up to 5 nM in both buffer and serum conditions. The sensitivity of this LFSA developed in this study was ranged from 0.137 to 25 nM in buffer and from 0.105 to 25 nM in spiked human serum, respectively. The limit of detection (LOD) of this LFSA was found to be 0.137 nM and 0.105 nM in buffer and spiked human serum condition, respectively. Therefore, this study has shown successful development of a simple yet effective LFSA for vaspin detection, and this development is not only limited to this target, but also other targets with a pair of aptamers available, without any special and laborious instrumentations. This system will be particularly useful as a screening tool for rapid on-site detection of any targets with a pair of aptamers generated.