Diffractometric detection of proteins using microbead-based rolling circle amplification.
Diffractometric detection of proteins using microbead-based rolling circle amplification.
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DOI：
10.1021/ac901716d
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发表时间：
2010-01-01
影响因子：
7.4
通讯作者：
Savran, Cagri A.
中科院分区：
文献类型：
--
作者：
Lee, Joonhyung;Icoz, Kutay;Roberts, Ana;Ellington, Andrew D.;Savran, Cagri A.
我们提出了一种健壮的、灵敏的、荧光的或无标记的自组装光学衍射生物传感器，它利用滚圆放大(RCA)和磁性微珠作为信号增强方法。以适体为基础的夹心法对15-μm宽交替排列的微接触印迹链霉亲和素进行检测，可以特异性地捕获和检测血小板衍生生长因子B链。适配子用作连接挂锁探针的模板，环状探针又可用作RCA的模板。串联的RCA产物与生物素化的寡核肽杂交，然后捕获链霉亲和素标记的磁珠。因此，来自捕获的PDGF-BB的信号通过串联的RCA产物被放大，并且印刷区域上的衍射光栅产生不同强度的衍射模。检测到的衍射强度和表面微珠的密度随PDGF-BB浓度的变化而变化。我们的结果证明了一个健壮的生物传感平台，它易于建造和使用，并且没有荧光显微镜。这种自组装的微珠图案既可以在普通的明场显微镜下对分子结合事件进行可视化分析，也可以作为衍射光栅生物传感器使用。
We present a robust, sensitive, fluorescent or radio label-free self-assembled optical diffraction biosensor that utilizes rolling circle amplification (RCA) and magnetic microbeads as a signal enhancement method. An aptamer-based sandwich assay was performed on microcontact-printed streptavidin arranged in 15-μm-wide alternating lines, and could specifically capture and detect platelet-derived growth factor B-chain (PDGF-BB). An aptamer served as a template for the ligation of a padlock probe and the circularized probe could in turn be used as a template for RCA. The concatameric RCA product hybridized to biotinylated oligonuclotides which then captured streptavidin-labeled magnetic beads. In consequence, the signal from the captured PDGF-BB was amplified via the concatameric RCA product, and the diffraction gratings on the printed areas produced varying intensities of diffraction modes. The detected diffraction intensity and the density of the microbeads on the surface varied as a function of PDGF-BB concentration. Our results demonstrate a robust biosensing platform that is easy to construct and use, and devoid of fluorescence microscopy. The self-assembled bead patterns allow both a visual analysis of the molecular binding events under an ordinary bright-field microscope and serve as a diffraction grating biosensor.
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影响因子：
7.4
作者：
Savran, CA;Knudsen, SM;Manalis, SR
通讯作者：
Manalis, SR
影响因子：
4
作者：
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通讯作者：
Chen, Teng-Ming
影响因子：
12.6
作者：
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通讯作者：
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影响因子：
7.4
作者：
Henne, Walter A.;Doorneweerd, Derek D.;Savran, Cagri
通讯作者：
Savran, Cagri
影响因子：
2.7
作者：
Schopf, Eric;Fischer, Nicholas O.;Tok, Jeffrey B. -H.
通讯作者：
Tok, Jeffrey B. -H.