Cleavable DNA-protein hybrid molecular beacon: A novel efficient signal translator for sensitive fluorescence anisotropy bioassay
Cleavable DNA-protein hybrid molecular beacon: A novel efficient signal translator for sensitive fluorescence anisotropy bioassay
复制标题
可切割的 DNA-蛋白质杂化分子信标：一种用于灵敏荧光各向异性生物测定的新型高效信号转换器
DOI：
10.1016/j.talanta.2015.10.003
复制
发表时间：
2016-01-15
期刊：
影响因子：
6.1
通讯作者：
Yang, Bin
中科院分区：
文献类型：
--
作者：
Hu, Pan;Yang, Bin
荧光各向异性(FA)分析因其灵敏度高、格式均一、不受荧光强度影响等特点，已成为寡核苷酸生物检测领域的研究热点。然而，到目前为止，大多数FA探头都需要精心定制结构设计，因此既不适用于不同的传感系统，也不能有效地获得足够的信号响应。为了解决这一问题，通过将分子信标独特的稳定结构与链霉亲和素的大分子质量相结合，成功地设计了一种可切割的DNA-蛋白质杂化分子信标用于信号放大FA生物检测。与单链DNA探针或常规分子信标相比，DNA-蛋白质杂化分子信标具有更高的FA值，有可能在传感过程中获得较高的信号背景比。作为原理验证，该新型DNA-蛋白质杂化分子信标以DNAzyme-Pb2+为模型传感系统，进一步应用于FA生物检测。该方法可选择性地检测缓冲液中低至0.5 nM的Pb2+，并可用于实际样品的分析，具有较好的回收率。分子信标与大多数寡核苷酸探针的设计和基于酶的信号放大策略相兼容，可以作为一种新型的信号转换器，扩大FA技术在各种生物检测中的应用前景。(C)2015爱思唯尔B.V.保留所有权利。
Due to its unique features such as high sensitivity, homogeneous format, and independence on fluorescent intensity, fluorescence anisotropy (FA) assay has become a hotspot of study in oligonucleotide-based bioassays. However, until now most FA probes require carefully customized structure designs, and thus are neither generalizable for different sensing systems nor effective to obtain sufficient signal response. To address this issue, a cleavable DNA-protein hybrid molecular beacon was successfully engineered for signal amplified FA bioassay, via combining the unique stable structure of molecular beacon and the large molecular mass of streptavidin. Compared with single DNA strand probe or conventional molecular beacon, the DNA-protein hybrid molecular beacon exhibited a much higher FA value, which was potential to obtain high signal-background ratio in sensing process. As proof-of-principle, this novel DNA-protein hybrid molecular beacon was further applied for FA bioassay using DNAzyme-Pb2+ as a model sensing system. This FA assay approach could selectively detect as low as 0.5 nM Pb2+ in buffer solution, and also be successful for real samples analysis with good recovery values. Compatible with most of oligonucleotide probes' designs and enzyme-based signal amplification strategies, the molecular beacon can serve as a novel signal translator to expand the application prospect of FA technology in various bioassays. (C) 2015 Elsevier B.V. All rights reserved.