Department of Chemistry and Nanoscience, Ewha Womans University, Seoul, 120-750, Korea.
Analyst. 2011 Aug 21;136(16):3384-8. doi: 10.1039/c0an00978d. Epub 2011 Jul 11.
A homogeneous assay of the protective antigen in anthrax toxin is reported using two new PA-specific aptamers for selective and sensitive detection, based on reduction in the fluorescence emission according to the formation of the aptamer-PA ternary complex. PA at 1 nM was readily detected using OliGreen as a fluorophore in HEPES buffer. We also demonstrated that the PA detection could be performed in blood serum. The binding interaction between the aptamer and PA was strong enough to dehybridize double-stranded DNA paired completely with 12 bases at room temperature. Moreover, this fluorescence study revealed that the binding sites of the two aptamers were located differently on the PA protein. We believe our approach may lay the groundwork for the real-time detection of PA.
本文报道了一种炭疽毒素保护性抗原的均相分析方法，该方法使用了两种新的 PA 特异性适体，通过形成适体-PA 三元复合物来进行选择性和灵敏检测，从而减少荧光发射。在 HEPES 缓冲液中，使用 OliGreen 作为荧光染料，可轻松检测到 1 nM 的 PA。我们还证明，PA 的检测可以在血清中进行。适体与 PA 之间的结合相互作用足够强，可以在室温下使完全配对 12 个碱基的双链 DNA 解链。此外，这项荧光研究表明，两个适体的结合位点在 PA 蛋白上的位置不同。我们相信，我们的方法可能为 PA 的实时检测奠定基础。
