School of Chemical Engineering, Sungkyunkwan University, Suwon 440-746, South Korea.
Biosens Bioelectron. 2011 Aug 15;26(12):4823-7. doi: 10.1016/j.bios.2011.05.040. Epub 2011 Jun 1.
Herein, we demonstrate three strategies of signal amplification for ultrasensitive detection of human immunoglobulin E (hIgE) based on poly(di-acetylene) supramolecules. To fabricate the ultrasensitive PDA biosensor, ethylenediamine as an interlinker and aptamer as a receptor were introduced into the chip fabrication process. Using the prepared PDA liposome biosensor, the hIgE could be detected up to below 1.0 ng/ml by a primary response. In order to accomplish more ultrasensitive detection of protein on a PDA biosensor, polyclonal hIgE antibody was employed as an external mechanical force for the inducement of a secondary response. As a result, a PDA liposome biosensor sensitivity as high as 0.01 ng/ml for the target hIgE was obtained, with a sensitivity which is one hundred times of that of the method without signal amplification. These results indicate that the proposed strategies were capable of ultrasensitive quantitative and qualitative analyses of biomolecules without non-specific binding of non-target proteins.
本文展示了基于聚二乙炔超分子的三种信号放大策略,用于超灵敏检测人免疫球蛋白 E(hIgE)。为了制备超灵敏的 PDA 生物传感器,在芯片制造过程中引入了乙二胺作为连接体和适体作为受体。使用制备的 PDA 脂质体生物传感器,通过初步反应可以检测到低至 1.0ng/ml 以下的 hIgE。为了在 PDA 生物传感器上实现更灵敏的蛋白质检测,多克隆 hIgE 抗体被用作外部机械力,以诱导二次反应。结果,获得了针对靶 hIgE 的高达 0.01ng/ml 的 PDA 脂质体生物传感器灵敏度,其灵敏度比没有信号放大的方法高 100 倍。这些结果表明,所提出的策略能够在没有非目标蛋白质非特异性结合的情况下,对生物分子进行超灵敏的定量和定性分析。