School of Electrical, Computer, and Energy Engineering, Arizona State University, Tempe, AZ, United States.
School of Electrical, Computer, and Energy Engineering, Arizona State University, Tempe, AZ, United States.
Biosens Bioelectron. 2014 Jul 15;57:179-85. doi: 10.1016/j.bios.2014.01.056. Epub 2014 Feb 7.
Heavy metal ions, i.e., Cu(2+), are harmful to the environment and our health. In order to detect them, and circumvent or alleviate the weaknesses of existing detecting technologies, we contrive a unique Surface Plasmon Resonance (SPR) biosensor combined with competitive adsorption of proteins, termed the Vroman effect. This approach adopts native proteins (albumin) as bio-receptors that interact with Cu(2+) to be denatured. Denaturation disrupts the conformation of albumin so that it weakens its affinity to adsorb on the sensing surface. Through the competitive adsorption between the denatured albumins and the native ones, the displacement occurs adjacent to the sensing surface, and this process is real-time monitored by SPR, a surface-sensitive label-free biosensor. The affinities of native albumin is significantly higher than that of denatured albumin, demonstrated by measured KD of native and denatured albumin to gold surafce, 5.8±0.2×10(-5) M and 5.4±0.1×10(-4) M, respectively. Using our biosensor, Cu(2+) with concentration down to 0.1mg/L is detected in PBS, tap water, deionized water, and bottled water. The SPR biosensor is characterized for 5 different heavy metal ions, Cu(2+), Fe(3+), Mn(2+), Pb(2+), and Hg(2+), most common heavy metal ions found in tap water. At the maximum contaminant level (MCL) suggested by the United States Environmental Protection Agency (EPA), the SPR biosensor produces 13.5±0.4, 1.5±0.4, 0, 0, and 0 mDeg, respectively, suggesting the biosensor may be used to detect Cu(2+) in tap water samples.
重金属离子,如 Cu(2+),对环境和健康有害。为了检测它们,并规避或缓解现有检测技术的弱点,我们设计了一种独特的表面等离子体共振(SPR)生物传感器,结合蛋白质的竞争吸附,称为 Vroman 效应。该方法采用天然蛋白质(白蛋白)作为生物受体,与 Cu(2+)相互作用而发生变性。变性破坏了白蛋白的构象,使其对吸附到传感表面的亲和力减弱。通过变性白蛋白和天然白蛋白之间的竞争吸附,在传感表面附近发生置换,这一过程通过 SPR(一种表面敏感的无标记生物传感器)实时监测。天然白蛋白的亲和力明显高于变性白蛋白,通过对天然白蛋白和变性白蛋白与金表面的 KD 值的测量可以证明,分别为 5.8±0.2×10(-5) M 和 5.4±0.1×10(-4) M。使用我们的生物传感器,可以在 PBS、自来水、去离子水和瓶装水中检测到浓度低至 0.1mg/L 的 Cu(2+)。SPR 生物传感器的特点是对 5 种不同的重金属离子,Cu(2+)、Fe(3+)、Mn(2+)、Pb(2+)和 Hg(2+),这些都是自来水中常见的重金属离子。在美国环境保护署(EPA)建议的最大污染物水平(MCL)下,SPR 生物传感器分别产生 13.5±0.4、1.5±0.4、0、0 和 0 mDeg,表明该生物传感器可用于检测自来水中的 Cu(2+)。