Tang C S, Dusseiller M, Makohliso S, Heuschkel M, Sharma S, Keller B, Vörös J
Swiss Federal Laboratories for Materials Testing and Research (EMPA), Dübendorf, Switzerland.
Anal Chem. 2006 Feb 1;78(3):711-7. doi: 10.1021/ac051244a.
Microarray technology is a powerful tool that provides a high throughput of bioanalytical information within a single experiment. These miniaturized and parallelized binding assays are highly sensitive and have found widespread popularity especially during the genomic era. However, as drug diagnostics studies are often targeted at membrane proteins, the current arraying technologies are ill-equipped to handle the fragile nature of the protein molecules. In addition, to understand the complex structure and functions of proteins, different strategies to immobilize the probe molecules selectively onto a platform for protein microarray are required. We propose a novel approach to create a (membrane) protein microarray by using an indium tin oxide (ITO) microelectrode array with an electronic multiplexing capability. A polycationic, protein- and vesicle-resistant copolymer, poly(l-lysine)-grafted-poly(ethylene glycol) (PLL-g-PEG), is exposed to and adsorbed uniformly onto the microelectrode array, as a passivating adlayer. An electronic stimulation is then applied onto the individual ITO microelectrodes resulting in the localized release of the polymer thus revealing a bare ITO surface. Different polymer and biological moieties are specifically immobilized onto the activated ITO microelectrodes while the other regions remain protein-resistant as they are unaffected by the induced electrical potential. The desorption process of the PLL-g-PEG is observed to be highly selective, rapid, and reversible without compromising on the integrity and performance of the conductive ITO microelectrodes. As such, we have successfully created a stable and heterogeneous microarray of biomolecules by using selective electronic addressing on ITO microelectrodes. Both pharmaceutical diagnostics and biomedical technology are expected to benefit directly from this unique method.
微阵列技术是一种强大的工具,可在单个实验中提供高通量的生物分析信息。这些小型化且并行化的结合测定具有高度敏感性,尤其在基因组时代已广受欢迎。然而,由于药物诊断研究通常针对膜蛋白,当前的阵列技术难以应对蛋白质分子的脆弱特性。此外,为了解蛋白质的复杂结构和功能,需要不同策略将探针分子选择性地固定到蛋白质微阵列平台上。我们提出一种新颖方法,通过使用具有电子复用能力的氧化铟锡(ITO)微电极阵列来创建(膜)蛋白质微阵列。一种聚阳离子、抗蛋白质和抗囊泡的共聚物,聚(L - 赖氨酸)接枝聚(乙二醇)(PLL - g - PEG),作为钝化吸附层暴露并均匀吸附到微电极阵列上。然后对各个ITO微电极施加电子刺激,导致聚合物局部释放,从而暴露出裸露的ITO表面。不同的聚合物和生物部分被特异性固定到活化的ITO微电极上,而其他区域由于不受感应电势影响而保持抗蛋白质特性。观察到PLL - g - PEG的解吸过程具有高度选择性、快速且可逆,同时不影响导电ITO微电极的完整性和性能。因此,我们通过在ITO微电极上使用选择性电子寻址成功创建了稳定且异质的生物分子微阵列。药物诊断和生物医学技术预计都将直接受益于这种独特方法。