Lee Minsu, Kang Dong-Ku, Yang Hyun-Kyu, Park Keun-Hyung, Choe Soo Young, Kang Changsoo, Chang Soo-Ik, Han Moon Hi, Kang In-Cheol
Protein Chip Research Center, Biotechnology Research Institute, Chungbuk National University, Cheongju, Korea.
Proteomics. 2006 Feb;6(4):1094-103. doi: 10.1002/pmic.200500392.
Protein nanoarrays are addressable ensembles of nano-scale protein domain on solid surfaces. This method can serve as a useful platform for ultraminiaturized bioanalysis. In this study, we investigated single molecular nanopatterning and molecular interaction of proteins that were immobilized on Prolinker surface of gold-coated silicon wafer by using dip-pen nanolithography (DPN) method. Contact force and humidity were optimized at 0.01 nN and 80%, respectively. The domain features of protein nanoarrays were developed at the contact time of 5 s. The optimized conditions for the nanoarray process were applied to create protein nanoarray using integrin alpha(v)beta3 and angiogenin. Constructed protein nanoarrays using integrin alpha(v)beta3 have single molecular monolayer with regular domain shape (height 15 +/- 5 nm). The changed height value due to the single molecular interaction between integrin alpha(v)beta3 and vitronectin was approximately 30 +/- 5 nm on Prolinker surface as measured with atomic force microscopy tip. Taken together, these results suggest that protein nanoarray on Prolinker surface fabricated by well-controlled DPN process can be used to analyze single molecular interaction of protein.
蛋白质纳米阵列是固体表面上纳米级蛋白质结构域的可寻址集合体。该方法可作为超小型生物分析的有用平台。在本研究中，我们使用蘸笔纳米光刻（DPN）方法研究了固定在金涂层硅片的Prolinker表面上的蛋白质的单分子纳米图案化和分子相互作用。接触力和湿度分别优化为0.01 nN和80%。蛋白质纳米阵列的结构特征在接触时间为5 s时形成。将纳米阵列过程的优化条件应用于使用整合素α(v)β3和血管生成素创建蛋白质纳米阵列。使用整合素α(v)β3构建的蛋白质纳米阵列具有规则结构形状（高度为15±5 nm）的单分子单层。用原子力显微镜尖端测量，在Prolinker表面上，由于整合素α(v)β3与玻连蛋白之间的单分子相互作用而导致的高度变化值约为30±5 nm。综上所述，这些结果表明，通过良好控制的DPN过程在Prolinker表面制备的蛋白质纳米阵列可用于分析蛋白质的单分子相互作用。
