Ressine Anton, Marko-Varga György, Laurell Thomas
Department of Electrical Measurements, Lund Institute of Technology, Lund University, P.O. Box 118, SE-221 00 Lund, Sweden.
Biotechnol Annu Rev. 2007;13:149-200. doi: 10.1016/S1387-2656(07)13007-6.
One attractive method for monitoring biomolecular interactions in a highly parallel fashion is the use of microarrays. Protein microarray technology is an emerging and promising tool for protein analysis, which ultimately may have a large impact in clinical diagnostics, drug discovery studies and basic protein research. This chapter is based upon several original papers presenting our effort in the development of new protein microarray chip technology. The work describes a novel 3D surface/platform for protein characterization based on porous silicon. The simple adjustment of pore morphology and geometry offers a convenient way to control wetting behavior of the microarray substrates. In this chapter, an interesting insight into the surface role in bioassays performance is made. The up-scaled fabrication of the novel porous chips is demonstrated and stability of the developed supports as well as the fluorescent bioassay reproducibility and data quality issues are addressed. We also describe the efforts made by our group to link protein microarrays to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), suggesting porous silicon as a convenient platform for fast on-surface protein digestion protocols linked to MS-readout. The fabrication of ultra- and superhydrophobic states on porous silicon is also described and the utilization of these water-repellent properties for a new microscaled approach to superhydrophobic MALDI-TOF MS target anchor chip is covered.
一种以高度并行方式监测生物分子相互作用的吸引人的方法是使用微阵列。蛋白质微阵列技术是一种新兴且有前景的蛋白质分析工具，最终可能会对临床诊断、药物发现研究和基础蛋白质研究产生重大影响。本章基于几篇原创论文，介绍了我们在开发新型蛋白质微阵列芯片技术方面所做的努力。这项工作描述了一种基于多孔硅的用于蛋白质表征的新型三维表面/平台。孔隙形态和几何形状的简单调整提供了一种控制微阵列基板润湿性的便捷方法。在本章中，对生物测定性能中表面作用有了有趣的见解。展示了新型多孔芯片的扩大规模制造，并讨论了所开发支撑体的稳定性以及荧光生物测定的重现性和数据质量问题。我们还描述了我们团队为将蛋白质微阵列与基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）相连接所做的努力，表明多孔硅是用于与质谱读出相连接的快速表面蛋白质消化方案的便捷平台。还描述了在多孔硅上制备超疏水和超亲水状态，以及利用这些疏水性能开发一种用于超疏水MALDI-TOF MS靶标锚定芯片的新型微尺度方法。
