IRIDIUM OXIDE PROTEIN MICROARRAYS

说明(由申请人提供)：本建议书旨在满足 用于大规模并行的稳定、经济的蛋白质微阵列的研制 蛋白质相互作用的筛选。这样的阵列将在 蛋白质表达水平的测定，疾病诊断，药物发现， 并筛选完成过程中产生的数百万个蛋白质相互作用 人类基因组的。这一提议的关键创新是使用了一种 电化学生长的三维水合基质固定化蛋白质和 它在微图形化工艺中的应用 微阵列。三维固定化矩阵的使用有望 减少在传统排列方法中可能发生的变性 蛋白质被固定在二维底物上。此外，它是 预期由多孔基质提供的更大表面积将 允许更高的蛋白质负载量，因此与 平面衬底。在第一阶段，含有固定化抗体的微阵列 将被制造出来，它们的稳定性被评估，并且它们在 基于荧光的平行免疫分析和蛋白质的测定 我们将演示其表达水平。在第二阶段，更高密度的蛋白质 适合集成到自动化、高通量微流控芯片中的阵列 将开发输送系统和荧光分析仪。

DESCRIPTION (provided by applicant): This proposal addresses the need for the development of stable, economical protein microarrays for massive parallel screening of protein interactions. Such arrays will be invaluable in determination of protein expression levels, disease diagnosis, drug discovery, and screening the millions of protein interactions generated by the completion of the human genome. The key innovation of this proposal is the use of an electrochemically grown 3-dimensional hydrous matrix to immobilize proteins and its application in a micro-patterning procedure to fabricate economical microarrays. The use of a 3-dimensional immobilization matrix is expected to reduce the denaturing that can occur in traditional arraying methods where proteins are immobilized on 2-dimensional substrates. In addition, it is anticipated that the larger surface area provided by the porous matrix will allow higher protein loadings, and therefore increased sensitivity, compared to planar substrates. In Phase I, microarrays containing immobilized antibodies will be fabricated, their stability evaluated, and their application in fluorescence based parallel immunoassays and determination of protein expression levels will be demonstrated. In Phase II, higher density protein arrays suitable for integration into automated, high-throughput microfluidic delivery systems and fluorescent analyzers will be developed.