Tools and Methods for Producing High Quality Functional Protein Microarrays for Biomarker Discovery

翻译后摘要：功能蛋白质微阵列发现高通量疾病生物标志物的应用 发现，了解翻译后修饰（PTM）的发生率和作用 疾病病理学，研究酶、蛋白质-蛋白质和蛋白质- 小分子相互作用，等等。核酸可编程蛋白质阵列（NAPPA） 技术是用于生产无细胞原位表达的功能性蛋白质的稳健方法 人裂解物中的微阵列，在测定时新鲜表达和展示， 在过去的十年中，它被广泛应用于疾病生物标志物发现研究。我们的目标是 开发一种先进的仪器，用于自动化生产NAPPA IPC（分离蛋白 捕获）载玻片上的高密度蛋白质微阵列。一旦仪器研制成功，我们 我希望NAPPA蛋白质微阵列在简单性、速度和质量方面也能有类似的改进 当DNA微阵列从手动转移到自动时， 工作站。除了临床生物标志物研究外，它还是一个理想的平台，用于分析 动物模型中的疾病生物标志物，然后可以监测其中的一个子集作为预后指标。 生物标志物，药物开发临床前试验终点的指标。

Abstract: Functional protein microarrays find applications in high throughput disease biomarker discovery, understanding the incidence and role of post translational modifications (PTMs) in disease pathology, investigating activity and function of enzymes, protein–protein and protein– small molecule interactions, among others. Nucleic Acid Programmable Protein Array (NAPPA) technology is a robust method for producing cell-free in situ expressed functional protein microarrays in human lysate, freshly expressed and displayed at the time of assay, and has been widely applied in disease biomarker discovery research over the last decade. We aim to develop an advanced instrument for automating the production of NAPPA IPC (Isolated Protein Capture) high density protein microarrays on glass slides. Once the instrument is developed, we expect similar improvements in simplicity, speed and quality of NAPPA protein microarray assays as was seen when DNA microarrays were transferred from manual to automated workstations. In addition to clinical biomarker research, it is an ideal platform for profiling for disease biomarkers in animal models, a subset of which can then be monitored as prognostic biomarkers, indicators of end points in drug development pre-clinical trials.