Novel Sensor Integrated Proteome on Chip (SPOC) platform for evaluating kinetic parameters of protein interactions in high throughput

摘要 蛋白质组学领域的一个关键瓶颈是无法同时以经济高效的方式 同时评估数千种功能蛋白质的相互作用，以获得动力学数据。虽然 传统的原位蛋白质芯片技术很发达，它使用的是荧光染料 仅提供定性或半定量信息的基于检测。目前有 目前还没有一种技术可以提供全长折叠蛋白质的无标记实时相互作用分析 高吞吐量格式。为了解决这一未得到满足的需求，INanoBio正在开发新型传感器 结合原位表达和捕获的集成蛋白质组芯片(SPOC)平台 生物传感器上的功能蛋白质，以实现对数千种 实时的蛋白质。在这个SBIR二期项目中，我们建议通过以下方式开发SPOC平台 将原位产生的蛋白质阵列与表面等离子共振成像(SPRI)相结合，通过 建立集成的ProSPR仪器，使SPRI能够同时确定定性和 高通量蛋白质生物分子相互作用的动力学信息。我们将发展， 验证和应用1000个蛋白质SPOC SPRI传感器芯片研究临床相关蛋白质- 动物模型中产生的新抗体的蛋白质相互作用和靶外结合试验 对抗人类蛋白质。

Abstract A critical bottleneck in the field of proteomics is the inability to simultaneously and cost-effectively evaluate thousands of functional protein interactions at once to obtain kinetic data. Although conventional in-situ protein microarray technology is well developed, it uses fluorescent dye based detection that provides only qualitative or semi-quantitative information. There is currently no technology that offers label-free real-time interaction assays of full-length folded proteins in high throughput format. To address this unmet need, INanoBio is developing novel sensor integrated proteome on chip (SPOC) platform that combines in situ expression and capture of functional proteins on biosensors, to allow instant high throughput analysis of thousands of proteins in real time. In this SBIR phase II project, we propose to develop SPOC platform by integrating in situ produced protein arrays with surface plasmon resonance imaging (SPRi), by building integrated ProSPR instrument that will enable SPRi determination of both qualitative and kinetic information of protein biomolecular interactions in high throughput. We will develop, validate and apply 1000 protein SPOC SPRi sensor chips to study clinically relevant protein- protein interactions and test off-target binding of new antibodies produced in animal models against human proteins.