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且经济有效地 一次评估数千种功能性蛋白质相互作用以获得动力学数据。虽然 传统的原位蛋白质微阵列技术已经很发达，它使用荧光染料 基于检测，仅提供定性或半定量信息。目前有 没有技术可以提供全长折叠蛋白的无标记实时相互作用测定 高吞吐量格式。为了解决这一未满足的需求，INanoBio 正在开发新型传感器 集成蛋白质组芯片（SPOC）平台，结合了原位表达和捕获 生物传感器上的功能蛋白，可对数千种蛋白质进行即时高通量分析 实时蛋白质。在这个SBIR第二阶段项目中，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) 集成，通过 构建集成的 ProSPR 仪器，使 SPRi 能够确定定性和 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、 验证并应用 1000 个蛋白质 SPOC SPRi 传感器芯片来研究临床相关蛋白质 - 蛋白质相互作用并测试动物模型中产生的新抗体的脱靶结合 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.