Multiplexed detection and imaging of protein phosphorylation based on soluble nan

描述（由申请人提供）：随着最近的技术进步，可能是人类恶性肿瘤原因的多种重要信号通路已被不断发现和剖析。这些信号通路中的绝大多数涉及可逆的蛋白质磷酸化，并且关于磷酸化的位置和动力学的信息提供了关于信号网络如何起作用和相互作用的重要机制。而转化研究逐渐从实验室模型转向临床样本，最终目标是 在鉴定癌症生物标志物方面，仍然缺少简单可靠的磷酸化测定方法，用于复杂和典型异质性临床样品中磷酸化的常规检测。通过这项NIH SBIR I期研究，我们将进一步开发一种新的磷酸化检测策略，称为pIMAGO（磷酸成像），该策略最近已被我们引入到商业产品中，用于简单的常规磷酸化检测。这种新颖的设计不仅利用了可溶性纳米颗粒的量子尺寸特性，而且利用了分子的多功能性质，允许在不使用放射性同位素或昂贵的磷酸特异性抗体的情况下对蛋白质磷酸化进行高度选择性、灵敏性和简单的定性和定量评估。由于其尺寸和独特的性质，它还提供了同时多重检测磷酸化和总蛋白量的能力。我们建议优化膜上磷蛋白检测技术在常规生物医学研究。此外，我们将开发一种新型的基于荧光的pIMAGO试剂，用于直接检测和多重实验，为生物医学研究和开发增加另一个维度。 公共卫生相关性：蛋白质磷酸化与许多癌症类型的发生和发展有关，高效的磷酸化分析技术对于 癌症研究。NIH SBIR将支持将创新技术开发成商业产品的努力，为研究人员提供强大的工具和新的方向来对抗这种毁灭性的疾病。

DESCRIPTION (provided by applicant): With recent technical advances, multiple important signaling pathways that may be the causes of human malignancy have continuously been discovered and dissected. The vast majority of these signaling pathways involve reversible protein phosphorylation, and the information on the location and dynamics of phosphorylation provides important mechanisms on how the signaling networks function and interact. While translational research gradually shifts from lab models to clinical samples, with the ultimate goal of identifying cancer biomarkers, a simple and reliable phosphorylation assay method is still missing for routine detection of phosphorylation in complex and typically heterogeneous clinical samples. Through this NIH SBIR Phase I study, we will further develop a novel strategy for phosphorylation assay, termed pIMAGO (phospho-imaging) that has recently been introduced by us, into commercial products for simple, routine phosphorylation assays. This novel design takes advantage of not only the quantum size properties of the soluble nanoparticles, but also of the multi-functionalized nature of the molecule, allowing for highly selective, sensitive and simple qualitative and quantitative assessment of protein phosphorylation without the use of either radioactive isotopes or expensive phosphospecific antibodies. Due to its size and unique properties, it also offers the capability for multiplexed detection of phosphorylation and total protein amount simultaneously. We propose to optimize the technology for on-membrane phosphoprotein detection in routine biomedical research. In addition, we will develop a novel fluorescence-based pIMAGO reagent for direct detection and multiplexed experiments to add another dimension for biomedical research and development. PUBLIC HEALTH RELEVANCE: Protein phosphorylation relates to the onset and development of many cancer types and a highly efficient technology for phosphorylation analysis is critical for cancer research. This NIH SBIR will support an effort to develop an innovative technology into commercial products that equip researchers with powerful tools and new directions to combat the devastating disease.