Novel soluble nanopolymers for enrichment of low abundant phosphoproteins

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第一阶段研究，我们将开发可溶性纳米聚合物基试剂，称为PolyMAC（聚合物基金属离子亲和捕获），用于高效分离磷酸肽的商业产品。这种新颖的设计不仅利用了多功能化纳米颗粒的特性，更重要的是利用了分子的可溶性，允许在溶液相中螯合有限量的磷酸肽，以获得最佳效率和最大产率。我们建议最终将试剂作为商业产品进行优化和可扩展性。此外，将开发用于全面磷酸化蛋白质组学分析的高通量格式，以满足许多癌症生物学和蛋白质组学研究实验室/设施的需求。

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 develop soluble nanopolymer-based reagents, termed PolyMAC (Polymer-based Metal Ion Affinity Capture), into commercial products for the highly efficient isolation of phosphopeptides. This novel design takes advantage of not only the properties of multifunctionalized nanoparticles, but more importantly, the soluble nature of the molecule, allowing for the chelation of a limited amount of phosphopeptides in the solution phase for optimum efficiency and maximum yield. We propose to finalize optimization and scalability of the reagents as commercial products. In addition, high throughput formats for comprehensive phosphoproteomic analyses will be developed to address needs of many cancer biology and proteomics research labs/facilities.