Combining phage display and localized surface plasmon resonance for rapid engineering of high affinity protein-protein interfaces

特异性和紧密结合的蛋白质-蛋白质界面的工程化具有许多应用，从开发新的药学重要分子到创建具有新功能的设计蛋白质。由于目前缺乏合理设计这种类型的蛋白质-蛋白质界面的知识，最有效的方法是使用随机诱变方法结合分子展示。这些非知情或最小知情方法的一个缺点是需要筛选数百种不同的蛋白质变体以获得所需的活性。表面等离子体共振（SPR）方法最近已经被提出来有效地允许这样的筛选，然而传统的仪器成本过高。Nicoya SPR平台是传统SPR仪器的一种经过验证的、具有成本效益的替代品，但尚未被验证为这种类型的分子展示方法的有效工具。拟议的研究将开发通用的优化方法，以验证Nicoya OpenSPR系统用于这种类型的分子显示方法，以优化和/或设计新型蛋白质-蛋白质界面。这项工作将使Nicoya能够通过制造SPR将其仪器的应用扩展到这一新兴市场

Engineering of specific and tight binding protein-protein interfaces has numerous applications ranging from the development of novel pharmaceutically important molecules to the creation of designer proteins of novel function. As the knowledge to rationally design protein-protein interfaces of this type is currently lacking, the most effective methodology is to use random mutagenesis approaches coupled with molecular display. One downside to these non-informed or minimally-informed approaches is the necessity to screen many hundreds of different protein variants for the desired activity. Surface plasmon resonance (SPR) approaches have been recently proposed to efficiently allow for such screening, however the traditional instruments are cost prohibitive. The Nicoya SPR platform is a proven, cost effective, alternative to traditional SPR instruments, but has not been validated as an effective tool for this type of molecular display approach. The proposed research will develop common, optimized methodology to validate the Nicoya OpenSPR system for this type of molecular display approach to optimize and/or engineer novel protein-protein interfaces. This work will allow Nicoya to expand the application of their instrument into this emerging market segmant by making the SPR