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

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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