Heterotrimer Nanopores for Protein Sensing

Stochastic sensing of proteins using genetically engineered protein nanopores is a promising approach to developing highly sensitive and specific biosensors for diagnostic purposes. Sensing takes place where a nanopore protein is inserted into a planar lipid bilayer and ionic current flows because of an applied transmembrane potential. The flow of ionic current may be disrupted where a protein binds to sensing elements of the nanopore and occludes the pore. Here, I will develop a protein nanopore scaffold for biosensor engineering using OmpF, a bacterial porin found as a homotrimer. Desired specificity and affinity for target proteins will be engineered using two adjacent loops of a monomer. Multiplexed sensing will be attempted with each of the three monomers containing a distinct binding sequence for separate proteins. In parallel, bacterial display libraries will be constructed for high-throughput screening of OmpF-based nano-sensors, which will be validated using single-channel recording. This project will contribute to the field of stochastic sensing using nanopores, to eventually develop a sensor capable of real-time, sensitive analysis of a wide range of proteins from complex samples such as serum.
This project addresses the BBSRC responsive mode priority of synthetic biology, particularly the area of bionanoscience. A cross-council priority addressed is that of nanoscience.