DISRUPTIVE TECHNOLOGY FOR KINETIC STUDIES OF PROTEIN-SMALL MOLECULE BINDING

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Kinetic studies of reversible protein-small molecule binding are pivotal to understanding molecular mechanisms of diseases and development of molecular diagnostics and drug leads. Biosensor-based methods are most established in such studies, but they require the immobilization of either protein or small molecule on the sensor. The immobilization often blocks the binding. Here we propose equipment that will uniquely enable immobilization-free kinetic studies of protein-small molecule binding. The proposed instrument will be used by our 2 labs (37 HQP and ~$0.8M/year operating funds) during a 3-year technology-development stage. During this stage, we will: (i) assemble a unique instrument for immobilization-free kinetic studies, (ii) develop algorithms for fast and accurate signal processing and data analysis, (iii) use the instrument for studying binding kinetics for small molecules from libraries generated by Yudin’s lab and GSK (industrial collaborator), (iv) design commercial versions of the instrument and software, (v) initiate commercialization of the technology, and (vi) design and implement a training protocol for use of the instrument by HQP. After the technology-development stage, the instrument will be placed in the York Bioanalytical Core Facility (YBioCore). It will then be used by trained HQP from 10 labs carrying out relevant research at York. It will also be accessible to external users. Krylov’s lab will hold annual 3-day workshops for HQP. The workshops will include classes as well as hands-on exercises. The instrument will be maintained by Facility technical staff but operated by trained HQP. HQP with such skills are highly desired by the pharma and biotech industries. When in YBioCore, the instrument will be used by biologists studying molecular mechanisms of diseases and chemists developing diagnostic probes and drug leads. The unique capabilities of the instrument will facilitate obtaining kinetic data that would be difficult or impossible without this instrument. The data obtained with the instrument will enable the development of molecular diagnostics and drug leads which, in turn, will benefit Canadians. If we are not able to build the proposed instrument in 2017, 2 important directions in our labs will be practically stalled. In Krylov’s lab it is the development of the LSTDLPF-based technology, which is currently carried out by 3 PDFs and 2 grad students. In Yudin’s lab, it is macrocycle-protein binding studies, which are currently run by 4 grad students. A delay in acquiring the instrument will, thus, significantly delay the completion of degree requirements by 6 grad students and the career training of 3 PDFs. A delay in obtaining the instrument will also have a much wider impact as it will also delay the transfer of our highly-enabling technology to a broad community of academic and industrial end-users for whom this technology is being developed.