Authors: Chad Hyer, Bradley Bundy, Tyler Free
Mentors: Bradley Bundy
Insitution: Brigham Young University
Current trends in healthcare suggest a movement towards point of care treatment and diagnosis. This transition towards local or even at-home testing necessitates dramatic decreases in costs of diagnostic methods. Cell-free protein synthesis (CFPS) systems prove a potential tool for bringing diagnostic solutions to patients by providing patients with inexpensive, easy to use, and accurate tests that can be stored for long periods of time at ambient temperature. These CFPS diagnostic methods rely on the production of biosensor proteins using isolated molecular machinery from bacterial cell lysates and necessary substrates. When combined with human body fluids, these biosensors can detect the presence of important biomarkers of disease, aiding in diagnosis.
CFPS systems, however, can prove difficult to use with human body fluids as CFPS systems rely on mRNA directed production of proteins, and human samples contain significant concentrations of RNAses which inhibit the production of biosensor proteins, preventing proper test results. Traditionally, this issue can be remedied in a CFPS system using commercially sourced RNAse inhibitors, but these inhibitors dramatically increase prices. Here we report on our production of RNAse inhibitors within a CFPS system to decrease costs of CFPS diagnostics by 90%. In our work, we report on the optimization of reaction conditions for producing RNAse inhibitors within a CFPS glutamine biosensor for use as a cancer treatment monitoring diagnostic. Using our methods, we expect to be able to dramatically reduce the cost of CFPS based diagnostic tools, helping empower the shift to affordable point of care healthcare.