Protease chain reactions for molecular analysis of cancer markers

DESCRIPTION provided by applicant Our objective is to develop a Protease Chain Reaction technology ProCR that will enable ultra sensitive and highly quantitative detection of cancer markers The basic element of a protease chain reaction is a protease inhibitor complex The protease is inactive when bound to the inhibitor but once freed is capable of cleaving the inhibitor and releasing additional free protease A trace of free protease initiates the chain reaction and eventually causes the release of all protease from the inhibitory complex The lag time preceding the full release of protease is determined by the initial concentration of free protease The Phase I project developed a first generation of self amplifying complexes that form the core of the ProCR detection technology All Phase I milestones were significantly exceeded In Phase II we will develop powerful molecular sensors based on self amplifying complexes A sensor consists of three elements a sensing element that responds to the presence of an analyte the processing computational element that amplifies and quantifies the signal from the detection element and the transducer element that produces an optical signal Through this combination extremely sophisticated enzymatic sensors will be built to detect cancer markers Accordingly the three experimental Aims are to Engineer sensing elements Engineer enhanced processing computational elements and Engineer signaling components Fundamentally a protein chain reaction is a powerful analogue computer with two key characteristics that greatly facilitate the detection of target molecules It can convert th concentration of a specific target molecule into a time signature It can create enormous signal amplification analogous to the amplification of DNA by a polymerase chain reaction PCR Thus detection is enabled because the final observable signal produced by a target molecule can be very large and the time lag until onset of the signal is precisely correlated with the concentration of target molecule The long term goal is to develop protease inhibitor complexes as enzymatic nano processors which can be combined to detect multiple signals and to control output with multiple logic gates PUBLIC HEALTH RELEVANCE The development of polymerase chain reaction PCR technology demonstrated the extraordinary power of harnessing an enzyme to perform novel programmable reactions Our objective is here to develop an analogous protease chain reaction technology ProCR which will enable ultra sensitive molecular detection The long term goal is to improve cancer detection and prevention by enabling accurate quantitation of multiple low abundance molecular markers