Synthetic protease receptors for Point of Care diagnostics
Nanobiotechnology, Health, Diagnostics, Proteases, Point of Care, Synthetic Biology, Cancer, Directed Evolution
Proteases are valuable cancer biomarkers. For instance Prostate–Specific Antigen (PSA) test has been widely used to screen men for prostate cancer. It is also used to monitor men who have been diagnosed with prostate cancer to see if their cancer has recurred after initial treatment or is responding to therapy. The clinically used diagnostic PSA tests measure the total amount of PSA protein in the blood. However PSA is a serine protease which exists in serum in three forms of which only one has proteolytic activity. Furthermore changes to additional proteases such as members of kallikrein and Matrix Metalloproteinase (MMP) families have been shown to have diagnostic and prognostic value for prostate cancer. The ability to measure a panel of prostate cancer specific proteases in biological fluid would provide a much more differentiated and predictive diagnostic test. We combined structure-based protein design and directed evolution to create a prototype of a protease sensor, based on highly specific potyvirus protease, TVMV. In this design, the protease was C-terminally extended with a sequence containing a protease recognition site and a sequence that specifically binds and inhibits the active TVMV’s active site. We demonstrate that such biosensor can specifically detect and amplify the proteolytic activity.We now extend the developed technology to create a point-of-care diagnostic test for the active forms of kallikrein as well and matrix metalloproteinase that have diagnostic value for prostate cancer.
|Lead investigator||Dr Viktor Stein
|Research group||Alexandrov Group|