Tuesday, 31 May 2022
West Ballroom B/C/D (Vancouver Convention Center)
Complex and aggressive diseases, such as pancreatic cancer, are often correlated to poor prognoses and high mortality rates due to difficulties associated with early diagnosis. A primary challenge of diagnosing such diseases is the lack of a single diagnostic biomarker that can easily be detected. Furthermore, diseases that can be correlated to a combination of biomarkers require the ability to quantify a variety of molecular targets that vary significantly in size and complexity. Consequently, there remains a need for biosensing platforms that can detect a large variety of targets, such as CA-19-9 antigen, lactate dehydrogenase, PSA, etc. Recently, the Woldring lab has demonstrated the small binding proteins that can act as highly modular biorecognition elements. These proteins (~10 KDa) can be efficiently engineered to selectively bind to a wide range of biological small molecules and proteins. As a result, they can serve as excellent biorecognition elements (binders) to target biomarkers for sensing applications. Starting from a library of known binders, we employed computational tools, such as homology modeling (Swiss-Model) and high-resolution docking (Rosetta), to identify sequences of strong fibronectin binders that could bind co-operatively to a single target. The leading binders were then encoded by DNA and transformed into cellular hosts for protein expression and were characterized via flow cytometry and chromatography. Though the small proteins have a strong binding affinity towards specific target biomarkers, there is no inherent mechanism to generate a signal during the binding event. Our current research is aimed at covalently modifying these binding proteins with redox probes to enable the generation of an electrochemical signal upon binding to a target biomarker. We will be discussing our recent progress in designing novel protein-electrode interfaces. These protein-electrode interfaces will further enable us in fabricating a tunable biosensor capable of detecting a diverse array of relevant molecular biomarkers.