Abstract

Novel Split Trehalase as a Versatile Detection Assay for a Wide Range of Analytes

Marija Drikic, PhD Student, University of Calgary

Biosensors have a potential to become universal diagnostic devices available Anywhere, Anytime, to Anyone and for the detection of any analyte (anything). Although different studies over the years aimed to develop such a diagnostic device, until today none have managed to meet all 4 criteria and enter into clinical practice.  The most successful class of biosensors currently on the market is the glucometer used to monitor blood glucose. Performance in clinical samples, sensitivity and specificity have largely been optimized, measuring devices have been miniaturized and production costs optimized. The aim of our study was to develop a novel technology that uses the existing biosensor but increases the type and number of analytes that can be detected. We report the periplasmic trehalase of E. coli as a novel split enzyme reporter capable of converting various analytes into glucose. We proved that conditional complementation of the trehalase fragments leading to trehalose hydrolysis and glucose production, can be used to detect antibodies, bacterial cells, small molecules, but also protein-protein interaction and protein aggregation. We demonstrated retention of activity of split TreA in undiluted clinical samples. Furthermore, we applied this technology to develop and validate a diagnostic tool for efficient quantification of the total amount of immunoglobulins in bovine colostrum and serum. We explored its applicability do develop detection assays in different animal species. In conclusion, the resulting trehalase-based biosensor assay offers a versatile and convenient method for point-of-care applications as it does requires minimal sample preparation and can be integrated with existing glucometers or sensors.