Screening ligand-binding domains for rapid biosensor design and testing

Biosensors are sensing devices engineered to measure biological or chemical reactions by generating a response signal which is relative to the concentration of an analyte in the reaction. A general biosensor device is made up of five different components: an analyte, the bioreceptor, a transducer, the electronic and the display. When presented together, they form a sensing system that can be tailored to detect specific analytes. Energy conversion events controlled by engineered components within the biosensor molecule, take advantage of the specificity in binding to trigger measurable signals that can be later modified into digital form in order to be understandable. Previous studies done at the Alexandrov’s group have shown that it is possible to engineer allosterically regulated protein switches by domain insertion in order to design more integrated receptor/transducer architecture. This architecture is referred to as “the two-component biosensor”, and it is designed to incorporate ligand-binding domains of every nature in order to potentially have tailored-made biosensors for the most relevant biomarkers found in the blood. However, there is a need to test and select the incredible myriad of ligand-binding proteins present up to date. The work done here tackles a challenge in biosensor design where a potentially infinite arrange of combinations and alternatives can happen between the analyte and the bioreceptor module. This thesis describes the path towards a screening methodology for ligand-binding domains that could accelerate the development of new biosensors and, we show the complete design and testing of a new biosensor for a relevant human biomarker for coronary events. Given the ever-growing interest in biosensor research fuelled by the contemporary concern in human diagnostics and the rise of point-of-care analysis, technologies are extending their applications not only to the pharmaceutical industry, but also human health, environmental monitoring, food safety, forensics, and biomedical research