Split Protein System to Detect a Target Using Luminescence — 34p — Noah Nnabuo, Ceylin Kizilkaya, Dr. Yun Seok Choi

Athletics, particularly football, pose significant risks to long-term brain health due to repeated minor impacts, which emerging evidence shows are more harmful to cognitive function. Untreated, these injuries could lead to diseases such as chronic traumatic encephalopathy. Proactive measures are imperative to mitigate these risks and safeguard athletes’ long-term well-being. One promising avenue involves developing a sensor capable of detecting trauma in patients upon impact. This sensor, intended for use as a point-of-care method for multiple diseases such as sepsis, multiple organ failure, burns as well as trauma, aims to leverage abundant proteins as a biomarker. By measuring these specific protein levels in bodily fluids such as urine, blood, or cerebrospinal fluid, our sensor aims to enable early detection of trauma-induced cellular damage and practical application for various diseases. Current measuring methods are costly, time-consuming, and require specialized expertise. Our sensor design utilizes two protein-binding domains that bind to our protein simultaneously.

To develop a luminescent sensor, we attached a pair of split-luminescent proteins to our protein-binding domains. This design ensures luminescence is generated only when the split-proteins bind to our protein in bodily fluids. We successfully tested the luminescence emitted by our sensor construct upon merging all components to their respective positions (Split-protein bound to respective protein binding domain).  Higher concentrations of the specific protein resulted in more light, while lower concentrations resulted in less light being emitted. These findings underscore the sensor’s potential for precise quantification of our protein’s concentrations, highlighting its promise for future applications in medical diagnosis of disease and biomedical research.

Black Hills State University
Dr. Yun Seok Choi