Each year, 30,000 patients obtain transplants. To prevent graft rejection, immunosuppressants such as tacrolimus are prescribed. Due to tacrolimus's narrow therapeutic range, a dose that is too low places patients at risk for transplant rejection, but too high of a dose leads to kidney failure. The de facto method for monitoring of transplant patient health is bimonthly blood draws, which are cumbersome, painful, and difficult to translate into urgently needed dosage changes in a timely manner. To improve long-term transplant survival rates, we propose a finger-prick sensor that will provide patients and healthcare providers with a measurement of tacrolimus, immune health (through IL-12), and kidney damage (through cystatin C) levels 100 times more frequently than the status quo. Additionally, patient quality of life will be improved due to reduction in time and pain associated with blood draws. Optimal binding frequencies for each marker were found. However, due to limitations with EIS, the integration of the detection of the three markers into one multimarker sensing platform has not yet been realized. To this end, impedance-time tests were run on each marker along with different antibodies, and optimal times of each marker were determined to be 17s, 6s, and 2s, for tacrolimus, cystatin c, and IL-12, respectively (n=6). The integration of impedance-time analysis with traditional EIS methodologies has the potential to enable multi-marker analysis by analyzing binding kinetics on a single electrode with respect to time. Thus, our results provide unique insight into possibilities to improve and facilitate detection of multiple markers not only for the sensor for solid organ transplant patients, but for the monitoring of patients with disease that also entail the observation of multiple markers. Furthermore, the use of impedance-time testing also provides the ability for another way to optimize accuracy/precision of marker detection because it specifies a particular time, in addition to a particular optimal binding frequency, at which to measure concentration.
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