UC Irvine

Numerous factors, such as genetics, medical history, environment, and lifestyle contribute to patient responses to disease conditions. Approaches in precision medicine focus on examining disease in the context of overall patient health, so that prevention and effectively tailored treatments result in improved outcomes. This thesis describes investigations of disease characterization and tool development geared towards uses in precision medicine. The COVID-19 pandemic has opened new avenues of investigation for the scientific community. The most important of which is understanding why patients have such diverse responses to infection. Here, we develop an M13-bacteriophage display-based screening platform to understand how antibody responses play a role in foreshadowing and driving disease severity in a subset of patients. We further characterize disease response in the context of co-morbidities and viral infection history and find that both factors can negatively impact disease severity in patients with a specific antibody response. Additionally, sensitivity of these patients to rising levels of cytokines suggest specific routes of pre-emptive treatments that may be beneficial. We, additionally, develop a platform for rapid (<5 mins) point-of-care detection, which can be used to assist in multi-analyte testing or in urgent triaging. Precision medicine is also important in addressing changes in disease response over time. Real-time automated monitoring of disease markers can help improve disease management and reduce adverse side effects. Using techniques in protein engineering, we harness the specific binding interactions of native proteins to develop biosensors for managing highly dynamic disease conditions. Firstly, we engineer the human insulin receptor towards developing an optical insulin biosensor to improve automated type-1-diabetes disease management. The investigation focuses on the development of soluble, compact insulin-binding variants. We further test various assay designs for detecting insulin-dependent optical signaling. Secondly, we engineer a photo-stable version of a previously developed FRET-based calcium sensor, Twitch 2B. The new version of this sensor demonstrates improved stability and performance within an optical-fiber-based implantable device. Such a sensor can be applied to early diagnosis of hypocalcemia during blood transfusions. Taken together, these investigations explore a wide range of protein engineering techniques across multiple expression platforms. By harnessing native protein-protein interactions, we build prognostic and disease management tools that can contribute to advancing the field of precision medicine. Additionally, we develop optical detection platforms and rapid antibody screening methods that can be used to dynamically monitor and explore crucial molecular interactions in disease.