Point-of-Care Portable 3D-Printed Multispectral Sensor for Real-Time Enzyme Activity Monitoring in Healthcare Applications

Abstract

Absorbance spectroscopy finds many biomedical and physical applications ranging from studying the atomic and molecular details of the analyte to determination of unknown biological species and their concentration or activity in the samples. Commercially available laboratory-based spectrometers are usually bulky and require high power and laborious manual processing, making them unsuitable to be deployed in portable and space-constrained environments, thereby further limiting their utility for real-time on-site monitoring. To address these challenges, here we developed a portable 3D-printed multispectral spectrophotometer based on absorbance spectroscopy for real-time monitoring of enzyme molecular activity. Monitoring enzyme (such as tyrosinase) activity is critical, as it quantifies its reaction rate, which is dependent on many factors such as the enzyme and substrate concentrations, temperature, pH, and other regulators such as inhibitors and effectors. Tyrosinase is a critical enzyme responsible for melanin synthesis in living beings and exhibits enzymatic browning in fruits and vegetables. It finds various commercial applications in the fields of healthcare (skin pigmentation, wound healing, etc.), forensics, and food processing. Here, tyrosinase activity was monitored using a 3D-printed spectral sensor at different rates and compared against measurements obtained from laboratory instruments. The enzyme activity was also studied using kojic acid (i.e., a commonly employed commercial tyrosinase inhibitor) while varying its molar and volume concentrations to control the reaction rate at discrete activity levels. For tyrosinase activity monitoring, the fabricated device has shown significant correlation (R2 = 0.9999) compared to measurements from the standard table-top spectrophotometer. We also provide a performance comparison between the 3D-printed and the laboratory spectrophotometer instruments by studying tyrosinase enzyme activity with and without the influence of an inhibitor. Such a device can be translated into various absorbance spectroscopy-based point-of-care biomedical and healthcare applications.