Intein-Mediated Protein Engineering for Cell-Based Biosensors

Abstract

Cell-based sensors provide a flexible platform for screening biologically active targets and for monitoring their interactions in live cells. Their applicability extends across a vast array of biological research and clinical applications. Particularly, cell-based sensors are becoming a potent tool in drug discovery and cell-signaling studies by allowing function-based screening of targets in biologically relevant environments and enabling the in vivo visualization of cellular signals in real-time with an outstanding spatiotemporal resolution. In this review, we aim to provide a clear view of current cell-based sensor technologies, their limitations, and how the recent improvements were using intein-mediated protein engineering. We first discuss the characteristics of cell-based sensors and present several representative examples with a focus on their design strategies, which differentiate cell-based sensors from in vitro analytical biosensors. We then describe the application of intein-mediated protein engineering technology for cell-based sensor fabrication. Finally, we explain the characteristics of intein-mediated reactions and present examples of how the intein-mediated reactions are used to improve existing methods and develop new approaches in sensor cell fabrication to address the limitations of current technologies.

Electrochemical Sensors, Biosensors, and Optical Sensors for the Detection of Opioids and Their Analogs: Pharmaceutical, Clinical, and Forensic Applications

“…Improvements in the biocompatibility and biostability of colorimetric sensor components ensure the sensor's reliable performance in biological samples. This is critical for maintaining the sensor's accuracy over extended periods, making it suitable for applications in therapeutic drug monitoring and clinical diagnostics [159]. The designed cell-based biosensor used for biological samples can screen for ligands specific to cell surface receptors using an opioid-binding opioid receptor.…”