Building Synthetic Biosensors Using Red Blood Cell Proteins

Area of Science:

• Biotechnology
• Cell Engineering
• Diagnostic Development

Background:

• Engineered cell therapies are expanding beyond cancer immunotherapy.
• Red blood cells (RBCs) offer unique advantages for in situ diagnostics due to long circulation times.
• Existing tools for engineering RBCs into biosensors are limited, partly due to membrane remodeling during maturation.

Purpose of the Study:

• To develop a novel biosensing architecture for red blood cells (RBCs).
• To create biosensors utilizing RBC membrane proteins retained through erythropoiesis.
• To establish a foundation for RBC-based biosensors for diagnostic applications.

Main Methods:

• Developed a biosensing architecture based on RBC membrane proteins.
• Utilized a mechanism where extracellular ligand binding triggers intracellular split protein reconstitution (fluorophore or enzyme).
• Evaluated various biosensor designs, linkers, scaffolds, and output signals for in vitro performance.

Main Results:

• Identified optimal biosensor designs and features conferring substantial ligand-induced signals in vitro.
• Demonstrated successful engineering of erythroid precursor cells with RBC-protein biosensors.
• Confirmed functionality of engineered RBC biosensors in vivo.

Conclusions:

• Established a robust platform for creating red blood cell (RBC)-based biosensors.
• The developed biosensors leverage retained RBC membrane proteins for effective sensing.
• This work lays the groundwork for noninvasive physiological monitoring and diagnostics using engineered RBCs.