Engineering erythrocytes to be erythrosensors: first steps

Molecules can be loaded into mammalian erythrocytes through a reversible lysis pore that forms in the membrane when placed in hypotonic media, the result being resealed red cell ghosts. Many studies on the sidedness of transport processes have utilized this approach. In addition, red cell ghosts encapsulated with enzymes have been used in patients to treat specific enzyme deficiencies, particularly when the substrate can cross the red cell membrane. Our long-term goal is to put fluorescent sensors inside erythrocytes, return the loaded red cell ghosts to the animal or patient, and then monitor the fluorescence non-invasively to follow changes in plasma analyte concentration. In this paper, we present a novel dialysis method for making the red cell ghosts. In addition, we present a theoretical analysis showing that it is not necessary that every loaded red cell ghost has the same dye concentration. Finally we discuss the constraints on the optimal affinity for the sensor/analyte interaction.