Sonochemically-induced spectral shift as a probe of green fluorescent protein release from nano capsules

Encapsulation in the form of micro and nanocapsules is an attractive route for controlling the delivery and release of active proteins and peptides. Many approaches exist to probe the morphology of such capsules as well as their mechanisms of formation. By contrast, the release of proteins from such components in a complex biological environment has been challenging to probe directly. In this paper we show that the spectral differences between green fluorescent protein (GFP) in capsules and in its free form can be used to monitor in situ the release of the protein from the confinement of capsules. These findings represent a new route towards engineering the spectral characteristics of GFP through physical rather than chemical means. We demonstrate the use of GFP protein capsules for monitoring in real time the release of protein in live cells by exposing rat L6 myotubes to protein capsules. The GFP spheres with a blue fluorescent signal dissociate inside the L6 myotubes to individual GFP molecules with a change in fluorescent signal from blue to green. These sensitive spectral characteristics enabled us to resolve the dissociation of capsules inside the cells in both time and space. We discuss the implications of our results for quantifying parameters crucial for the delivery of proteins in biological environments.