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Protein-based nanobubbles such as halophilic archaeabacterial gas vesicles (GVs) represent a new class of stable, homogeneous nanoparticles with acoustic properties that allow them to be visualized by ultrasound (US) waves. To design GVs as theranostic agents, we modified them to respond to light, with a view to locally generate reactive oxygen species that can kill cancer cells. Specifically, up to 60,000 photoreactive chlorin  e 6 (C e 6) molecules were chemically attached to lysine ε-amino groups present on the surface of each purified  Halobacterium sp . NRC-1 GV. The resulting fluorescent NRC-1 C e 6-GVs have dimensions comparable to that of native GVs and were efficiently taken up by human breast [MCF-7] and human hypopharyngeal [FaDu-GFP] cancer cells as monitored by confocal microscopy and flow cytometry. When exposed to light, internalized C e 6-GVs were 200-fold more effective on a molar basis than free C e 6 at killing cells. These results demonstrate the potential of C e 6-GVs as novel and promising nanomaterials for image-guided photodynamic therapy.

Coupling Chlorin e6 to the surface of Nanoscale Gas Vesicles strongly enhances their intracellular delivery and photodynamic killing of cancer cells