Our POC grant aimed to further develop our membrane voltage nanosensors to the level of commercial reagents. The goals were: (i) identify leading applications in neuroscience and brain research, (ii) improve current technology to meet the applications demands by fine tuning the control over size and shape of the particle and testing various types of coating, testing insertion into different lipid compositions in bilayers and cells, and by testing non-specific internalization, (iii) improve the stability and performance reproducibility of the sensors, (iv) develop methods for upscaling the synthesis of these NPs, (v) and formulate GLP protocols required for commercialization.
We encountered difficulties with scaling-up the production of peptide-coated voltage nanosensors. This forced us to change strategies:
(i) we changed the coating technology from peptides to mixture of lipids. This technology is simpler and easier to up-scale. We obtained good results for staining membranes with lipid-coated nanosensors. We are currently investigating their membrane potential sensitivity. The commercialization of lipid-coated nanosensors is therefore delayed.
(ii) we developed an alternative approach for localized voltage sensing based on a 2-component (FRET-based) sensors. This technology is very promising and we will pursue to protect IP rights.
