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Bean, Andrew
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document
Biomimetic Metal-Organic Frameworks as Protective Scaffolds for Live-virus Encapsulation and Vaccine Stabilisation – TEM Staining Considerations.
Abstract
The stabilisation of vaccines during transport and storage is essential to their efficacy. We report the biomimetic-mineralisation of live-viral vaccines using metal-organic frameworks (MOFs), zeolitic imidazole framework-8 (ZIF-8) to enhance the storage stability at RT or 370C, from days to months [1]. Negatively charged proteins increase the local concentrations of positively charged metal ions facilitating reversible composite formation of the ZIF-8 around the biomacromolecule, in this case the virusZIF-8 and aluminium fumarate (Alfum), Newcastle Disease Virus (NDV) V4 strain and Influenza A WSN strain were encapsulated with retention of their viral titre. Live-virus recovery was determined using a tissue culture infectious dose (TCID50) assay. The composites were further stabilised with trehalose (T) and skim milk (SM) before freeze-drying. The thermal stability of the MOF@Vaccine composite, ZIF-8@NDV+T/SM was compared to control NDV and non-encapsulated, freeze-dried NDV+T/SM composite at 40C and 370C over a period of 12 weeks.Using transmission electron microscopy (TEM) we observed the sequence of events in ZIF-8@WSN and Alfum@WSN disintegration using release buffer and confirmed the release of structurally intact WSN virions from the composites. Phosphotungstic acid (PTA) negative stain could not be employed during the release phase of the reaction as PTA has been reported to precipitate onto ZIF-8 further stabilising the composite [2]. Composites freeze dried with T and SM released morphologically intact virus after 12 weeks at 370C indicating much improved vaccine storage conditions.Figure. 1. Transmission Electron Microscopy visualization of ZIF-8 encapsulation and release of the NDV live viral vaccine. The NDV virus (a) is encapsulated using the ZIF-8 MOF forming an amorphous composite (b). The electron density image of the composite correlates with its SEM image in inset b. A sodium citrate buffer was used to chelate the zinc ions causing MOF disintegration (c) releasing structurally intact NDV virions (d).