Thermal resilience of ensilicated lysozyme via calorimetric and in vivo analysis (2020)


A. Doekhie,*a M. N. Slade, a L. Cliff,a L. Weaver,a R. Castaing,b J. Paulin,c Y.-C. Chen,a K. J. Edler, a F. Koumanov, d K. J. Marchbank, c J. M. H. van den Elsene  and  A. Sartbaevaa

*Corresponding author

aDepartment of Chemistry, University of Bath, Claverton Down, Bath, UK

bMaterial and Chemical Characterisation Facility, University of Bath, Claverton Down, Bath, UK

cThe Medical School, Framlington Place, Newcastle University, Newcastle upon Tyne, UK

dDepartment for Health, University of Bath, Claverton Down, Bath, UK

eDepartment of Biology and Biochemistry, University of Bath, Claverton Down, Bath, UK

Ensilication is a novel method of protein thermal stabilisation using silica. It uses a modified sol–gel process which tailor fits a protective silica shell around the solvent accessible protein surface. This, electrostatically attached, shell has been found to protect the protein against thermal influences and retains its native structure and function after release. Here, we report the calorimetric analysis of an ensilicated model protein, hen egg-white lysozyme (HEWL) under several ensilication conditions. DSC, TGA-DTA-MS, CD, were used to determine unfolding temperatures of native, released and ensilicated lysozyme to verify the thermal resilience of the ensilicated material. Our findings indicate that ensilication protects against thermal fluctuations even at low concentrations of silica used for ensilication. Secondly, the thermal stabilisation is comparable to lyophilisation, and in some cases is even greater than lyophilisation. Additionally, we performed a mouse in vivo study using lysozyme to demonstrate the antigenic retention over long-term storage. The results suggest that protein is confined within the ensilicated material, and thus is unable to unfold and denature but is still functional after long-term storage.

DOI: https://doi.org/10.1039/D0RA06412B