Stability of crystallised and spray-dried lysozyme

Moisture and temperature promote protein degradation. The stabilities of commercial, crystallised and spray-dried lysozyme, a model protein, were assessed under these stresses to explore whether a crystalline protein had better storage stability than a conventionally produced one. Samples were maintained at different relative humidities (RH) and temperatures for 20 weeks and stabilities estimated in solid and aqueous states. Differential scanning calorimetry (DSC) and thermogravimetry (TGA) characterised solid samples. Fourier transform Raman (FT-Raman) spectroscopy analysed solid material and aqueous solutions. High sensitivity differential scanning calorimetry (HSDSC) and enzymatic assays were used to monitor solutions. DSC and HSDSC data revealed that crystals maintained thermal stability at high RH; spray drying appreciably changed melting characteristics. These results correlated with enzymatic assays that demonstrated good activity retention for crystals but less so for spray-dried material (e.g. 95 and 87% relative to fresh samples after 20 weeks at 40 degrees C/75% RH). FT-Raman analysis showed that crystallised lysozyme better-maintained protein conformational integrity compared to spray-dried samples in accelerated stability studies. Based on TGA data, spray-dried protein absorbed water on storage under humid conditions, which induced instability. Thus, crystallisation enhanced storage stability of lysozyme with negligible loss of activity.