Stable sugar-based protein formulations by supercritical fluid drying

Nataša Jovanović, Andréanne Bouchard, Marc Sutter, Michiel Van Speybroeck, Gerard W. Hofland, Geert Jan Witkamp, Daan J.A. Crommelin, Wim Jiskoot*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

35 Scopus citations


The aim of this work was to produce stable, sugar-containing protein formulations by supercritical fluid (SCF) drying. Lysozyme solutions with and without added sucrose or trehalose were dried by spraying them in an SCF composed of CO2 and ethanol or CO2 only. The protein-to-sugar ratio was varied between 1:0 and 1:10 (w/w). Dried formulations were stored at 4 °C for three months and analyzed by Karl Fischer titration, scanning electron microscopy, X-ray powder diffraction, differential scanning calorimetry and Fourier transform infrared spectroscopy. Lysozyme stability after reconstitution was determined by an enzymatic activity assay, UV/Vis spectroscopy, and SDS-PAGE. Smooth, spherical particles of 1-25 μm size were obtained. All formulations were initially amorphous. Crystallization during storage only occurred with a protein-to-sugar ratio of 1:10 and could be avoided by performing SCF drying without ethanol. Absence of residual ethanol in dried trehalose formulations increased the glass transition temperature up to 120 °C. Lysozyme in dried formulations was structurally stable, with exception of the 1:0 and 1:1 protein-to-sugar ratios, where reversible protein aggregation occurred. The results show that by avoiding ethanol, which up to now has been considered mandatory for efficient drying of aqueous solutions, and by choosing the proper protein-to-sugar ratio, it is possible to obtain stable, sugar-based protein formulations through SCF drying.

Original languageEnglish (US)
Pages (from-to)102-108
Number of pages7
JournalInternational Journal of Pharmaceutics
Issue number1-2
StatePublished - Jan 4 2008
Externally publishedYes


  • FTIR
  • Lysozyme
  • Protein stabilization
  • Protein structure
  • Sucrose
  • Supercritical fluid drying
  • Trehalose

ASJC Scopus subject areas

  • Pharmaceutical Science


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