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Stable sugar-based protein formulations by supercritical fluid drying.

Natasa Jovanović1, Andréanne Bouchard, Marc Sutter

  • 1Department of Pharmaceutical Sciences, Utrecht Institute for Pharmaceutical Sciences (UIPS), 3508 TB Utrecht, The Netherlands.

International Journal of Pharmaceutics
|July 31, 2007
PubMed
Summary

Supercritical fluid (SCF) drying successfully produced stable, amorphous protein formulations using sugars like trehalose. Optimizing the protein-to-sugar ratio and avoiding ethanol improved stability and prevented crystallization during storage.

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Area of Science:

  • Biotechnology
  • Materials Science
  • Pharmaceutical Sciences

Background:

  • Producing stable, sugar-containing protein formulations is crucial for pharmaceutical applications.
  • Supercritical fluid (SCF) drying offers a potential method for creating these formulations.

Purpose of the Study:

  • To develop stable, sugar-containing protein formulations using supercritical fluid (SCF) drying.
  • To investigate the impact of different protein-to-sugar ratios and drying conditions on formulation stability.

Main Methods:

  • Lysozyme solutions with sucrose or trehalose were spray-dried in a supercritical CO(2) and ethanol mixture or CO(2) only.
  • Formulations were analyzed for particle morphology, crystallinity, thermal properties, and structural integrity.
  • Lysozyme stability was assessed post-reconstitution using enzymatic activity assays, spectroscopy, and SDS-PAGE.

Main Results:

  • Smooth, spherical particles (1-25 microm) were obtained, initially amorphous.
  • Crystallization during storage was observed only at a 1:10 protein-to-sugar ratio and was preventable by excluding ethanol.
  • Excluding ethanol in trehalose formulations increased the glass transition temperature to 120°C.
  • Lysozyme remained structurally stable, except at low sugar ratios (1:0, 1:1), where reversible aggregation occurred.

Conclusions:

  • Stable, sugar-based protein formulations can be produced via SCF drying by optimizing the protein-to-sugar ratio and omitting ethanol.
  • This method avoids ethanol, previously considered essential, enabling the creation of high-quality protein formulations.