Pharmaceutical Research

, Volume 10, Issue 8, pp 1232–1237

Decreased Protein-Stabilizing Effects of Cryoprotectants Due to Crystallization

  • Ken-ichi Izutsu
  • Sumie Yoshioka
  • Tadao Terao
Article

DOI: 10.1023/A:1018988823116

Cite this article as:
Izutsu, K., Yoshioka, S. & Terao, T. Pharm Res (1993) 10: 1232. doi:10.1023/A:1018988823116

Abstract

The stabilizing effects of various additives against inactivation of an enzyme (β-galactosidase from Aspergillus oryzae) during freeze-drying were studied, with a focus on their crystallinity. The crystalline morphology of mannitol and inositol in freeze-dried cakes depended on the solute concentrations before freezing and the freeze-drying method used. The additives in their amorphous state showed concentration-dependent stabilization of the enzyme, whereas additive crystallization during freeze-drying decreased their effects. Heat treatment before freeze-drying also caused crystallization and diminished the stabilizing effects. Noncovalent soluble aggregates were observed in the inactivated enzyme solution. These results show the importance of maintaining the amorphous state of additives used as stabilizing agents during freeze-drying.

freeze-drying protein-stabilizing effects cryoprotectants crystallization β-galactosidase mannitol 

Copyright information

© Plenum Publishing Corporation 1993

Authors and Affiliations

  • Ken-ichi Izutsu
    • 1
  • Sumie Yoshioka
    • 1
  • Tadao Terao
    • 1
  1. 1.National Institute of Hygienic SciencesSeta-gaya-ku, TokyoJapan

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