Lyoprotection of aviscumine with low molecular weight dextrans.
The aim of this research was to ascertain whether dextrans with low molecular weight will stabilize aviscumine. During freeze-drying increasing concentrations of dextran T1 (MW 1000) stabilized aviscumine. Eight percent of dextran resulted in a nearly 100% recovery of the activity and in addition a complete amorphous structure of the solid phase was obtained. By decreasing the molecular weight of the dextran from 75 to 1 kDa, the protein activity was increased by 20% in the lyophilisate. Combinations of dextran with either trehalose or mannitol showed no additional effects on stability. The improved stabilization of aviscumine using low molecular weight dextrans is explained by an increased interaction between the protein and the dextran molecules (like hydrogen bonds), whereas they are sterically hindered if larger dextran molecules are used. When the protein concentration was increased from 10 to 100 microg/ml (in formulas with 8% dextran T1), no influence on the protein activity could be found. With regard to the carbohydrate-binding activity of the protein, it was shown that the optimal content of residual water in the lyophilisate should be about 2%. Above and below this percentage a destabilization of the protein was observed. The often discussed failure of dextran as a stabilizing excipient in the freeze-drying of proteins seems to be a question of the selection of the correct molecular weight.
Gloger
O
O
Department of Pharmaceutics and Biopharmaceutics, Christian Albrecht University Kiel, Gutenbergstrasse 76, D-24118 Kiel, Germany.
Witthohn
K
K
Müller
B W
BW
D000970:Antineoplastic Agents / Q000737:chemistry*; D002152:Calorimetry, Differential Scanning; D003460:Crystallization; D003911:Dextrans / Q000737:chemistry*; D004355:Drug Stability; D004797:Enzyme-Linked Immunosorbent Assay; D005079:Excipients / Q000737:chemistry; D005612:Freeze Drying; D006358:Hot Temperature; D008353:Mannitol / Q000737:chemistry; D008970:Molecular Weight; D028321:Plant Preparations / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry*; D054790:Ribosome Inactivating Proteins, Type 2; D014118:Toxins, Biological / Q000737:chemistry*; D014199:Trehalose / Q000737:chemistry; D014867:Water / Q000737:chemistry; D014961:X-Ray Diffraction
D000970:Antineoplastic Agents; D003911:Dextrans; D005079:Excipients; D028321:Plant Preparations; D010940:Plant Proteins; D054790:Ribosome Inactivating Proteins, Type 2; D014118:Toxins, Biological; C009674:ribosome inactivating protein, Viscum; D014867:Water; D008353:Mannitol; D014199:Trehalose