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Hydrogen Peroxide Induced Protein Oxidation During Storage and Lyophilization Process
Weiqiang Cheng1, Xiaoyang Zheng1, Mark Yang1
1Biopharmaceutics Development, Sanofi Genzyme, Framingham, Massachusetts 01701.
Journal of Pharmaceutical Sciences
|May 31, 2016
Summary
Hydrogen peroxide (HP) significantly impacts protein oxidation and aggregation during lyophilization, even in frozen states. Lyophilization effectively removes HP, but protein damage still occurs.
Area of Science:
- Protein chemistry
- Biopharmaceutical formulation
- Lyophilization technology
Background:
- The effects of hydrogen peroxide (HP) on proteins in liquid solutions are well-documented.
- The impact of HP on proteins during the lyophilization process remains largely unexplored.
Purpose of the Study:
- To investigate the effects of HP on proteins during lyophilization.
- To assess HP removal efficiency during the lyophilization process.
Main Methods:
- Protein formulations and placebos were spiked with HP (up to 5.0 ppm) and lyophilized.
- HP concentration, protein oxidation, and aggregation were monitored before, during, and after lyophilization, and during storage at 25°C.
Main Results:
- Lyophilization removed an average of 94.1% of HP from protein formulations, compared to 72.5% from placebos.
- Significant increases in protein oxidation and aggregation were observed post-lyophilization.
- Protein oxidation correlated with decreased HP concentration across all temperatures studied.
Conclusions:
- Hydrogen peroxide significantly contributes to protein oxidation and aggregation during lyophilization.
- Protein oxidation and subsequent aggregation can occur even when the formulation is frozen.
- Oxidized proteins exhibit increased susceptibility to aggregation during the lyophilization cycle.
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