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Peroxide formation in polysorbate 80 and protein stability.
Emily Ha1, Wei Wang, Y John Wang
1Analytics & Formulation Department, Process Sciences, Bayer Biotechnology, 800 Dwight Way, Berkeley, California 94701, USA.
Journal of Pharmaceutical Sciences
|September 13, 2002
Summary
Peroxides form easily in polysorbate 80, especially in solution with air and light. Higher peroxide levels in polysorbate 80 accelerate protein oxidation, but glutathione can inhibit this effect.
Area of Science:
- Pharmaceutical Science
- Biochemistry
- Materials Science
Background:
- Nonionic surfactants like polysorbate 80 are crucial excipients in protein drug formulation.
- Residual peroxides in surfactants can compromise the stability of oxidation-sensitive protein therapeutics.
- Understanding peroxide formation and its impact is vital for ensuring drug efficacy and safety.
Purpose of the Study:
- To investigate peroxide generation in polysorbate 80 under various storage conditions.
- To evaluate the oxidative potential of polysorbate 80 peroxides on a model protein, IL-2 mutein.
- To identify strategies for mitigating peroxide-induced protein oxidation.
Main Methods:
- Incubation of neat and aqueous polysorbate 80 under different conditions (temperature, air, light).
- Quantification of peroxide levels in polysorbate 80.
- Assessment of IL-2 mutein oxidation in liquid and solid states using polysorbate 80 with varying peroxide levels.
- Evaluation of glutathione's protective effect in lyophilized formulations.
- Analysis of oxidation during lyophilization annealing.
Main Results:
- Peroxides readily form in neat polysorbate 80 at elevated temperatures with air.
- Aqueous polysorbate 80 shows faster peroxide formation, accelerated by light.
- Peroxide formation is inhibited by excluding air/oxygen.
- Higher peroxide levels in polysorbate 80 significantly accelerate IL-2 mutein oxidation in both liquid and solid states.
- Glutathione effectively inhibits peroxide-induced oxidation in lyophilized IL-2 mutein.
- High peroxide levels cause immediate oxidation during lyophilization annealing.
Conclusions:
- Polysorbate 80 is susceptible to peroxide formation, influenced by storage conditions.
- The level of peroxides in polysorbate 80 directly impacts the oxidation of sensitive proteins.
- Strategies to minimize peroxide formation and the use of antioxidants like glutathione are critical for stabilizing protein pharmaceuticals.
- Careful consideration of lyophilization process steps, such as annealing, is necessary when using polysorbate 80 with oxidation-sensitive proteins.