Assessment of the effects of sterilization methods on protein drug stability by elucidating decomposition mechanism and material analysis

Journal of Pharmaceutical Sciences

Krueger

Abe

et al. 2017

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The effects of sterilization methods on the storage stability of erythropoietin (EPO) in polymer-based syringes were assessed by quantifying protein oxidation, aggregation, and particle formation. Micro-particle counting and size exclusion chromatography coupled with a multi-angle light scattering detector demonstrated much lower levels of protein particles and aggregates for EPO stored for 12 weeks in steam-sterilized than in radiation (Rad)-sterilized syringes. Intermediate levels of damage were observed for EPO stored in ethylene oxide-sterilized syringes. HPLC analysis documented that the Rad-sterilized syringes caused increased oxidation of the protein during storage. In contrast, in the steam- and ethylene oxide-sterilized syringes EPO oxidation did not change. Analysis with electron spin resonance revealed that only Rad-sterilized syringes formed radicals in the syringe body, which persisted over the 12-week storage period. These results demonstrated that Rad-sterilization generated radicals in the syringes which in turn caused increased EPO oxidation, particle formation, and protein aggregation. Therefore, steam sterilization was shown to be a preferable sterilization method for the polymer-based syringe system when using biopharmaceutical drugs highly sensitive to oxidation, and particle formation and aggregation.

Section: Methodsmentioning

confidence: 99%

Section: Sterilization Of P-pfsmentioning

confidence: 99%

Section: Electron Spin Resonancementioning

confidence: 99%

Section: Analysis Of Methionine and Oxidized Methionine In Epomentioning

confidence: 99%

Section: Introductionmentioning

confidence: 97%

See 3 more Smart Citations

Impact of Sterilization Method on Protein Aggregation and Particle Formation in Polymer-Based Syringes

Journal of Pharmaceutical Sciences

Krueger

Abe

et al. 2017

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Electron beam sterilization of cyclo olefin polymer leads to polymer degradation and production of alkyl radicals

J of Applied Polymer Sci

Imae

Takahashi

et al. 2016

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We investigated the effects of electron-beam (EB) sterilization on syringe barrels manufactured from cyclo olefin polymer (COP). The chemical structure of the polymer was determined by interpreting the 13 C NMR and DEPT-135 spectra of the COP resin. The antioxidants in the resin were identified by analyzing the liquid chromatography-photo diode array-mass spectrometry (LC-PDA-MS) data for the methanol extract of the resin and the gas chromatography-mass spectrometry (GC-MS) data for the supercritical methanol degradation products of the extract. NMR and LC-PDA-MS analyses revealed that EB sterilization produces degradation products in the COP main chain and reduces the quantity of the antioxidants in the COP resin. ESR spectra of the EB-sterilized syringe barrels indicated the presence and location of alkyl radicals, which were generated in the COP main chain by EB sterilization. ESR analyses also indicated that the quantity of alkyl radicals decreased over time.

Carbon radicals generated by solid polymers: Electron spin resonance spectroscopy for detection of species in water

Mochizuki

Ono

J of Applied Polymer Sci

et al. 2019

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Radicals generated in plastic medical devices (solid phase) by γ-rays or electron-beam irradiation during sterilization are known to cause oxidation of protein drugs, resulting in a loss or reduction in drug efficacy. The generation of radical species in water by the radical species in solid polymers has not been proved. Using electron spin resonance (ESR) spectroscopy, we confirm the generation of new radicals in water by γ-ray irradiated cyclic olefin polymers (COP). ESR measurements are obtained using 4-hydroxy-2,2, 6,6-tetramethylpiperidine-1-oxyl (TEMPOL) as a spin probe and 5-(2,2-dimethyl-1,3-propoxycyclophosphoryl)-5-methyl-1-pyrroline Noxide (CYPMPO) as a spin trap, in which the irradiated COP was immersed. The ESR signals indicate the TEMPOL radicals decline over time, suggesting the generation of new radicals. Conversely, the characteristic ESR signals of the adduct formed by the reaction between CYPMPO and the hydroxyl radical are observed. Thus, hydroxyl radicals are generated because of the migration of the radicals from COP to water.