Effect of Technically Relevant X-Ray Doses on the Structure and Function of Alcohol Dehydrogenase and Hen Egg-White Lysozyme
| Autor: | Karl Lohner, Lisa-Marie Schaden, Peter Laggner, Johannes Poms, Stephan Sacher, Sharareh Salar-Behzadi, Johannes Khinast, Miriam Wimmer-Teubenbacher |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Solid-state Pharmaceutical Science 02 engineering and technology Radiation Dosage 03 medical and health sciences chemistry.chemical_compound Scattering Small Angle Animals Pharmacology (medical) Irradiation Alcohol dehydrogenase Pharmacology Chromatography biology Small-angle X-ray scattering X-Rays Organic Chemistry X-ray Alcohol Dehydrogenase 021001 nanoscience & nanotechnology Enzyme assay Structure and function 030104 developmental biology chemistry biology.protein Molecular Medicine Muramidase Lysozyme 0210 nano-technology Biotechnology |
| Zdroj: | Pharmaceutical research. 35(7) |
| ISSN: | 1573-904X |
| Popis: | The effect of different irradiation doses on the structure and activity of lyophilized powders of Hen Egg-White Lysozyme (HEWL) and alcohol dehydrogenase (ADH) was investigated using these substances as models for robust and sensitive proteins, respectively. Three doses were selected to cover the ranges of radio-sterilization (25kGy), treatment of blood products (25Gy) and annual background radiation dose (approximately 2mGy). The results offer an initial screening of different irradiation doses and support the development of X-ray imaging methods as non-destructive process analytical technology (PAT) tools for detecting the visible particulate matters in such products. HEWL and ADH were exposed to X-rays in the solid state. The effect of irradiation was determined directly after irradiation and after storage. Structural changes and degradation were investigated using SAXS, SDS-PAGE and HPLC-MS. Protein functionality was assessed via activity assays. Lower irradiation doses of 25Gy and 2mGy had no significant impact on the structure and enzyme activity. The dose of 25kGy caused a significant decrease in the enzyme activity and structural changes immediately after irradiation of ADH and after storage of irradiated HEWL at −20°C. The results emphasize the importance of careful selection of radiation doses for development of X-ray imaging methods as PAT tools inspection of solid biopharmaceutical products. |
| Databáze: | OpenAIRE |
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