Research Center Pharmaceutical Engineering (RCPE) GmbH, Graz, Austria.
Institute of Molecular Biosciences, Biophysics Division, University of Graz, Graz, Austria.
Pharm Res. 2018 May 7;35(7):135. doi: 10.1007/s11095-018-2417-2.
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.
使用卵清溶菌酶（HEWL）和醇脱氢酶（ADH）这两种模型药物，分别考察不同辐照剂量对冻干粉末结构和活性的影响。选择了三种剂量，涵盖了辐照灭菌（25kGy）、血液制品处理（25Gy）和年背景辐射剂量（约 2mGy）的范围。结果为不同辐照剂量提供了初步筛选，并支持将 X 射线成像方法作为无损过程分析技术（PAT）工具，用于检测此类产品中的可见颗粒物质。
HEWL 和 ADH 在固态下暴露于 X 射线。辐照后的直接和储存后的影响均进行了测定。使用小角 X 射线散射（SAXS）、SDS-PAGE 和 HPLC-MS 研究结构变化和降解。通过活性测定评估蛋白质功能。
较低剂量 25Gy 和 2mGy 对结构和酶活性没有显著影响。25kGy 的剂量会导致 ADH 辐照后和 -20°C 储存后辐照 HEWL 的酶活性显著下降和结构发生变化。
这些结果强调了在开发 X 射线成像方法作为 PAT 工具检查固体生物制药产品时，仔细选择辐射剂量的重要性。
