BioPharmaceutical Development and Marketed Product Support, Centocor R&D, Inc., Schaffhausen, Switzerland.
J Pharm Sci. 2010 Aug;99(8):3343-61. doi: 10.1002/jps.22123.
The development of a Microflow digital imaging (MDI) method to detect and monitor protein particulates in a high concentration IgG1 monoclonal antibody formulation is presented. The MDI assay was optimized and qualified as a characterization assay in terms of accuracy and precision of particle size and number using polystyrene standards (5-300 microm) and protein particles (2 to >100 microm). The stability profile of a 90 mg/mL IgG1 formulation stored at 2-8 degrees C and -70 degrees C for up to 18 months was then investigated. The MDI assay results showed improved sensitivity to detect subvisible particulates (>or=5 microm) compared to conventional light obscuration detection, presumably due to the translucent nature of the protein particles. For evaluation of visible protein particles (>70 microm), a good overall correlation was observed for MDI, inverted microscopy and visual assessments. Long-term stability data for a high concentration IgG1 monoclonal antibody formulation demonstrated an accumulation of protein particles in certain size categories with a concomitant increase in the overall particle size distribution over time. The weight amount of protein particulates in the IgG1 formulation was measured experimentally as approximately 0.022% (approximately 20 microg/mL) after storage at 2-8 degrees C for 16 months. Similar results were obtained by calculation from the MDI particle data indicating a low level of protein particulates by weight. The nature and composition of the protein particulates formed during storage were further characterized by a combination of inverted microscopy, FTIR microscopy, and SEM-EDX. Particulates were identified as protein with silicone, although some particles also contained other elements such as aluminum. The combination of MDI results and protein characterization studies have provided an enhanced understanding of protein particulate formation during long-term storage of a high concentration IgG1 monoclonal antibody formulation.
介绍了一种微流数字成像（MDI）方法的开发，用于检测和监测高浓度 IgG1 单克隆抗体制剂中的蛋白质颗粒。使用聚苯乙烯标准品（5-300 微米）和蛋白质颗粒（2 至>100 微米），对 MDI 测定法进行了优化和鉴定，以确保在粒径和数量方面的准确性和精密度。然后研究了 90mg/mL IgG1 制剂在 2-8°C 和-70°C 下储存长达 18 个月的稳定性。MDI 测定结果显示，与传统的光遮挡检测相比，检测亚可见颗粒（>或=5 微米）的灵敏度有所提高，这可能是由于蛋白质颗粒的半透明性质所致。对于评估可见蛋白质颗粒（>70 微米），MDI、倒置显微镜和目视评估之间观察到了良好的总体相关性。高浓度 IgG1 单克隆抗体制剂的长期稳定性数据表明，在某些粒径类别中蛋白质颗粒的积累随时间推移而增加，同时总体粒径分布也随之增加。在 2-8°C 下储存 16 个月后，通过实验测量 IgG1 制剂中的蛋白质颗粒重量约为 0.022%（约 20μg/mL）。通过从 MDI 颗粒数据计算得出了相似的结果，表明按重量计蛋白质颗粒的含量较低。通过倒置显微镜、FTIR 显微镜和 SEM-EDX 的组合，进一步对形成的蛋白质颗粒的性质和组成进行了表征。颗粒被鉴定为蛋白质和硅酮，尽管一些颗粒还含有其他元素，如铝。MDI 结果和蛋白质特征研究的结合，提供了对高浓度 IgG1 单克隆抗体制剂在长期储存过程中蛋白质颗粒形成的增强理解。
