Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, Kawasaki, Kanagawa 210-9501, Japan.
Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, Kawasaki, Kanagawa 210-9501, Japan.
J Pharm Sci. 2021 Mar;110(3):1189-1196. doi: 10.1016/j.xphs.2020.10.022. Epub 2020 Oct 15.
Pre-filled syringes (PFS) have been in widespread use as an administration device for therapeutic antibodies in recent decades. Generally, the inner barrel and syringe of PFS are coated with silicone oil (SO) for lubrication. Multiple studies have focused on the fact that the SO adsorbs denatured antibody molecules, and induces antibody aggregation. Aggregated antibodies are recognized as a potential risk for evoking immunogenic responses in patients. The characteristics of the aggregated antibody-SO complexes, including their concentration, population, shape, three-dimensional (3D) image, and Fcγ Receptors (FcγRs) activation have been obscurely acknowledged so far. In the present work, we prepared aggregated antibody-SO complexes by agitation and analyzed using multifaceted techniques such as flow imaging, confocal fluorescence microscopy, and cell-based assays for FcγRs activation. The results emphasized that the SO accelerates the increase in sub-visible particles and antibody aggregation. The confocal fluorescence microscopy analysis revealed the high-resolution 3D images of aggregated antibody-SO complexes. The FcγRs reporter cell assay clarified that the pre-mixed and agitated Ab + SO have higher FcγRs activation capability compared to the agitated Ab. Overall, this study advances the view that SO has an effect to increase the risk of agitation-induced aggregated antibody particles.
预充式注射器（PFS）作为一种治疗性抗体的给药装置，在近几十年得到了广泛应用。通常，PFS 的内筒和注射器都涂有硅油（SO）以实现润滑。多项研究集中于 SO 会吸附变性抗体分子并诱导抗体聚集这一事实。聚集的抗体被认为是在患者中引发免疫原性反应的潜在风险。到目前为止，聚集的抗体-SO 复合物的特性，包括其浓度、群体、形状、三维（3D）图像和 Fcγ 受体（FcγRs）的激活，仍未得到明确的认识。在本工作中，我们通过搅拌制备了聚集的抗体-SO 复合物，并使用流式成像、共聚焦荧光显微镜和基于细胞的 FcγRs 激活测定等多种技术进行了分析。结果强调了 SO 加速了亚可见颗粒和抗体聚集的增加。共聚焦荧光显微镜分析揭示了聚集的抗体-SO 复合物的高分辨率 3D 图像。FcγRs 报告细胞测定表明，与单独搅拌的抗体相比，预先混合和搅拌的 Ab+SO 具有更高的 FcγRs 激活能力。总的来说，这项研究表明 SO 会增加搅拌诱导的聚集抗体颗粒的风险。
