Coriolis Pharma, Fraunhoferstrasse 18 b, 82152 Martinsried, Germany; Division of BioTherapeutics, Leiden Academic Centre for Drug Research, Leiden University, the Netherlands.
Inserm, Inflammation, Microbiome and Immunosurveillance, Université Paris-Saclay, Châtenay-Malabry, France.
J Pharm Sci. 2021 Feb;110(2):952-958. doi: 10.1016/j.xphs.2020.11.011. Epub 2020 Nov 19.
Sucrose is a commonly used stabilizing excipient in protein formulations. However, recent studies have indicated the presence of nanoparticulate impurities (NPIs) in the size range of 100-200 nm in pharmaceutical-grade sucrose. Furthermore, isolated NPIs have been shown to induce protein aggregation when added to monoclonal antibody formulations. Moreover, nanoparticles are popular vaccine delivery systems used to increase the immunogenicity of antigens. Therefore, we hypothesized that NPIs may have immunostimulatory properties. In this study, we evaluated the immunomodulatory effects of NPIs in presence and absence of trastuzumab in vitro with monocyte-derived dendritic cells (moDCs). Exposure of trastuzumab, the model IgG used in this study, to NPIs led to an increase in concentration of proteinaceous particles in the sub-micron range. When added to moDCs, the NPIs alone or in presence of trastuzumab did not affect cell viability or cytotoxicity. Moreover, no significant effect on the expression of surface markers, and cytokine and chemokine production was observed. Our findings showed, surprisingly, no evidence of any immunomodulatory activity of NPIs. As this study was limited to a single IgG formulation and to in vitro immunological read-outs, further work is required to fully understand the immunogenic potential of NPIs.
蔗糖是蛋白质制剂中常用的稳定赋形剂。然而，最近的研究表明，药用级蔗糖中存在粒径在 100-200nm 之间的纳米颗粒杂质（NPIs）。此外，当添加到单克隆抗体制剂中时，分离出的 NPIs 已被证明会诱导蛋白质聚集。此外，纳米颗粒是常用的疫苗递送系统，用于提高抗原的免疫原性。因此，我们假设 NPIs 可能具有免疫刺激特性。在这项研究中，我们评估了 NPIs 在存在和不存在曲妥珠单抗的情况下对单核细胞衍生的树突状细胞（moDCs）的体外免疫调节作用。曲妥珠单抗，即本研究中使用的模型 IgG，暴露于 NPIs 会导致亚微米范围内蛋白质颗粒的浓度增加。当添加到 moDCs 中时，NPIs 单独或与曲妥珠单抗一起使用不会影响细胞活力或细胞毒性。此外，表面标志物的表达以及细胞因子和趋化因子的产生也没有明显变化。令人惊讶的是，我们的研究结果表明，NPIs 没有任何免疫调节活性的证据。由于这项研究仅限于单一 IgG 制剂和体外免疫学检测，因此需要进一步的工作来充分了解 NPIs 的免疫原性潜力。
