Department of Pharmacy-Pharmaceutical Technology and Biopharmaceutics, Ludwig-Maximilians-Universität München, 81377 Munich, Germany.
Biostabilization Laboratory, Department of Mechanical Engineering, University of Minnesota, Minneapolis, MN 55455, USA.
Eur J Pharm Biopharm. 2019 Jan;134:185-189. doi: 10.1016/j.ejpb.2018.12.001. Epub 2018 Dec 4.
In this communication, the effect of mannitol and trehalose crystallization on the unfolding of IgG, a monoclonal antibody, in the frozen state with repeat freeze/thaw under different pH conditions was explored. Formulations were annealed at -20 °C for 20 h five times (interrupted by freeze/thaw). This was done to induce excipient crystallization. Characterization of the frozen-thawed samples was performed by circular dichroism, particle analysis, and size exclusion chromatography. At a pH of 3, formation of insoluble and soluble aggregates was observed however, these could be reduced by the use of a surfactant. Cryoprotectant free formulations showed higher monomer content after freeze/thaw. At pH5, a single freeze/thaw cycle did not result in a significant increase in particle numbers. At pH range of 4-7 however, aggregate formation in the size range of 1-25 µm was observed after 5freeze/thaw cycles.
在本研究中，探索了甘露醇和海藻糖结晶对单克隆抗体 IgG 在不同 pH 值条件下反复冻融状态下展开的影响。制剂在-20°C 下退火 20 小时，共进行五次（冻融中断）。这是为了诱导赋形剂结晶。通过圆二色性、颗粒分析和尺寸排阻色谱法对冷冻-解冻样品进行了表征。在 pH 值为 3 时，观察到形成不溶性和可溶性聚集体，但通过使用表面活性剂可以减少这些聚集体。无冷冻保护剂的制剂在冻融后显示出更高的单体含量。在 pH5 时，单次冻融循环不会导致颗粒数显著增加。然而，在 pH 值为 4-7 的范围内，在经过 5 次冻融循环后，在 1-25µm 的大小范围内观察到了聚集体的形成。
