Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
J Pharm Sci. 2011 Jul;100(7):2526-42. doi: 10.1002/jps.22493. Epub 2011 Jan 25.
Understanding antibody aggregation is of great significance for the pharmaceutical industry. We studied the aggregation of five different therapeutic monoclonal antibodies (mAbs) with size-exclusion chromatography-high-performance liquid chromatography (SEC-HPLC), fluorescence spectroscopy, electron microscopy, and light scattering methods at various temperatures with the aim of gaining insight into the aggregation process and developing models of it. In particular, we find that the kinetics can be described by a second-order model and are non-Arrhenius. Thus, we develop a non-Arrhenius model to connect accelerated aggregation experiments at high temperature to long-term storage experiments at low temperature. We evaluate our model by predicting mAb aggregation and comparing it with long-term behavior. Our results suggest that the number of monomers and mAb conformations within aggregates vary with the size and age of the aggregates, and that only certain sizes of aggregates are populated in the solution. We also propose a kinetic model based on conformational changes of proteins and monomer peak loss kinetics from SEC-HPLC. This model could be employed for a detail analysis of mAb aggregation kinetics.
理解抗体聚集对于制药行业具有重要意义。我们使用尺寸排阻色谱-高效液相色谱（SEC-HPLC）、荧光光谱法、电子显微镜和光散射法，在不同温度下研究了五种不同治疗性单克隆抗体（mAb）的聚集情况，旨在深入了解聚集过程并建立其模型。特别是，我们发现动力学可以用二级模型来描述，并且是非阿仑尼乌斯的。因此，我们开发了一个非阿仑尼乌斯模型，将高温下的加速聚集实验与低温下的长期储存实验联系起来。我们通过预测 mAb 聚集并将其与长期行为进行比较来评估我们的模型。结果表明，聚集物内的单体数量和 mAb 构象随聚集物的大小和年龄而变化，并且仅在溶液中存在某些大小的聚集物。我们还提出了一个基于蛋白质构象变化和 SEC-HPLC 中单体峰损失动力学的动力学模型。该模型可用于对 mAb 聚集动力学进行详细分析。
