Impurity Profiling in Chromatographic Purification of Monoclonal Antibodies

由美国国家科学基金会化学和生物分离项目颁发的这一奖项，支持对中国仓鼠卵巢（CHO）细胞培养产生的单克隆抗体（mab）纯化过程中要去除的主要杂质进行表征的工作，这种纯化在工业实践中普遍使用。我们将鉴定在产物流中持续存在的杂质蛋白，主要来源是制造单克隆抗体的细胞产生的蛋白质，因此称为宿主细胞蛋白（HCP）。要表征的另一类杂质是产品相关杂质，即与单抗产品结合的蛋白质，因此在纯化步骤中难以从工艺流中去除。这些分析用于确定在给定的单抗过程中要去除的主要杂质，并可以作为优化从单抗产品中分离它们的策略的基础。与表征这两类杂质互补的是表征它们在涉及液相色谱的重要分离类中的行为。了解不同蛋白质（单克隆抗体产物和杂质）在不同条件下的色谱行为，将有助于确定最适合从产品流中去除杂质的条件。结果将应用于优化特定mAb分离的代表性工作。在这个项目中开发的能力可以帮助简化大型和不断增长的单克隆抗体治疗类的工业过程开发方法。研究也将作为学生教育和研究训练的有效工具，通过学生将进行大部分的研究。

0966644LenhoffThis NSF award by the Chemical and Biological Separations program supports work to characterize the principal impurities to be removed in the purification of monoclonal antibodies (mAbs) produced in cell culture by Chinese hamster ovary (CHO) cells, which are used ubiquitously in industrial practice. We will identify the impurity proteins that persist in the product stream, with the main source being proteins produced by the cells that make the mAbs, hence the name host cell protein (HCP). An additional class of impurities to be characterized are product-associated impurities, i.e., proteins that bind to the mAb product and are therefore difficult to remove from the process stream during the purification steps. These analyses to identify the principal impurities to be removed in a given mAb process and can serve as the basis for optimizing the strategies for separating them from the mAb product. Complementary to characterizing these two classes of impurities is characterization of their behavior in the important class of separations involving liquid chromatography. Knowledge of the chromatographic behavior of the different proteins, both mAb product and impurity, under different conditions will allow identification of the conditions most suitable for removing the impurities from the product stream. The results will be applied to representative efforts to optimize particular mAb separations. Capabilities such as those to be developed in this project can be instrumental in streamlining industrial process development methods for the large and growing class of mAb therapeutics. The research will also serve as an effective vehicle for student education and research training, via the students who will perform most of the research.