Impurity Profiling in Chromatographic Purification of Monoclonal Antibodies

0966644 Lenhoff化学和生物分离项目授予的NSF奖项支持在纯化中国仓鼠卵巢（CHO）细胞培养物中产生的单克隆抗体（mAb）过程中去除主要杂质的工作，这些单克隆抗体在工业实践中广泛使用。我们将鉴定出在产品流中持续存在的杂质蛋白，主要来源是由制造mAb的细胞产生的蛋白质，因此命名为宿主细胞蛋白（HCP）。另一类待表征的杂质是产品相关杂质，即，与mAb产物结合并因此难以在纯化步骤期间从工艺流中除去的蛋白质。这些分析用于鉴别在给定mAb工艺中待去除的主要杂质，并可作为优化从mAb产品中分离杂质的策略的基础。对这两类杂质进行表征的补充是对它们在涉及液相色谱的重要分离类别中的行为进行表征。了解不同条件下不同蛋白质（mAb产品和杂质）的色谱行为，将有助于鉴别最适合从产物流中除去杂质的条件。结果将应用于代表性工作，以优化特定mAb分离。在这个项目中开发的能力，可以简化工业过程开发方法的大型和不断增长的单克隆抗体治疗方法。该研究还将作为学生教育和研究培训的有效工具，通过学生谁将执行大部分的研究。

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.