The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.
The Novo Nordisk Foundation Center for Biosustainability at the University of California, San Diego, School of Medicine, La Jolla, CA, 92093, USA.
Nat Commun. 2020 Apr 20;11(1):1908. doi: 10.1038/s41467-020-15866-w.
Host cell proteins (HCPs) are process-related impurities generated during biotherapeutic protein production. HCPs can be problematic if they pose a significant metabolic demand, degrade product quality, or contaminate the final product. Here, we present an effort to create a "clean" Chinese hamster ovary (CHO) cell by disrupting multiple genes to eliminate HCPs. Using a model of CHO cell protein secretion, we predict that the elimination of unnecessary HCPs could have a non-negligible impact on protein production. We analyze the HCP content of 6-protein, 11-protein, and 14-protein knockout clones. These cell lines exhibit a substantial reduction in total HCP content (40%-70%). We also observe higher productivity and improved growth characteristics in specific clones. The reduced HCP content facilitates purification of a monoclonal antibody. Thus, substantial improvements can be made in protein titer and purity through large-scale HCP deletion, providing an avenue to increased quality and affordability of high-value biopharmaceuticals.
宿主细胞蛋白（HCPs）是生物治疗性蛋白生产过程中产生的与工艺相关的杂质。如果 HCPs 对代谢有较大需求、降低产品质量或污染最终产品，它们就会成为问题。在这里，我们通过破坏多个基因来消除 HCPs，从而努力创建一个“清洁”的中国仓鼠卵巢（CHO）细胞。利用 CHO 细胞蛋白分泌模型，我们预测消除不必要的 HCPs 可能会对蛋白生产产生不可忽视的影响。我们分析了 6 蛋白、11 蛋白和 14 蛋白敲除克隆的 HCP 含量。这些细胞系的总 HCP 含量（40%-70%）显著降低。我们还观察到特定克隆中的生产力提高和生长特性改善。减少的 HCP 含量有利于单克隆抗体的纯化。因此，通过大规模 HCP 缺失可以显著提高蛋白滴度和纯度，为提高高价值生物制药的质量和降低成本提供了途径。
