Proteomic Analysis of Host Cell Protein Dynamics in the Culture Supernatants of Antibody-Producing CHO Cells.
Proteomic Analysis of Host Cell Protein Dynamics in the Culture Supernatants of Antibody-Producing CHO Cells.
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DOI：
10.1038/srep44246
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发表时间：
2017-03-10
影响因子：
4.6
通讯作者：
Lee GM
中科院分区：
文献类型：
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作者：
Park JH;Jin JH;Lim MS;An HJ;Kim JW;Lee GM
中国仓鼠卵巢 (CHO) 细胞是用于生产治疗性蛋白质（包括单克隆抗体 (mAb)）的最常见细胞系。裂解细胞分泌和释放的宿主细胞蛋白 (HCP) 在重组 CHO (rCHO) 细胞培养过程中在细胞外积累，可能会损害产品质量。为了在培养过程中保持良好的 mAb 质量，通过纳流液相色谱-串联质谱法对产 mAb 的 rCHO 细胞系的分批和补料分批培养物中细胞外积累的 HCP 进行了鉴定和定量，然后进行了基因本体论和功能分析。由于细胞浓度较高且培养时间较长，补料分批培养中鉴定和定量的 HCP（鉴定出 2145 个蛋白质，定量出 1673 个蛋白质）比分批培养（鉴定出 1934 个蛋白质，定量出 1486 个蛋白质）更多。 HCP 的聚类分析表明，影响 mAb 质量的 HCP 浓度分布（Lgmn、Ctsd、Gbl1 和 B4galt1）与培养过程中 mAb 质量属性（例如聚集、电荷变体和 N-糖基化）的变化相关。总而言之，本研究中获得的 HCP 数据集为确定在培养和纯化步骤中要去除的适当靶蛋白提供了见解，以确保良好的 mAb 质量。
Chinese hamster ovary (CHO) cells are the most common cell line used for the production of therapeutic proteins including monoclonal antibodies (mAbs). Host cell proteins (HCPs), secreted and released from lysed cells, accumulate extracellularly during the cultures of recombinant CHO (rCHO) cells, potentially impairing product quality. In an effort to maintain good mAb quality during the cultures, HCPs accumulated extracellularly in batch and fed-batch cultures of a mAb-producing rCHO cell line were identified and quantified by nanoflow liquid chromatography-tandem mass spectrometry, followed by their gene ontology and functional analysis. Due to higher cell concentration and longer culture duration, more HCPs were identified and quantitated in fed-batch culture (2145 proteins identified and 1673 proteins quantified) than in batch culture (1934 proteins identified and 1486 proteins quantified). Clustering analysis of HCPs showed that the concentration profiles of HCPs affecting mAb quality (Lgmn, Ctsd, Gbl1, and B4galt1) correlated with changes in mAb quality attributes such as aggregation, charge variants, and N-glycosylation during the cultures. Taken together, the dataset of HCPs obtained in this study provides insights into determining the appropriate target proteins to be removed during both the cultures and purification steps for ensuring good mAb quality.