Host cell protein dynamics in recombinant CHO cells Impacts from harvest to purification and beyond
Host cell protein dynamics in recombinant CHO cells Impacts from harvest to purification and beyond
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DOI：
10.4161/bioe.23382
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发表时间：
2013-09-01
期刊：
影响因子：
4.9
通讯作者：
Smales, C. Mark
中科院分区：
文献类型：
--
作者：
Hogwood, Catherine E. M.;Bracewell, Daniel G.;Smales, C. Mark
在生产重组蛋白产品（如单克隆抗体）期间，制造商必须证明在临床使用之前将宿主细胞杂质和污染物清除到适当水平。这些包括宿主细胞DNA和RNA，产品相关污染物，如聚集体，以及重要的宿主细胞蛋白（HCPs）。尽管去除HCP很重要，但在细胞培养和下游加工（DSP）过程中，这些蛋白质的特性和动态在很大程度上是未知的。SELDI-TOF质谱法等技术的改进以及ELISA的金标准技术已经允许对现有的总hcp进行半定量。然而，直到最近才利用技术来识别存在的hcp，并将其与监测hcp在发酵和下游加工过程中的动态的方法发展相结合。为了在提高HCP清除率方面做出基于知识的决策，在细胞系和特定产品的基础上识别潜在的问题HCP是至关重要的。了解HCP动力学将有助于在未来提供一个平台，合理操纵和设计和/或选择合适的重组CHO细胞系和下游加工步骤，以限制有问题的HCP。
During the production of recombinant protein products, such as monoclonal antibodies, manufacturers must demonstrate clearance of host cell impurities and contaminants to appropriate levels prior to use in the clinic. These include host cell DNA and RNA, product related contaminants such as aggregates, and importantly host cell proteins (HCPs). Despite the importance of HCP removal, the identity and dynamics of these proteins during cell culture and downstream processing (DSP) are largely unknown. Improvements in technologies such as SELDI-TOF mass spectrometry alongside the gold standard technique of ELISA has allowed semi-quantification of the total HCPs present. However, only recently have techniques been utilized in order to identify those HCPs present and align this with the development of approaches to monitor the dynamics of HCPs during both fermentation and downstream processing. In order to enable knowledge based decisions with regards to improving HCP clearance it is vital to identify potential problematic HCPs on a cell line and product specific basis. Understanding the HCP dynamics will in the future help provide a platform to rationally manipulate and engineer and/or select suitable recombinant CHO cell lines and downstream processing steps to limit problematic HCPs.