title: Host cell protein dynamics in recombinant CHO cells: impacts from harvest to purification and beyond.
abstract: 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.
authors: Centre for Molecular Processing and School of Biosciences; University of Kent; Canterbury, Kent, UK; Advanced Centre for Biochemical Engineering; Department of Biochemical Engineering; University College London; Torrington Place, London, UK.
authors: Catherine Em
authors: CE
authors: Hogwood
authors: Daniel G
authors: DG
authors: Bracewell
authors: C Mark
authors: CM
authors: Smales
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D016466:CHO Cells; D002846:Chromatography, Affinity; D002853:Chromatography, Liquid; D003412:Cricetulus; D004340:Drug Contamination / Q000517:prevention & control*; D015180:Electrophoresis, Gel, Two-Dimensional; D005818:Genetic Engineering / Q000379:methods; D007141:Immunoglobulin Fc Fragments / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D013058:Mass Spectrometry; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D013205:Staphylococcal Protein A
chemical_list: D000911:Antibodies, Monoclonal; D007141:Immunoglobulin Fc Fragments; D011994:Recombinant Proteins; D013205:Staphylococcal Protein A
keywords: Chinese hamster ovary (CHO); downstream processing; host cell protein; mammalian cell culture; monoclonal antibody; protein A chromatography; proteomics