Tait, Andrew S. and Hogwood, Catherine E.M. and Smales, C. Mark and Bracewell, Daniel G. (2011) Host cell protein dynamics in the supernatant of a mAb producing CHO cell line. Biotechnology and Bioengineering, 109 (4). pp. 971-982. ISSN 00063592. (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided)
The characterization of host cell protein (HCP) content during the production of therapeutic recombinant proteins is an important aspect in the drug development process. Despite this, key components of the HCP profile and how this changes with processing has not been fully investigated. Here we have investigated the supernatant HCP profile at different times throughout culture of a null and model GS-CHO monoclonal antibody producing mammalian cell line grown in fed-batch mode. Using 2D-PAGE and LC-MS/MS we identify a number of intracellular proteins (e.g., protein disulfide isomerise; elongation factor 2; calreticulin) that show a significant change in abundance relative to the general increase in HCP concentration observed with progression of culture. Those HCPs that showed a significant change in abundance across the culture above the general increase were dependent on the cell line examined. Further, our data suggests that the majority of HCPs in the supernatant of the cell lines investigated here arise through lysis or breakage of cells, associated with loss in viability, and are not present due to the secretion of protein material from within the cell. SELDI-TOF and principal components analysis were also investigated to enable rapid monitoring of changes in the HCP profile. SELDI-TOF analysis showed the same trends in the HCP profile as observed by 2D-PAGE analysis and highlighted biomarkers that could be used for process monitoring. These data further our understanding of the relationship between the HCP profile and cell viability and may ultimately enable a more directed development of purification strategies and the development of cell lines based upon their HCP profile.
|Uncontrolled keywords:||host cell protein;Chinese hamster ovary cells;mammalian cell culture;downstream processing;monoclonal antibody;proteomics|
|Divisions:||Faculties > Science Technology and Medical Studies > School of Biosciences|
|Depositing User:||Sue Davies|
|Date Deposited:||27 Mar 2012 13:32|
|Last Modified:||22 May 2014 09:26|
|Resource URI:||https://kar.kent.ac.uk/id/eprint/29205 (The current URI for this page, for reference purposes)|