Smales, Christopher Mark, Dinnis, Diane M., Stansfield, Scott H., Alete, Daniel E., Sage, E.A., Birch, John R., Racher, Andrew J., Marshall, Carol T., James, David C. (2004) Comparative proteomic analysis of GS-NSO murine myeloma cell lines with varying recombinant monoclonal antibody production rate. Biotechnology and Bioengineering, 88 (4). pp. 474-488. ISSN 0006-3592. (doi:10.1002/bit.20272) (Access to this publication is currently restricted. You may be able to access a copy if URLs are provided) (KAR id:60)
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Official URL: http://dx.doi.org/10.1002/bit.20272 |
Abstract
We have employed an inverse engineering strategy based on quantitative proteome analysis to identify changes in intracellular protein abundance that correlate with increased specific recombinant monoclonal antibody production (qMab) by engineered murine myeloma (NS0) cells. Four homogeneous NS0 cell lines differing in qMab were isolated from a pool of primary transfectants. The proteome of each stably transfected cell line was analyzed at mid-exponential growth phase by two-dimensional gel electrophoresis (2D-PAGE) and individual protein spot volume data derived from digitized gel images were compared statistically. To identify changes in protein abundance associated with qMab datasets were screened for proteins that exhibited either a linear correlation with cell line qMab or a conserved change in abundance specific only to the cell line with highest qMab. Several proteins with altered abundance were identified by mass spectrometry. Proteins exhibiting a significant increase in abundance with increasing qMab included molecular chaperones known to interact directly with nascent immunoglobulins during their folding and assembly (e.g., BiP, endoplasmin, protein disulfide isomerase). 2D-PAGE analysis showed that in all cell lines Mab light chain was more abundant than heavy chain, indicating that this is a likely prerequisite for efficient Mab production. In summary, these data reveal both the adaptive responses and molecular mechanisms enabling mammalian cells in culture to achieve high-level recombinant monoclonal antibody production.
Item Type: | Article |
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DOI/Identification number: | 10.1002/bit.20272 |
Uncontrolled keywords: | NSO murine myeloma cells; monoclonal antibody; proteomics |
Subjects: | Q Science |
Divisions: | Divisions > Division of Natural Sciences > Biosciences |
Depositing User: | Mark Smales |
Date Deposited: | 19 Dec 2007 17:51 UTC |
Last Modified: | 16 Nov 2021 09:38 UTC |
Resource URI: | https://kar.kent.ac.uk/id/eprint/60 (The current URI for this page, for reference purposes) |
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