Comparative proteomic analysis of GS-NSO murine myeloma cell lines with varying recombinant monoclonal antibody production rate

Smales, CM, Dinnis, DM, Stansfield, SH, Alete, D, Sage, EA, Birch, JR, Racher, AJ, Marshall, CT and James, DC (2004) Comparative proteomic analysis of GS-NSO murine myeloma cell lines with varying recombinant monoclonal antibody production rate. Biotechnology And Bioengineering, 88 4: 474-488. doi:10.1002/bit.20272


Author Smales, CM
Dinnis, DM
Stansfield, SH
Alete, D
Sage, EA
Birch, JR
Racher, AJ
Marshall, CT
James, DC
Title Comparative proteomic analysis of GS-NSO murine myeloma cell lines with varying recombinant monoclonal antibody production rate
Journal name Biotechnology And Bioengineering   Check publisher's open access policy
ISSN 0006-3592
Publication date 2004-01-01
Sub-type Article (original research)
DOI 10.1002/bit.20272
Open Access Status Not Open Access
Volume 88
Issue 4
Start page 474
End page 488
Total pages 15
Editor E Swanson
Place of publication USA
Publisher John Wiley & Sons, Inc.
Language eng
Subject C1
290602 Process Control and Simulation
670706 Organic industrial chemicals not elsewhere classified
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 (NSO) cells. Four homogeneous NSO 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 clatasets 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. (C) 2004 Wiley Periodicals, Inc.
Keyword Biotechnology & Applied Microbiology
Nso Murine Myeloma Cells
Monoclonal Antibody
Proteomics
Chinese-hamster Ovary
Glutamine-synthetase Gene
High-level Expression
Messenger-rna Levels
Cho-cells
Endoplasmic-reticulum
Batch Culture
Insect Cells
Saccharomyces-cerevisiae
Subclone Development
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collections: Faculty of Engineering, Architecture and Information Technology Publications
2005 Higher Education Research Data Collection
 
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Created: Wed, 15 Aug 2007, 13:14:31 EST