Premium
Proteome analysis of antibody‐producing CHO cell lines with different metabolic profiles
Author(s) -
Pascoe Deborah E.,
Arnott David,
Papoutsakis Eleftherios T.,
Miller William M.,
Andersen Dana C.
Publication year - 2007
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.21460
Subject(s) - cell culture , proteome , pyruvate kinase , biochemistry , biology , chinese hamster ovary cell , intracellular , antibody , cell , metabolism , two dimensional gel electrophoresis , glycolysis , proteomics , chemistry , immunology , genetics , gene
Two‐dimensional gel electrophoresis and tandem mass spectrometry were used to identify proteins associated with a metabolic shift during fed‐batch cultures of two recombinant antibody‐producing CHO cell lines. The first cell line underwent a marked change in lactate metabolism during culture, initially producing lactate and then consuming it, while the second cell line produced lactate for a similar duration but did not later consume it. The first cell line displayed a declining specific antibody productivity during culture, correlating to the 2‐D gel results and the intracellular antibody concentration determined by HPLC. Several statistical analysis methods were compared during this work, including a fixed fold‐change criterion and t ‐tests using standard deviations determined in several ways from the raw data and mathematically transformed data. Application of a variance‐stabilizing transformation enabled the use of a global empirical standard deviation in the t ‐tests. Most of the protein spots changing in each cell line did not change significantly in the other cell line. A substantial fraction of the changing proteins were glycolytic enzymes; others included proteins related to antibody production, protein processing, and cell structure. Enolase, pyruvate kinase, BiP/GRP78, and protein disulfide isomerase were found in spots that changed over time in both cell lines, and some protein changes differed from previous reports. These data provide a foundation for future investigation of metabolism in industrially relevant mammalian cell culture processes, and suggest that along with differences between cell types, the proteins expressed in cultures with low lactate concentrations may depend on how those conditions were generated. Biotechnol. Bioeng. 2007;98: 391–410. © 2007 Wiley Periodicals, Inc.