Efficient long-term and high-yielded production of a recombinant proteoglycan in eukaryotic HEK293 cells using a membrane-based bioreactor

被引:10
作者
Adam, Estelle [1 ,2 ]
Sarrazin, Stephane [2 ,3 ]
Landolfi, Cindy [2 ]
Motte, Vincent [2 ]
Lortat-Jacob, Hugues [3 ]
Lassalle, Philippe [1 ]
Delehedde, Maryse [1 ,2 ]
机构
[1] Inst Pasteur, INSERM, U774, F-59019 Lille, France
[2] Endotis Pharma, F-59120 Loos, France
[3] Univ Grenoble 1, CNRS, Inst Biol Struct, UMR 5075 CEA, F-38027 Grenoble, France
关键词
bioreactor CELLine (TM); high density cell culture; endocan; proteoglycan; glycosylation;
D O I
10.1016/j.bbrc.2008.01.141
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Standard culture systems of eukaryotic cells generally failed to deliver sufficient amounts of recombinant proteins without increasing the costs of production. We here showed that membrane-based bioreactors, initially developed for the production of monoclonal antibodies, can be very useful for the production using engineered HEK293 cells, of a recombinant proteoglycan called endocan, with achievement of high level expression and efficient long-term production. When compared to standard procedures, the growth in suspension and at high density of these cells in one bioreactor promoted a 60-fold increase of the concentration of the soluble recombinant endocan. These culture conditions did not affect cell viability, stable expression, recognition by specific monoclonal antibodies or electrophoretic profile of the recombinant endocan. Such an easy to scale up system to produce recombinant protein should open soon new opportunities to study structure and functions of endocan or any other glycosylated cell products newly investigated. (c) 2008 Elsevier Inc. All rights reserved.
引用
收藏
页码:297 / 302
页数:6
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