The use of UCOE vectors in combination with a preadapted serum free, suspension cell line allows for rapid production of large quantities of protein

被引:65
作者
Benton, T [1 ]
Chen, T [1 ]
McEntee, M [1 ]
Fox, B [1 ]
King, D [1 ]
Crombie, R [1 ]
Thomas, TC [1 ]
Bebbington, C [1 ]
机构
[1] Corixa Corp, Biol Proc Dev & Mol Biol, San Francisco, CA 94080 USA
关键词
preadapted serum-free cell culture; rapid protein production; UCOE;
D O I
10.1023/A:1021141712344
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
UCOE vectors contain non-tissue specific chromatin-opening-elements that permit rapid expression of a protein in an integration independent manner. Efficient expression can be derived from a single copy of an integrated gene site resulting in a higher percentage of cells expressing the marker gene in the selected pool in comparison to standard non-UCOE containing vectors. This, in combination with the utilization of a serum-free, suspension adapted parent cell line allows for rapid production of large quantities of protein in a short period of time. Utilizing this system more than 300 mg of a recombinant antibody has been produced in less than 1 month from transfection pools in shake flask. Selected subclones have been scaled into small bioreactors in less than 2 months, producing significant quantities of monoclonal antibody using a protocol generic for the parent cell line. The increased efficiency obtained with the UCOE vector reduces the number of transfectants which need to be screened in order to obtain high productivity subclones. Transfection of a standard host cell line, preadapted to grow in a large-scale setting, allows for rapid cell line development decreasing the transition time from research into development and manufacturing. Alternatively, the traditional approach of using a parent cell line which requires serum-free and suspension adaptation after transfection further increases the need for screening a large number of subclones, because many of the subclones will not be able to grow under conditions that allow large-scale protein production. The use of a preadapted cell line can reduce the time required to develop a cell line from months to weeks.
引用
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页码:43 / 46
页数:4
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