EXPRESSION OF HUMAN INTERFERON-GAMMA GENE IN HUMAN TISSUE CULTURE CELLS

被引:6
|
作者
Leister, Peter [1 ]
Tileva, Milena [2 ]
Krachmarova, Elena [2 ]
Nacheva, Genoveva [2 ]
机构
[1] InVivo Biotech Serv GmbH, Hennigsdorf, Germany
[2] Bulgarian Acad Sci, Inst Mol Biol Roumen Tsanev, Sofia, Bulgaria
关键词
human interferon-gamma; hIFN gamma; eukaryotic expression; transient expression; CAP-T cell line; HEKF cell line; CHEMICALLY SYNTHESIZED GENE; HUMAN IMMUNE INTERFERON; ESCHERICHIA-COLI; N-GLYCOSYLATION; BATCH CULTURE; JAK-STAT; PROTEINS; GROWTH; ACTIVATION; CYTOKINES;
D O I
10.5504/BBEQ.2012.0128
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
Interferon-gamma is a cytokine secreted by natural killer cells, CD4 T(H)1 cells and CD8 cytotoxic lymphocytes in response to mitogenic and antigenic stimuli. It is endowed with multiple biological activities and because of that is one of the first targets of recombinant DNA technology. Human interferon-gamma (hIFN gamma) is expressed so far in Escherichia coli and various eukaryotic hosts. The disadvantages of the bacterial hIFN gamma and unsatisfactory results from the eukaryotic expression were the reason to employ transient expression of the hIFN gamma gene in CAP-T and HEKF cells, widely used for industrial production of human proteins. The obtained results demonstrated that (unlike with other human genes): i) cell viability decreased very fast and reached the critical level of 50 % after five days only; ii) cell density did not exceed 0.7x10(5) cells/mL and sharply decreased after four days of cultivation; iii) the yield of hIFN gamma secreted in the supernatant did not exceed 5.8 mg/L. Biotechnol. & Biotechnol. Eq. 2013, 27(1), 3573-3576
引用
收藏
页码:3573 / 3576
页数:4
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