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Titolo:
A novel autoregulated proliferation-controlled production process using recombinant CHO cells
Autore:
Mazur, X; Eppenberger, HM; Bailey, JE; Fussenegger, M;
Indirizzi:
ETH Zurich, Inst Biotechnol, CH-8093 Zurich, Switzerland ETH Zurich Zurich Switzerland CH-8093 chnol, CH-8093 Zurich, Switzerland
Titolo Testata:
BIOTECHNOLOGY AND BIOENGINEERING
fascicolo: 2, volume: 65, anno: 1999,
pagine: 144 - 150
SICI:
0006-3592(19991020)65:2<144:ANAPPP>2.0.ZU;2-1
Fonte:
ISI
Lingua:
ENG
Soggetto:
HAMSTER OVARY CELLS; TETRACYCLINE-REGULATED SYSTEM; CONTROLLED GENE-EXPRESSION; GLUTAMINE-SYNTHETASE GENE; HIGH-LEVEL EXPRESSION; PROTEIN-FREE MEDIUM; MAMMALIAN-CELLS; BATCH CULTURE; PRODUCTION KINETICS; INTERFERON-GAMMA;
Keywords:
bioprocesses; heterologous protein; proliferation;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
53
Recensione:
Indirizzi per estratti:
Indirizzo: Fussenegger, M ETH Zurich, Inst Biotechnol, CH-8093 Zurich, Switzerland ETH Zurich Zurich Switzerland CH-8093 Zurich, Switzerland
Citazione:
X. Mazur et al., "A novel autoregulated proliferation-controlled production process using recombinant CHO cells", BIOTECH BIO, 65(2), 1999, pp. 144-150

Abstract

Controlled proliferation bioprocesses have shown great enhancement of heterologous protein production. This novel technology has been implemented here using a multicistronic expression unit encoding the product gene and a cytostatic cell-cycle-arresting gene (p27) under control of a single tetracycline-repressible (tet(off)) promoter. The strict genetic linkage of both genes allows the dissection of the production process into a nonproductive growth phase (dicistronic expression unit repressed) followed by a proliferation-inhibited production phase (dicistronic expression unit induced) when the cells have reached an optimal cell density. Based on rapid degradation of the external repressible agents tetracycline (tet) and doxycycline (dox) in the cell culture medium, we developed a self-regulated process for transition from the growth phase to the production phase in a fashion that is dependent only on the starting cell population and the initial concentration of the tetracyclines. With this process, no change in medium is required toaccomplish the transition from growth to production phase. The two-phase bioprocess achieved here by tet switch-controlled proliferation is reliable and allows a growth-arrested production phase of at least 7 days, during which cells remain in a well-defined, highly viable physiological state and show enhanced heterologous protein production. This Tet(SWITCH) process is readily adaptable to a variety of industrial processes designed for production of difficult-to-express protein pharmaceuticals. (C) 1999 John Wiley & Sons, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 23/09/20 alle ore 21:06:42