Protein expression in baculovirus vector expression system

A baculovirus, recombinant baculovirus technology, applied in the field of enhancing protein or polypeptide expression, can solve the problem of not considering DNA and RNA accumulation and infection cycle, etc.

Inactive Publication Date: 2006-11-08
STICHTING INST VOOR DIERHOUDERIJ & DIERGEZONDHEID +1
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  • Summary
  • Abstract
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Problems solved by technology

To a large extent, when viewed as black-box systems, they generalize our current quantitative understanding of the interaction between baculoviruses and insect cells, without accounting for DNA and RNA accumulation and infection cycles,

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  • Protein expression in baculovirus vector expression system
  • Protein expression in baculovirus vector expression system
  • Protein expression in baculovirus vector expression system

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[0035] specific implementation plan

[0036] Pestiviruses of the Flaviviridae family generally include classical swine fever virus (CSFV), border disease virus (BDV), and bovine viral diarrhea virus (BVDV). The genomes of several BVDV and CSFV strains have been sequenced (Renard et al., 1987, European Patent Application 0208672; Collett et al., 1988, Virology 165, 191-199; Deng and Brock, 1992, Virology 1991, 865- 679; Meyers et al., 1989, Virology 171, 555-567; Moormann et al., 1990, Virology 177, 184-188). For BDV, only incomplete genome nucleotide sequences were available. The pestivirus genome is a positive-strand RNA molecule of about 12.5 kb containing a large open reading frame. The open reading frame translates into a hypothetical polyprotein of approximately 4,000 amino acids, which is processed by viral and cellular-encoded proteases. The open reading frame is flanked by two small, highly conserved untranslated regions that may be involved in genome replication. ...

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Abstract

The present invention relates to methods of enhancing protein expression in baculovirus expression systems. The present invention provides a method for producing a recombinant protein in insect cell culture comprising selecting a recombinant baculovirus expressing said protein, growing insect cells in a sufficient volume of culture vessel containing at least 2 liters of growth medium, and using at least An inoculum of baculovirus infects cells at a density of 1 x 10 at a multiplicity of infection <0.01 5 to 5×10 6 cells / ml of cells. The present invention further provides for the production of recombinant pestiviruses E2 or E in insect cell cultures having a final concentration of said protein (fragment) in the growth medium at harvest of at least 100 μg / ml ms protein or fragment method. The present invention further provides a method for producing recombinant follicle stimulating hormone, its alpha subunit and / or beta subunit or complexes and fragments thereof, at a concentration of 15 micrograms / ml in the growth medium at harvest.

Description

technical field [0001] The present invention relates to a method for enhancing protein or polypeptide expression in a baculovirus vector expression system. When recombinant DNA technology was developed, it was expected to use genetically modified bacteria to produce proteins on a large scale. However, most commercially valuable proteins require post-translational modifications before they can become biologically active. So, now more often than not, animal cells are used for the production of recombinant proteins. Background technique [0002] Among animal cells, insect cells have become increasingly important in the production of recombinant proteins. A convenient and versatile baculovirus vector system using insect cells has now been developed. However, the physiological information of insect cells is quite scarce, and vaccines produced by baculovirus recombinant technology are generally accepted. One example is the gp160 envelope protein expressed by the baculovirus of...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63C12N7/00C12P21/00A61K39/12C12N15/40C07K14/185C12N15/09A61K39/00A61K39/187A61P31/12A61P31/14C07K14/18C12N15/866C12P21/02
CPCA61K39/00C07K14/005C12N15/86C12N2710/14143C12N2770/24322C12N2770/24334A61P15/00A61P31/12A61P31/14C12N15/63
Inventor 迪特马尔·克雷茨多恩迪尔克·弗朗西斯库斯·马里纳斯·范德维尔亚伯拉罕·约翰尼斯·德斯米特罗伯特茨·雅各布斯·玛丽亚·莫尔曼埃里克·谢斯·阿曼
Owner STICHTING INST VOOR DIERHOUDERIJ & DIERGEZONDHEID
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