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Plasmid copy number and plasmid stability.

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1
Universit├Ąt Bielefeld AG, Fermentationstechnik, Technische Fakult├Ąt, 33501 Bielefeld, Germany. karl.friehs@uni-bielefeld.de

Abstract

Many expression systems in research and industry use plasmids as vectors for the production of recombinant proteins or non-proteinous recombinant substances. Plasmids have an essential impact on productivity. Related factors are plasmid copy number, structural plasmid stability and segregational plasmid stability. Plasmid copy number determines the gene dosage accessible for expression and many plasmids lead generally to a high productivity. To analyze an expression system the quantification of plasmid copy number is very helpful. Therefore, different methods for the determination of plasmid copy number are described. Structural plasmid stability exists, when all generated plasmids have the correct base sequence. The analysis of structural instabilities is not trivial and some methods are reported. When all daughter cells get at least one plasmid during cell division, the culture is segregational stable. The development of plasmid free cells can lead to a significant loss in productivity. Different methods for lab scale and industrial scale help to avoid segregational instability. Since plasmids are used as pharmaceuticals, additional aspects of stability have to be taken into account. These include stability during downstream processing, stability after application and stability during storage and shipping.

PMID:
15088763
[Indexed for MEDLINE]
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