Production of recombinant RNase Ba and its application in downstream processing of plasmid DNA for pharmaceutical use

Voß C, Lindau D, Flaschel E (2006)
BIOTECHNOLOGY PROGRESS 22(3): 737-744.

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Zeitschriftenaufsatz | Veröffentlicht | Englisch
Abstract / Bemerkung
The demand for new strategies in downstream processing of biopharmaceutical plasmid DNA has increased in response to the importance of nucleic acids as active pharmaceutical ingredients (API) in gene therapy and genetic vaccination. Led by the problematic usage of animal-derived proteins for producing reagents of clinical applications, we present an opportunity of removing RNA prior to chromatographic steps by using a recombinant RNase Ba (barnase of Bacillus amyloliquefaciens) as an alternative to bovine RNase A. An expression vector for RNase Ba production was constructed enabling periplasmic localization of the recombinant protein. Cultivation of the RNase-producing clone showed stable activity (3.6 kU mL(-1) during stationary phase) throughout the cultivation process. After purification the RNase activity was tested and compared to that of commercially available RNase A. RNase Ba showed no DNase activity even after prolonged incubation with plasmid DNA. Thus, it is a suitable substitute for bovine RNase A in pharmaceutical purification processes.
Erscheinungsjahr
Zeitschriftentitel
BIOTECHNOLOGY PROGRESS
Band
22
Zeitschriftennummer
3
Seite
737-744
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Voß C, Lindau D, Flaschel E. Production of recombinant RNase Ba and its application in downstream processing of plasmid DNA for pharmaceutical use. BIOTECHNOLOGY PROGRESS. 2006;22(3):737-744.
Voß, C., Lindau, D., & Flaschel, E. (2006). Production of recombinant RNase Ba and its application in downstream processing of plasmid DNA for pharmaceutical use. BIOTECHNOLOGY PROGRESS, 22(3), 737-744. doi:10.1021/bp050417e
Voß, C., Lindau, D., and Flaschel, E. (2006). Production of recombinant RNase Ba and its application in downstream processing of plasmid DNA for pharmaceutical use. BIOTECHNOLOGY PROGRESS 22, 737-744.
Voß, C., Lindau, D., & Flaschel, E., 2006. Production of recombinant RNase Ba and its application in downstream processing of plasmid DNA for pharmaceutical use. BIOTECHNOLOGY PROGRESS, 22(3), p 737-744.
C. Voß, D. Lindau, and E. Flaschel, “Production of recombinant RNase Ba and its application in downstream processing of plasmid DNA for pharmaceutical use”, BIOTECHNOLOGY PROGRESS, vol. 22, 2006, pp. 737-744.
Voß, C., Lindau, D., Flaschel, E.: Production of recombinant RNase Ba and its application in downstream processing of plasmid DNA for pharmaceutical use. BIOTECHNOLOGY PROGRESS. 22, 737-744 (2006).
Voß, Carsten, Lindau, D, and Flaschel, Erwin. “Production of recombinant RNase Ba and its application in downstream processing of plasmid DNA for pharmaceutical use”. BIOTECHNOLOGY PROGRESS 22.3 (2006): 737-744.

7 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Microbial ribonucleases (RNases): production and application potential
Hameş EE, Demir T., World J Microbiol Biotechnol 31(12), 2015
PMID: IND604418260
Barnase and binase: twins with distinct fates.
Ulyanova V, Vershinina V, Ilinskaya O., FEBS J 278(19), 2011
PMID: 21824291
On the stability of plasmid DNA vectors during cell culture and purification.
Freitas SS, Azzoni AR, Santos JA, Monteiro GA, Prazeres DM., Mol Biotechnol 36(2), 2007
PMID: 17914194
Reverse micellar extraction systems for the purification of pharmaceutical grade plasmid DNA.
Streitner N, Voss C, Flaschel E., J Biotechnol 131(2), 2007
PMID: 17673324

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