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.

Journal Article | Published | English

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Abstract
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.
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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.
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.
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5 Citations in Europe PMC

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