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Immobilisation of a repressor protein for binding of plasmid DNA |
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| Authors: | Hasche Anja Voss Carsten |
| Affiliation: | Department of Fermentation Engineering, Faculty of Technology, Bielefeld University, Universitaetsstrasse 25, P.O. Box 100131, D-33501 Bielefeld, Germany. |
| Abstract: | The use of plasmid DNA in gene therapy and genetic vaccination has increased the need for scalable and sustainable production processes. One key challenge for bioprocess engineering is the separation of plasmid DNA from structurally related impurities. Affinity purification procedures allow a highly selective capturing of the target molecule. In this paper, we present the isolation of a his-tagged lac repressor, its non-covalent immobilisation to different matrices and binding of DNA, thus enabling us to screen for combinations of ligands and stationary phases by using a building block principle. |
| Keywords: | ADA adenosine deaminase LacI lac repressor LB Luria Bertani DTT dithiothreitol PMSF phenylmethylsulfonylfluoride SDS-PAGE sodiumdodecylsulfate-polyacrylamide gel electrophoresis IPTG Plasmid DNA Gene therapy Genetic vaccination Protein–DNA interaction Building block principle Plasmid DNA purification |
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