Production of supercoiled multimeric plasmid DNA for biopharmaceutical application

Production of nucleic acids as an active pharmaceutical ingredient (API) in gene therapy and genetic vaccination is gaining more and more importance. Non-viral vectors like plasmid DNA are currently investigated in various clinical trials. Supercoiled multimeric plasmids are of particular interest f...

Full description

Saved in:
Bibliographic Details
Published in:Journal of biotechnology 2003-11, Vol.105 (3), p.205-213
Main Authors: Voß, Carsten, Schmidt, Torsten, Schleef, Martin, Friehs, Karl, Flaschel, Erwin
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biopharmaceutical
Biopharmaceutics
Bioreactors
Biotechnology
Biotechnology - methods
Cloning, Molecular
Concatemer
Cultivation
Culture Media
DNA Replication
DNA, Superhelical - chemistry
DNA, Superhelical - genetics
DNA, Superhelical - metabolism
Escherichia coli - genetics
Escherichia coli - growth & development
Fundamental and applied biological sciences. Psychology
Gene therapy
Genetic vaccination
Genetic Vectors
nucleic acids
Plasmid
Plasmids
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Production of nucleic acids as an active pharmaceutical ingredient (API) in gene therapy and genetic vaccination is gaining more and more importance. Non-viral vectors like plasmid DNA are currently investigated in various clinical trials. Supercoiled multimeric plasmids are of particular interest for pharmaceutical purpose because they contain multiple copies of a therapeutic gene and can therefore be more efficient vectors. A process for the preparation of Escherichia coli strains replicating dimers, trimers, and tetramers of a 4.6 kb plasmid is presented. Cultivation of these clones on semi-defined glycerol medium in a 7 l bioreactor shows structural stability of dimers and trimers during the whole cultivation process. Plasmid concentrations and selectivities are compared to the corresponding cultivation with the plasmid monomer. Cultivation of the tetramer replicating strain shows a disintegration of the plasmid multimer and reconstitution of the monomer and smaller multimers.
ISSN:0168-1656
1873-4863
DOI:10.1016/j.jbiotec.2003.07.004