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Off‐line quantitative monitoring of plasmid copy number in bacterial fermentation by capillary electrophoresis
Author(s) -
Breuer Susanne,
Marzban Gorji,
CserjanPuschman Monika,
Dürrschmid Eberhard,
Bayer Karl
Publication year - 1998
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150191420
Subject(s) - capillary electrophoresis , plasmid , chromatography , gel electrophoresis , dna , electrophoresis , resolution (logic) , chemistry , analytical chemistry (journal) , biology , microbiology and biotechnology , biochemistry , computer science , artificial intelligence
Capillary electrophoresis (CE) is an effective instrumental alternative to conventional slab gel electrophoresis in the determination of plasmid copy number during recombinant protein formation processes. This analytical setup provides efficient separation of different species of linearized plasmid molecules and quantification by UV detection. Both fused silica and gel‐filled capillaries are assessed with respect to peak resolution and reproducibility. The application of coated capillaries eliminates the electroosmatic flow to a large extent, resulting in excellent separation of DNA fragments. The application of UV detection enables the analysis of linearized plasmid DNA with a conventional laboratory CE device. All investigated plasmids show good peak resolution due to their significant differences in molecular size, which is essential for sufficient separation of individual DNA molecules.