Off-line quantitative monitoring of plasmid copy number in bacterial fermentation by capillary electrophoresis.
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.
Breuer
S
S
Institute of Applied Microbiology, University of Agricultural Sciences, Vienna, Austria.
Marzban
G
G
Cserjan-Puschman
M
M
Dürrschmid
E
E
Bayer
K
K
D001419:Bacteria / Q000378:metabolism*; D010582:Bacteriophage lambda / Q000235:genetics; D002021:Buffers; D004279:DNA, Viral / Q000302:isolation & purification; D015247:Deoxyribonuclease HindIII / Q000378:metabolism; D019075:Electrophoresis, Capillary / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics; D005285:Fermentation; D005782:Gels; D008970:Molecular Weight; D010957:Plasmids / Q000032:analysis* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis*; D012822:Silicon Dioxide
D002021:Buffers; D004279:DNA, Viral; D005782:Gels; D011994:Recombinant Proteins; D012822:Silicon Dioxide; D015247:Deoxyribonuclease HindIII