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Degradation of supercoiled plasmid DNA within a capillary device
Author(s) -
Meacle F.J.,
Zhang H.,
Papantoniou I.,
Ward J.M.,
TitchenerHooker N.J.,
Hoare M.
Publication year - 2006
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.21275
Subject(s) - capillary action , plasmid , breakage , laminar flow , shear stress , chemistry , degradation (telecommunications) , dna supercoil , dna , drop (telecommunication) , chromatography , materials science , mechanics , composite material , dna replication , biochemistry , physics , telecommunications , computer science
Supercoiled plasmid DNA is susceptible to fluid stress in large‐scale manufacturing processes. A capillary device was used to generate controlled shear conditions and the effects of different stresses on plasmid DNA structure were investigated. Computational fluid dynamics (CFD) analysis was employed to characterize the flow environment in the capillary device and different analytical techniques were used to quantify the DNA breakage. It was found that the degradation of plasmid DNA occurred at the entrance of the capillary and that the shear stress within the capillary did not affect the DNA structure. The degradation rate of plasmids was well correlated with the average elongational strain rate or the pressure drop at the entrance region. The conclusion may also be drawn that laminar shear stress does not play a significant role in plasmid DNA degradation. Biotechnol. Bioeng. 2007; 97: 1148–1157. © 2006 Wiley Periodicals, Inc.