Degradation of supercoiled plasmid DNA within a capillary device

被引:14
|
作者
Meacle, F. J.
Zhang, H.
Papantoniou, I.
Ward, J. M.
Titchener-Hooker, N. J.
Hoare, M.
机构
[1] UCL, Adv Ctr Biochem Engn, London WC1E 7JE, England
[2] Merck Res Labs, West Point, PA USA
[3] UCL, Dept Biochem & Mol Biol, London WC1E 6BT, England
基金
英国工程与自然科学研究理事会;
关键词
plasmid DNA; capillary device; shear rate; breakage; computational fluid dynamics;
D O I
10.1002/bit.21275
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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.
引用
收藏
页码:1148 / 1157
页数:10
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