Meacle F J, Zhang H, Papantoniou I, Ward J M, Titchener-Hooker N J, Hoare M
The Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London, United Kingdom.
Biotechnol Bioeng. 2007 Aug 1;97(5):1148-57. doi: 10.1002/bit.21275.
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.
在大规模生产过程中,超螺旋质粒DNA易受流体应力影响。使用毛细管装置产生可控的剪切条件,并研究不同应力对质粒DNA结构的影响。采用计算流体动力学(CFD)分析来表征毛细管装置内的流动环境,并使用不同的分析技术来量化DNA断裂情况。研究发现,质粒DNA的降解发生在毛细管入口处,且毛细管内的剪切应力不影响DNA结构。质粒的降解速率与入口区域的平均伸长应变率或压降密切相关。还可以得出层流剪切应力在质粒DNA降解中不起重要作用的结论。