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
相关论文
共 50 条
  • [31] Transgene expression of transfected supercoiled plasmid DNA concatemers in mammalian cells
    Escoffre, Jean-Michel
    Bellard, Elisabeth
    Golzio, Muriel
    Teissie, Justin
    Rols, Marie-Pierre
    JOURNAL OF GENE MEDICINE, 2009, 11 (11): : 1071 - 1073
  • [32] Transgene expression of transfected supercoiled plasmid DNA concatemers in mammalian cells
    Maucksch, Christof
    Bohla, Alexander
    Hoffmann, Florian
    Schleef, Martin
    Aneja, Manish Kumar
    Elfinger, Markus
    Hartl, Dominik
    Rudolph, Carsten
    JOURNAL OF GENE MEDICINE, 2009, 11 (05): : 444 - 453
  • [33] Isolation of ultrapure supercoiled plasmid-DNA using preparative electrophoresis
    Roeder, T
    Görich, D
    Roeder, NP
    Bruchhaus, I
    ELECTROPHORESIS, 1998, 19 (10) : 1575 - 1576
  • [34] PARTIALLY SUPERCOILED REPLICATION INTERMEDIATES OF R PLASMID DNA ARE RESISTANT TO RELAXATION
    WOMBLE, DD
    WARREN, RL
    ROWND, RH
    NATURE, 1976, 264 (5587) : 676 - 678
  • [35] Variety of molecular conformation of plasmid pUC18 DNA and solenoidally supercoiled DNA
    Huang, XT
    Wang, ZQ
    Wu, YW
    Fan, TY
    Wang, SR
    Wang, XK
    SCIENCE IN CHINA SERIES C-LIFE SCIENCES, 1996, 39 (06): : 571 - 583
  • [36] Capillary electrophoresis of supercoiled DNA in dilute polymer solutions.
    Hammond, RW
    Oana, H
    Schwinefus, JJ
    Wang, SC
    Bonadio, J
    Levy, RJ
    Morris, MD
    ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 1997, 213 : 3 - ANYL
  • [37] Biorecognition of supercoiled plasmid DNA isoform in lysine-affinity chromatography
    Sousa, A.
    Sousa, F.
    Queiroz, J. A.
    JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, 2009, 877 (27): : 3257 - 3260
  • [38] INTERACTION INVITRO OF NONEPITHELIAL INTERMEDIATE FILAMENT PROTEINS WITH SUPERCOILED PLASMID DNA
    KUHN, S
    VORGIAS, CE
    TRAUB, P
    JOURNAL OF CELL SCIENCE, 1987, 87 : 543 - 554
  • [39] Separation and purification of supercoiled plasmid DNA by mixed-mode chromatography
    Zhang P.
    Tan Y.
    Sun Y.
    Zhang Q.
    Yao S.
    Lin D.
    Gao Xiao Hua Xue Gong Cheng Xue Bao/Journal of Chemical Engineering of Chinese Universities, 2023, 37 (05): : 806 - 812
  • [40] A chromatographic method for determination of supercoiled plasmid DNA concentration in complex solutions
    Bennemo, Mia
    Blom, Hans
    Emilsson, Anna
    Lemmens, Raf
    JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, 2009, 877 (24): : 2530 - 2536