Salt-induced changes in plasmid DNA transmission through ultrafiltration membranes.

Abstract:

:Recent studies have demonstrated the feasibility of using ultrafiltration for the purification of plasmid DNA, but there is still little understanding of the factors governing DNA transmission. Experimental data were obtained for the transmission of a 3.0 kbp supercoiled plasmid DNA through composite regenerated cellulose ultrafiltration membranes as a function of solution ionic environment in a stirred ultrafiltration cell. The dependence on salt concentration was quite dramatic, with the sieving coefficient increasing by more than 80-fold as the NaCl concentration increased from 1 to 150 mM at a fixed filtrate flux. At the same total ionic strength, the sieving coefficient in an MgCl2 solution was significantly larger than that evaluated in NaCl. The sieving results are consistent with independent studies showing a reduction in the effective plasmid size due to salt specific shielding of intramolecular electrostatic interactions. DNA transmission was also a strong function of the filtrate flux, with negligible transmission below a critical value of the flux. The predicted values of the critical filtrate flux determined using a modified elongational flow model were in excellent agreement with the experimental data. These results clearly demonstrate that salt-induced changes in plasmid DNA structure have a significant effect on plasmid DNA transmission through ultrafiltration membranes.

journal_name

Biotechnol Bioeng

authors

Latulippe DR,Zydney AL

doi

10.1002/bit.21575

subject

Has Abstract

pub_date

2008-02-01 00:00:00

pages

390-8

issue

2

eissn

0006-3592

issn

1097-0290

journal_volume

99

pub_type

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