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|Title:||Prediction of shear damage of plasmid DNA in pump and centrifuge operations using an ultra scale-down device|
|Citation:||Biotechnology Progress, 2007; 23(4):858-865|
|Publisher:||Amer Chemical Soc|
|Hu Zhang, Simyee Kong, Andrew Booth, Rihab Boushaba, M. Susana Levy, Michael Hoare|
|Abstract:||Supercoiled circular (SC) plasmid DNA is often subjected to fluid stress in large-scale manufacturing processes. It is thus important to characterize the engineering environment within a particular unit operation as well as within the associated ancillary equipment during process design for plasmid DNA manufacture so as to avoid shear-induced degradation of the SC isoform, which would compromise product efficacy in therapeutic applications. In the past few years, ultra scale-down (USD) tools were developed within our laboratory to mimic the engineering environments experienced by biomolecules within a range of manufacturing-scale ancillary, primary recovery, and purification operations, using milliliter quantities of material. Through the use of a USD shear device, the effect of elongational strain rate on SC plasmid DNA degradation was studied in this paper, and from that, the impact of a centrifugal pump, a Mono pump, and a disk-stack centrifuge feed zone on SC plasmid DNA degradation was predicted and experimentally verified at scale. Model predictions, over the range of conditions studied, were in good agreement with experimental values, demonstrating the potential of the USD approach as a decisional tool during bioprocess design.|
Electrophoresis, Agar Gel
|Appears in Collections:||Aurora harvest 5|
Chemical Engineering publications
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