Prediction of Shear Damage of Plasmid DNA in Pump and Centrifuge Operations Using an Ultra Scale-Down Device

详细信息    查看全文

• 作者：Hu Zhang ; Simyee Kong ; Andrew Booth ; Rihab Boushaba ; M. Susana Levy ; Michael Hoare
• 刊名：Biotechnology Progress
• 出版年：2007
• 出版时间：August 2007
• 年：2007
• 卷：23
• 期：4
• 页码：858 - 865
• 全文大小：249K
• 年卷期：v.23,no.4(August 2007)
• ISSN：1520-6033

文摘

Supercoiled circular (SC) plasmid DNA is often subjected to fluid stress in large-scalemanufacturing processes. It is thus important to characterize the engineering environment withina particular unit operation as well as within the associated ancillary equipment during processdesign 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 engineeringenvironments experienced by biomolecules within a range of manufacturing-scale ancillary,primary recovery, and purification operations, using milliliter quantities of material. Throughthe use of a USD shear device, the effect of elongational strain rate on SC plasmid DNAdegradation was studied in this paper, and from that, the impact of a centrifugal pump, a Monopump, and a disk-stack centrifuge feed zone on SC plasmid DNA degradation was predictedand experimentally verified at scale. Model predictions, over the range of conditions studied,were in good agreement with experimental values, demonstrating the potential of the USDapproach as a decisional tool during bioprocess design.