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Large-scale purification of pharmaceutical-grade plasmid DNA using tangential flow filtration and multi-step chromatography

Paper ID Volume ID Publish Year Pages File Format Full-Text
20756 43190 2013 6 PDF Available
Title
Large-scale purification of pharmaceutical-grade plasmid DNA using tangential flow filtration and multi-step chromatography
Abstract

The demand for pharmaceutical-grade plasmid DNA in vaccine applications and gene therapy has been increasing in recent years. In the present study, a process consisting of alkaline lysis, tangential flow filtration, purification by anion exchange chromatography, hydrophobic interaction chromatography and size exclusion chromatography was developed. The final product met the requirements for pharmaceutical-grade plasmid DNA. The chromosomal DNA content was <1 μg/mg plasmid DNA, and RNA was not detectable by agarose gel electrophoresis. Moreover, the protein content was <2 μg/mg plasmid DNA, and the endotoxin content was <10 EU/mg plasmid DNA. The process was scaled up to yield 800 mg of pharmaceutical-grade plasmid DNA from approximately 2 kg of bacterial cell paste. The overall yield of the final plasmid DNA reached 48%. Therefore, we have established a rapid and efficient production process for pharmaceutical-grade plasmid DNA.

Keywords
DNA vaccine; Purification; Gene therapy; Chromatography; Alkaline lysis
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Large-scale purification of pharmaceutical-grade plasmid DNA using tangential flow filtration and multi-step chromatography
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 116, Issue 3, September 2013, Pages 281–286
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
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Any Questions? feel free to contact us