Removal of RNA impurities by tangential flow filtration in an RNase-free plasmid DNA purification process

Eon‐Duval, Alex; MacDuff, Robert H; Fisher, C. A.; Harris, Mark; Brook, Chris

doi:10.1016/s0003-2697(03)00050-2

Cited by 63 publications

(48 citation statements)

References 19 publications

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“…These therapeutic applications require high amounts of highly pure pDNA. There is a variety of purification strategies available for the purification of pDNA using conventional methods, including tangential flow filtration (TFF) [59][60][61], membrane adsorption [62][63][64] and chromatography [65,66]. Monolithic columns with different functionalities have also been tested for the purification of pDNA.…”

Section: Nucleic Acid Purificationmentioning

confidence: 99%

Monoliths in Bioprocess Technology

2015

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Monolithic columns are a special type of chromatography column, which can be used for the purification of different biomolecules. They have become popular due to their high mass transfer properties and short purification times. Several articles have already discussed monolith manufacturing, as well as monolith characteristics. In contrast, this review focuses on the applied aspect of monoliths and discusses the most relevant biomolecules that can be successfully purified by them. We describe success stories for viruses, nucleic acids and proteins and compare them to conventional purification methods. Furthermore, the advantages of monolithic columns over particle-based resins, as well as the limitations of monoliths are discussed. With a compilation of commercially available monolithic columns, this review aims at serving as a 'yellow pages' for bioprocess engineers who face the challenge of purifying a certain biomolecule using monoliths.

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Section: Nucleic Acid Purificationmentioning

confidence: 99%

Monoliths in Bioprocess Technology

2015

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“…However, some studies (Eon-Duval et al 2003) have shown that by combining the precipitating effect of calcium chloride salt on high-molecular weight RNA with the clearance of low-molecular-weight RNA by tangential flow filtration (TFF), it is possible to reduce RNA to undetectable levels. Despite these promising results, plasmid size and degree of supercoiling may affect the performance of TFF and therefore conditions may have to be reassessed individually for each plasmid.…”

Section: Primary Isolationmentioning

confidence: 99%

Biomedical application of plasmid DNA in gene therapy: A new challenge for chromatography

Sousa

Passarinha²,

Queiroz³

2009

Biotechnology and Genetic Engineering Reviews

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Gene therapy and DNA vaccination are clinical fields gradually emerging in the last few decades, in particular after the discovery of some gene-related diseases. The increased relevance of biomedical applications of plasmid DNA (pDNA) to induce therapeutic effects has had a great impact on biopharmaceutical research and industry. Although there are several steps involved in the pDNA manufacturing process, the several unit operations must be designed and integrated into a global process. After the plasmid has been designed according to the requirements for clinical administeration to humans, it is biosynthesised mainly by an E. coli host. The overriding priority of the production process is to improve plasmid quantity -the production conditions need to be optimised to guarantee pDNA stability and biological activity.The complexity and diversity of biomolecules present on the pDNA-containing extracts represent the main concern and limitation to achieve pure and biologically active pDNA. There has been a recent intenstification of the improvement of existing purification procedures or the establishment of novel schemes for plasmid purification.

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“…By using calcium chloride as a precipitation agent, the authors were able to remove a large quantity of the contaminating RNA and keep loss of product low. Subsequent ultrafiltration using a 100 kDa membrane resulted in plasmid DNA that was essentially free of RNA [30]. The cationic surfactant cetyltrimethylammoniumbromide (CTAB) was used for selective precipitation of plasmid DNA by Lander et al [31].…”

Section: Separation Of Plasmid Dna and Rnamentioning

confidence: 99%

Downstream Processing of Plasmid DNA for Gene Therapy and Genetic Vaccination

Voss

2008

Chem Eng & Technol

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Plasmid DNA is used as a cloning vector to deliver recombinant genetic information into microorganisms. Since the 1990s, this principle has also been applied for the delivery of therapeutic genes in gene therapy and genetic vaccination. This non-viral gene delivery is afflicted with fewer safety concerns in comparison to viral systems. Processes for the production of high-quality plasmid DNA at multiand kilogram scale are necessary to meet the needs of clinical trials as well as future therapeutics. Cell disruption, the separation of structurally-related impurities and analytical techniques for process and quality control are the main challenges for bioengineering. This review summarizes the development in these fields over the past recent years.

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Removal of RNA impurities by tangential flow filtration in an RNase-free plasmid DNA purification process

Cited by 63 publications

References 19 publications

Monoliths in Bioprocess Technology

Monoliths in Bioprocess Technology

Biomedical application of plasmid DNA in gene therapy: A new challenge for chromatography

Downstream Processing of Plasmid DNA for Gene Therapy and Genetic Vaccination

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