Abstract
Purification of high-quality plasmid DNA in large quantities is a crucial step in its production for therapeutic use and is usually conducted by different chromatographic techniques. Large-scale preparations require the optimization of yield and homogeneity, while maximizing removal of contaminants and preserving molecular integrity. The advantages of Convective Interaction Media® (CIM®) monolith stationary phases, including low backpressure, fast separation of macromolecules, and flow-rate-independent resolution qualified them to be used effectively in separation of plasmid DNA on laboratory as well as on large scale. A development and scale-up of plasmid DNA downstream process based on chromatographic monoliths is described and discussed below. Special emphasis is put on the introduction of process analytical technology principles and tools for optimization and control of a downstream process.
Key words
- Chromatography
- Column
- Downstream process
- Large-scale
- Monolith
- Pharmaceutical grade
- Plasmid DNA manufacturing
- Plasmid DNA purification
- Process analytical control
- Scale-up
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Acknowledgments
We thank the research and application team of BIA Separations, Ajdovščina, Slovenia, for contributing work and discussions, especially Franci Smrekar, Boštjan Gabor, Nika Lendero Krajnc, Miloš Barut, and Aleš Podgornik for their crucial roles in pDNA DSP development; Tomas Kostelec for his support with figures and text revision.
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Černigoj, U., Štrancar, A. (2021). Scale-Up of Plasmid DNA Downstream Process Based on Chromatographic Monoliths. In: Sousa, Â. (eds) DNA Vaccines. Methods in Molecular Biology, vol 2197. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0872-2_9
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DOI: https://doi.org/10.1007/978-1-0716-0872-2_9
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