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Plasmid pVAX1-NH36 purification by membrane and bead perfusion chromatography
- Source :
- Bioprocess and biosystems engineering. 40(3)
- Publication Year :
- 2016
-
Abstract
- The demand for plasmid DNA (pDNA) has increased in response to the rapid advances in vaccines applications to prevent and treat infectious diseases caused by virus, bacteria or parasites, such as Leishmania species. The immunization protocols require large amounts of supercoiled plasmid DNA (sc-pDNA) challenging the development of efficient and profitable processes for capturing and purified pDNA molecules from large volumes of lysates. A typical bioprocess involves four steps: fermentation, primary recovery, intermediate recovery and final purification. Ion-exchange chromatography is one of the key operations in the purification schemes of pDNA owing the chemical structure of these macromolecules. The goal of this research was to compare the performance of the final purification step of pDNA using ion-exchange chromatography on columns packed with Mustang Q membranes or perfusive beads POROS 50 HQ. The experimental results showed that both matrixes could separate the plasmid pVAX1-NH36 (3936 bp) from impurities in clarified Escherichia coli lysates with an adequate resolution. In addition, a 24- and 21-fold global purification factor was obtained. An 88 and 63% plasmid recuperation was achieved with ion-exchange membranes and perfusion beads, respectively. A better understanding of perfusion-based matrices for the purification of pDNA was developed in this research.
- Subjects :
- 0106 biological sciences
0301 basic medicine
Macromolecular Substances
Bioengineering
medicine.disease_cause
01 natural sciences
Chromatography, Affinity
03 medical and health sciences
chemistry.chemical_compound
Industrial Microbiology
Plasmid
010608 biotechnology
medicine
Escherichia coli
Bioprocess
Chromatography, High Pressure Liquid
Electrophoresis, Agar Gel
Leishmania
Vaccines
Chromatography
biology
Membranes, Artificial
General Medicine
DNA
biology.organism_classification
Chromatography, Ion Exchange
Perfusion
030104 developmental biology
Membrane
Biochemistry
chemistry
Fermentation
Adsorption
Industrial and production engineering
Bacteria
Biotechnology
Plasmids
Subjects
Details
- ISSN :
- 16157605
- Volume :
- 40
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- Bioprocess and biosystems engineering
- Accession number :
- edsair.doi.dedup.....11d78524c4c72642fac6b3500463dd75