Citation: Miladys LIMONTA, Lourdes ZUMALACÁRREGUI, Urska VIDIC, Nika LENDERO KRAJNC. Comparison of CIM^® C4-HLD monolithic column with Sartobind phenyl membrane column for pIDKE2 purification[J]. Chinese Journal of Chromatography, ;2017, 35(10): 1028-1036. doi: 10.3724/SP.J.1123.2017.05005 shu

Comparison of CIM^® C4-HLD monolithic column with Sartobind phenyl membrane column for pIDKE2 purification

1.
Center for Genetic Engineering and Biotechnology, Habana 10600, Cuba;
2.
Technical University of Havana "José Antonio Echeverría", Havana 19390, Cuba;
3.
BIA Separations, Ajdovscina SI-5270, Slovenia

Received Date: 8 May 2017

Fund Project: Center for Genetic Engineering and Biotechnology and BIA Separations Fund.

The main component of the Center for Genetic Engineering and Biotechnology (CIGB) candidate vaccine against Hepatitis C virus (HCV) is the pIDKE2 plasmid.The current designed downstream process for the production of pIDKE2 fulfils all regulatory requirements and renders the required quantities of pharmaceutical-grade plasmid DNA (pDNA) with 95% purity.The advantages of this procedure include high plasmid purity and the elimination of undesirable additives,such as toxic organic extractants and animal-derived enzymes.However,yields and consequently the productivity of the process are low.Previous work demonstrated that the most critical step of the process is the reverse phase chromatography,where conventional porous particle resins are used.Therefore,to increase the process productivity,alternative technologies such as membranes and chromatographic monoliths were tested as alternative options for this critical step.Here,a comparison between the behaviors of CIM^® C4-HLD and Sartobind phenyl matrices was performed.To obtain higher productivities and purities,the dynamic binding capacities and selectivities were evaluated.The results showed that both matrices had a similar capacity for pIDKE2 plasmid,but the separation of pDNA isoforms using CIM technology was much better than that with Sartobind.Additionally,the optimal conditions for loading plasmid DNA on a CIM^® C4-HLD 800-mL monolithic column in a real production process were determined.These optimizations will allow production levels to satisfy the high plasmid consumption demanded by clinical trials.
- membrane,
- monolith,
- downstream processing,
- plasmid DNA (pDNA),
- breakthrough,
- supercoiled (SC) plasmid,
- open circular (OC) plasmid
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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Comparison of CIM® C4-HLD monolithic column with Sartobind phenyl membrane column for pIDKE2 purification

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通讯作者: 陈斌, bchen63@163.com

Comparison of CIM^® C4-HLD monolithic column with Sartobind phenyl membrane column for pIDKE2 purification