The development of a monolith-based purification process for Orthopoxvirus vaccinia virus Lister strain
| Autor: | Yuhong Zhou, Petra Kramberger, David Vincent, Yaohe Wang, Aleš Štrancar, Ajoy Velayudhan, Rosana Hudej |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
viruses Vaccinia virus 01 natural sciences Biochemistry Virus Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound Virology Orthopoxvirus Monolith geography Nuclease Chromatography geography.geographical_feature_category biology Strain (chemistry) Chemistry 010401 analytical chemistry Organic Chemistry General Medicine DNA biology.organism_classification 0104 chemical sciences Oncolytic virus 030104 developmental biology Cell culture biology.protein Vaccinia |
| Zdroj: | Journal of chromatography. A. 1524 |
| ISSN: | 1873-3778 |
| Popis: | The purification of large viruses remains an important field of research and development. The development of efficient purification trains is restricted by limited analytical methods, as well as by the complexity of large viruses, as well as the high variability in starting material from cell culture. Vaccinia virus holds great potential as an oncolytic and immunotherapeutic vaccine against a broad spectrum of cancers. In this work, monolith-based capture and polishing chromatographic steps for vaccinia virus Lister strain has been developed. Virus produced in CV-1 cells was harvested and passed through a 0.8μm pre-filter before loading onto CIEX, AIEX and HIC CIM monoliths. Without the need for nuclease treatment, up to 99% of the total DNA loaded can be removed from the vaccinia feed stream by the CIM OH monolith, which also reduces the total protein concentration in the product pool to LLOQ levels, and achieves infectious virus recoveries of 90%. Binding capacities of greater than 1×109pfu of vaccinia per mL of matrix were obtained on both CIM SO3 and CIM OH monoliths. Multiple orthogonal analytical methods have been used to develop process knowledge and understanding. |
| Databáze: | OpenAIRE |
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