| DOI | Resolve DOI: https://doi.org/10.1016/j.jviromet.2010.01.008 |
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| Author | Search for: Dormond, Edwige1; Search for: Chahal, Parminder1; Search for: Bernier, Alice1; Search for: Tran, Rosa1; Search for: Perrier, Michel; Search for: Kamen, Amine1 |
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| Affiliation | - National Research Council of Canada. NRC Biotechnology Research Institute
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| Format | Text, Article |
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| Subject | Helper-dependent adenoviral vector; Purification; Iodixanol; Ultracentrifugation; Chromatography |
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| Abstract | The preparation of large amount of purified helper-dependent adenoviral vector material is hampered by the lack of development of downstream processes with proven records on separation and recovery efficiencies. In order to facilitate the use of clinical-grade helper-dependent virus material for large-scale in vivo studies, a three-step purification scheme consisting of (1) an anion-exchange chromatography for initial capturing of virus, (2) a shallow iodixanol density gradient ultracentrifugation for the removal of helper virus from helper-dependent virus, and (3) a size-exclusion chromatography for the removal of iodixanol and residual protein contaminants as a polishing step was developed. The use of a fast iodixanol density ultracentrifugation step was highly effective in separating infectious helper-dependent virus from contaminating helper virus. The overall downstream processing scheme gave 80% infectious particle yield. The contamination ratio of helper virus in the helper-dependent virus preparation are reduced from 2.57 to 0.03% corresponding to a reduction of helper virus by factors of 85 by two iodixanol purification steps. It was also demonstrated that size-exclusion chromatography is an excellent step for the removal of iodixanol and polishing of the final helper-dependent virus preparation |
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| Publication date | 2010-04-01 |
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| In | |
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| Language | English |
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| Peer reviewed | Yes |
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| NPARC number | 16225326 |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction (opens in a new tab) |
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| Record identifier | 9defe678-fa80-452e-a6d5-2a8094ad5593 |
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| Record created | 2010-11-05 |
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| Record modified | 2020-04-17 |
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