| Download | - View accepted manuscript: Development and validation of a HPLC method for the quantification of baculovirus particles (PDF, 819 KiB)
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| DOI | Resolve DOI: https://doi.org/10.1016/j.jchromb.2010.11.011 |
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| Author | Search for: Transfiguracion, Julia1; Search for: Mena, Jimmy A.2; Search for: Aucoin, Marc G.; Search for: Kamen, Amine A.1 |
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| Affiliation | - National Research Council of Canada. NRC Biotechnology Research Institute
- National Research Council of Canada
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| Format | Text, Article |
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| Subject | Baculovirus; HPLC; Quantification; Total virus particle; Viral genome labeling |
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| Abstract | A HPLC method using an anion exchange column was developed for the quantification of baculovirus particles. To properly detect the virus eluting from the column, a nucleic acid dye was used to amplify the signal projected by the virus. The viral genome was labeled by incubating the virus with SYBR Green I at 37-¦C for a minimum of 1h. The virus was specifically eluted from the contaminants in 8.9min at a NaCl concentration of 480mM NaCl (in 20mM Tris-HCl, pH 7.5). The total run time of the method was 25min. The method resulted in a linear response from 1+ù10<sup>8</sup> to 5.0+ù10<sup>10</sup>viral particles (VP/ml). The detection limit was 3.0+ù10<sup>7</sup> and the quantification limit was 1+ù10<sup>8</sup>VP/ml. The intra-assay precision was <10% for both purified and crude virus preparations whereas the inter-assay precisions were <5% and <10% for purified and crude virus preparations, respectively. The recovery/accuracy of the method ranged from 78 to 101%. This method is a robust monitoring tool to facilitate research activities with baculovirus vector and accelerate development of baculovirus-based processes for manufacturing of biologics. |
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| Publication date | 2011-01-01 |
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| In | |
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| Language | English |
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| Peer reviewed | Yes |
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| NRC number | NRCC 52769 |
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| NPARC number | 16616684 |
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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 | b0bd7562-494c-4518-bcd4-ef48f0593528 |
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| Record created | 2011-03-31 |
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| Record modified | 2020-04-21 |
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