Téléchargement | - Voir le manuscrit accepté : Development and validation of a HPLC method for the quantification of baculovirus particles (PDF, 819 Kio)
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DOI | Trouver le DOI : https://doi.org/10.1016/j.jchromb.2010.11.011 |
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Auteur | Rechercher : Transfiguracion, Julia1; Rechercher : Mena, Jimmy A.2; Rechercher : Aucoin, Marc G.; Rechercher : Kamen, Amine A.1 |
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Affiliation | - Conseil national de recherches du Canada. Institut de recherche en biotechnologie du CNRC
- Conseil national de recherches du Canada
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Format | Texte, Article |
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Sujet | Baculovirus; HPLC; Quantification; Total virus particle; Viral genome labeling |
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Résumé | 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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Date de publication | 2011-01-01 |
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Dans | |
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Langue | anglais |
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Publications évaluées par des pairs | Oui |
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Numéro du CNRC | NRCC 52769 |
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Numéro NPARC | 16616684 |
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Exporter la notice | Exporter en format RIS |
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Signaler une correction | Signaler une correction (s'ouvre dans un nouvel onglet) |
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Identificateur de l’enregistrement | b0bd7562-494c-4518-bcd4-ef48f0593528 |
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Enregistrement créé | 2011-03-31 |
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Enregistrement modifié | 2020-04-21 |
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