Author | dc.contributor.author | Lucero, Alicia T. | |
Author | dc.contributor.author | Mercado, Sergio A. | |
Author | dc.contributor.author | Sánchez, Anamaria C. | |
Author | dc.contributor.author | Contador, Carolina A. | |
Author | dc.contributor.author | Andrews Farrow, Bárbara | |
Author | dc.contributor.author | Asenjo de Leuze, Juan | |
Admission date | dc.date.accessioned | 2018-07-12T14:00:06Z | |
Available date | dc.date.available | 2018-07-12T14:00:06Z | |
Publication date | dc.date.issued | 2017 | |
Cita de ítem | dc.identifier.citation | Journal of Chemical Technology and Biotechnology, 92 (9): 2445-2452 | es_ES |
Identifier | dc.identifier.other | 10.1002/jctb.5255 | |
Identifier | dc.identifier.uri | https://repositorio.uchile.cl/handle/2250/149781 | |
Abstract | dc.description.abstract | BACKGROUND: Gene therapy is a potent alternative for long-lasting inhibition of alcohol consumption. This study compares the purification of a recombinant adenoviral vector serotype 5 (rAdV5) for use in gene therapy against alcoholism using two anion-exchange methods. RESULTS: Two anion-exchange chromatography methods using fast protein liquid chromatography were compared using a packed-bed column (Q-Sepharose (TM) XL) and two monolithic columns (CIM (TM) QA-1 and CIM (TM) DEAE-1). An improved and reproducible separation of recombinant adenovirus type 5 from cell lysate contaminants was achieved using the two strong anion-exchange columns in a two-step gradient chromatography. Higher adenovirus yields were achieved using the CIM QA-1 tube monolithic column at sample volumes of both 1 and 10 mL compared with the Q-Sepharose XL column. At higher flow rates, the CIM QA-1 tube monolithic column achieved better separation of the target fraction. Process recovery was improved from 28% using the Q-Sepharose XL column to 34% with the CIM QA-1 tube monolithic column quantified as vector genome. Analysis by SDS-PAGE demonstrated a purity of 70% for purified adenovirus using the CIM QA-1 tube monolithic column. CONCLUSION: This study indicated that the use of a CIM QA-1 tube monolithic column is a better alternative than Q-Sepharose XL, and CIM DEAE-1 tube monolithic columns for the primary purification process of rAdV5 carrying the human aldehyde dehydrogenase-2 antisense gene. This purification strategy has been used as a basis to scale-up a GLP process for the production of material at the National Research Council of Canada to be used in preclinical trials of this gene therapy against alcoholism. | es_ES |
Patrocinador | dc.description.sponsorship | University of Chile FONDEF D08I1051 CONICYT Millennium Scientific Initiative 'Institute for Cell Dynamics and Biotechnology' Millenium Institute CONICYT FB0001 CeBiB | es_ES |
Lenguage | dc.language.iso | en | es_ES |
Publisher | dc.publisher | Wiley | es_ES |
Source | dc.source | Journal of Chemical Technology and Biotechnology | es_ES |
Keywords | dc.subject | Adenovirus serotype 5 | es_ES |
Keywords | dc.subject | Alcoholism | es_ES |
Keywords | dc.subject | Anion exchange | es_ES |
Keywords | dc.subject | Columns | es_ES |
Keywords | dc.subject | Gene therapy | es_ES |
Keywords | dc.subject | Purification | es_ES |
Título | dc.title | Purification of adenoviral vector serotype 5 for gene therapy against alcoholism using anion exchange chromatography | es_ES |
Document type | dc.type | Artículo de revista | |
dcterms.accessRights | dcterms.accessRights | Acceso a solo metadatos | es_ES |
Cataloguer | uchile.catalogador | tjn | es_ES |
Indexation | uchile.index | Artículo de publicación ISI | es_ES |