Large-Scale Production of High-Quality Helper-Dependent Adenoviral Vectors Using Adherent Cells in Cell Factories
The most efficient and widely used system for generating helper-dependent adenoviral vectors (HDAds) is the Cre/loxP system developed by Graham and co-workers (Parks, R.J., Chen, L., Anton, M., Sankar, U., Rudnicki, M.A., and Graham, F.L. [ 1996 ]. Proc. Natl. Acad. Sci. U. S. A. 93, 13565-13570). A...
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description | The most efficient and widely used system for generating helper-dependent adenoviral vectors (HDAds) is the Cre/loxP system developed by Graham and co-workers (Parks, R.J., Chen, L., Anton, M., Sankar, U., Rudnicki, M.A., and Graham, F.L. [ 1996 ]. Proc. Natl. Acad. Sci. U. S. A. 93, 13565-13570). Alternative systems have been developed for HDAd production, but all are limited by the technical complexity of a three-component vector production system for reproducibly generating large quantities of adenovirus with high infectivity and low helper virus (HV) contamination. Recently, these problems were addressed by Ng and co-workers (Palmer, D., and Ng, P. [ 2003 ]. Mol Ther. 8, 846-852), who developed an improved system that combines the use of a suspension-adapted producer cell line expressing high levels of Cre recombinase, a HV resistant to mutation, and a refined purification protocol. With this system, >1 x 10(13) highly infectious vector particles are easily produced without vector genome rearrangements and having very low HV contamination levels. However, the Ng system incorporates a spinner flask culture system that involves considerable time, effort, and tissue culture medium to produce HDAds. We have an alternative system to obtain comparable quantities with equivalent quality to the spinner flask approach but requiring reduced labor and lower volumes of medium. This method utilizes a 10-chamber cell factory with adherent cells to produce high infectivity of HDAds with minimal HV contamination while improving yield and reducing technical complexity, effort, and medium requirements. This system is easily translatable to the production of clinical-grade HDAds for human trials. |
doi_str_mv | 10.1089/hum.2009.096 |
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[ 1996 ]. Proc. Natl. Acad. Sci. U. S. A. 93, 13565-13570). Alternative systems have been developed for HDAd production, but all are limited by the technical complexity of a three-component vector production system for reproducibly generating large quantities of adenovirus with high infectivity and low helper virus (HV) contamination. Recently, these problems were addressed by Ng and co-workers (Palmer, D., and Ng, P. [ 2003 ]. Mol Ther. 8, 846-852), who developed an improved system that combines the use of a suspension-adapted producer cell line expressing high levels of Cre recombinase, a HV resistant to mutation, and a refined purification protocol. With this system, >1 x 10(13) highly infectious vector particles are easily produced without vector genome rearrangements and having very low HV contamination levels. However, the Ng system incorporates a spinner flask culture system that involves considerable time, effort, and tissue culture medium to produce HDAds. We have an alternative system to obtain comparable quantities with equivalent quality to the spinner flask approach but requiring reduced labor and lower volumes of medium. This method utilizes a 10-chamber cell factory with adherent cells to produce high infectivity of HDAds with minimal HV contamination while improving yield and reducing technical complexity, effort, and medium requirements. This system is easily translatable to the production of clinical-grade HDAds for human trials.</description><identifier>ISSN: 1043-0342</identifier><identifier>EISSN: 1557-7422</identifier><identifier>DOI: 10.1089/hum.2009.096</identifier><identifier>PMID: 19719388</identifier><identifier>CODEN: HGTHE3</identifier><language>eng</language><publisher>Larchmont, NY: Liebert</publisher><subject>Adenoviridae - growth & development ; Adenovirus ; Adenoviruses ; Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy ; Applied cell therapy and gene therapy ; beta-Galactosidase - metabolism ; Biological and medical sciences ; Biotechnology ; Brief Reports ; Cell Adhesion ; Cell Culture Techniques - methods ; Cell Line ; Fundamental and applied biological sciences. Psychology ; Gene therapy ; Genetic aspects ; Genetic vectors ; Genetic Vectors - biosynthesis ; Health aspects ; Health. Pharmaceutical industry ; Helper Viruses - growth & development ; Humans ; Immune response ; Industrial applications and implications. Economical aspects ; Medical sciences ; Transfusions. Complications. Transfusion reactions. Cell and gene therapy</subject><ispartof>Human gene therapy, 2010-01, Vol.21 (1), p.120-126</ispartof><rights>2015 INIST-CNRS</rights><rights>COPYRIGHT 2010 Mary Ann Liebert, Inc.</rights><rights>Copyright 2010, Mary Ann Liebert, Inc.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c511t-e48d539d8470682e8373cabfaf92196fdb183f57286aa86658633caca53f25663</citedby><cites>FETCH-LOGICAL-c511t-e48d539d8470682e8373cabfaf92196fdb183f57286aa86658633caca53f25663</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=22428638$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19719388$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>SUZUKI, Masataka</creatorcontrib><creatorcontrib>CELA, Racel</creatorcontrib><creatorcontrib>CLARKE, Christian</creatorcontrib><creatorcontrib>BERTIN, Terry K</creatorcontrib><creatorcontrib>MOURINO, Susana</creatorcontrib><creatorcontrib>LEE, Brendan</creatorcontrib><title>Large-Scale Production of High-Quality Helper-Dependent Adenoviral Vectors Using Adherent Cells in Cell Factories</title><title>Human gene therapy</title><addtitle>Hum Gene Ther</addtitle><description>The most efficient and widely used system for generating helper-dependent adenoviral vectors (HDAds) is the Cre/loxP system developed by Graham and co-workers (Parks, R.J., Chen, L., Anton, M., Sankar, U., Rudnicki, M.A., and Graham, F.L. [ 1996 ]. Proc. Natl. Acad. Sci. U. S. A. 93, 13565-13570). Alternative systems have been developed for HDAd production, but all are limited by the technical complexity of a three-component vector production system for reproducibly generating large quantities of adenovirus with high infectivity and low helper virus (HV) contamination. Recently, these problems were addressed by Ng and co-workers (Palmer, D., and Ng, P. [ 2003 ]. Mol Ther. 8, 846-852), who developed an improved system that combines the use of a suspension-adapted producer cell line expressing high levels of Cre recombinase, a HV resistant to mutation, and a refined purification protocol. With this system, >1 x 10(13) highly infectious vector particles are easily produced without vector genome rearrangements and having very low HV contamination levels. However, the Ng system incorporates a spinner flask culture system that involves considerable time, effort, and tissue culture medium to produce HDAds. We have an alternative system to obtain comparable quantities with equivalent quality to the spinner flask approach but requiring reduced labor and lower volumes of medium. This method utilizes a 10-chamber cell factory with adherent cells to produce high infectivity of HDAds with minimal HV contamination while improving yield and reducing technical complexity, effort, and medium requirements. This system is easily translatable to the production of clinical-grade HDAds for human trials.</description><subject>Adenoviridae - growth & development</subject><subject>Adenovirus</subject><subject>Adenoviruses</subject><subject>Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy</subject><subject>Applied cell therapy and gene therapy</subject><subject>beta-Galactosidase - metabolism</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Brief Reports</subject><subject>Cell Adhesion</subject><subject>Cell Culture Techniques - methods</subject><subject>Cell Line</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene therapy</subject><subject>Genetic aspects</subject><subject>Genetic vectors</subject><subject>Genetic Vectors - biosynthesis</subject><subject>Health aspects</subject><subject>Health. Pharmaceutical industry</subject><subject>Helper Viruses - growth & development</subject><subject>Humans</subject><subject>Immune response</subject><subject>Industrial applications and implications. Economical aspects</subject><subject>Medical sciences</subject><subject>Transfusions. Complications. Transfusion reactions. Cell and gene therapy</subject><issn>1043-0342</issn><issn>1557-7422</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kttrFDEUhwdRbK2--SwDor44ay6T24uwbK0rLKhofQ3ZzMlsZGayTWYK_e-b6S7VgkggOeR858qvKF5itMBIqg-7qV8QhNQCKf6oOMWMiUrUhDzONqpphWhNTopnKf1GCFPGxdPiBCuBFZXytLjamNhC9cOaDspvMTSTHX0YyuDKtW931ffJdH68KdfQ7SFW57CHoYFhLJf5Dtc-mq78BXYMMZWXyQ9tduwgzsQKui6VfrgzygszQx7S8-KJM12CF8f3rLi8-PRzta42Xz9_WS03lWUYjxXUsmFUNbIWiEsCkgpqzdYZpwhW3DVbLKljgkhujOScSU4zYA2jjjDO6Vnx8ZB3P217aGxuKTer99H3Jt7oYLx-6Bn8TrfhWhNJVM1QTvDumCCGqwnSqHufbJ7FDBCmpAWlAiEkcCbf_pckmCKF7np6fQDbvG7tBxdyZTvDekkoJQpzVmdq8Q8qnwZ6b8MAzuf_BwHvDwE2hpQiuPspMdKzSHQWiZ5ForNIMv7q7838gY-qyMCbI2BSFoaLZrA-3XOE1HnrVNJbu9PEQA</recordid><startdate>20100101</startdate><enddate>20100101</enddate><creator>SUZUKI, Masataka</creator><creator>CELA, Racel</creator><creator>CLARKE, Christian</creator><creator>BERTIN, Terry K</creator><creator>MOURINO, Susana</creator><creator>LEE, Brendan</creator><general>Liebert</general><general>Mary Ann Liebert, Inc</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>7U7</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20100101</creationdate><title>Large-Scale Production of High-Quality Helper-Dependent Adenoviral Vectors Using Adherent Cells in Cell Factories</title><author>SUZUKI, Masataka ; CELA, Racel ; CLARKE, Christian ; BERTIN, Terry K ; MOURINO, Susana ; LEE, Brendan</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c511t-e48d539d8470682e8373cabfaf92196fdb183f57286aa86658633caca53f25663</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Adenoviridae - growth & development</topic><topic>Adenovirus</topic><topic>Adenoviruses</topic><topic>Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy</topic><topic>Applied cell therapy and gene therapy</topic><topic>beta-Galactosidase - metabolism</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Brief Reports</topic><topic>Cell Adhesion</topic><topic>Cell Culture Techniques - methods</topic><topic>Cell Line</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene therapy</topic><topic>Genetic aspects</topic><topic>Genetic vectors</topic><topic>Genetic Vectors - biosynthesis</topic><topic>Health aspects</topic><topic>Health. Pharmaceutical industry</topic><topic>Helper Viruses - growth & development</topic><topic>Humans</topic><topic>Immune response</topic><topic>Industrial applications and implications. Economical aspects</topic><topic>Medical sciences</topic><topic>Transfusions. Complications. Transfusion reactions. Cell and gene therapy</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>SUZUKI, Masataka</creatorcontrib><creatorcontrib>CELA, Racel</creatorcontrib><creatorcontrib>CLARKE, Christian</creatorcontrib><creatorcontrib>BERTIN, Terry K</creatorcontrib><creatorcontrib>MOURINO, Susana</creatorcontrib><creatorcontrib>LEE, Brendan</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Human gene therapy</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>SUZUKI, Masataka</au><au>CELA, Racel</au><au>CLARKE, Christian</au><au>BERTIN, Terry K</au><au>MOURINO, Susana</au><au>LEE, Brendan</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Large-Scale Production of High-Quality Helper-Dependent Adenoviral Vectors Using Adherent Cells in Cell Factories</atitle><jtitle>Human gene therapy</jtitle><addtitle>Hum Gene Ther</addtitle><date>2010-01-01</date><risdate>2010</risdate><volume>21</volume><issue>1</issue><spage>120</spage><epage>126</epage><pages>120-126</pages><issn>1043-0342</issn><eissn>1557-7422</eissn><coden>HGTHE3</coden><abstract>The most efficient and widely used system for generating helper-dependent adenoviral vectors (HDAds) is the Cre/loxP system developed by Graham and co-workers (Parks, R.J., Chen, L., Anton, M., Sankar, U., Rudnicki, M.A., and Graham, F.L. [ 1996 ]. Proc. Natl. Acad. Sci. U. S. A. 93, 13565-13570). Alternative systems have been developed for HDAd production, but all are limited by the technical complexity of a three-component vector production system for reproducibly generating large quantities of adenovirus with high infectivity and low helper virus (HV) contamination. Recently, these problems were addressed by Ng and co-workers (Palmer, D., and Ng, P. [ 2003 ]. Mol Ther. 8, 846-852), who developed an improved system that combines the use of a suspension-adapted producer cell line expressing high levels of Cre recombinase, a HV resistant to mutation, and a refined purification protocol. With this system, >1 x 10(13) highly infectious vector particles are easily produced without vector genome rearrangements and having very low HV contamination levels. However, the Ng system incorporates a spinner flask culture system that involves considerable time, effort, and tissue culture medium to produce HDAds. We have an alternative system to obtain comparable quantities with equivalent quality to the spinner flask approach but requiring reduced labor and lower volumes of medium. This method utilizes a 10-chamber cell factory with adherent cells to produce high infectivity of HDAds with minimal HV contamination while improving yield and reducing technical complexity, effort, and medium requirements. This system is easily translatable to the production of clinical-grade HDAds for human trials.</abstract><cop>Larchmont, NY</cop><pub>Liebert</pub><pmid>19719388</pmid><doi>10.1089/hum.2009.096</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Adenoviridae - growth & development Adenovirus Adenoviruses Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy Applied cell therapy and gene therapy beta-Galactosidase - metabolism Biological and medical sciences Biotechnology Brief Reports Cell Adhesion Cell Culture Techniques - methods Cell Line Fundamental and applied biological sciences. Psychology Gene therapy Genetic aspects Genetic vectors Genetic Vectors - biosynthesis Health aspects Health. Pharmaceutical industry Helper Viruses - growth & development Humans Immune response Industrial applications and implications. Economical aspects Medical sciences Transfusions. Complications. Transfusion reactions. Cell and gene therapy |
title | Large-Scale Production of High-Quality Helper-Dependent Adenoviral Vectors Using Adherent Cells in Cell Factories |
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