A replication-competent adenovirus assay for E1-deleted Ad35 vectors produced in PER.C6 cells
Abstract The presence of replication-competent adenovirus (RCA) is a safety concern for biologics based on recombinant adenoviruses and RCA testing is therefore mandatory for release of clinical material. RCA, which arises from homologous recombination between Ad5 vectors and HEK-293 cells, can be e...
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| Veröffentlicht in: | Vaccine 2007-03, Vol.25 (12), p.2228-2237 |
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| creator | Marzio, G Kerkvliet, E Bogaards, J.A Koelewijn, S De Groot, A Gijsbers, L Weverling, G.J Vogels, R Havenga, M Custers, J Pau, M.G Guichoux, J.Y Lewis, J Goudsmit, J |
| description | Abstract The presence of replication-competent adenovirus (RCA) is a safety concern for biologics based on recombinant adenoviruses and RCA testing is therefore mandatory for release of clinical material. RCA, which arises from homologous recombination between Ad5 vectors and HEK-293 cells, can be eliminated by the use of PER.C6 cells in combination with a matched vector. However, little is known on RCA formation with vectors based on adenovirus serotypes other than Ad5 and reliable RCA assays to test them are generally lacking. Here we report on the development and qualification of a sensitive RCA assay for Ad35, a promising alternative to Ad5 vectors. The assay is able to detect 1 RCA in 3 × 1010 vector particles with 95% confidence, thus meeting current FDA requirements, and can discriminate between RCA and other rare CPE-causing entities, including helper dependent E1 positive particles (HDEP). Using this assay, the first batches of Ad35 vectors produced in PER.C6 cells were analysed and found to be free of RCA and HDEP. Based on the statistical model used, we anticipate that our approach to RCA assay development can be broadly applicable to other adenoviral vectors. |
| doi_str_mv | 10.1016/j.vaccine.2006.12.011 |
| format | Article |
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RCA, which arises from homologous recombination between Ad5 vectors and HEK-293 cells, can be eliminated by the use of PER.C6 cells in combination with a matched vector. However, little is known on RCA formation with vectors based on adenovirus serotypes other than Ad5 and reliable RCA assays to test them are generally lacking. Here we report on the development and qualification of a sensitive RCA assay for Ad35, a promising alternative to Ad5 vectors. The assay is able to detect 1 RCA in 3 × 1010 vector particles with 95% confidence, thus meeting current FDA requirements, and can discriminate between RCA and other rare CPE-causing entities, including helper dependent E1 positive particles (HDEP). Using this assay, the first batches of Ad35 vectors produced in PER.C6 cells were analysed and found to be free of RCA and HDEP. Based on the statistical model used, we anticipate that our approach to RCA assay development can be broadly applicable to other adenoviral vectors.</description><identifier>ISSN: 0264-410X</identifier><identifier>EISSN: 1873-2518</identifier><identifier>DOI: 10.1016/j.vaccine.2006.12.011</identifier><identifier>PMID: 17250936</identifier><identifier>CODEN: VACCDE</identifier><language>eng</language><publisher>Oxford: Elsevier Ltd</publisher><subject>Adenoviridae - genetics ; Adenoviridae - growth & development ; Adenovirus ; Adenovirus E1 Proteins - genetics ; Adenoviruses ; Allergy and Immunology ; Applied microbiology ; Assay ; Biological and medical sciences ; Biological Assay - methods ; Cell Line ; Cell Line, Tumor ; Confidence intervals ; Fundamental and applied biological sciences. Psychology ; Gene Deletion ; Genetic Vectors - genetics ; HDEP ; Humans ; Microbiology ; Miscellaneous ; PER.C6 ; Polymerase Chain Reaction ; RCA ; Reproducibility of Results ; Statistical models ; Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects) ; Virology ; Virus Replication</subject><ispartof>Vaccine, 2007-03, Vol.25 (12), p.2228-2237</ispartof><rights>Elsevier Ltd</rights><rights>2006 Elsevier Ltd</rights><rights>2007 INIST-CNRS</rights><rights>Copyright Elsevier Limited Mar 8, 2007</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c507t-347011cf1fb17eb0fc58a3390e2842fe4b5529c5cb7e2c9bfd87e0810dbdd6303</citedby><cites>FETCH-LOGICAL-c507t-347011cf1fb17eb0fc58a3390e2842fe4b5529c5cb7e2c9bfd87e0810dbdd6303</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0264410X06013314$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=18550925$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17250936$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Marzio, G</creatorcontrib><creatorcontrib>Kerkvliet, E</creatorcontrib><creatorcontrib>Bogaards, J.A</creatorcontrib><creatorcontrib>Koelewijn, S</creatorcontrib><creatorcontrib>De Groot, A</creatorcontrib><creatorcontrib>Gijsbers, L</creatorcontrib><creatorcontrib>Weverling, G.J</creatorcontrib><creatorcontrib>Vogels, R</creatorcontrib><creatorcontrib>Havenga, M</creatorcontrib><creatorcontrib>Custers, J</creatorcontrib><creatorcontrib>Pau, M.G</creatorcontrib><creatorcontrib>Guichoux, J.Y</creatorcontrib><creatorcontrib>Lewis, J</creatorcontrib><creatorcontrib>Goudsmit, J</creatorcontrib><title>A replication-competent adenovirus assay for E1-deleted Ad35 vectors produced in PER.C6 cells</title><title>Vaccine</title><addtitle>Vaccine</addtitle><description>Abstract The presence of replication-competent adenovirus (RCA) is a safety concern for biologics based on recombinant adenoviruses and RCA testing is therefore mandatory for release of clinical material. RCA, which arises from homologous recombination between Ad5 vectors and HEK-293 cells, can be eliminated by the use of PER.C6 cells in combination with a matched vector. However, little is known on RCA formation with vectors based on adenovirus serotypes other than Ad5 and reliable RCA assays to test them are generally lacking. Here we report on the development and qualification of a sensitive RCA assay for Ad35, a promising alternative to Ad5 vectors. The assay is able to detect 1 RCA in 3 × 1010 vector particles with 95% confidence, thus meeting current FDA requirements, and can discriminate between RCA and other rare CPE-causing entities, including helper dependent E1 positive particles (HDEP). Using this assay, the first batches of Ad35 vectors produced in PER.C6 cells were analysed and found to be free of RCA and HDEP. Based on the statistical model used, we anticipate that our approach to RCA assay development can be broadly applicable to other adenoviral vectors.</description><subject>Adenoviridae - genetics</subject><subject>Adenoviridae - growth & development</subject><subject>Adenovirus</subject><subject>Adenovirus E1 Proteins - genetics</subject><subject>Adenoviruses</subject><subject>Allergy and Immunology</subject><subject>Applied microbiology</subject><subject>Assay</subject><subject>Biological and medical sciences</subject><subject>Biological Assay - methods</subject><subject>Cell Line</subject><subject>Cell Line, Tumor</subject><subject>Confidence intervals</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Deletion</subject><subject>Genetic Vectors - genetics</subject><subject>HDEP</subject><subject>Humans</subject><subject>Microbiology</subject><subject>Miscellaneous</subject><subject>PER.C6</subject><subject>Polymerase Chain Reaction</subject><subject>RCA</subject><subject>Reproducibility of Results</subject><subject>Statistical models</subject><subject>Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects)</subject><subject>Virology</subject><subject>Virus Replication</subject><issn>0264-410X</issn><issn>1873-2518</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>BENPR</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNqFklur1DAUhYMonvHoT1AKom-teydNLy_KMIwXOKB4AV8kpMkuZOykY9IOzL83ZQYGzst5CoRvr6yVtRl7iVAgYPVuVxy1Mc5TwQGqAnkBiI_YCpta5Fxi85itgFdlXiL8vmHPYtwBgBTYPmU3WHMJrahW7M86C3QYnNGTG31uxv2BJvJTpi358ejCHDMdoz5l_RiyLeaWhgTYbG2FzI5kpjHE7BBGO5t063z2bfu92FSZoWGIz9mTXg-RXlzOW_br4_bn5nN-9_XTl836LjcS6ikXZZ3Mmx77DmvqoDey0UK0QLwpeU9lJyVvjTRdTdy0XW-bmqBBsJ21lQBxy96edZORfzPFSe1dXBxoT-McVdUCyvTIgyAHUZeylAl8fQ_cjXPwKYRalLCSvGwSJc-UCWOMgXp1CG6vw0khqKUmtVOXmtRSk0KuUtI09-qiPnd7stepSy8JeHMBdDR66IP2xsUr18jE8cXmhzNH6XePjoKKxpFPVbiQulF2dA9aeX9PwQzOp30Y_tKJ4jW1imlA_Vh2alkpqACFwFL8Bysyxfc</recordid><startdate>20070308</startdate><enddate>20070308</enddate><creator>Marzio, G</creator><creator>Kerkvliet, E</creator><creator>Bogaards, J.A</creator><creator>Koelewijn, S</creator><creator>De Groot, A</creator><creator>Gijsbers, L</creator><creator>Weverling, G.J</creator><creator>Vogels, R</creator><creator>Havenga, M</creator><creator>Custers, J</creator><creator>Pau, M.G</creator><creator>Guichoux, J.Y</creator><creator>Lewis, J</creator><creator>Goudsmit, J</creator><general>Elsevier Ltd</general><general>Elsevier</general><general>Elsevier Limited</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>3V.</scope><scope>7QL</scope><scope>7RV</scope><scope>7T2</scope><scope>7T5</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88C</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8G5</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>GUQSH</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9-</scope><scope>K9.</scope><scope>KB0</scope><scope>LK8</scope><scope>M0R</scope><scope>M0S</scope><scope>M0T</scope><scope>M1P</scope><scope>M2O</scope><scope>M7N</scope><scope>M7P</scope><scope>MBDVC</scope><scope>NAPCQ</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20070308</creationdate><title>A replication-competent adenovirus assay for E1-deleted Ad35 vectors produced in PER.C6 cells</title><author>Marzio, G ; Kerkvliet, E ; Bogaards, J.A ; Koelewijn, S ; De Groot, A ; Gijsbers, L ; Weverling, G.J ; Vogels, R ; Havenga, M ; Custers, J ; Pau, M.G ; Guichoux, J.Y ; Lewis, J ; Goudsmit, J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c507t-347011cf1fb17eb0fc58a3390e2842fe4b5529c5cb7e2c9bfd87e0810dbdd6303</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Adenoviridae - genetics</topic><topic>Adenoviridae - growth & development</topic><topic>Adenovirus</topic><topic>Adenovirus E1 Proteins - genetics</topic><topic>Adenoviruses</topic><topic>Allergy and Immunology</topic><topic>Applied microbiology</topic><topic>Assay</topic><topic>Biological and medical sciences</topic><topic>Biological Assay - methods</topic><topic>Cell Line</topic><topic>Cell Line, Tumor</topic><topic>Confidence intervals</topic><topic>Fundamental and applied biological sciences. 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Academic</collection><jtitle>Vaccine</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Marzio, G</au><au>Kerkvliet, E</au><au>Bogaards, J.A</au><au>Koelewijn, S</au><au>De Groot, A</au><au>Gijsbers, L</au><au>Weverling, G.J</au><au>Vogels, R</au><au>Havenga, M</au><au>Custers, J</au><au>Pau, M.G</au><au>Guichoux, J.Y</au><au>Lewis, J</au><au>Goudsmit, J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A replication-competent adenovirus assay for E1-deleted Ad35 vectors produced in PER.C6 cells</atitle><jtitle>Vaccine</jtitle><addtitle>Vaccine</addtitle><date>2007-03-08</date><risdate>2007</risdate><volume>25</volume><issue>12</issue><spage>2228</spage><epage>2237</epage><pages>2228-2237</pages><issn>0264-410X</issn><eissn>1873-2518</eissn><coden>VACCDE</coden><abstract>Abstract The presence of replication-competent adenovirus (RCA) is a safety concern for biologics based on recombinant adenoviruses and RCA testing is therefore mandatory for release of clinical material. RCA, which arises from homologous recombination between Ad5 vectors and HEK-293 cells, can be eliminated by the use of PER.C6 cells in combination with a matched vector. However, little is known on RCA formation with vectors based on adenovirus serotypes other than Ad5 and reliable RCA assays to test them are generally lacking. Here we report on the development and qualification of a sensitive RCA assay for Ad35, a promising alternative to Ad5 vectors. The assay is able to detect 1 RCA in 3 × 1010 vector particles with 95% confidence, thus meeting current FDA requirements, and can discriminate between RCA and other rare CPE-causing entities, including helper dependent E1 positive particles (HDEP). Using this assay, the first batches of Ad35 vectors produced in PER.C6 cells were analysed and found to be free of RCA and HDEP. Based on the statistical model used, we anticipate that our approach to RCA assay development can be broadly applicable to other adenoviral vectors.</abstract><cop>Oxford</cop><pub>Elsevier Ltd</pub><pmid>17250936</pmid><doi>10.1016/j.vaccine.2006.12.011</doi><tpages>10</tpages></addata></record> |
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| identifier | ISSN: 0264-410X |
| ispartof | Vaccine, 2007-03, Vol.25 (12), p.2228-2237 |
| issn | 0264-410X 1873-2518 |
| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Adenoviridae - genetics Adenoviridae - growth & development Adenovirus Adenovirus E1 Proteins - genetics Adenoviruses Allergy and Immunology Applied microbiology Assay Biological and medical sciences Biological Assay - methods Cell Line Cell Line, Tumor Confidence intervals Fundamental and applied biological sciences. Psychology Gene Deletion Genetic Vectors - genetics HDEP Humans Microbiology Miscellaneous PER.C6 Polymerase Chain Reaction RCA Reproducibility of Results Statistical models Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects) Virology Virus Replication |
| title | A replication-competent adenovirus assay for E1-deleted Ad35 vectors produced in PER.C6 cells |
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