{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T02:15:35Z","timestamp":1789352135051,"version":"build-2803163510"},"reference-count":17,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2015,9,24]],"date-time":"2015-09-24T00:00:00Z","timestamp":1443052800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100000046","name":"National Research Council Canada","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100000046","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100008658","name":"Deutsches Krebsforschungszentrum","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100008658","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotechnology Journal"],"published-print":{"date-parts":[[2016,2]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Viral vectors used for gene and oncolytic therapy belong to the most promising biological products for future therapeutics. Clinical success of recombinant adeno\u2010associated virus (rAAV) based therapies raises considerable demand for viral vectors, which cannot be met by current manufacturing strategies. Addressing existing bottlenecks, we improved a plasmid system termed rep\/cap split packaging and designed a minimal plasmid encoding adenoviral helper function. Plasmid modifications led to a 12\u2010fold increase in rAAV vector titers compared to the widely used pDG standard system. Evaluation of different production approaches revealed superiority of processes based on anchorage\u2010 and serum\u2010dependent HEK293T cells, exhibiting about 15\u2010fold higher specific and volumetric productivity compared to well\u2010established suspension cells cultivated in serum\u2010free medium. As for most other viral vectors, classical stirred\u2010tank bioreactor production is thus still not capable of providing drug product of sufficient amount. We show that manufacturing strategies employing classical surface\u2010providing culture systems can be successfully transferred to the new fully\u2010controlled, single\u2010use bioreactor system Integrity<jats:sup>TM<\/jats:sup> iCELLis<jats:sup>TM<\/jats:sup>. In summary, we demonstrate substantial bioprocess optimizations leading to more efficient and scalable production processes suggesting a promising way for flexible large\u2010scale rAAV manufacturing.<\/jats:p>","DOI":"10.1002\/biot.201500176","type":"journal-article","created":{"date-parts":[[2015,8,18]],"date-time":"2015-08-18T17:05:40Z","timestamp":1439917540000},"page":"290-297","source":"Crossref","is-referenced-by-count":58,"title":["Rational plasmid design and bioprocess optimization to enhance recombinant adeno\u2010associated virus (AAV) productivity in mammalian cells"],"prefix":"10.1002","volume":"11","author":[{"given":"Verena V.","family":"Emmerling","sequence":"first","affiliation":[{"name":"Department of Gene Therapy University of Ulm  Ulm Baden-W\u00fcrttemberg Germany"},{"name":"Development Rentschler Biotechnologie GmbH  Laupheim Baden-W\u00fcrttemberg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Antje","family":"Pegel","sequence":"additional","affiliation":[{"name":"Development Rentschler Biotechnologie GmbH  Laupheim Baden-W\u00fcrttemberg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ernest G.","family":"Milian","sequence":"additional","affiliation":[{"name":"Human Health Therapeutics Portfolio National Research Council of Canada  Montreal QC Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alina","family":"Venereo\u2010Sanchez","sequence":"additional","affiliation":[{"name":"Human Health Therapeutics Portfolio National Research Council of Canada  Montreal QC Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marion","family":"Kunz","sequence":"additional","affiliation":[{"name":"Development Rentschler Biotechnologie GmbH  Laupheim Baden-W\u00fcrttemberg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica","family":"Wegele","sequence":"additional","affiliation":[{"name":"Development Rentschler Biotechnologie GmbH  Laupheim Baden-W\u00fcrttemberg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amine A.","family":"Kamen","sequence":"additional","affiliation":[{"name":"Department of Bioengineering McGill University  Montreal Quebec Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan","family":"Kochanek","sequence":"additional","affiliation":[{"name":"Department of Gene Therapy University of Ulm  Ulm Baden-W\u00fcrttemberg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Markus","family":"Hoerer","sequence":"additional","affiliation":[{"name":"Development Rentschler Biotechnologie GmbH  Laupheim Baden-W\u00fcrttemberg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2015,9,24]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0912-807"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1272\/jnms.79.394"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-61779-370-7_1"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2013.10.038"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.2174\/156652310793797685"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biotechadv.2007.09.001"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1098\/rstb.2014.0040"},{"key":"e_1_2_7_9_1","unstructured":"Hoerer M. 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