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During the COVID\u201019 pandemic, it gained renewed attention, but at the same time, it raised concerns due to side effects observed with some of the resulting vaccines administered to patients. It has been indicated that these side effects might be attributed to impurities present in the final product. Therefore, constant enhancement of the vaccine purity and further improvement of impurity detection methods are needed. In this work, we showcase an example of industry\u2010relevant adenovirus bioprocess optimization. Our data show the effect of upstream parameters on the bioburden introduced to the downstream process. We provide an example of process optimization using a combination of the PATfix analytical method, ddPCR, infectivity, total DNA, and total protein analyses to optimize cell density, multiplicity of infection, and length of production. Additionally, we provide data illustrating the robustness of the convective interaction media quaternary amine monolithic chromatography step. This anion exchange strategy was shown to remove over 99% of protein and DNA impurities, including those unable to be addressed by tangential flow filtration, while maintaining high adenovirus recoveries.<\/jats:p>","DOI":"10.1002\/elps.202300131","type":"journal-article","created":{"date-parts":[[2023,12,7]],"date-time":"2023-12-07T23:30:27Z","timestamp":1701991827000},"page":"369-379","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Bridging upstream and downstream for improved adenovirus 5 bioprocess"],"prefix":"10.1002","volume":"45","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-2603-3731","authenticated-orcid":false,"given":"Ivana","family":"Petrovi\u0107 Koshmak","sequence":"first","affiliation":[{"name":"Sartorius BIA Separations Mirce  Ajdov\u0161\u010dina Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-1459-0126","authenticated-orcid":false,"given":"Hana","family":"Jug","sequence":"additional","affiliation":[{"name":"Sartorius BIA Separations Mirce  Ajdov\u0161\u010dina Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Katja","family":"Vrabec","sequence":"additional","affiliation":[{"name":"Sartorius BIA Separations Mirce  Ajdov\u0161\u010dina Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana","family":"Mavri","sequence":"additional","affiliation":[{"name":"Sartorius BIA Separations Mirce  Ajdov\u0161\u010dina Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Valentina","family":"Novak","sequence":"additional","affiliation":[{"name":"Sartorius BIA Separations Mirce  Ajdov\u0161\u010dina Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Petra","family":"Dekleva","sequence":"additional","affiliation":[{"name":"Sartorius BIA Separations Mirce  Ajdov\u0161\u010dina Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Veronika","family":"Fujs","sequence":"additional","affiliation":[{"name":"Sartorius BIA Separations Mirce  Ajdov\u0161\u010dina Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9633-2066","authenticated-orcid":false,"given":"Maja","family":"Leskovec","sequence":"additional","affiliation":[{"name":"Sartorius BIA Separations Mirce  Ajdov\u0161\u010dina Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ale\u0161","family":"\u0160trancar","sequence":"additional","affiliation":[{"name":"Sartorius BIA Separations Mirce  Ajdov\u0161\u010dina Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2023,12,7]]},"reference":[{"key":"e_1_2_9_2_1","first-page":"1257","article-title":"Achieve significantly increased adenovirus yield with density gradient ultracentrifugation: a comparative study","volume":"8","author":"Sternisha S","year":"2022","journal-title":"Cell Gene Ther Insights"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12026-023-09357-5"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41541-021-00356-x"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/S2352-3026(21)00306-9"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.3324\/haematol.2021.280154"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2020.568513"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.7554\/ELIFE.78513"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2005.03.227"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.coche.2021.100780"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1002\/jctb.5255"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/gt.2012.52"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2005.10.002"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1080\/21645515.2015.1009817"},{"key":"e_1_2_9_15_1","volume-title":"PATfix analytical method for adenovirus sample characterization in upstream process development","author":"Petrovic Koshmak I","year":"2022"},{"key":"e_1_2_9_16_1","unstructured":"SeigneuretF EngaM RoblinA PennorsM ToublancE DoutheS et\u00a0al.CIM monolithic matrix as an efficient and polyvalent tool to prepare and analyze a cGMP compliant batch of recombinant Adenovirus serotype 5. 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