{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T19:35:42Z","timestamp":1790969742465,"version":"4.1.0"},"reference-count":51,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2025,4,22]],"date-time":"2025-04-22T00:00:00Z","timestamp":1745280000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Biotech &amp; Bioengineering"],"published-print":{"date-parts":[[2025,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Adenoviral vectors (AdVs) are gaining prominence in cancer therapy and vaccine development, posing the need for a modern AdV manufacturing platform. Current AdV purification by ion\u2010exchange chromatography indeed struggles to achieve the product's yield and purity of processes that employ affinity technologies. Addressing these challenges, this study presents the first affinity\u2010based process that delivers high product yield and clearance of host cell proteins and DNA (HCPs and hcDNA) in two chromatography steps. The affinity capture utilizes resins functionalized with peptide ligands that target AdV hexon proteins (AEFFIWNA and TNDGPDYSSPLTGSG), and provide high capacity (&gt;\u20095\u00b710\n                    <jats:sup>10<\/jats:sup>\n                    \u2009vp\/mL of resin) and yield under mild elution conditions (~50% at pH 8.0). Peptide\u2010functionalized adsorbents prepared using different matrices (polymethylmethacrylate vs. agarose) were initially tested to compare the purification performance. AEFFIWNA\u2010SulfoLink resin was selected for its yield of cell\u2010transducing AdVs (~50%) and removal of HCPs and hcDNA (144\u2010fold and 56\u2010fold). Similarly, TNDGPDYSSPLTGSG\u2010Toyopearl resin afforded ~50% yield and &gt;\u200950\u2010fold reduction of impurities. Additional gains in product purity were achieved by optimizing the washing step, which removed free hexon proteins and additional HCPs. All peptide\u2010functionalized resins maintained their purification performance for 10 cycles upon regeneration at pH ~2.0. The purification process was assembled to include clarification, affinity capture in bind\u2010and\u2010elute mode using AEFFIWNA\u2010SulfoLink resin, and polishing in flow\u2010through mode using mixed\u2010mode resins. The optimized process provided a yield ~50% of cell\u2010infecting units (IFU) and a product titer ~10\n                    <jats:sup>7<\/jats:sup>\n                    IFU\/mL, along with residual HCP and hcDNA levels (8.76\u2009ng\/mL and 44\u2009ng per dose, respectively) that meet clinical requirements.\n                  <\/jats:p>","DOI":"10.1002\/bit.29006","type":"journal-article","created":{"date-parts":[[2025,4,23]],"date-time":"2025-04-23T00:50:43Z","timestamp":1745369443000},"page":"1780-1792","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Integrating Affinity Chromatography in the Platform Process for Adenovirus Purification"],"prefix":"10.1002","volume":"122","author":[{"given":"Yuxuan","family":"Wu","sequence":"first","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering North Carolina State University Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eduardo","family":"Barbieri","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering North Carolina State University Raleigh North Carolina USA"},{"name":"LigaTrap Technologies LLC Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William K.","family":"Smith","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering North Carolina State University Raleigh North Carolina USA"},{"name":"Biomanufacturing Training and Education Center (BTEC) Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arianna","family":"Minzoni","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering North Carolina State University Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryan E.","family":"Kilgore","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering North Carolina State University Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenning","family":"Chu","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering North Carolina State University Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael A.","family":"Daniele","sequence":"additional","affiliation":[{"name":"Joint Department of Biomedical Engineering North Carolina State University and University of North Carolina at Chapel Hill Raleigh North Carolina USA"},{"name":"Department of Electrical and Computer Engineering North Carolina State University Raleigh North Carolina USA"},{"name":"North Carolina Viral Vector Initiative in Research and Learning (NC\u2010VVIRAL) North Carolina State University Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5633-434X","authenticated-orcid":false,"given":"Stefano","family":"Menegatti","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering North Carolina State University Raleigh North Carolina USA"},{"name":"LigaTrap Technologies LLC Raleigh North Carolina USA"},{"name":"Biomanufacturing Training and Education Center (BTEC) Raleigh North Carolina USA"},{"name":"North Carolina Viral Vector Initiative in Research and Learning (NC\u2010VVIRAL) North Carolina State University Raleigh North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,4,22]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/mtm.2016.30"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2014.10.003"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.28594"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0301-0082(98)00028-8"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2005.10.005"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1525-0016(02)90719-7"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.gt.3302611"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chroma.2022.463363"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.28495"},{"key":"e_1_2_9_11_1","first-page":"OF1","article-title":"FDA Approval Summary: Nadofaragene Firadenovec\u2010Vncg for Bacillus Calmette\u2013Gu\u00e9rin\u2013Unresponsive Non\u2013Muscle\u2010Invasive Bladder Cancer","author":"Colbert L.","year":"2025","journal-title":"Clinical Cancer Research"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2022.11.015"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1002\/jgm.1383"},{"key":"e_1_2_9_14_1","article-title":"Cloning and Large\u2010Scale Production of High\u2010Capacity Adenoviral Vectors Based on the Human Adenovirus Type 5","volume":"107","author":"Ehrke\u2010Schulz E.","year":"2016","journal-title":"Journal of Visualized Experiments"},{"key":"e_1_2_9_15_1","unstructured":"FDA's Drug Information Soundcast in Clinical Oncology (FDA DISCO).2022. 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