{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T05:44:17Z","timestamp":1792043057797,"version":"4.4.0"},"reference-count":29,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T00:00:00Z","timestamp":1791936000000},"content-version":"vor","delay-in-days":13,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T00:00:00Z","timestamp":1790812800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotechnology Journal"],"published-print":{"date-parts":[[2026,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Recombinant adeno\u2010associated virus (rAAV) vectors are essential for gene therapy, but their production generates empty and partial capsids as product\u2010related impurities that can compromise efficacy and safety. While empty capsids have been partially characterized, the isolation and study of partial capsids remain hindered by a lack of robust separation methods. Here, we present an optimized iodixanol density gradient ultracentrifugation strategy enabling high\u2010resolution separation and enrichment of empty, partial, and full AAV capsids. Key innovations include expanding the 40% iodixanol layer to create a broader intermediate\u2010density zone and implementing controlled bottom\u2010puncture fractionation. Applied to a rAAV2 preparation, this method enriched partial capsids from 18.9% in the starting material to approximately 94.9% in the target fraction (determined by AUC integration under the equal response assumption). For a rAAV2 variant with a low baseline of partial capsids (1.3%), over 20\u2010fold enrichment was achieved, reaching 30.1% purity as determined by AUC integration under the equal response assumption. This approach provides partial\u2010capsid\u2010enriched fractions, enabling detailed characterization of partial capsids, supporting critical quality attribute assessment, and facilitating the development of safer, more effective rAAV\u2010based therapeutics.<\/jats:p>","DOI":"10.1002\/biot.70319","type":"journal-article","created":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T04:46:07Z","timestamp":1792039567000},"source":"Crossref","is-referenced-by-count":0,"title":["Efficient and High\u2010Resolution Separation of Partial, Empty, and Full AAV Capsids by Optimized Iodixanol Density Gradient Ultracentrifugation"],"prefix":"10.1002","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-4061-1284","authenticated-orcid":false,"given":"Shan","family":"Fang","sequence":"first","affiliation":[{"name":"InnoStellar Biotherapeutics Co., Ltd.  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun","family":"Wu","sequence":"additional","affiliation":[{"name":"InnoStellar Biotherapeutics Co., Ltd.  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanfang","family":"Shen","sequence":"additional","affiliation":[{"name":"InnoStellar Biotherapeutics Co., Ltd.  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dandan","family":"Qi","sequence":"additional","affiliation":[{"name":"InnoStellar Biotherapeutics Co., Ltd.  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhuoyu","family":"Ni","sequence":"additional","affiliation":[{"name":"InnoStellar Biotherapeutics Co., Ltd.  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fenghua","family":"Wang","sequence":"additional","affiliation":[{"name":"InnoStellar Biotherapeutics Co., Ltd.  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,10,14]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00867\u201013"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.162250899"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biotechadv.2021.107764"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41392\u2010024\u201001780\u2010w"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2021.02.010"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtn.2022.08.027"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/biot.70020"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1089\/hgtb.2019.088"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.3390\/biomedicines2010080"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1002\/cpmo.56"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1089\/hgtb.2017.192"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1089\/hgtb.2015.051"},{"key":"e_1_2_10_14_1","doi-asserted-by":"crossref","first-page":"1235","DOI":"10.3390\/v16081235","article-title":"In\u2010Depth Comparison of Adeno\u2010Associated Virus Containing Fractions After CsCl Ultracentrifugation Gradient Separation","volume":"16","author":"Janc M.","year":"2024","journal-title":"Viruses"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1007\/978\u20101\u20104939\u20103271\u20109_7"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.3791\/66460"},{"key":"e_1_2_10_17_1","unstructured":"F.Wu Y.Wang Z.Ni andF.Wang Engineered Adeno\u2010associated Virus Capsid Proteins and Vectors and Their Uses CN120842333A (2025)."},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2019.01570"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1089\/hgtb.2015.048"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00249\u2010023\u201001646\u2010z"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41434\u2010024\u201000444\u20102"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1089\/hum.2023.169"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2024.101382"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2021.08.002"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.4c05229"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtn.2025.102582"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.xphs.2025.01.005"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2025.101530"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.xphs.2021.06.031"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.xphs.2025.104150"}],"container-title":["Biotechnology Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/biot.70319","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/biot.70319","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/biot.70319","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T04:46:15Z","timestamp":1792039575000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/biot.70319"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,10]]},"references-count":29,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2026,10]]}},"alternative-id":["10.1002\/biot.70319"],"URL":"https:\/\/doi.org\/10.1002\/biot.70319","archive":["Portico"],"relation":{},"ISSN":["1860-6768","1860-7314"],"issn-type":[{"value":"1860-6768","type":"print"},{"value":"1860-7314","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10]]},"article-number":"e70319"}}