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Although there is a very efficacious vaccine since the 1930s, YF still causes 29,000\u201360,000 annual deaths. During recent YF outbreaks there were issues of vaccine shortage of the current egg\u2010derived vaccine; rare but fatal vaccine adverse effects occurred; and cases were imported to Asia, where the circulating mosquito vector could potentially start local transmission. Here we investigated the production of YF virus\u2010like particles (VLPs) using stably transfected HEK293 cells. Process intensification was achieved by combining sequential FACS (fluorescence\u2010activated cell sorting) rounds to enrich the stable cell pool in terms of high producers and the use of perfusion processes. At shaken\u2010tube scale, FACS enrichment of cells allowed doubling VLP production, and pseudoperfusion cultivation (with daily medium exchange) further increased VLP production by 9.3\u2010fold as compared to batch operation mode. At perfusion bioreactor scale, the use of an inclined settler as cell retention device showed operational advantages over an ATF system. A one\u2010step steric exclusion chromatography purification allowed significant removal of impurities and is a promising technique for future integration of upstream and downstream operations. Characterization by different techniques confirmed the identity and 3D\u2010structure of the purified VLPs.<\/jats:p>","DOI":"10.1002\/bit.27864","type":"journal-article","created":{"date-parts":[[2021,6,18]],"date-time":"2021-06-18T11:18:53Z","timestamp":1624015133000},"page":"3581-3592","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Process intensification for the production of yellow fever virus\u2010like particles as potential recombinant vaccine antigen"],"prefix":"10.1002","volume":"118","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3260-5525","authenticated-orcid":false,"given":"Renata G. F.","family":"Alvim","sequence":"first","affiliation":[{"name":"COPPE, PEQ, Cell Culture Engineering Laboratory (LECC) Federal University of Rio de Janeiro (UFRJ)  Rio de Janeiro RJ Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1003-1938","authenticated-orcid":false,"given":"T\u00falio M.","family":"Lima","sequence":"additional","affiliation":[{"name":"COPPE, PEQ, Cell Culture Engineering Laboratory (LECC) Federal University of Rio de Janeiro (UFRJ)  Rio de Janeiro RJ Brazil"},{"name":"School of Chemistry (EQ) Federal University of Rio de Janeiro (UFRJ)  Rio de Janeiro RJ Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9523-9441","authenticated-orcid":false,"given":"Jerson L.","family":"Silva","sequence":"additional","affiliation":[{"name":"Institute of Medical Biochemistry Leopoldo de Meis (IBqM) Federal University of Rio de Janeiro (UFRJ)  Rio de Janeiro RJ Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0063-5888","authenticated-orcid":false,"given":"Guilherme A. P.","family":"de Oliveira","sequence":"additional","affiliation":[{"name":"Institute of Medical Biochemistry Leopoldo de Meis (IBqM) Federal University of Rio de Janeiro (UFRJ)  Rio de Janeiro RJ Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5904-410X","authenticated-orcid":false,"given":"Leda R.","family":"Castilho","sequence":"additional","affiliation":[{"name":"COPPE, PEQ, Cell Culture Engineering Laboratory (LECC) Federal University of Rio de Janeiro (UFRJ)  Rio de Janeiro RJ Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2021,7]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2019.05.064"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biotechadv.2018.04.011"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/9783527699902.ch2"},{"key":"e_1_2_9_5_1","first-page":"115","volume-title":"Continuous Processing in Pharmaceutical Manufacturing","author":"Castilho","year":"2015"},{"key":"e_1_2_9_6_1","unstructured":"Centers for Disease Control and Prevention.\u00a0(2019). Yellow fever.https:\/\/www.cdc.gov\/yellowfever\/index.html"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1002\/btpr.1704"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2019.06.005"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.3389\/fbioe.2020.00672"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/btpr.2915"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1026192228471"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2015.03.088"},{"key":"e_1_2_9_13_1","first-page":"77","article-title":"The yellow fever vaccine: A history","volume":"83","author":"Frierson J. 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Yellow fever global vaccine stockpile in emergencies.https:\/\/www.who.int\/news-room\/feature-stories\/detail\/yellow-fever-global-vaccine-stockpile-in-emergencies#:~:text=The%20response%20to%20the%20Angola an%20outbreak%20in%201%20country"}],"container-title":["Biotechnology and Bioengineering"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/bit.27864","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/bit.27864","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/bit.27864","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T21:55:08Z","timestamp":1760046908000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/bit.27864"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,7]]},"references-count":37,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2021,9]]}},"alternative-id":["10.1002\/bit.27864"],"URL":"https:\/\/doi.org\/10.1002\/bit.27864","archive":["Portico"],"relation":{},"ISSN":["0006-3592","1097-0290"],"issn-type":[{"value":"0006-3592","type":"print"},{"value":"1097-0290","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,7]]},"assertion":[{"value":"2020-10-06","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2021-06-14","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2021-07-01","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}