{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,10]],"date-time":"2026-04-10T05:55:16Z","timestamp":1775800516104,"version":"3.50.1"},"reference-count":58,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2026,3,4]],"date-time":"2026-03-04T00:00:00Z","timestamp":1772582400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0"},{"start":{"date-parts":[[2026,4,10]],"date-time":"2026-04-10T00:00:00Z","timestamp":1775779200000},"content-version":"vor","delay-in-days":37,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0"}],"funder":[{"name":"Novo Nordisk Foundation Postdoctoral Fellowship","award":["NNF22OC0078741"],"award-info":[{"award-number":["NNF22OC0078741"]}]},{"name":"Marie Sk\u0142odowska-Curie Actions (MSCA) Postdoctoral Fellowship","award":["101105465"],"award-info":[{"award-number":["101105465"]}]},{"name":"Plan General del Conocimiento, Ministerio de Ciencia, Innovaci\u00f3n y Universidades, Gobierno de Espa\u00f1a","award":["PID2022-139019OB-I00"],"award-info":[{"award-number":["PID2022-139019OB-I00"]}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"DOI":"10.1038\/s41598-026-41596-y","type":"journal-article","created":{"date-parts":[[2026,3,4]],"date-time":"2026-03-04T08:36:58Z","timestamp":1772613418000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of a scalable production bioprocess for HIV-1 virus-like particles coupling continuous VLP harvesting with end-to-end downstream processing"],"prefix":"10.1038","volume":"16","author":[{"given":"Elianet","family":"Lorenzo","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jes\u00fas","family":"Lavado-Garc\u00eda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pol","family":"P\u00e9rez-Rubio","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maurizio","family":"Cattaneo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesc","family":"G\u00f2dia","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Laura","family":"Cervera","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2026,3,4]]},"reference":[{"key":"41596_CR1","doi-asserted-by":"publisher","first-page":"174","DOI":"10.1016\/j.nbt.2017.07.010","volume":"39","author":"J Fuenmayor","year":"2017","unstructured":"Fuenmayor, J., G\u00f2dia, F. & Cervera, L. Production of virus-like particles for vaccines. New Biotechnol. 39, 174\u2013180. https:\/\/doi.org\/10.1016\/j.nbt.2017.07.010 (2017).","journal-title":"New Biotechnol."},{"key":"41596_CR2","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s12951-021-00806-7","volume":"19","author":"S Nooraei","year":"2021","unstructured":"Nooraei, S. et al. Virus-like particles: preparation, immunogenicity and their roles as nanovaccines and drug nanocarriers. J. Nanobiotechnology 19, 1\u201327 (2021).","journal-title":"J. Nanobiotechnology"},{"key":"41596_CR3","doi-asserted-by":"publisher","first-page":"3122","DOI":"10.1128\/JVI.80.6.3122-3125.2006","volume":"80","author":"R Zu\u00f1iga","year":"2006","unstructured":"Zu\u00f1iga, R. et al. Relative Dominance of Gag p24-Specific Cytotoxic T Lymphocytes Is Associated with Human Immunodeficiency Virus Control. J. Virol. 80, 3122\u20133125 (2006).","journal-title":"J. Virol."},{"key":"41596_CR4","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/1423-0127-16-69","volume":"16","author":"CD Lee","year":"2009","unstructured":"Lee, C. D., Yan, Y. P., Liang, S. M. & Wang, T. F. Production of FMDV virus-like particles by a SUMO fusion protein approach in Escherichia coli. J. Biomed. Sci. 16, 1\u20137 (2009).","journal-title":"J. Biomed. Sci."},{"key":"41596_CR5","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2019.07.057","author":"A Venereo-S\u00e1nchez","year":"2019","unstructured":"Venereo-S\u00e1nchez, A. et al. Characterization of influenza H1N1 Gag virus-like particles and extracellular vesicles co-produced in HEK-293SF. Vaccine https:\/\/doi.org\/10.1016\/j.vaccine.2019.07.057 (2019).","journal-title":"Vaccine"},{"key":"41596_CR6","doi-asserted-by":"publisher","first-page":"3043","DOI":"10.1016\/j.vaccine.2011.01.100","volume":"29","author":"BM Giles","year":"2011","unstructured":"Giles, B. M. & Ross, T. M. A computationally optimized broadly reactive antigen (COBRA) based H5N1 VLP vaccine elicits broadly reactive antibodies in mice and ferrets. Vaccine 29, 3043\u20133054 (2011).","journal-title":"Vaccine"},{"key":"41596_CR7","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/1743-422X-10-129","volume":"10","author":"AJ Chua","year":"2013","unstructured":"Chua, A. J. et al. A novel platform for virus-like particle-display of flaviviral envelope domain III: Induction of Dengue and West Nile virus neutralizing antibodies. Virol. J. 10, 1\u201318 (2013).","journal-title":"Virol. J."},{"key":"41596_CR8","doi-asserted-by":"publisher","first-page":"4060","DOI":"10.1128\/jvi.69.7.4060-4068.1995","volume":"69","author":"L Garnier","year":"1995","unstructured":"Garnier, L. et al. Incorporation of pseudorabies virus gD into human immunodeficiency virus type 1 Gag particles produced in baculovirus-infected cells. J. Virol. 69, 4060\u20134068 (1995).","journal-title":"J. Virol."},{"key":"41596_CR9","doi-asserted-by":"publisher","DOI":"10.3390\/vaccines9030251","author":"D Fontana","year":"2021","unstructured":"Fontana, D. et al. Chimeric vlps based on hiv-1 gag and a fusion rabies glycoprotein induce specific antibodies against rabies and foot-and-mouth disease virus. Vaccines https:\/\/doi.org\/10.3390\/vaccines9030251 (2021).","journal-title":"Vaccines"},{"key":"41596_CR10","doi-asserted-by":"publisher","first-page":"500","DOI":"10.1006\/viro.1995.1181","volume":"208","author":"N Osterrieder","year":"1995","unstructured":"Osterrieder, N., Wagner, R., Brandm\u00fcller, C., Schmidt, P. & Wolf, H. Protection against EHV-1 challenge infection in the murine model after vaccination with various formulations of recombinant glycoprotein gp14 (gB). Virology 208, 500\u2013510. https:\/\/doi.org\/10.1006\/viro.1995.1181 (1995).","journal-title":"Virology"},{"key":"41596_CR11","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1016\/j.virol.2009.09.012","volume":"395","author":"P Di Bonito","year":"2009","unstructured":"Di Bonito, P. et al. Anti-tumor CD8+ T cell immunity elicited by HIV-1-based virus-like particles incorporating HPV-16 E7 protein. Virology 395, 45\u201355 (2009).","journal-title":"Virology"},{"key":"41596_CR12","first-page":"1","volume":"10","author":"A Boix-Besora","year":"2022","unstructured":"Boix-Besora, A., Lorenzo, E., Lavado-Garc\u00eda, J., G\u00f2dia, F. & Cervera, L. Optimization, production, purification and characterization of HIV-1 GAG-based virus-like particles functionalized with SARS-CoV-2. Vaccines (Basel). 10, 1\u201318 (2022).","journal-title":"Vaccines (Basel)."},{"key":"41596_CR13","doi-asserted-by":"publisher","DOI":"10.1002\/wnan.1579","author":"MO Mohsen","year":"2020","unstructured":"Mohsen, M. O., Speiser, D. E., Knuth, A. & Bachmann, M. F. Virus-like particles for vaccination against cancer. Wiley Interdiscip. Rev. Nanomed. Nanobiotechnol. https:\/\/doi.org\/10.1002\/wnan.1579 (2020).","journal-title":"Wiley Interdiscip. Rev. Nanomed. Nanobiotechnol."},{"key":"41596_CR14","doi-asserted-by":"publisher","first-page":"918","DOI":"10.1080\/07388551.2017.1419459","volume":"38","author":"S Guti\u00e9rrez-Granados","year":"2018","unstructured":"Guti\u00e9rrez-Granados, S., Cervera, L., Kamen, A. A. & G\u00f2dia, F. Advancements in mammalian cell transient gene expression (TGE) technology for accelerated production of biologics. Crit. Rev. Biotechnol. 38, 918\u2013940. https:\/\/doi.org\/10.1080\/07388551.2017.1419459 (2018).","journal-title":"Crit. Rev. Biotechnol."},{"key":"41596_CR15","doi-asserted-by":"publisher","first-page":"1328","DOI":"10.1016\/j.biotechadv.2018.04.011","volume":"36","author":"JM Bielser","year":"2018","unstructured":"Bielser, J. M., Wolf, M., Souquet, J., Broly, H. & Morbidelli, M. Perfusion mammalian cell culture for recombinant protein manufacturing \u2013 A critical review. Biotechnol. Adv. 36, 1328\u20131340 (2018).","journal-title":"Biotechnol. Adv."},{"key":"41596_CR16","doi-asserted-by":"publisher","DOI":"10.1016\/j.biotechadv.2022.108017","author":"J Lavado-Garc\u00eda","year":"2022","unstructured":"Lavado-Garc\u00eda, J., P\u00e9rez-Rubio, P., Cervera, L. & G\u00f2dia, F. The cell density effect in animal cell-based bioprocessing: Questions, insights and perspectives. Biotechnol. Adv. https:\/\/doi.org\/10.1016\/j.biotechadv.2022.108017 (2022).","journal-title":"Biotechnol. Adv."},{"key":"41596_CR17","doi-asserted-by":"publisher","DOI":"10.1002\/btpr.2831","author":"J Hong","year":"2019","unstructured":"Hong, J. et al. Development of an alternating tangential flow (ATF) perfusion-based transient gene expression (TGE) bioprocess for universal influenza vaccine. Biotechnol. Prog. https:\/\/doi.org\/10.1002\/btpr.2831 (2019).","journal-title":"Biotechnol. Prog."},{"key":"41596_CR18","doi-asserted-by":"publisher","first-page":"617","DOI":"10.3389\/fbioe.2020.00617","volume":"8","author":"J Lavado-Garc\u00eda","year":"2020","unstructured":"Lavado-Garc\u00eda, J., Cervera, L. & G\u00f2dia, F. An alternative perfusion approach for the intensification of virus-like particle production in HEK293 cultures. Front. Bioeng. Biotechnol. 8, 617 (2020).","journal-title":"Front. Bioeng. Biotechnol."},{"key":"41596_CR19","doi-asserted-by":"publisher","first-page":"5947","DOI":"10.1007\/s00253-023-12689-9","volume":"107","author":"MD Hein","year":"2023","unstructured":"Hein, M. D. et al. Production of retroviral vectors in continuous high cell density culture. Appl. Microbiol. Biotechnol. 107, 5947\u20135961 (2023).","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"41596_CR20","doi-asserted-by":"publisher","DOI":"10.1016\/j.jip.2011.05.004","author":"T Vicente","year":"2011","unstructured":"Vicente, T., Rold\u00e3o, A., Peixoto, C., Carrondo, M. J. T. & Alves, P. M. Large-scale production and purification of VLP-based vaccines. J. Invertebr. Pathol. https:\/\/doi.org\/10.1016\/j.jip.2011.05.004 (2011).","journal-title":"J. Invertebr. Pathol."},{"key":"41596_CR21","doi-asserted-by":"publisher","first-page":"2376","DOI":"10.1002\/jctb.6821","volume":"97","author":"M Mittal","year":"2022","unstructured":"Mittal, M., Banerjee, M., Lua, L. H. L. & Rathore, A. S. Current status and future challenges in transitioning to continuous bioprocessing of virus-like particles. J. Chem. Technol. Biotechnol. 97, 2376\u20132385 (2022).","journal-title":"J. Chem. Technol. Biotechnol."},{"key":"41596_CR22","doi-asserted-by":"publisher","first-page":"4532","DOI":"10.1021\/acs.jproteome.0c00581","volume":"19","author":"J Lavado-Garc\u00eda","year":"2020","unstructured":"Lavado-Garc\u00eda, J. et al. Molecular characterization of the coproduced extracellular vesicles in HEK293 during virus-like particle production. J. Proteome Res. 19, 4532 (2020).","journal-title":"J. Proteome Res."},{"key":"41596_CR23","doi-asserted-by":"publisher","first-page":"654","DOI":"10.1016\/j.tibtech.2013.09.002","volume":"31","author":"Q Zhao","year":"2013","unstructured":"Zhao, Q., Li, S., Yu, H., Xia, N. & Modis, Y. Virus-like particle-based human vaccines: Quality assessment based on structural and functional properties. Trends Biotechnol. 31, 654\u2013663 (2013).","journal-title":"Trends Biotechnol."},{"key":"41596_CR24","first-page":"1","volume":"00","author":"MG Moleirinho","year":"2019","unstructured":"Moleirinho, M. G., Silva, R. J. S., Alves, P. M., Carrondo, M. J. T. & Peixoto, C. Current challenges in biotherapeutic particles manufacturing. Expert Opin. Biol. Ther. 00, 1\u201315 (2019).","journal-title":"Expert Opin. Biol. Ther."},{"key":"41596_CR25","first-page":"1","volume":"9","author":"I Gonz\u00e1lez-Dom\u00ednguez","year":"2021","unstructured":"Gonz\u00e1lez-Dom\u00ednguez, I. et al. A four-step purification process for gag vlps: From culture supernatant to high-purity lyophilized particles. Vaccines (Basel). 9, 1\u201319 (2021).","journal-title":"Vaccines (Basel)."},{"key":"41596_CR26","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.chroma.2020.461378","volume":"1627","author":"P Pereira Aguilar","year":"2020","unstructured":"Pereira Aguilar, P. et al. Capture and purification of Human Immunodeficiency Virus-1 virus-like particles: Convective media vs porous beads. J. Chromatogr. A 1627, 1\u201311 (2020).","journal-title":"J. Chromatogr. A"},{"key":"41596_CR27","doi-asserted-by":"publisher","DOI":"10.1002\/jssc.201600765","author":"P Steppert","year":"2017","unstructured":"Steppert, P. et al. Separation of HIV-1 gag virus-like particles from vesicular particles impurities by hydroxyl-functionalized monoliths. J. Sep. Sci. https:\/\/doi.org\/10.1002\/jssc.201600765 (2017).","journal-title":"J. Sep. Sci."},{"key":"41596_CR28","doi-asserted-by":"publisher","first-page":"93","DOI":"10.1016\/j.chroma.2016.05.053","volume":"1455","author":"P Steppert","year":"2016","unstructured":"Steppert, P. et al. Purification of HIV-1 gag virus-like particles and separation of other extracellular particles. J. Chromatogr. A 1455, 93\u2013101 (2016).","journal-title":"J. Chromatogr. A"},{"key":"41596_CR29","doi-asserted-by":"publisher","first-page":"2672","DOI":"10.1002\/bit.28419","volume":"120","author":"E Lorenzo","year":"2023","unstructured":"Lorenzo, E., Miranda, L., G\u00f2dia, F. & Cervera, L. Downstream process design for Gag HIV-1 based virus-like particles. Biotechnol. Bioeng. 120, 2672\u20132684 (2023).","journal-title":"Biotechnol. Bioeng."},{"key":"41596_CR30","doi-asserted-by":"publisher","first-page":"1149","DOI":"10.1586\/erv.10.115","volume":"9","author":"A Rold\u00e3o","year":"2010","unstructured":"Rold\u00e3o, A., Mellado, M. C. M., Castilho, L. R., Carrondo, M. J. T. & Alves, P. M. Virus-like particles in vaccine development. Expert Rev. Vaccines 9, 1149\u20131176. https:\/\/doi.org\/10.1586\/erv.10.115 (2010).","journal-title":"Expert Rev. Vaccines"},{"key":"41596_CR31","doi-asserted-by":"publisher","DOI":"10.1016\/j.jpba.2019.113060","author":"\u00c9 Szab\u00f3","year":"2020","unstructured":"Szab\u00f3, \u00c9., Baranyai, L. Z., S\u00fct\u0151, Z., Salg\u00f3, A. & Gergely, S. Attenuated total reflection fourier transform infrared spectroscopy based methods for identification of chromatography media formulations used in downstream processes. J. Pharm. Biomed. Anal. https:\/\/doi.org\/10.1016\/j.jpba.2019.113060 (2020).","journal-title":"J. Pharm. Biomed. Anal."},{"key":"41596_CR32","doi-asserted-by":"publisher","first-page":"237","DOI":"10.1016\/j.biologicals.2014.05.007","volume":"42","author":"OS Kumru","year":"2014","unstructured":"Kumru, O. S. et al. Vaccine instability in the cold chain: Mechanisms, analysis and formulation strategies. Biologicals 42, 237\u2013259 (2014).","journal-title":"Biologicals"},{"key":"41596_CR33","doi-asserted-by":"publisher","first-page":"191","DOI":"10.1023\/B:PHAM.0000016234.73023.75","volume":"21","author":"XC Tang","year":"2004","unstructured":"Tang, X. C. & Pikal, M. J. Design of freeze-drying processes for pharmaceuticals: Practical advice. Pharm. Res. 21, 191\u2013200 (2004).","journal-title":"Pharm. Res."},{"key":"41596_CR34","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/pharmaceutics10030131","volume":"10","author":"F Emami","year":"2018","unstructured":"Emami, F., Vatanara, A., Park, E. J. & Na, D. H. Drying technologies for the stability and bioavailability of biopharmaceuticals. Pharmaceutics 10, 1\u201322 (2018).","journal-title":"Pharmaceutics"},{"key":"41596_CR35","doi-asserted-by":"publisher","first-page":"1085","DOI":"10.1021\/acs.jproteome.9b00601","volume":"19","author":"J Lavado-Garc\u00eda","year":"2020","unstructured":"Lavado-Garc\u00eda, J., Jorge, I., Cervera, L., V\u00e1zquez, J. & G\u00f2dia, F. Multiplexed quantitative proteomic analysis of HEK293 provides insights into molecular changes associated with the cell density effect, transient transfection, and virus-like particle production. J. Proteome Res. 19, 1085\u20131099 (2020).","journal-title":"J. Proteome Res."},{"key":"41596_CR36","doi-asserted-by":"publisher","DOI":"10.1002\/bit.27679","author":"J Lavado-Garc\u00eda","year":"2021","unstructured":"Lavado-Garc\u00eda, J. et al. Metabolic engineering of HEK293 cells to improve transient transfection and cell budding of HIV\u20101 virus\u2010like particles. Biotechnol. Bioeng. https:\/\/doi.org\/10.1002\/bit.27679 (2021).","journal-title":"Biotechnol. Bioeng."},{"key":"41596_CR37","doi-asserted-by":"publisher","first-page":"130","DOI":"10.1016\/j.bej.2016.07.003","volume":"114","author":"MM Buffo","year":"2016","unstructured":"Buffo, M. M. et al. Influence of dual-impeller type and configuration on oxygen transfer, power consumption, and shear rate in a stirred tank bioreactor. Biochem. Eng. J. 114, 130\u2013139 (2016).","journal-title":"Biochem. Eng. J."},{"key":"41596_CR38","doi-asserted-by":"publisher","first-page":"2672","DOI":"10.1002\/bit.28419","volume":"120","author":"E Lorenzo","year":"2023","unstructured":"Lorenzo, E., Miranda, L., G\u00f2dia, F. & Cervera, L. Downstream process design for Gag HIV - 1 based virus - like particles. Biotechnol. Bioeng. 120, 2672\u20132684 (2023).","journal-title":"Biotechnol. Bioeng."},{"key":"41596_CR39","doi-asserted-by":"publisher","first-page":"2660","DOI":"10.1002\/bit.27786","volume":"118","author":"J Lavado-Garc\u00eda","year":"2021","unstructured":"Lavado-Garc\u00eda, J. et al. Characterization of HIV-1 virus-like particles and determination of Gag stoichiometry for different production platforms. Biotechnol. Bioeng. 118, 2660\u20132675 (2021).","journal-title":"Biotechnol. Bioeng."},{"key":"41596_CR40","doi-asserted-by":"publisher","first-page":"934","DOI":"10.1002\/bit.25503","volume":"112","author":"L Cervera","year":"2015","unstructured":"Cervera, L., Guti\u00e9rrez-Granados, S., Berrow, N. S., Segura, M. M. & G\u00f2dia, F. Extended gene expression by medium exchange and repeated transient transfection for recombinant protein production enhancement. Biotechnol. Bioeng. 112, 934\u2013946 (2015).","journal-title":"Biotechnol. Bioeng."},{"key":"41596_CR41","doi-asserted-by":"publisher","first-page":"3040","DOI":"10.1002\/bit.27470","volume":"117","author":"A Nikolay","year":"2020","unstructured":"Nikolay, A., de Grooth, J., Genzel, Y., Wood, J. A. & Reichl, U. Virus harvesting in perfusion culture: Choosing the right type of hollow fiber membrane. Biotechnol. Bioeng. 117, 3040\u20133052 (2020).","journal-title":"Biotechnol. Bioeng."},{"key":"41596_CR42","doi-asserted-by":"publisher","first-page":"107","DOI":"10.1016\/j.coche.2018.09.009","volume":"22","author":"S Guti\u00e9rrez-Granados","year":"2018","unstructured":"Guti\u00e9rrez-Granados, S., G\u00f2dia, F. & Cervera, L. Continuous manufacturing of viral particles. Curr. Opin. Chem. Eng. 22, 107\u2013114 (2018).","journal-title":"Curr. Opin. Chem. Eng."},{"key":"41596_CR43","doi-asserted-by":"publisher","first-page":"81","DOI":"10.1016\/j.seppur.2019.02.040","volume":"218","author":"SB Carvalho","year":"2019","unstructured":"Carvalho, S. B. et al. Efficient filtration strategies for the clarification of influenza virus-like particles derived from insect cells. Sep. Purif. Technol. 218, 81\u201388 (2019).","journal-title":"Sep. Purif. Technol."},{"key":"41596_CR44","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1016\/j.biologicals.2017.11.002","volume":"51","author":"NT Lan","year":"2018","unstructured":"Lan, N. T. et al. Stability of virus-like particles of red-spotted grouper nervous necrosis virus in the aqueous state, and the vaccine potential of lyophilized particles. Biologicals 51, 25\u201331 (2018).","journal-title":"Biologicals"},{"key":"41596_CR45","doi-asserted-by":"publisher","DOI":"10.1016\/j.bej.2020.107868","volume":"167","author":"RM Comisel","year":"2021","unstructured":"Comisel, R. M., Kara, B., Fiesser, F. H. & Farid, S. S. Lentiviral vector bioprocess economics for cell and gene therapy commercialization. Biochem. Eng. J. 167, 107868 (2021).","journal-title":"Biochem. Eng. J."},{"key":"41596_CR46","doi-asserted-by":"publisher","first-page":"7251","DOI":"10.1007\/s00253-021-11561-y","volume":"105","author":"MD Hein","year":"2021","unstructured":"Hein, M. D. et al. Cell culture\u2013based production of defective interfering influenza A virus particles in perfusion mode using an alternating tangential flow filtration system. Appl. Microbiol. Biotechnol. 105, 7251\u20137264 (2021).","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"41596_CR47","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2024.101190","volume":"32","author":"P P\u00e9rez-Rubio","year":"2024","unstructured":"P\u00e9rez-Rubio, P. et al. Extracellular vesicle depletion and UGCG overexpression mitigate the cell density effect in HEK293 cell culture transfection. Mol. Ther. Methods Clin. Dev. 32, 101190 (2024).","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"41596_CR48","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1016\/j.chroma.2018.12.035","volume":"1588","author":"K Reiter","year":"2019","unstructured":"Reiter, K. et al. Separation of virus-like particles and extracellular vesicles by flow-through and heparin affinity chromatography. J. Chromatogr. A 1588, 77\u201384 (2019).","journal-title":"J. Chromatogr. A"},{"key":"41596_CR49","doi-asserted-by":"publisher","first-page":"7070","DOI":"10.1016\/j.vaccine.2019.07.001","volume":"37","author":"P Pereira Aguilar","year":"2019","unstructured":"Pereira Aguilar, P. et al. Polymer-grafted chromatography media for the purification of enveloped virus-like particles, exemplified with HIV-1 gag VLP. Vaccine 37, 7070\u20137080 (2019).","journal-title":"Vaccine"},{"key":"41596_CR50","doi-asserted-by":"publisher","first-page":"366","DOI":"10.2174\/1566523218666181109141257","volume":"18","author":"MG Moleirinho","year":"2018","unstructured":"Moleirinho, M. G. et al. Clinical-grade oncolytic adenovirus purification using polysorbate 20 as an alternative for cell lysis. Curr. Gene Ther. 18, 366\u2013374 (2018).","journal-title":"Curr. Gene Ther."},{"key":"41596_CR51","doi-asserted-by":"publisher","first-page":"203","DOI":"10.1016\/j.biologicals.2007.11.005","volume":"36","author":"I Knezevic","year":"2008","unstructured":"Knezevic, I., Stacey, G. & Petricciani, J. WHO study group on cell substrates for production of biologicals, Geneva, Switzerland, 11\u201312 June 2007. Biologicals 36, 203\u2013211 (2008).","journal-title":"Biologicals"},{"key":"41596_CR52","first-page":"1","volume":"11","author":"PL Ferreira","year":"2023","unstructured":"Ferreira, P. L. et al. Evaluation of novel chromatographic prototypes for supercoiled plasmid DNA polishing. Front. Bioeng. Biotechnol. 11, 1\u201313 (2023).","journal-title":"Front. Bioeng. Biotechnol."},{"key":"41596_CR53","doi-asserted-by":"publisher","first-page":"7061","DOI":"10.1016\/j.vaccine.2019.05.066","volume":"37","author":"TM Lima","year":"2019","unstructured":"Lima, T. M., Souza, M. O. & Castilho, L. R. Purification of flavivirus VLPs by a two-step chomatographic process. Vaccine 37, 7061\u20137069 (2019).","journal-title":"Vaccine"},{"key":"41596_CR54","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.jchromb.2017.05.029","volume":"1060","author":"MA Al-ghobashy","year":"2017","unstructured":"Al-ghobashy, M. A., Mostafa, M. M., Abed, H. S., Fathalla, F. A. & Salem, M. Y. Correlation between Dynamic Light Scattering and Size Exclusion High Performance Liquid Chromatography for monitoring the effect of pH on stability of biopharmaceuticals Correspondence\u202f: Aggregate formation is a major problem affecting both safety and eff. J. Chromatogr. B 1060, 1\u20139 (2017).","journal-title":"J. Chromatogr. B"},{"key":"41596_CR55","doi-asserted-by":"publisher","first-page":"1143","DOI":"10.1016\/j.vaccine.2015.01.031","volume":"33","author":"Y Yang","year":"2015","unstructured":"Yang, Y. et al. Size-exclusion HPLC provides a simple, rapid, and versatile alternative method for quality control of vaccines by characterizing the assembly of antigens. Vaccine 33, 1143\u20131150 (2015).","journal-title":"Vaccine"},{"key":"41596_CR56","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1007\/s00253-024-13389-8","volume":"109","author":"A D\u00edaz-Maneh","year":"2025","unstructured":"D\u00edaz-Maneh, A. et al. Targeted knockdown of ATM, ATR, and PDE\u03b4 increases Gag HIV-1 VLP production in HEK293 cells. Appl. Microbiol. Biotechnol. 109, 1\u201318 (2025).","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"41596_CR57","doi-asserted-by":"publisher","first-page":"1207","DOI":"10.1002\/bit.28051","volume":"119","author":"J Lavado-Garc\u00eda","year":"2022","unstructured":"Lavado-Garc\u00eda, J., Zhang, T., Cervera, L., G\u00f2dia, F. & Wuhrer, M. Differential N- and O-glycosylation signatures of HIV-1 Gag virus-like particles and coproduced extracellular vesicles. Biotechnol. Bioeng. 119, 1207 (2022).","journal-title":"Biotechnol. Bioeng."},{"key":"41596_CR58","doi-asserted-by":"publisher","first-page":"5851","DOI":"10.1021\/acs.iecr.8b04717","volume":"58","author":"O Yang","year":"2019","unstructured":"Yang, O., Prabhu, S. & Ierapetritou, M. Comparison between batch and continuous monoclonal antibody production and economic analysis. Ind. Eng. Chem. Res. 58, 5851\u20135863 (2019).","journal-title":"Ind. Eng. Chem. Res."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-026-41596-y","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-026-41596-y.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-026-41596-y.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,10]],"date-time":"2026-04-10T05:02:09Z","timestamp":1775797329000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-026-41596-y"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,4]]},"references-count":58,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,12]]}},"alternative-id":["41596"],"URL":"https:\/\/doi.org\/10.1038\/s41598-026-41596-y","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,4]]},"assertion":[{"value":"9 October 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"20 February 2026","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"4 March 2026","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"MC is the co-founder and a shareholder of Artemis Biosystems. Inc, supplier of the VHU-1 module. However, this author had no involvement in the study design, data collection, analysis, or interpretation of results. All other authors declare no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"12009"}}