{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T02:18:30Z","timestamp":1789957110332,"version":"4.0.1"},"reference-count":86,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2021,7,26]],"date-time":"2021-07-26T00:00:00Z","timestamp":1627257600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000038","name":"Natural Sciences and Engineering Research Council of Canada","doi-asserted-by":"publisher","award":["doctoral scholarship"],"award-info":[{"award-number":["doctoral scholarship"]}],"id":[{"id":"10.13039\/501100000038","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001804","name":"Canada Research Chairs","doi-asserted-by":"publisher","award":["CRC\/240394"],"award-info":[{"award-number":["CRC\/240394"]}],"id":[{"id":"10.13039\/501100001804","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>Viral vectors and viral vaccines are invaluable tools in prevention and treatment of diseases. Many infectious diseases are controlled using vaccines designed from subunits or whole viral structures, whereas other genetic diseases and cancers are being treated by viruses used as vehicles for delivering genetic material in gene therapy or as therapeutic agents in virotherapy protocols. Viral vectors and vaccines are produced in different platforms, from traditional embryonated chicken eggs to more advanced cell cultures. All these expression systems, like most cells and cellular tissues, are known to spontaneously release extracellular vesicles (EVs). EVs share similar sizes, biophysical characteristics and even biogenesis pathways with enveloped viruses, which are currently used as key ingredients in a number of viral vectors and licensed vaccine products. Herein, we review distinctive features and similarities between EVs and enveloped viruses as we revisit the downstream processing steps and analytical technologies currently implemented to produce and document viral vector and vaccine products. Within a context of well-established viral vector and vaccine safety profiles, this review provides insights on the likely presence of EVs in the final formulation of enveloped virus products and discusses the potential to further resolve and document these components.<\/jats:p>","DOI":"10.3390\/vaccines9080823","type":"journal-article","created":{"date-parts":[[2021,7,26]],"date-time":"2021-07-26T09:25:52Z","timestamp":1627291552000},"page":"823","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":43,"title":["Critical Assessment of Purification and Analytical Technologies for Enveloped Viral Vector and Vaccine Processing and Their Current Limitations in Resolving Co-Expressed Extracellular Vesicles"],"prefix":"10.3390","volume":"9","author":[{"given":"Aline","family":"Minh","sequence":"first","affiliation":[{"name":"Department of Bioengineering, McGill University, Montreal, QC H3A 0E9, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9110-8815","authenticated-orcid":false,"given":"Amine A.","family":"Kamen","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, McGill University, Montreal, QC H3A 0E9, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,7,26]]},"reference":[{"key":"ref_1","unstructured":"(2021, June 07). U.S. Food & Drug Administration. Vaccines Licensed for Use in the United States|FDA, Available online: https:\/\/www.fda.gov\/vaccines-blood-biologics\/vaccines\/vaccines-licensed-use-united-states."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"e3108","DOI":"10.1002\/jgm.3108","article-title":"Clinical translation of gene medicine","volume":"21","author":"Wang","year":"2019","journal-title":"J. Gene Med."},{"key":"ref_3","unstructured":"(2021, May 19). GTCT. Gene Therapy Clinical Trials Worldwide. Available online: http:\/\/www.abedia.com\/wiley."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1080\/14712598.2020.1693541","article-title":"Current challenges in biotherapeutic particles manufacturing","volume":"20","author":"Moleirinho","year":"2020","journal-title":"Expert Opin. Biol. Ther."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1016\/0264-410X(84)90045-8","article-title":"Vaccination with a live RA 27\/3 strain rubella vaccine (Rudivax), a follow-up study","volume":"2","author":"Suni","year":"1984","journal-title":"Vaccine"},{"key":"ref_6","unstructured":"(2021, June 07). U.S. Food & Drug Administration. Approved Cellular and Gene Therapy Products|FDA, Available online: https:\/\/www.fda.gov\/vaccines-blood-biologics\/cellular-gene-therapy-products\/approved-cellular-and-gene-therapy-products."},{"key":"ref_7","unstructured":"(2021, June 07). European Medicines Agency. Zynteglo. Available online: https:\/\/www.ema.europa.eu\/en\/medicines\/human\/EPAR\/zynteglo."},{"key":"ref_8","unstructured":"(2021, June 07). European Medicines Agency. Strimvelis. Available online: https:\/\/www.ema.europa.eu\/en\/medicines\/human\/EPAR\/strimvelis."},{"key":"ref_9","unstructured":"(2021, June 07). European Medicines Agency. Zalmoxis. Available online: https:\/\/www.ema.europa.eu\/en\/medicines\/human\/EPAR\/zalmoxis."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1083\/jcb.201211138","article-title":"Extracellular vesicles: Exosomes, microvesicles, and friends","volume":"200","author":"Raposo","year":"2013","journal-title":"J. Cell Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1038\/s41580-019-0158-7","article-title":"Extracellular vesicles: A new communication paradigm?","volume":"20","author":"Raposo","year":"2019","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Phillips, W., Willms, E., and Hill, A.F. (2021). Understanding extracellular vesicle and nanoparticle heterogeneity: Novel methods and considerations. Proteomics, e2000118.","DOI":"10.1002\/pmic.202000118"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1535750","DOI":"10.1080\/20013078.2018.1535750","article-title":"Minimal information for studies of extracellular vesicles 2018 (MISEV2018): A position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines","volume":"7","author":"Thery","year":"2018","journal-title":"J. Extracell. Vesicles"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"442","DOI":"10.3389\/fimmu.2014.00442","article-title":"Tetraspanins in extracellular vesicle formation and function","volume":"5","author":"Andreu","year":"2014","journal-title":"Front. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1016\/j.ceb.2014.05.004","article-title":"Biogenesis and secretion of exosomes","volume":"29","author":"Kowal","year":"2014","journal-title":"Curr. Opin. Cell Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"4146","DOI":"10.1073\/pnas.1200448109","article-title":"Formation and release of arrestin domain-containing protein 1-mediated microvesicles (ARMMs) at plasma membrane by recruitment of TSG101 protein","volume":"109","author":"Nabhan","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"e02344-17","DOI":"10.1128\/mBio.02344-17","article-title":"Nef Secretion into Extracellular Vesicles or Exosomes Is Conserved across Human and Simian Immunodeficiency Viruses","volume":"9","author":"McNamara","year":"2018","journal-title":"mBio"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1251","DOI":"10.1074\/jbc.M115.662171","article-title":"Exosomes from HIV-1-infected Cells Stimulate Production of Pro-inflammatory Cytokines through Trans-activating Response (TAR) RNA","volume":"291","author":"Sampey","year":"2016","journal-title":"J. Biol. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"10592","DOI":"10.1073\/pnas.1831413100","article-title":"The Trojan exosome hypothesis","volume":"100","author":"Gould","year":"2003","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"9155","DOI":"10.1073\/pnas.1605146113","article-title":"Extracellular vesicles and viruses: Are they close relatives?","volume":"113","author":"Cremer","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1186\/s12985-018-1073-9","article-title":"Mass spectrometry-based investigation of measles and mumps virus proteome","volume":"15","author":"Sviben","year":"2018","journal-title":"Virol. J."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Do Minh, A., Star, A.T., Stupak, J., Fulton, K.M., Haqqani, A.S., G\u00e9linas, J.-F., Li, J., Twine, S.M., and Kamen, A.A. (2021). Characterization of Extracellular Vesicles Secreted in Lentiviral Producing HEK293SF Cell Cultures. Viruses, 13.","DOI":"10.20944\/preprints202103.0696.v1"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1107","DOI":"10.1128\/JVI.01909-07","article-title":"Identification of host proteins associated with retroviral vector particles by proteomic analysis of highly purified vector preparations","volume":"82","author":"Segura","year":"2008","journal-title":"J. Virol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1016\/j.jconrel.2015.07.030","article-title":"Using exosomes, naturally-equipped nanocarriers, for drug delivery","volume":"219","author":"Batrakova","year":"2015","journal-title":"J. Control. Release"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Konadu, K.A., Huang, M.B., Roth, W., Armstrong, W., Powell, M., Villinger, F., and Bond, V. (2016). Isolation of Exosomes from the Plasma of HIV-1 Positive Individuals. J. Vis. Exp.","DOI":"10.3791\/53495-v"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.jim.2008.07.007","article-title":"Discrimination between exosomes and HIV-1: Purification of both vesicles from cell-free supernatants","volume":"338","author":"Cantin","year":"2008","journal-title":"J. Immunol. Methods"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"DeMarino, C., Barclay, R.A., Pleet, M.L., Pinto, D.O., Branscome, H., Paul, S., Lepene, B., El-Hage, N., and Kashanchi, F. (2019). Purification of High Yield Extracellular Vesicle Preparations Away from Virus. J. Vis. Exp.","DOI":"10.3791\/59876-v"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.biotechadv.2015.11.005","article-title":"Clarification of vaccines: An overview of filter based technology trends and best practices","volume":"34","author":"Besnard","year":"2016","journal-title":"Biotechnol. Adv."},{"key":"ref_29","first-page":"6","article-title":"Single-Use Depth Filters: Application in Clarifying Industrial Cell Cultures\u2014BioProcess International","volume":"16","author":"Schmidt","year":"2017","journal-title":"BioProcess Int."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1002\/bit.20301","article-title":"A novel purification strategy for retrovirus gene therapy vectors using heparin affinity chromatography","volume":"90","author":"Segura","year":"2005","journal-title":"Biotechnol. Bioeng."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"500","DOI":"10.1128\/aem.39.3.500-504.1980","article-title":"Simple method for the concentration of influenza virus from allantoic fluid on microporous filters","volume":"39","author":"Goyal","year":"1980","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_32","unstructured":"U.S. Food & Drug Administration (2019). Package Insert\u2014JYNNEOS, Bavarian Nordic A\/S."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"e2000019","DOI":"10.1002\/biot.202000019","article-title":"Advances in Lentivirus Purification","volume":"16","author":"Moreira","year":"2021","journal-title":"Biotechnol. J."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1389","DOI":"10.1128\/JCM.03109-12","article-title":"Development of a pseudotyped-lentiviral-vector-based neutralization assay for chikungunya virus infection","volume":"51","author":"Kishishita","year":"2013","journal-title":"J. Clin. Microbiol."},{"key":"ref_35","unstructured":"Turksen, K. (2019). High-Grade Purification of Third-Generation HIV-Based Lentiviral Vectors by Anion Exchange Chromatography for Experimental Gene and Stem Cell Therapy Applications. Skin Stem Cells: Methods and Protocols, Springer."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1016\/j.jbiotec.2006.07.028","article-title":"Purification of retroviral vectors for clinical application: Biological implications and technological challenges","volume":"127","author":"Rodrigues","year":"2007","journal-title":"J. Biotechnol."},{"key":"ref_37","unstructured":"U.S. Food & Drug Administration (2018). Package Insert and Patient Information\u2014IXIARO, Valneva Austria GmbH."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1002\/0471143030.cb0322s30","article-title":"Isolation and characterization of exosomes from cell culture supernatants and biological fluids","volume":"30","author":"Thery","year":"2006","journal-title":"Curr. Protoc. Cell Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2061","DOI":"10.1016\/j.nano.2017.03.011","article-title":"Higher functionality of extracellular vesicles isolated using size-exclusion chromatography compared to ultracentrifugation","volume":"13","author":"Mol","year":"2017","journal-title":"Nanomed. Nanotechnol. Biol. Med."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Van Deun, J., Mestdagh, P., Sormunen, R., Cocquyt, V., Vermaelen, K., Vandesompele, J., Bracke, M., De Wever, O., and Hendrix, A. (2014). The impact of disparate isolation methods for extracellular vesicles on downstream RNA profiling. J. Extracell. Vesicles, 3.","DOI":"10.3402\/jev.v3.24858"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Perry, C., and Rayat, A. (2021). Lentiviral Vector Bioprocessing. Viruses, 13.","DOI":"10.3390\/v13020268"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"846","DOI":"10.1002\/ceat.200800118","article-title":"Downstream Processing: From Egg to Cell Culture-Derived Influenza Virus Particles","volume":"31","author":"Wolff","year":"2008","journal-title":"Chem. Eng. Technol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1002\/jgm.1021","article-title":"Scaleable purification process for gene therapy retroviral vectors","volume":"9","author":"Rodrigues","year":"2007","journal-title":"J. Gene Med."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1299","DOI":"10.1002\/jgm.778","article-title":"Upscaling of lentiviral vector production by tangential flow filtration","volume":"7","author":"Geraerts","year":"2005","journal-title":"J. Gene Med."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"27031","DOI":"10.3402\/jev.v4.27031","article-title":"Optimized exosome isolation protocol for cell culture supernatant and human plasma","volume":"4","author":"Lobb","year":"2015","journal-title":"J. Extracell. Vesicles"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1016\/j.biotechadv.2013.01.007","article-title":"Recent advances in bioprocessing application of membrane chromatography","volume":"31","author":"Orr","year":"2013","journal-title":"Biotechnol. Adv."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Zimmermann, K., Scheibe, O., Kocourek, A., Muelich, J., Jurkiewicz, E., and Pfeifer, A. (2011). Highly efficient concentration of lenti- and retroviral vector preparations by membrane adsorbers and ultrafiltration. BMC Biotechnol., 11.","DOI":"10.1186\/1472-6750-11-55"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1016\/j.aca.2018.09.021","article-title":"Monolith columns for liquid chromatographic separations of intact proteins: A review of recent advances and applications","volume":"1046","author":"Lynch","year":"2019","journal-title":"Anal. Chim. Acta"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1010","DOI":"10.1080\/21645515.2015.1009817","article-title":"Downstream processing and chromatography based analytical methods for production of vaccines, gene therapy vectors, and bacteriophages","volume":"11","author":"Kramberger","year":"2015","journal-title":"Hum. Vaccines Immunother."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"981","DOI":"10.1016\/j.jchromb.2011.03.012","article-title":"Concentration and purification of rubella virus using monolithic chromatographic support","volume":"879","author":"Forcic","year":"2011","journal-title":"J. Chromatogr. B Anal. Technol. Biomed. Life Sci."},{"key":"ref_51","unstructured":"Hansen, S.P., Rene, F., Udo, R., Michael, W., and Gram, A.P. (2011). Purification of Vaccinia Viruses Using Hydrophobic Interaction Chromatography. (US20110306114A1), U.S. Patent."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/0166-0934(91)90186-4","article-title":"Purification of measles virus by affinity chromatography and by ultracentrifugation: A comparative study","volume":"32","author":"Njayou","year":"1991","journal-title":"J. Virol. Methods"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.chroma.2016.04.022","article-title":"Nonspecific native elution of proteins and mumps virus in immunoaffinity chromatography","volume":"1447","author":"Brgles","year":"2016","journal-title":"J. Chromatogr. A"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"939","DOI":"10.1016\/j.vaccine.2006.08.043","article-title":"Lectin-affinity chromatography for downstream processing of MDCK cell culture derived human influenza A viruses","volume":"25","author":"Opitz","year":"2007","journal-title":"Vaccine"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"564","DOI":"10.1128\/jvi.38.2.564-570.1981","article-title":"Unusual lectin-binding properties of a herpes simplex virus type 1-specific glycoprotein","volume":"38","author":"Olofsson","year":"1981","journal-title":"J. Virol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1016\/j.jviromet.2009.06.025","article-title":"Purification of cell culture-derived influenza virus A\/Puerto Rico\/8\/34 by membrane-based immobilized metal affinity chromatography","volume":"161","author":"Opitz","year":"2009","journal-title":"J. Virol. Methods"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"8994","DOI":"10.1128\/JVI.78.17.8994-9006.2004","article-title":"Immobilized Cobalt Affinity Chromatography Provides a Novel, Efficient Method for Herpes Simplex Virus Type 1 Gene Vector Purification","volume":"78","author":"Jiang","year":"2004","journal-title":"J. Virol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"9820","DOI":"10.1128\/JVI.78.18.9820-9827.2004","article-title":"Tagging Retrovirus Vectors with a Metal Binding Peptide and One-Step Purification by Immobilized Metal Affinity Chromatography","volume":"78","author":"Ye","year":"2004","journal-title":"J. Virol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"2705","DOI":"10.1016\/j.chroma.2008.08.029","article-title":"Immobilized metal affinity chromatography of histidine-tagged lentiviral vectors using monolithic adsorbents","volume":"1216","author":"Cheeks","year":"2009","journal-title":"J. Chromatogr. A"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"5491","DOI":"10.1016\/j.vaccine.2018.02.090","article-title":"Affinity chromatography for vaccines manufacturing: Finally ready for prime time?","volume":"37","author":"Zhao","year":"2019","journal-title":"Vaccine"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1002\/bit.10388","article-title":"Purification of a functional gene therapy vector derived from Moloney murine leukaemia virus using membrane filtration and ceramic hydroxyapatite chromatography","volume":"80","author":"Kuiper","year":"2002","journal-title":"Biotechnol. Bioeng."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"5730","DOI":"10.1038\/s41598-018-24163-y","article-title":"Rapid isolation and enrichment of extracellular vesicle preparations using anion exchange chromatography","volume":"8","author":"Heath","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1016\/j.omtm.2021.03.016","article-title":"Development of a scalable and robust AEX method for enriched rAAV preparations in genome-containing VCs of serotypes 5, 6, 8, and 9","volume":"21","author":"Joshi","year":"2021","journal-title":"Mol. Methods Clin. Dev."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"461773","DOI":"10.1016\/j.chroma.2020.461773","article-title":"Modern isolation and separation techniques for extracellular vesicles","volume":"1636","author":"Liangsupree","year":"2021","journal-title":"J. Chromatogr. A"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1089\/hgtb.2019.076","article-title":"Toward a Scalable Purification Protocol of GaLV-TR-Pseudotyped Lentiviral Vectors","volume":"30","author":"Boudeffa","year":"2019","journal-title":"Hum. Gene Ther. Methods"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"3721","DOI":"10.1016\/j.vaccine.2014.04.033","article-title":"Efficient chromatographic reduction of ovalbumin for egg-based influenza virus purification","volume":"32","author":"Blom","year":"2014","journal-title":"Vaccine"},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"11561","DOI":"10.1038\/s41598-017-10646-x","article-title":"Reproducible and scalable purification of extracellular vesicles using combined bind-elute and size exclusion chromatography","volume":"7","author":"Corso","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"1560808","DOI":"10.1080\/20013078.2018.1560808","article-title":"TEM ExosomeAnalyzer: A computer-assisted software tool for quantitative evaluation of extracellular vesicles in transmission electron microscopy images","volume":"8","author":"Ilkovics","year":"2019","journal-title":"J. Extracell. Vesicles"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"1555419","DOI":"10.1080\/20013078.2018.1555419","article-title":"Quality of extracellular vesicle images by transmission electron microscopy is operator and protocol dependent","volume":"8","author":"Rikkert","year":"2019","journal-title":"J. Extracell. Vesicles"},{"key":"ref_70","first-page":"1","article-title":"Droplet Digital Polymerase Chain Reaction Method for Absolute Quantification of Influenza A Viruses","volume":"33","author":"Feng","year":"2017","journal-title":"Bing Du Xue Bao"},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1089\/hgtb.2017.198","article-title":"Determination of Lentiviral Infectious Titer by a Novel Droplet Digital PCR Method","volume":"29","author":"Wang","year":"2018","journal-title":"Hum. Gene Ther. Methods"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"100806","DOI":"10.1016\/j.mex.2020.100806","article-title":"Titration methods for rVSV-based vaccine manufacturing","volume":"7","author":"Kiesslich","year":"2020","journal-title":"MethodsX"},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"7","DOI":"10.3389\/fmicb.2016.01500","article-title":"Quantification of Virus Particles Using Nanopore-Based Resistive-Pulse Sensing Techniques","volume":"7","author":"Yang","year":"2016","journal-title":"Front. Microbiol."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"589","DOI":"10.1016\/j.jviromet.2013.07.026","article-title":"Quantitation of influenza virus using field flow fractionation and multi-angle light scattering for quantifying influenza A particles","volume":"193","author":"Bousse","year":"2013","journal-title":"J. Virol. Methods"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"2596","DOI":"10.1111\/j.1538-7836.2010.04074.x","article-title":"Optical and non-optical methods for detection and characterization of microparticles and exosomes","volume":"8","author":"Hoekstra","year":"2010","journal-title":"J Thromb Haemost"},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"3377","DOI":"10.1016\/j.vaccine.2011.02.090","article-title":"Optimization and qualification of a quantitative reversed-phase HPLC method for hemagglutinin in influenza preparations and its comparative evaluation with biochemical assays","volume":"29","author":"Lorbetskie","year":"2011","journal-title":"Vaccine"},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1016\/j.jchromb.2004.09.034","article-title":"High-performance liquid chromatographic total particles quantification of retroviral vectors pseudotyped with vesicular stomatitis virus-G glycoprotein","volume":"813","author":"Transfiguracion","year":"2004","journal-title":"J. Chromatogr. B Anal. Technol. Biomed. Life Sci."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"803","DOI":"10.1016\/j.omtm.2020.08.005","article-title":"Rapid In-Process Monitoring of Lentiviral Vector Particles by High-Performance Liquid Chromatography","volume":"18","author":"Transfiguracion","year":"2020","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"ref_79","doi-asserted-by":"crossref","unstructured":"Joshi, P.R.H., Bernier, A., Chahal, P.S., and Kamen, A. (2021). Development and Validation of an Anion-Exchange High-Performance Liquid Chromatography Method for Analysis of Empty Capsids and Capsids Encapsidating Genetic Material in a Purified Preparation of Recombinant Adeno-Associated Virus Serotype 5. Hum. Gene Ther.","DOI":"10.1089\/hum.2020.317"},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"857","DOI":"10.3390\/v7030857","article-title":"Evaluation of ViroCyt\u00ae Virus Counter for rapid filovirus quantitation","volume":"7","author":"Rossi","year":"2015","journal-title":"Viruses"},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.virol.2017.08.005","article-title":"Droplet digital PCR for rapid enumeration of viral genomes and particles from cells and animals infected with orthopoxviruses","volume":"511","author":"Americo","year":"2017","journal-title":"Virology"},{"key":"ref_82","doi-asserted-by":"crossref","unstructured":"Lippe, R. (2018). Flow Virometry: A Powerful Tool To Functionally Characterize Viruses. J. Virol., 92.","DOI":"10.1128\/JVI.01765-17"},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"e00320-17","DOI":"10.1128\/JVI.00320-17","article-title":"Quantitative Evaluation of Protein Heterogeneity within Herpes Simplex Virus 1 Particles","volume":"91","author":"Duron","year":"2017","journal-title":"J. Virol."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"5082","DOI":"10.1016\/j.vaccine.2016.08.074","article-title":"Single-particle characterization of oncolytic vaccinia virus by flow virometry","volume":"34","author":"Tang","year":"2016","journal-title":"Vaccine"},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"17769","DOI":"10.1038\/s41598-017-18227-8","article-title":"Single-Particle Discrimination of Retroviruses from Extracellular Vesicles by Nanoscale Flow Cytometry","volume":"7","author":"Tang","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_86","doi-asserted-by":"crossref","unstructured":"Hawley, T.S., and Hawley, R.G. (2018). Analysis of Individual Extracellular Vesicles by Flow Cytometry. Flow Cytometry Protocols, Springer.","DOI":"10.1007\/978-1-4939-7346-0"}],"container-title":["Vaccines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-393X\/9\/8\/823\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:34:57Z","timestamp":1760164497000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-393X\/9\/8\/823"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,7,26]]},"references-count":86,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2021,8]]}},"alternative-id":["vaccines9080823"],"URL":"https:\/\/doi.org\/10.3390\/vaccines9080823","relation":{},"ISSN":["2076-393X"],"issn-type":[{"value":"2076-393X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,7,26]]}}}