{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T14:18:38Z","timestamp":1771337918517,"version":"3.50.1"},"reference-count":80,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2021,12,10]],"date-time":"2021-12-10T00:00:00Z","timestamp":1639094400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Strengthening the capacity of the Scientific Centre of Excellence CerVirVac for research in viral immunology and vaccinology financed by the European Regional Development Fund","award":["KK.01.1.1.01.0006"],"award-info":[{"award-number":["KK.01.1.1.01.0006"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>There is a large unmet need for a prophylactic vaccine against human cytomegalovirus (HCMV) to combat the ubiquitous infection that is ongoing with this pathogen. A vaccination against HCMV could protect immunocompromised patients and prevent birth defects caused by congenital HCMV infections. Moreover, cytomegalovirus (CMV) has a number of features that make it a very interesting vector platform for gene therapy. In both cases, preparation of a highly purified virus is a prerequisite for safe and effective application. Murine CMV (MCMV) is by far the most studied model for HCMV infections with regard to the principles that govern the immune surveillance of CMVs. Knowledge transfer from MCMV and mice to HCMV and humans could be facilitated by better understanding and characterization of the biological and biophysical properties of both viruses. We carried out a detailed investigation of HCMV and MCMV growth kinetics as well as stability under the influence of clarification and different storage conditions. Further, we investigated the possibilities to concentrate and purify both viruses by ultracentrifugation and ion-exchange chromatography. Defective enveloped particles were not separately analyzed; however, the behavior of exosomes was examined during all experiments. The effectiveness of procedures was monitored using CCID50 assay, Nanoparticle tracking analysis, ELISA for host cell proteins, and quantitative PCR for host cell DNA. MCMV generally proved to be more robust in handling. Despite its greater sensitivity, HCMV was efficiently (100% recovery) purified and concentrated by anion-exchange chromatography using QA monolithic support. The majority of the host genomic DNA as well as most of the host cell proteins were removed by this procedure.<\/jats:p>","DOI":"10.3390\/v13122481","type":"journal-article","created":{"date-parts":[[2021,12,13]],"date-time":"2021-12-13T03:14:29Z","timestamp":1639365269000},"page":"2481","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Production- and Purification-Relevant Properties of Human and Murine Cytomegalovirus"],"prefix":"10.3390","volume":"13","author":[{"given":"Sanda","family":"Ravli\u0107","sequence":"first","affiliation":[{"name":"Centre for Research and Knowledge Transfer in Biotechnology, University of Zagreb, 10000 Zagreb, Croatia"},{"name":"Center of Excellence for Viral Immunology and Vaccines, CERVirVac, 10000 Zagreb, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6080-8808","authenticated-orcid":false,"given":"Marija","family":"Brgles","sequence":"additional","affiliation":[{"name":"Centre for Research and Knowledge Transfer in Biotechnology, University of Zagreb, 10000 Zagreb, Croatia"},{"name":"Center of Excellence for Viral Immunology and Vaccines, CERVirVac, 10000 Zagreb, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lea","family":"Hir\u0161l","sequence":"additional","affiliation":[{"name":"Center of Excellence for Viral Immunology and Vaccines, CERVirVac, 10000 Zagreb, Croatia"},{"name":"Croatia Center for Proteomics, Faculty of Medicine, University of Rijeka, 51000 Rijeka, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stipan","family":"Jonji\u0107","sequence":"additional","affiliation":[{"name":"Center of Excellence for Viral Immunology and Vaccines, CERVirVac, 10000 Zagreb, Croatia"},{"name":"Croatia Center for Proteomics, Faculty of Medicine, University of Rijeka, 51000 Rijeka, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7370-0997","authenticated-orcid":false,"given":"Beata","family":"Halassy","sequence":"additional","affiliation":[{"name":"Centre for Research and Knowledge Transfer in Biotechnology, University of Zagreb, 10000 Zagreb, Croatia"},{"name":"Center of Excellence for Viral Immunology and Vaccines, CERVirVac, 10000 Zagreb, Croatia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,12,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1301","DOI":"10.1099\/vir.0.79888-0","article-title":"Genetic content of wild-type human cytomegalovirus","volume":"85","author":"Dolan","year":"2004","journal-title":"J. Gen. Virol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1088","DOI":"10.1126\/science.1227919","article-title":"Decoding human cytomegalovirus","volume":"338","author":"Weisburd","year":"2012","journal-title":"Science"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1007\/s00430-007-0066-x","article-title":"The history of cytomegalovirus and its diseases","volume":"197","author":"Ho","year":"2008","journal-title":"Med. Microbiol. Immunol."},{"key":"ref_4","unstructured":"Reddehase, M.J. (2013). Synopsis of Clinical Aspects of Human Cytomegalovirus Disease. Cytomegaloviruses: From Molecular Pathogenesis to Intervention, Caister Academic Press."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1366","DOI":"10.1056\/NEJM200105033441804","article-title":"Intrauterine Transmission of Cytomegalovirus to Infants of Women with Preconceptional Immunity","volume":"344","author":"Boppana","year":"2001","journal-title":"N. Engl. J. Med."},{"key":"ref_6","unstructured":"Reddehase, M.J. (2013). The Epidemiology and Public Health Impact of Congenital Cytomegalovirus Infection. Cytomegaloviruses: From Molecular Pathogenesis to Intervention, Caister Academic Press."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1128\/CMR.00034-08","article-title":"Immunobiology of Human Cytomegalovirus: From Bench to Bedside","volume":"22","author":"Tania","year":"2009","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1128\/CMR.00062-12","article-title":"The \u201cSilent\u201d Global Burden of Congenital Cytomegalovirus","volume":"26","author":"Sheetal","year":"2013","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2022","DOI":"10.4049\/jimmunol.170.4.2022","article-title":"Memory Inflation: Continuous Accumulation of Antiviral CD8+ T Cells Over Time","volume":"170","author":"Karrer","year":"2003","journal-title":"J. Immunol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"911","DOI":"10.1016\/S0531-5565(03)00134-7","article-title":"Age-associated accumulation of CMV-specific CD8+ T cells expressing the inhibitory killer cell lectin-like receptor G1 (KLRG1)","volume":"38","author":"Ouyang","year":"2003","journal-title":"Exp. Gerontol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1113","DOI":"10.1002\/eji.200425534","article-title":"Evolution of diverse antiviral CD8+ T cell populations after murine cytomegalovirus infection","volume":"35","author":"Sierro","year":"2005","journal-title":"Eur. J. Immunol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"673","DOI":"10.1084\/jem.20050882","article-title":"Broadly targeted human cytomegalovirus-specific CD4+ and CD8+ T cells dominate the memory compartments of exposed subjects","volume":"202","author":"Sylwester","year":"2005","journal-title":"J. Exp. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"650","DOI":"10.1016\/j.immuni.2008.07.017","article-title":"Memory Inflation during Chronic Viral Infection Is Maintained by Continuous Production of Short-Lived, Functional T Cells","volume":"29","author":"Snyder","year":"2008","journal-title":"Immunity"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Smith, C.J., Turula, H., and Snyder, C.M. (2014). Systemic Hematogenous Maintenance of Memory Inflation by MCMV Infection. PLoS Pathog., 10.","DOI":"10.1371\/journal.ppat.1004233"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1128\/JVI.76.1.151-164.2002","article-title":"Two Antigenic Peptides from Genes m123 and m164 of Murine Cytomegalovirus Quantitatively Dominate CD8 T-Cell Memory in the H-2d Haplotype","volume":"76","author":"Rafaela","year":"2002","journal-title":"J. Virol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1007\/s00430-019-00597-7","article-title":"Vaccine vectors: The bright side of cytomegalovirus","volume":"208","year":"2019","journal-title":"Med. Microbiol. Immunol."},{"key":"ref_17","first-page":"52","article-title":"CD8+ T cell programming by cytomegalovirus vectors: Applications in prophylactic and therapeutic vaccination","volume":"47","year":"2017","journal-title":"Physiol. Behav."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1038\/nature12519","article-title":"Immune clearance of highly pathogenic SIV infection","volume":"502","author":"Hansen","year":"2013","journal-title":"Nature"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1038\/nm.4473","article-title":"Prevention of tuberculosis in rhesus macaques by a cytomegalovirus-based vaccine","volume":"24","author":"Hansen","year":"2018","journal-title":"Nat. Med."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Hansen, S.G., Womack, J., Scholz, I., Renner, A., Edgel, K.A., Xu, G., Ford, J.C., Grey, M., St Laurent, B., and Turner, J.M. (2019). Cytomegalovirus vectors expressing Plasmodium knowlesi antigens induce immune responses that delay parasitemia upon sporozoite challenge. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0210252"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1430","DOI":"10.1038\/s41591-018-0130-7","article-title":"Early antiretroviral therapy limits SIV reservoir establishment to delay or prevent post-treatment viral rebound","volume":"24","author":"Okoye","year":"2018","journal-title":"Nat. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"7730","DOI":"10.1128\/JVI.02696-09","article-title":"A Spread-Deficient Cytomegalovirus for Assessment of First-Target Cells in Vaccination","volume":"84","author":"Andreas","year":"2010","journal-title":"J. Virol."},{"key":"ref_23","unstructured":"Yurochko, A.D., and Miller, W.E. (2014). Use of Recombinant Approaches to Construct Human Cytomegalovirus Mutants BT\u2013Human Cytomegaloviruses: Methods and Protocols, Humana Press."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1126\/science.1185350","article-title":"Evasion of CD8+ T cells is critical for superinfection by cytomegalovirus","volume":"328","author":"Hansen","year":"2010","journal-title":"Science"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"536","DOI":"10.1158\/2326-6066.CIR-14-0044","article-title":"Cytomegalovirus-Based Vaccine Expressing a Modified Tumor Antigen Induces Potent Tumor-Specific CD8+ T-cell Response and Protects Mice from Melanoma","volume":"3","author":"Qiu","year":"2015","journal-title":"Cancer Immunol. Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2306","DOI":"10.4049\/jimmunol.1302523","article-title":"A novel murine cytomegalovirus vaccine vector protects against Mycobacterium tuberculosis","volume":"193","author":"Beverley","year":"2014","journal-title":"J. Immunol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1097\/CJI.0b013e3182585d50","article-title":"A cytomegalovirus-based vaccine expressing a single tumor-specific CD8+ T-cell epitope delays tumor growth in a murine model of prostate cancer","volume":"35","author":"Klyushnenkova","year":"2012","journal-title":"J. Immunother."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2261","DOI":"10.1016\/j.vaccine.2015.03.029","article-title":"A cytomegalovirus-based vaccine provides long-lasting protection against lethal Ebola virus challenge after a single dose","volume":"33","author":"Tsuda","year":"2015","journal-title":"Vaccine"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1038\/mi.2016.48","article-title":"Intranasal administration of RSV antigen-expressing MCMV elicits robust tissue-resident effector and effector memory CD8+ T cells in the lung","volume":"10","author":"Morabito","year":"2017","journal-title":"Mucosal Immunol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1016\/j.bbrc.2013.06.068","article-title":"Cytomegalovirus-based cancer vaccines expressing TRP2 induce rejection of melanoma in mice","volume":"437","author":"Xu","year":"2013","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"S196","DOI":"10.1093\/cid\/cit587","article-title":"Cytomegalovirus Vaccines","volume":"57","author":"McVoy","year":"2013","journal-title":"Clin. Infect. Dis."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1625","DOI":"10.1007\/s40265-016-0653-5","article-title":"Cytomegalovirus Vaccines: Current Status and Future Prospects","volume":"76","author":"Anderholm","year":"2016","journal-title":"Drugs"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1086\/421999","article-title":"Vaccine Development to Prevent Cytomegalovirus Disease: Report from the National Vaccine Advisory Committee","volume":"39","author":"Modlin","year":"2004","journal-title":"Clin. Infect. Dis."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"859","DOI":"10.1021\/bp000078j","article-title":"Effects of culture parameters on the production of retroviral vectors by a human packaging cell line","volume":"16","author":"McTaggart","year":"2000","journal-title":"Biotechnol. Prog."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"3261","DOI":"10.1016\/j.vaccine.2006.01.019","article-title":"Serum-free influenza virus production avoiding washing steps and medium exchange in large-scale microcarrier culture","volume":"24","author":"Genzel","year":"2006","journal-title":"Vaccine"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"854","DOI":"10.1021\/bp000110+","article-title":"Application of a serum-free medium for the growth of Vero cells and the production of reovirus","volume":"16","author":"Butler","year":"2000","journal-title":"Biotechnol. Prog."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"4456","DOI":"10.1016\/j.vaccine.2007.03.017","article-title":"Yield increases in intact influenza vaccine virus from chicken allantoic fluid through isolation from insoluble allantoic debris","volume":"25","author":"Hughes","year":"2007","journal-title":"Vaccine"},{"key":"ref_38","first-page":"38","article-title":"Impact of cell density and viability on primary clarification of mammalian cell broth","volume":"5","author":"Iammarino","year":"2007","journal-title":"Bioprocess Int."},{"key":"ref_39","first-page":"54","article-title":"Use of membrane technology in bioprocessing therapeutic proteins from inclusion bodies of E. coli","volume":"7","author":"Pattnaik","year":"2009","journal-title":"Bioprocess Int."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"30","DOI":"10.12665\/J111.Raghunath","article-title":"Best Practices for Optimization and Scale-Up of Microfiltration TFF Processes","volume":"11","author":"Raghunath","year":"2012","journal-title":"Bioprocess. J."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1016\/j.biotechadv.2005.12.001","article-title":"Downstream processing of oncoretroviral and lentiviral gene therapy vectors","volume":"24","author":"Kamen","year":"2006","journal-title":"Biotechnol. Adv."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1016\/j.biologicals.2014.05.007","article-title":"Vaccine instability in the cold chain: Mechanisms, analysis and formulation strategies","volume":"42","author":"Kumru","year":"2014","journal-title":"Biologicals"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1455","DOI":"10.1007\/s00705-016-2801-3","article-title":"Stability, biophysical properties and effect of ultracentrifugation and diafiltration on measles virus and mumps virus","volume":"161","author":"Sviben","year":"2016","journal-title":"Arch. Virol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1016\/j.jchromb.2013.02.004","article-title":"Fast separation of large biomolecules using short monolithic columns","volume":"927","author":"Podgornik","year":"2013","journal-title":"J. Chromatogr. B"},{"key":"ref_45","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_46","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.jchromb.2017.04.015","article-title":"Recovery of infective virus particles in ion-exchange and hydrophobic interaction monolith chromatography is influenced by particle charge and total-to-infective particle ratio","volume":"1054","author":"Sviben","year":"2017","journal-title":"J. Chromatogr. B Anal. Technol. Biomed. Life Sci."},{"key":"ref_47","first-page":"1591","article-title":"Chromatographic Purification of Virus Particles","volume":"m","author":"Flickinger","year":"2009","journal-title":"Encycl. Ind. Biotechnol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"2725","DOI":"10.1016\/j.chroma.2008.11.010","article-title":"Rapid high-performance liquid chromatographic analysis of adenovirus type 5 particles with a prototype anion-exchange analytical monolith column","volume":"1216","author":"Whitfield","year":"2009","journal-title":"J. Chromatogr. A"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.chroma.2012.11.058","article-title":"Fast purification of the filamentous Potato virus Y using monolithic chromatographic supports","volume":"1272","author":"Rupar","year":"2013","journal-title":"J. Chromatogr. A"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1016\/j.jchromb.2011.10.044","article-title":"A monolith purification process for virus-like particles from yeast homogenate","volume":"880","author":"Burden","year":"2012","journal-title":"J. Chromatogr. B"},{"key":"ref_51","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_52","doi-asserted-by":"crossref","first-page":"1451","DOI":"10.1586\/erv.11.111","article-title":"Downstream processing of cell culture-derived virus particles","volume":"10","author":"Wolf","year":"2011","journal-title":"Expert Rev. Vaccines"},{"key":"ref_53","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":"Hoen","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"766","DOI":"10.1128\/jvi.10.4.766-775.1972","article-title":"Nonviral microbodies with viral antigenicity produced in cytomegalovirus-infected cells","volume":"10","author":"Craighead","year":"1972","journal-title":"J. Virol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1016\/0042-6822(83)90122-8","article-title":"Isolation and characterization of a noninfectious virion-like particle released from cells infected with human strains of cytomegalovirus","volume":"130","author":"Irmiere","year":"1983","journal-title":"Virology"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"1887","DOI":"10.1016\/j.vaccine.2009.11.049","article-title":"Comparisons of mumps virus potency estimates obtained by 50% cell culture infective dose assay and plaque assay","volume":"28","author":"Forcic","year":"2010","journal-title":"Vaccine"},{"key":"ref_57","unstructured":"Barker, K. (2020, February 01). Phenol-Chloroform Isoamyl Alcohol (PCI) DNA Extraction. Bench 1998. Available online: http:\/\/hosted.usf.edu\/ecoimmunology\/wpcontent\/uploads\/2014\/07\/PCI-extraction.pdf."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1023\/A:1008133717035","article-title":"Media formulation options and manufacturing process controls to safeguard against introduction of animal origin contaminants in animal cell culture","volume":"33","author":"Jayme","year":"2000","journal-title":"Cytotechnology"},{"key":"ref_59","first-page":"255","article-title":"Replication of poliovirus (LSc 2 ab) in perfused cultures of MRC-5 diploid cells","volume":"37","author":"Mann","year":"1976","journal-title":"Dev. Biol. Stand."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1016\/0264-410X(93)90204-B","article-title":"Preparation and immunogenicity of an inactivated hepatitis A vaccine","volume":"11","author":"Pellegrini","year":"1993","journal-title":"Vaccine"},{"key":"ref_61","first-page":"97","article-title":"Human diploid cell strains (HDCS) viral vaccines","volume":"93","author":"Fletcher","year":"1998","journal-title":"Dev. Biol. Stand."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/0092-1157(76)90018-4","article-title":"The status of human diploid cell strain MRC-5 as an approved substrate for the production of viral vaccines","volume":"4","author":"Jacobs","year":"1976","journal-title":"J. Biol. Stand."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"2867","DOI":"10.1099\/0022-1317-80-11-2867","article-title":"Modification of Human Cytomegalovirus Tropism through Propagation In Vitro Is Associated with Changes in the Viral Genome","volume":"80","author":"Sinzger","year":"1999","journal-title":"J. Gen. Virol."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1099\/vir.0.83286-0","article-title":"Cloning and sequencing of a highly productive, endotheliotropic virus strain derived from human cytomegalovirus TB40\/E","volume":"89","author":"Sinzger","year":"2008","journal-title":"J. Gen. Virol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"914","DOI":"10.1128\/am.22.5.914-918.1971","article-title":"Propagation and purification of high-titer human cytomegalovirus","volume":"22","author":"Chambers","year":"1971","journal-title":"Appl. Microbiol."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1186\/1743-422X-11-71","article-title":"Resolving the titer of murine cytomegalovirus by plaque assay using the M2-10B4 cell line and a low viscosity overlay","volume":"11","author":"Zurbach","year":"2014","journal-title":"Virol. J."},{"key":"ref_67","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_68","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_69","doi-asserted-by":"crossref","first-page":"eaam6892","DOI":"10.1126\/science.aam6892","article-title":"Atomic structure of the human cytomegalovirus capsid with its securing tegument layer of pp150","volume":"356","author":"Yu","year":"2017","journal-title":"Science"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1159\/000150509","article-title":"Structure and assembly of the virion","volume":"39","author":"Gibson","year":"1996","journal-title":"Intervirology"},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"6696","DOI":"10.1016\/j.vaccine.2019.09.027","article-title":"Stabilization and formulation of a recombinant Human Cytomegalovirus vector for use as a candidate HIV-1 vaccine","volume":"37","author":"Kumru","year":"2019","journal-title":"Vaccine"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"10960","DOI":"10.1128\/JVI.78.20.10960-10966.2004","article-title":"Identification of proteins in human cytomegalovirus (HCMV) particles: The HCMV proteome","volume":"78","author":"Varnum","year":"2004","journal-title":"J. Virol."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"6997","DOI":"10.1128\/JVI.72.9.6997-7004.1998","article-title":"Identification of persistent RNA-DNA hybrid structures within the origin of replication of human cytomegalovirus","volume":"72","author":"Prichard","year":"1998","journal-title":"J. Virol."},{"key":"ref_74","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_75","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1186\/s12985-016-0463-0","article-title":"Identification of mumps virus protein and lipid composition by mass spectrometry","volume":"13","author":"Brgles","year":"2016","journal-title":"Virol. J."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"843","DOI":"10.1002\/bit.25545","article-title":"Improved virus purification processes for vaccines and gene therapy","volume":"112","author":"Nestola","year":"2015","journal-title":"Biotechnol. Bioeng."},{"key":"ref_77","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_78","doi-asserted-by":"crossref","unstructured":"Gomez, P.L., and Robinson, J.M. (2018). Vaccine Manufacturing. Plotkin\u2019s Vaccines, Elsevier. [7th ed.].","DOI":"10.1016\/B978-0-323-35761-6.00005-5"},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1128\/MMBR.00063-15","article-title":"Extracellular vesicles exploit viral entry routes for cargo delivery","volume":"80","author":"Masoumi","year":"2016","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.virol.2018.08.008","article-title":"Human cytomegalovirus-infected cells release extracellular vesicles that carry viral surface proteins","volume":"524","author":"Zicari","year":"2018","journal-title":"Virology"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/12\/2481\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:45:17Z","timestamp":1760168717000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/12\/2481"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,12,10]]},"references-count":80,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["v13122481"],"URL":"https:\/\/doi.org\/10.3390\/v13122481","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,12,10]]}}}