{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T02:36:22Z","timestamp":1791426982541,"version":"4.3.3"},"reference-count":44,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2021,11,9]],"date-time":"2021-11-09T00:00:00Z","timestamp":1636416000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","award":["APP1139754"],"award-info":[{"award-number":["APP1139754"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","award":["APP1178896"],"award-info":[{"award-number":["APP1178896"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>Alternative delivery systems such as the high-density microarray patch (HD-MAP) are being widely explored due to the variety of benefits they offer over traditional vaccine delivery methods. As vaccines are dry coated onto the HD-MAP, there is a need to ensure the stability of the vaccine in a solid state upon dry down. Other challenges faced are the structural stability during storage as a dried vaccine and during reconstitution upon application into the skin. Using a novel live chimeric virus vaccine candidate, BinJ\/DENV2-prME, we explored a panel of pharmaceutical excipients to mitigate vaccine loss during the drying and storage process. This screening identified human serum albumin (HSA) as the lead stabilizing excipient. When bDENV2-coated HD-MAPs were stored at 4 \u00b0C for a month, we found complete retention of vaccine potency as assessed by the generation of potent virus-neutralizing antibody responses in mice. We also demonstrated that HD-MAP wear time did not influence vaccine deposition into the skin or the corresponding immunological outcomes. The final candidate formulation with HSA maintained ~100% percentage recovery after 6 months of storage at 4 \u00b0C.<\/jats:p>","DOI":"10.3390\/vaccines9111301","type":"journal-article","created":{"date-parts":[[2021,11,9]],"date-time":"2021-11-09T21:39:07Z","timestamp":1636493947000},"page":"1301","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Developing a Stabilizing Formulation of a Live Chimeric Dengue Virus Vaccine Dry Coated on a High-Density Microarray Patch"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4886-9437","authenticated-orcid":false,"given":"Jovin J. Y.","family":"Choo","sequence":"first","affiliation":[{"name":"Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5316-0986","authenticated-orcid":false,"given":"Christopher L. D.","family":"McMillan","sequence":"additional","affiliation":[{"name":"Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3260-0548","authenticated-orcid":false,"given":"Germain J. P.","family":"Fernando","sequence":"additional","affiliation":[{"name":"Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD 4072, Australia"},{"name":"Translational Research Institute, Vaxxas Pty Ltd., 37 Kent Street, Brisbane, QLD 4102, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roy A.","family":"Hall","sequence":"additional","affiliation":[{"name":"Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2040-5190","authenticated-orcid":false,"given":"Paul R.","family":"Young","sequence":"additional","affiliation":[{"name":"Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0139-9829","authenticated-orcid":false,"given":"Jody","family":"Hobson-Peters","sequence":"additional","affiliation":[{"name":"Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4375-7890","authenticated-orcid":false,"given":"David A.","family":"Muller","sequence":"additional","affiliation":[{"name":"Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD 4072, Australia"},{"name":"Translational Research Institute, Vaxxas Pty Ltd., 37 Kent Street, Brisbane, QLD 4102, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,11,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1038\/nature12060","article-title":"The global distribution and burden of dengue","volume":"496","author":"Bhatt","year":"2013","journal-title":"Nature"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Holbrook, M.R. (2017). Historical Perspectives on Flavivirus Research. Viruses, 9.","DOI":"10.3390\/v9050097"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"16055","DOI":"10.1038\/nrdp.2016.55","article-title":"Dengue infection","volume":"2","author":"Guzman","year":"2016","journal-title":"Nat. Rev. Dis. Prim."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"326","DOI":"10.1007\/s12250-015-3624-1","article-title":"Dengue epidemiology and pathogenesis: Images of the future viewed through a mirror of the past","volume":"30","author":"Islam","year":"2015","journal-title":"Virol. Sin."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1016\/j.tim.2013.12.011","article-title":"Global spread of dengue virus types: Mapping the 70 year history","volume":"22","author":"Messina","year":"2014","journal-title":"Trends Microbiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"7112","DOI":"10.1016\/j.vaccine.2015.11.022","article-title":"Development of a recombinant, chimeric tetravalent dengue vaccine candidate","volume":"33","author":"Osorio","year":"2015","journal-title":"Vaccine"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1519","DOI":"10.1016\/j.clinthera.2017.07.001","article-title":"Progress and Works in Progress: Update on Flavivirus Vaccine Development","volume":"39","author":"Collins","year":"2017","journal-title":"Clin. Ther."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"957","DOI":"10.1093\/infdis\/jis936","article-title":"A Single Dose of Any of Four Different Live Attenuated Tetravalent Dengue Vaccines Is Safe and Immunogenic in Flavivirus-naive Adults: A Randomized, Double-blind Clinical Trial","volume":"207","author":"Durbin","year":"2013","journal-title":"J. Infect. Dis."},{"key":"ref_9","unstructured":"Limited, T.P.C. (2021, October 24). Takeda Begins Regulatory Submissions for Dengue Vaccine Candidate in EU and Dengue-Endemic Countries. Available online: https:\/\/www.takeda.com\/newsroom\/newsreleases\/2021\/takeda-begins-regulatory-submissions-for-dengue-vaccine-candidate-in-eu--and-dengue-endemic-countries\/."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"811","DOI":"10.4161\/hv.7.8.16274","article-title":"Transcutaneous and intradermal vaccination","volume":"7","author":"Combadiere","year":"2011","journal-title":"Hum. Vaccines"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1028","DOI":"10.1189\/jlb.0208105","article-title":"Viral replication and paracrine effects result in distinct, functional responses of dendritic cells following infection with dengue 2 virus","volume":"84","author":"Nightingale","year":"2008","journal-title":"J. Leukoc. Biol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"8833","DOI":"10.1016\/j.vaccine.2007.10.020","article-title":"Evaluation of the clinical performance of a new intradermal vaccine administration technique and associated delivery system","volume":"25","author":"Laurent","year":"2007","journal-title":"Vaccine"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"991","DOI":"10.1080\/21645515.2015.1010871","article-title":"Intradermal vaccination using the novel microneedle device MicronJet600: Past, present, and future","volume":"11","author":"Levin","year":"2015","journal-title":"Hum. Vaccines Immunother."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2361","DOI":"10.1038\/jid.2014.174","article-title":"Colocalization of Cell Death with Antigen Deposition in Skin Enhances Vaccine Immunogenicity","volume":"134","author":"Depelsenaire","year":"2014","journal-title":"J. Investig. Dermatol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1016\/j.jconrel.2012.01.030","article-title":"Nanopatch targeted delivery of both antigen and adjuvant to skin synergistically drives enhanced antibody responses","volume":"159","author":"Fernando","year":"2012","journal-title":"J. Control. Release"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Fernando, G., Chen, X., Prow, T., Crichton, M., Fairmaid, E.J., Roberts, M., Frazer, I., Brown, L., and Kendall, M.A.F. (2010). Potent Immunity to Low Doses of Influenza Vaccine by Probabilistic Guided Micro-Targeted Skin Delivery in a Mouse Model. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0010266"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Forster, A.H., Witham, K., Depelsenaire, A.C.I., Veitch, M., Wells, J.W., Wheatley, A., Pryor, M., Lickliter, J.D., Francis, B., and Rockman, S. (2020). Safety, tolerability, and immunogenicity of influenza vaccination with a high-density microarray patch: Results from a randomized, controlled phase I clinical trial. PLoS Med., 17.","DOI":"10.1371\/journal.pmed.1003024"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Muller, D.A., Depelsenaire, A.C.I., Shannon, A.E., Watterson, D., Corrie, S.R., Owens, N.S., Agyei-Yeboah, C., Cheung, S.T.M., Zhang, J., and Fernando, G.J.P. (2019). Efficient Delivery of Dengue Virus Subunit Vaccines to the Skin by Microprojection Arrays. Vaccines, 7.","DOI":"10.3390\/vaccines7040189"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-017-13011-0","article-title":"High-density microprojection array delivery to rat skin of low doses of trivalent inactivated poliovirus vaccine elicits potent neutralising antibody responses","volume":"7","author":"Muller","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"22094","DOI":"10.1038\/srep22094","article-title":"Inactivated poliovirus type 2 vaccine delivered to rat skin via high density microprojection array elicits potent neutralising antibody responses","volume":"6","author":"Muller","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41541-019-0134-4","article-title":"Microprojection arrays applied to skin generate mechanical stress, induce an inflammatory transcriptome and cell death, and improve vaccine-induced immune responses","volume":"4","author":"Ng","year":"2019","journal-title":"npj Vaccines"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"29368","DOI":"10.1038\/srep29368","article-title":"Potent response of QS-21 as a vaccine adjuvant in the skin when delivered with the Nanopatch, resulted in adjuvant dose sparing","volume":"6","author":"Ng","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1016\/j.jconrel.2012.07.024","article-title":"Induction of potent CD8+ T cell responses through the delivery of subunit protein vaccines to skin antigen-presenting cells using densely packed microprojection arrays","volume":"162","author":"Ng","year":"2012","journal-title":"J. Control. Release"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"3779","DOI":"10.1016\/j.vaccine.2018.05.053","article-title":"Safety, tolerability, acceptability and immunogenicity of an influenza vaccine delivered to human skin by a novel high-density microprojection array patch (Nanopatch\u2122)","volume":"36","author":"Fernando","year":"2018","journal-title":"Vaccine"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-020-75169-4","article-title":"Innate local response and tissue recovery following application of high density microarray patches to human skin","volume":"10","author":"Muller","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"McMillan, C.L.D., Choo, J.J.Y., Idris, A., Supramaniam, A., Modhiran, N., Amarilla, A.A., Isaacs, A., Cheung, S.T.M., Liang, B., and Bielefeldt-Ohmann, H. (2021). Complete protection by a single-dose skin patch\u2013delivered SARS-CoV-2 spike vaccine. Sci. Adv., 7.","DOI":"10.1126\/sciadv.abj8065"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41541-021-00328-1","article-title":"A chimeric dengue virus vaccine candidate delivered by high density microarray patches protects against infection in mice","volume":"6","author":"Choo","year":"2021","journal-title":"npj Vaccines"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1007\/s13346-018-00608-9","article-title":"Development of a thermostable microneedle patch for polio vaccination","volume":"9","author":"Kolluru","year":"2018","journal-title":"Drug Deliv. Transl. Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"740","DOI":"10.1002\/jps.24283","article-title":"Development of a Thermostable Microneedle Patch for Influenza Vaccination","volume":"104","author":"Mistilis","year":"2015","journal-title":"J. Pharm. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2501","DOI":"10.1080\/21645515.2021.1887692","article-title":"Formulation Development and Improved Stability of a Combination Measles and Rubella Live-Viral Vaccine Dried for Use in the NanopatchTM Microneedle Delivery System","volume":"17","author":"Wan","year":"2021","journal-title":"Hum. Vaccines Immunother."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1540","DOI":"10.1016\/j.xphs.2018.01.027","article-title":"Development of Stabilizing Formulations of a Trivalent Inactivated Poliovirus Vaccine in a Dried State for Delivery in the Nanopatch\u2122 Microprojection Array","volume":"107","author":"Wan","year":"2018","journal-title":"J. Pharm. Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-018-26800-y","article-title":"Molecular basis for dengue virus broad cross-neutralization by humanized monoclonal antibody 513","volume":"8","author":"Wong","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.str.2017.11.017","article-title":"Structural and Functional Characterization of a Cross-Reactive Dengue Virus Neutralizing Antibody that Recognizes a Cryptic Epitope","volume":"26","author":"Li","year":"2017","journal-title":"Structure"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1093\/oxfordjournals.aje.a118408","article-title":"A Simple Method of Estimating Fifty per Cent Endpoints12","volume":"27","author":"Reed","year":"1938","journal-title":"Am. J. Epidemiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"4712","DOI":"10.1016\/j.vaccine.2015.02.074","article-title":"A microneedle patch containing measles vaccine is immunogenic in non-human primates","volume":"33","author":"Edens","year":"2015","journal-title":"Vaccine"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1038\/ni.3058","article-title":"A new class of highly potent, broadly neutralizing antibodies isolated from viremic patients infected with dengue virus","volume":"16","author":"Dejnirattisai","year":"2014","journal-title":"Nat. Immunol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1017\/S002217240003936X","article-title":"The measurement of the D-antigen in poliovirus preparations","volume":"60","author":"Beale","year":"1962","journal-title":"J. Hyg."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"e10127","DOI":"10.1002\/btm2.10127","article-title":"Microneedle-based intradermal delivery of stabilized dengue virus","volume":"4","author":"Turvey","year":"2019","journal-title":"Bioeng. Transl. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"930","DOI":"10.1093\/infdis\/jiv532","article-title":"Stability of a Vesicular Stomatitis Virus\u2013Vectored Ebola Vaccine","volume":"213","author":"Arnemo","year":"2015","journal-title":"J. Infect. Dis."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s12887-019-1425-7","article-title":"Safety and immunogenicity of a varicella vaccine without human serum albumin (HSA) versus a HSA-containing formulation administered in the second year of life: A phase III, double-blind, randomized study","volume":"19","author":"Faust","year":"2019","journal-title":"BMC Pediatr."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2132","DOI":"10.1016\/j.vaccine.2015.03.017","article-title":"M-M-R\u00aeII manufactured using recombinant human albumin (rHA) and M-M-R\u00aeII manufactured using human serum albumin (HSA) exhibit similar safety and immunogenicity profiles when administered as a 2-dose regimen to healthy children","volume":"33","author":"Wiedmann","year":"2015","journal-title":"Vaccine"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"7456","DOI":"10.1016\/j.vaccine.2011.07.054","article-title":"Novel formulations enhance the thermal stability of live-attenuated flavivirus vaccines","volume":"29","author":"Wiggan","year":"2011","journal-title":"Vaccine"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"544","DOI":"10.3109\/10837450.2012.751408","article-title":"Handbook of Pharmaceutical Excipients\u20147th Edition","volume":"18","author":"Raymond","year":"2013","journal-title":"Pharm. Dev. Technol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/j.jconrel.2012.12.010","article-title":"Depth-resolved characterization of diffusion properties within and across minimally-perturbed skin layers","volume":"166","author":"Raphael","year":"2013","journal-title":"J. Control. 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