{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T12:11:47Z","timestamp":1792843907474,"version":"4.5.0"},"reference-count":38,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2023,12,29]],"date-time":"2023-12-29T00:00:00Z","timestamp":1703808000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"International Development Research Center (IDRC)","award":["109618"],"award-info":[{"award-number":["109618"]}]},{"name":"International Development Research Center (IDRC)","award":["2021-00032"],"award-info":[{"award-number":["2021-00032"]}]},{"name":"Canada Research Chair\u2014CRC","award":["109618"],"award-info":[{"award-number":["109618"]}]},{"name":"Canada Research Chair\u2014CRC","award":["2021-00032"],"award-info":[{"award-number":["2021-00032"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>Newcastle disease (ND) remains a critical disease affecting poultry in sub-Saharan Africa. In some countries, repeated outbreaks have a major impact on local economies and food security. Recently, we developed an adenovirus-vectored vaccine encoding the Fusion protein from an Ethiopian isolate of Newcastle disease virus (NDV). The adenoviral vector was designed, and a manufacturing process was developed in the context of the Livestock Vaccine Innovation Fund initiative funded by the International Development Research Centre (IDRC) of Canada. The industrially relevant recombinant vaccine technology platform is being transferred to the National Veterinary Institute (Ethiopia) for veterinary applications. Here, a manufacturing process using HEK293SF suspension cells cultured in stirred-tank bioreactors for the vaccine production is proposed. Taking into consideration supply chain limitations, options for serum-free media selection were evaluated. A streamlined downstream process including a filtration, an ultrafiltration, and a concentration step was developed. With high volumetric yields (infectious titers up to 5 \u00d7 109 TCID50\/mL) in the culture supernatant, the final formulations were prepared at 1010 TCID50\/mL, either in liquid or lyophilized forms. The liquid formulation was suitable and safe for mucosal vaccination and was stable for 1 week at 37 \u00b0C. Both the liquid and lyophilized formulations were stable after 6 months of storage at 4 \u00b0C. We demonstrate that the instillation of the adenoviral vector through the nasal cavity can confer protection to chickens against a lethal challenge with NDV. Overall, a manufacturing process for the adenovirus-vectored vaccine was developed, and protective doses were determined using a convenient route of delivery. Formulation and storage conditions were established, and quality control protocols were implemented.<\/jats:p>","DOI":"10.3390\/vaccines12010041","type":"journal-article","created":{"date-parts":[[2023,12,29]],"date-time":"2023-12-29T11:19:21Z","timestamp":1703848761000},"page":"41","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Intranasally Delivered Adenoviral Vector Protects Chickens against Newcastle Disease Virus: Vaccine Manufacturing and Stability Assessments for Liquid and Lyophilized Formulations"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1295-8323","authenticated-orcid":false,"given":"Omar","family":"Farn\u00f3s","sequence":"first","affiliation":[{"name":"Viral Vectors and Vaccines Bioprocessing Group, Department of Bioengineering, McGill University, Montreal, QC H3A 0G4, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6002-7972","authenticated-orcid":false,"given":"Barbara Cristina","family":"Martins Fernandes Paes","sequence":"additional","affiliation":[{"name":"Viral Vectors and Vaccines Bioprocessing Group, Department of Bioengineering, McGill University, Montreal, QC H3A 0G4, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4573-2913","authenticated-orcid":false,"given":"Belayneh","family":"Getachew","sequence":"additional","affiliation":[{"name":"Research and Development Directorate, National Veterinary Institute, Bishoftu P.O. Box 19, Ethiopia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4853-9422","authenticated-orcid":false,"given":"Samia","family":"Rourou","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Microbiology, Vaccinology and Biotechnology Development, Group of Biotechnology Development, Institut Pasteur de Tunis, Universit\u00e9 Tunis El Manar, 13, Place Pasteur. BP.74., Tunis 1002, Tunisia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ameni","family":"Chaabene","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Microbiology, Vaccinology and Biotechnology Development, Group of Biotechnology Development, Institut Pasteur de Tunis, Universit\u00e9 Tunis El Manar, 13, Place Pasteur. BP.74., Tunis 1002, Tunisia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7329-8632","authenticated-orcid":false,"given":"Esayas","family":"Gelaye","sequence":"additional","affiliation":[{"name":"Research and Development Directorate, National Veterinary Institute, Bishoftu P.O. Box 19, Ethiopia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takele A.","family":"Tefera","sequence":"additional","affiliation":[{"name":"Research and Development Directorate, National Veterinary Institute, Bishoftu P.O. Box 19, Ethiopia"}],"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":"Viral Vectors and Vaccines Bioprocessing Group, Department of Bioengineering, McGill University, Montreal, QC H3A 0G4, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,12,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Grace, D., Roesel, K., and Lore, T. (2014). Poverty and Gender Aspects of Food Safety and Informal Markets in Sub-Saharan Africa, ILRI (aka ILCA and ILRAD).","DOI":"10.4324\/9781315745046"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1016\/j.vetmic.2016.12.019","article-title":"Newcastle disease vaccines\u2014A solved problem or a continuous challenge?","volume":"206","author":"Dimitrov","year":"2017","journal-title":"Vet. Microbiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"7867","DOI":"10.1128\/AEM.02716-13","article-title":"Genetic Diversity of Newcastle Disease Virus in Wild Birds and Pigeons in West Africa","volume":"79","author":"Snoeck","year":"2013","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1089931","DOI":"10.3389\/fvets.2022.1089931","article-title":"Sero-prevalence of Newcastle disease and associated risk factors in chickens at backyard chicken production Kindo Koisha, Wolaita zone, Southern Ethiopia","volume":"9","author":"Wodajo","year":"2023","journal-title":"Front Vet. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1017\/S0043933910000528","article-title":"Poultry production and performance in the Federal Democratic Republic of Ethiopia","volume":"66","author":"Wilson","year":"2010","journal-title":"World Poult. Sci. J."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Tekelemariam, T.H., Walkden-Brown, S., Atire, F.A., Tefera, D.A., Alemayehu, D.H., and Gerber, P.F. (2022). Detection of Chicken Respiratory Pathogens in Live Markets of Addis Ababa, Ethiopia, and Epidemiological Implications. Vet. Sci., 9.","DOI":"10.3390\/vetsci9090503"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1016\/j.dci.2013.06.007","article-title":"Effects of Newcastle Disease Virus Vaccine Antibodies on the Shedding and Transmission of Challenge Viruses","volume":"41","author":"Miller","year":"2013","journal-title":"Dev. Comp. Immunol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2540","DOI":"10.1056\/NEJMp0803650","article-title":"Vaccine preparedness\u2014Are we ready for the next influenza pandemic?","volume":"358","author":"Wright","year":"2008","journal-title":"N. Engl. J. Med."},{"key":"ref_9","unstructured":"(2023). The WOAH Manual of Diagnostic Tests and Vaccines for Terrestrial Animals, OIE. [12th ed.]. Chapter 3.3.14. WOAH Terrestrial Manual 2021."},{"key":"ref_10","unstructured":"Allan, W.H., Lancaster, J.E., and Toth, B. (1978). Newcastle Disease Vaccines\u2014Their Production and Use, FAO."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Farn\u00f3s, O., Gelaye, E., Trabelsi, K., Bernier, A., Subramani, K., Kallel, H., Yami, M., and Kamen, A.A. (2020). Establishing a Robust Manufacturing Platform for Recombinant Veterinary Vaccines: An Adenovirus-Vector Vaccine to Control Newcastle Disease Virus Infections of Poultry in Sub-Saharan Africa. Vaccines, 8.","DOI":"10.3390\/vaccines8020338"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1002\/bit.27945","article-title":"Manufacturing a chimpanzee adenovirus-vectored SARS-CoV-2 vaccine to meet global needs","volume":"119","author":"Joe","year":"2022","journal-title":"Biotechnol. Bioeng."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1209","DOI":"10.1080\/21645515.2015.1016674","article-title":"Progress on adenovirus-vectored universal influenza vaccines","volume":"11","author":"Xiang","year":"2015","journal-title":"Hum. Vaccines Immunother."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"741","DOI":"10.1002\/biot.201400390","article-title":"Large-scale adenovirus and poxvirus-vectored vaccine manufacturing to enable clinical trials","volume":"10","author":"Kallel","year":"2015","journal-title":"Biotechnol. J."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"300","DOI":"10.1016\/j.vaccine.2011.10.095","article-title":"Process optimization and scale-up for production of rabies vaccine live adenovirus vector (AdRG1.3)","volume":"30,","author":"Shen","year":"2012","journal-title":"Vaccine"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"984","DOI":"10.1016\/j.vaccine.2013.12.015","article-title":"Oral vaccination and protection of red foxes (Vulpes vulpes) against rabies using ONRAB, an adenovirus-rabies recombinant vaccine","volume":"32","author":"Brown","year":"2014","journal-title":"Vaccine"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"790","DOI":"10.7589\/2017-09-242","article-title":"Field Trials of Ontario Rabies Vaccine Bait in the Northeastern USA, 2012\u20132014","volume":"54","author":"Gilbert","year":"2018","journal-title":"J. Wildl. Dis."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1111\/tbed.12724","article-title":"Safety profile of a replication-deficient human adenovirus-vectored foot-and-mouth disease virus serotype A24 subunit vaccine in cattle","volume":"65","author":"Barrera","year":"2018","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"6221","DOI":"10.1016\/j.vaccine.2019.08.017","article-title":"Duration of protection and humoral immunity induced by an adenovirus-vectored subunit vaccine for foot-and-mouth disease (FMD) in Holstein steers","volume":"37","author":"Sitt","year":"2019","journal-title":"Vaccine"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.virol.2019.07.022","article-title":"A chimpanzee adenoviral vector-based rabies vaccine protects beagle dogs from lethal rabies virus challenge","volume":"536","author":"Wang","year":"2019","journal-title":"Virology"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1002\/(SICI)1097-0290(19980905)59:5<567::AID-BIT6>3.0.CO;2-8","article-title":"Serum-free production of recombinant proteins and adenoviral vectors by 293SF-3F6 cells","volume":"59","author":"Cote","year":"1998","journal-title":"Biotechnol. Bioeng."},{"key":"ref_22","first-page":"2051","article-title":"Replication and supercoiling of simian virus 40 DNA in cell extracts from human cells","volume":"5","author":"Stillman","year":"1985","journal-title":"Mol. Cell. Biol."},{"key":"ref_23","first-page":"493","article-title":"A simple method of estimating fifty-percent endpoints","volume":"27","author":"Reed","year":"1938","journal-title":"Am. J. Hyg."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Berg, A., Wright, D., Dulal, P., Stedman, A., Fedosyuk, S., Francis, M.J., Charleston, B., Warimwe, G.M., and Douglas, A.D. (2021). Stability of Chimpanzee Adenovirus Vectored Vaccines (ChAdOx1 and ChAdOx2) in Liquid and Lyophilised Formulations. Vaccines, 9.","DOI":"10.3390\/vaccines9111249"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1016\/j.ijpharm.2011.12.056","article-title":"Investigation on formulation and preparation of adenovirus encoding human endostatin lyophilized powders","volume":"427","author":"Chen","year":"2012","journal-title":"Int. J. Pharm."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"6951","DOI":"10.1016\/j.vaccine.2019.04.056","article-title":"Simian adenovirus vector production for early-phase clinical trials: A simple method applicable to multiple serotypes and using entirely disposable product-contact components","volume":"37","author":"Fedosyuk","year":"2019","journal-title":"Vaccine"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2458","DOI":"10.1002\/jps.20157","article-title":"Development of stable liquid formulations for adenovirus-based vaccines","volume":"93","author":"Evans","year":"2004","journal-title":"J. Pharm. Sci."},{"key":"ref_28","unstructured":"Evans, R., Volkin, D., and Isopi, L. (2008). Adenovirus Formulations. (U.S. Patent 7456009B2)."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1043","DOI":"10.1002\/jps.23907","article-title":"Modulation of protein adsorption by poloxamer 188 in relation to polysorbates 80 and 20 at solid surfaces","volume":"103","author":"Kim","year":"2014","journal-title":"J. Pharm. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1264","DOI":"10.1016\/j.xphs.2018.10.057","article-title":"Solution stability of poloxamer 188 under stress conditions","volume":"108","author":"Wang","year":"2019","journal-title":"J. Pharm. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1186\/s41120-022-00055-4","article-title":"The degradation of poloxamer 188 in buffered formulation conditions","volume":"8","author":"Chen","year":"2022","journal-title":"AAPS Open"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"373","DOI":"10.3109\/10837459809009865","article-title":"Factors that influence stability of recombinant adenoviral preparations for human gene therapy","volume":"3","author":"Croyle","year":"1998","journal-title":"Pharm. Dev. Technol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1007\/s13346-012-0085-z","article-title":"Strategies for intranasal delivery of vaccines","volume":"3","author":"Zaman","year":"2013","journal-title":"Drug Deliv. Transl. Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1796","DOI":"10.4049\/jimmunol.168.4.1796","article-title":"Nasal-associated lymphoid tissue is a mucosal inductive site for virus-specific humoral and cellular immune responses","volume":"168","author":"Zuercher","year":"2002","journal-title":"J. Immunol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Kang, H., Yan, M., Yu, Q., and Yang, Q. (2013). Characteristics of nasal-associated lymphoid tissue (NALT) and nasal absorption capacity in chicken. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0084097"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Wang, J., Lan, Q., Zong, X., Zhu, G., Yang, R., Yang, G., Jiang, Y., Yang, W., Huang, H., and Shi, C. (2023). Protection against genotype VII Newcastle disease virus by a mucosal subunit vaccination based on bacterium-like particles bearing the F or HN antigen. Int. J. Biol. Macromol., 244.","DOI":"10.1016\/j.ijbiomac.2023.125293"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"896","DOI":"10.1016\/j.cell.2022.02.005","article-title":"Respiratory mucosal delivery of next-generation COVID-19 vaccine provides robust protection against both ancestral and variant strains of SARS-CoV-2","volume":"185","author":"Afkhami","year":"2022","journal-title":"Cell"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1038\/d41586-022-02851-0","article-title":"China and India approve nasal COVID vaccines\u2014Are they a game changer?","volume":"609","author":"Waltz","year":"2022","journal-title":"Nature"}],"container-title":["Vaccines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-393X\/12\/1\/41\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:44:22Z","timestamp":1760132662000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-393X\/12\/1\/41"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,12,29]]},"references-count":38,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2024,1]]}},"alternative-id":["vaccines12010041"],"URL":"https:\/\/doi.org\/10.3390\/vaccines12010041","relation":{},"ISSN":["2076-393X"],"issn-type":[{"value":"2076-393X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,12,29]]}}}