{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T04:54:51Z","timestamp":1768280091266,"version":"3.49.0"},"reference-count":33,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2024,7,23]],"date-time":"2024-07-23T00:00:00Z","timestamp":1721692800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100012399","name":"Sanofi","doi-asserted-by":"publisher","award":["W15QKN-16-9-1002"],"award-info":[{"award-number":["W15QKN-16-9-1002"]}],"id":[{"id":"10.13039\/100012399","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100012399","name":"Sanofi","doi-asserted-by":"publisher","award":["W15QKN-16-9-1002"],"award-info":[{"award-number":["W15QKN-16-9-1002"]}],"id":[{"id":"10.13039\/100012399","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100012399","name":"Biomedical Advanced Research and Development Authority","doi-asserted-by":"publisher","award":["W15QKN-16-9-1002"],"award-info":[{"award-number":["W15QKN-16-9-1002"]}],"id":[{"id":"10.13039\/100012399","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100012399","name":"Biomedical Advanced Research and Development Authority","doi-asserted-by":"publisher","award":["W15QKN-16-9-1002"],"award-info":[{"award-number":["W15QKN-16-9-1002"]}],"id":[{"id":"10.13039\/100012399","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>The purpose of this study was to develop a formulation for a recombinant prefusion spike protein vaccine against SARS-CoV-2. It was found that the spike protein was susceptible to aggregation due to mechanical stress. Therefore, formulation studies were initiated focused on screening pharmaceutical excipients capable of preventing this. The screening of a panel of potential stabilizing conditions found that Tween 20 could inhibit mechanically induced aggregation. A concentration-dependent study indicated that a higher concentration of Tween 20 (0.2% v\/v) was required to prevent conformational changes in the trimer. The conformational changes induced by mechanical stress were characterized by size exclusion chromatography (SEC) and hydrogen\u2013deuterium exchange mass spectrometry (HDX-MS), indicating the formation of an extended trimeric conformation that was also unable to bind to antibodies directed to the S2 domain. Long-term stability modeling, using advanced kinetic analysis, indicated that the formulation containing 0.2% (v\/v) Tween 20 at a neutral pH was predicted to be stable for at least two years at 2 \u00b0C to 8 \u00b0C. Additional stabilizer screening conducted by thermal shift assay indicated that sucrose and glycerol were able to significantly increase the spike protein melting temperature (Tm) and improve the overall thermostability of the spike protein in a short-term stability study. Thus, while 0.2% (v\/v) Tween 20 was sufficient to prevent aggregation and to maintain spike protein stability under refrigeration, the addition of sucrose further improved vaccine thermostability. Altogether, our study provides a systematic approach to the formulation of protein-based COVID-19 vaccine and highlights the impact of mechanical stress on the conformation of the spike protein and the significance of surfactants and stabilizers in maintaining the structural and functional integrity of the spike protein.<\/jats:p>","DOI":"10.3390\/vaccines12080830","type":"journal-article","created":{"date-parts":[[2024,7,23]],"date-time":"2024-07-23T14:26:50Z","timestamp":1721744810000},"page":"830","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Formulation Development of a COVID-19 Recombinant Spike Protein-Based Vaccine"],"prefix":"10.3390","volume":"12","author":[{"given":"Emily","family":"Xiao","sequence":"first","affiliation":[{"name":"Global Vaccine Drug Product Development, Sanofi, 1755 Steeles Avenue West, Toronto, ON M2R 3T4, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cl\u00e9mentine","family":"Mirabel","sequence":"additional","affiliation":[{"name":"Global Vaccine Drug Product Development, Sanofi, 1755 Steeles Avenue West, Toronto, ON M2R 3T4, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8395-9430","authenticated-orcid":false,"given":"Didier","family":"Cl\u00e9net","sequence":"additional","affiliation":[{"name":"Global Vaccine Drug Product Development, Sanofi, 1541 Avenue Marcel M\u00e9rieux, 69280 Marcy-L\u2019\u00c9toile, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shaolong","family":"Zhu","sequence":"additional","affiliation":[{"name":"Analytical Sciences, Sanofi, 1755 Steeles Avenue West, Toronto, ON M2R 3T4, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9069-6819","authenticated-orcid":false,"given":"Andrew","family":"James","sequence":"additional","affiliation":[{"name":"External Research and Development, Sanofi, 1755 Steeles Avenue West, Toronto, ON M2R 3T4, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luciano","family":"Ettorre","sequence":"additional","affiliation":[{"name":"Analytical Sciences, Sanofi, 1755 Steeles Avenue West, Toronto, ON M2R 3T4, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Trevor","family":"Williams","sequence":"additional","affiliation":[{"name":"Analytical Sciences, Sanofi, 1755 Steeles Avenue West, Toronto, ON M2R 3T4, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jason","family":"Szeto","sequence":"additional","affiliation":[{"name":"Analytical Sciences, Sanofi, 1755 Steeles Avenue West, Toronto, ON M2R 3T4, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nausheen","family":"Rahman","sequence":"additional","affiliation":[{"name":"Global Vaccine Drug Product Development, Sanofi, 1755 Steeles Avenue West, Toronto, ON M2R 3T4, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7001-7615","authenticated-orcid":false,"given":"Salvador Fernando","family":"Ausar","sequence":"additional","affiliation":[{"name":"Global Vaccine Drug Product Development, Sanofi, 1755 Steeles Avenue West, Toronto, ON M2R 3T4, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,7,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1016\/j.cell.2020.02.058","article-title":"Structure, Function, and Antigenicity of the SARS-CoV-2 Spike Glycoprotein","volume":"181","author":"Walls","year":"2020","journal-title":"Cell"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1016\/j.coviro.2021.02.006","article-title":"Coronavirus entry: How we arrived at SARS-CoV-2","volume":"47","author":"Whittaker","year":"2021","journal-title":"Curr. 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