{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T22:21:22Z","timestamp":1788387682758,"version":"build-2803163510"},"reference-count":57,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2026,2,5]],"date-time":"2026-02-05T00:00:00Z","timestamp":1770249600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NIFDC Fund for Key Technology Research","award":["GJJS-2022-1-4"],"award-info":[{"award-number":["GJJS-2022-1-4"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Background\/Objectives: Virus-like particles (VLPs) are effective vaccine platforms but are susceptible to degradation, which compromises stability and immunogenicity. A key challenge is the lack of sensitive early indicators of instability. This study aimed to systematically evaluate the stability of an aluminum-free recombinant hepatitis E virus VLP vaccine under various stresses and identify predictive markers of instability. Methods: The VLP vaccine was subjected to thermal stress (4 \u00b0C, 25 \u00b0C, 37 \u00b0C, 56 \u00b0C for up to 28 d), repeated freeze\u2013thaw cycles (up to 30 cycles), and mechanical agitation (orbital shaking at 100 and 300 rpm for up to 12 d). Stability was assessed using a multi-parameter panel monitoring critical quality attributes: conformational and colloidal stability, formation of high-molecular-weight species, mean particle size, polydispersity index, charge heterogeneity, and antigen content. Results: Changes in charge heterogeneity were the earliest indicator of instability, detectable within 3 days at 25 \u00b0C, 8 h at 37 \u00b0C, and 4 h at 56 \u00b0C, preceding losses in structural integrity or antigen-binding function. The VLPs remained stable at 25 \u00b0C for 28 d. Freeze\u2013thaw cycles induced a basic shift in charge variants without compromising structure or function, while high-intensity shaking (300 rpm) caused aggregation after 3\u20136 d. The effects of common excipients were also characterized. Conclusions: Charge-variant analysis serves as a sensitive and predictive marker for VLP vaccine instability. The study delineates the distinct impacts of different stress factors and provides critical data for optimizing formulation design and storage strategies to enhance VLP vaccine stability.<\/jats:p>","DOI":"10.3390\/ph19020269","type":"journal-article","created":{"date-parts":[[2026,2,5]],"date-time":"2026-02-05T10:35:37Z","timestamp":1770287737000},"page":"269","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Stability Under Different Stress Treatments of a Virus-like Particle Vaccine Based on a Recombinant Hepatitis E Vaccine"],"prefix":"10.3390","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-2062-9994","authenticated-orcid":false,"given":"Zhiyun","family":"Qi","sequence":"first","affiliation":[{"name":"School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110016, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sha","family":"Guo","sequence":"additional","affiliation":[{"name":"WHO Collaborating Centre for Standardization and Evaluation of Biologicals, State Key Laboratory of Drug Regulatory Science, NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, NMPA Key Laboratory for Quality Research and Evaluation of Biological Products, National Institutes for Food and Drug Control, Beijing 102629, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanhan","family":"Li","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110016, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xijie","family":"Xia","sequence":"additional","affiliation":[{"name":"WHO Collaborating Centre for Standardization and Evaluation of Biologicals, State Key Laboratory of Drug Regulatory Science, NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, NMPA Key Laboratory for Quality Research and Evaluation of Biological Products, National Institutes for Food and Drug Control, Beijing 102629, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuangshuang","family":"Qi","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110016, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Enlian","family":"Tang","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110016, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-6444-0183","authenticated-orcid":false,"given":"Zhenhao","family":"Zhou","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110016, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yiping","family":"Wang","sequence":"additional","affiliation":[{"name":"WHO Collaborating Centre for Standardization and Evaluation of Biologicals, State Key Laboratory of Drug Regulatory Science, NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, NMPA Key Laboratory for Quality Research and Evaluation of Biological Products, National Institutes for Food and Drug Control, Beijing 102629, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chuanfei","family":"Yu","sequence":"additional","affiliation":[{"name":"WHO Collaborating Centre for Standardization and Evaluation of Biologicals, State Key Laboratory of Drug Regulatory Science, NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, NMPA Key Laboratory for Quality Research and Evaluation of Biological Products, National Institutes for Food and Drug Control, Beijing 102629, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xing","family":"Wu","sequence":"additional","affiliation":[{"name":"WHO Collaborating Centre for Standardization and Evaluation of Biologicals, State Key Laboratory of Drug Regulatory Science, NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, NMPA Key Laboratory for Quality Research and Evaluation of Biological Products, National Institutes for Food and Drug Control, Beijing 102629, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2837-6459","authenticated-orcid":false,"given":"Hao","family":"Wu","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110016, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1186\/s44280-025-00080-4","article-title":"Hepatitis E Virus: A Foodborne Pathogen of Concern in China","volume":"3","author":"Wang","year":"2025","journal-title":"One Health Adv."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"288","DOI":"10.1093\/cid\/ciab919","article-title":"Hepatitis E Virus Species C Infection in Humans, Hong Kong","volume":"75","author":"Sridhar","year":"2022","journal-title":"Clin. 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