{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T06:46:27Z","timestamp":1791009987050,"version":"4.1.0"},"reference-count":49,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T00:00:00Z","timestamp":1760486400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The recent SARS-CoV-2 pandemic has highlighted the need for quickly adaptable technologies in vaccine manufacturing. This can be achieved through virus-like particles (VLPs) as presentation platforms for target antigens. In this study, we investigated the purification of VLPs of the Hepatitis B Core antigen (HBc) and the SplitCore (SplCo) technology. The outer surface protein C (OspC) of Borrelia burgdorferi was genetically fused to HBc and its N-terminal SplCo protein. Product solubility in E. coli increased from 40% for HBc-OspC to 90% for SplCo-OspC. This could not be reproduced with similar SARS-CoV-2 receptor-binding domain fusions due to inclusion body formation. Hydrophobicity was found to be significantly lowered for the OspC fusions, in particular for the SplCo variant. Pre-purified samples were generated by precipitating soluble cell lysate. Subsequently, solubilized precipitates were subjected to anion exchange chromatography (AEX), and the elution fractions obtained contained VLPs, albeit with low purity. The VLPs were also disassembled prior to AEX for dissociative purification, but a subsequent reassembly could not be achieved for both fusion variants. A novel HBc variant was constructed for post-translational modification via click-chemistry. The solubility and hydrophobicity of this HBc variant remained high, but native AEX resulted in complete product loss. By contrast, a yield of 84% VLPs was obtained for the modified HBc after dissociative AEX. The surface-exposed azide group on the particles, introduced for click-chemistry, enabled coupling to fluorophores without compromising VLP stability. Conjugation efficiencies of up to 59% were obtained. These results suggest the potential of combining HBc and click-chemistry for future applications, e.g., the presentation of immunogenic epitopes or antigens. This underlines that for every antigen, both the optimal scaffold-decoration strategy and the subsequent manufacturing process should be carefully selected.<\/jats:p>","DOI":"10.3390\/ijms262010036","type":"journal-article","created":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T11:57:42Z","timestamp":1760529462000},"page":"10036","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Modular Virus-like Particles for Antigen Presentation: Comparing Genetic Fusion and Click-Chemistry for Purification"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-5085-6204","authenticated-orcid":false,"given":"Karsten","family":"Balbierer","sequence":"first","affiliation":[{"name":"ARTES Biotechnology GmbH, 40764 Langenfeld, Germany"},{"name":"Institute of Pharmaceutical and Medicinal Chemistry, University of M\u00fcnster, 48149 M\u00fcnster, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Volker","family":"Jenzelewski","sequence":"additional","affiliation":[{"name":"ARTES Biotechnology GmbH, 40764 Langenfeld, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fabian C.","family":"Herrmann","sequence":"additional","affiliation":[{"name":"Institute of Pharmaceutical Biology and Phytochemistry, University of M\u00fcnster, 48149 M\u00fcnster, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Piontek","sequence":"additional","affiliation":[{"name":"ARTES Biotechnology GmbH, 40764 Langenfeld, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0666-2676","authenticated-orcid":false,"given":"Joachim","family":"Jose","sequence":"additional","affiliation":[{"name":"Institute of Pharmaceutical and Medicinal Chemistry, University of M\u00fcnster, 48149 M\u00fcnster, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,10,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"S18","DOI":"10.1016\/0264-410X(90)90209-5","article-title":"Hepatitis B: Global importance and need for control","volume":"8","author":"Maynard","year":"1990","journal-title":"Vaccine"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1002\/bip.22759","article-title":"Different applications of virus-like particles in biology and medicine: Vaccination and delivery systems","volume":"105","author":"Shirbaghaee","year":"2016","journal-title":"Biopolymers"},{"key":"ref_3","first-page":"112","article-title":"Engineering Virus-like Particles for Antigen and Drug Delivery","volume":"19","author":"Hill","year":"2018","journal-title":"Curr. 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