{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,1]],"date-time":"2025-11-01T08:42:57Z","timestamp":1761986577948,"version":"build-2065373602"},"reference-count":82,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2024,10,10]],"date-time":"2024-10-10T00:00:00Z","timestamp":1728518400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Bayerischer Forschungsverbund FOR-COVID","award":["F.2-F2412.32\/1\/45"],"award-info":[{"award-number":["F.2-F2412.32\/1\/45"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The emergence of hitherto unknown viral pathogens presents a great challenge for researchers to develop effective therapeutics and vaccines within a short time to avoid an uncontrolled global spread, as seen during the coronavirus disease 2019 (COVID-19) pandemic. Therefore, rapid and simple methods to identify immunogenic antigens as potential therapeutical targets are urgently needed for a better pandemic preparedness. To address this problem, we chose the well-characterized Modified Vaccinia virus Ankara (MVA)-T7pol expression system to establish a workflow to identify immunogens when a new pathogen emerges, generate candidate vaccines, and test their immunogenicity in an animal model. By using this system, we detected severe acute respiratory syndrome (SARS) coronavirus 2 (SARS-CoV-2) nucleoprotein (N)-, and spike (S)-specific antibodies in COVID-19 patient sera, which is in line with the current literature and our observations from previous immunogenicity studies. Furthermore, we detected antibodies directed against the SARS-CoV-2-membrane (M) and -ORF3a proteins in COVID-19 patient sera and aimed to generate recombinant MVA candidate vaccines expressing either the M or ORF3a protein. When testing our candidate vaccines in a prime-boost immunization regimen in humanized HLA-A2.1-\/HLA-DR1-transgenic H-2 class I-\/class II-knockout mice, we were able to demonstrate M- and ORF3a-specific cellular and humoral immune responses. Hence, the established workflow using the MVA-T7pol expression system represents a rapid and efficient tool to identify potential immunogenic antigens and provides a basis for future development of candidate vaccines.<\/jats:p>","DOI":"10.3390\/ijms252010898","type":"journal-article","created":{"date-parts":[[2024,10,10]],"date-time":"2024-10-10T07:53:05Z","timestamp":1728546785000},"page":"10898","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Implementation of an Immunoassay Based on the MVA-T7pol-Expression System for Rapid Identification of Immunogenic SARS-CoV-2 Antigens: A Proof-of-Concept Study"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7921-5746","authenticated-orcid":false,"given":"Satendra","family":"Kumar","sequence":"first","affiliation":[{"name":"Division of Virology, Department of Veterinary Sciences, Ludwig Maximilians University Munich (LMU Munich), 85764 Oberschlei\u00dfheim, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7680-3630","authenticated-orcid":false,"given":"Liangliang","family":"Nan","sequence":"additional","affiliation":[{"name":"Division of Virology, Department of Veterinary Sciences, Ludwig Maximilians University Munich (LMU Munich), 85764 Oberschlei\u00dfheim, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-5468-2695","authenticated-orcid":false,"given":"Georgia","family":"Kalodimou","sequence":"additional","affiliation":[{"name":"Division of Virology, Department of Veterinary Sciences, Ludwig Maximilians University Munich (LMU Munich), 85764 Oberschlei\u00dfheim, Germany"},{"name":"German Center for Infection Research, Partner Site Munich, 85764 Oberschlei\u00dfheim, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sylvia","family":"Jany","sequence":"additional","affiliation":[{"name":"Division of Virology, Department of Veterinary Sciences, Ludwig Maximilians University Munich (LMU Munich), 85764 Oberschlei\u00dfheim, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Astrid","family":"Freudenstein","sequence":"additional","affiliation":[{"name":"Division of Virology, Department of Veterinary Sciences, Ludwig Maximilians University Munich (LMU Munich), 85764 Oberschlei\u00dfheim, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christine","family":"Brandm\u00fcller","sequence":"additional","affiliation":[{"name":"Division of Virology, Department of Veterinary Sciences, Ludwig Maximilians University Munich (LMU Munich), 85764 Oberschlei\u00dfheim, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2890-7515","authenticated-orcid":false,"given":"Katharina","family":"M\u00fcller","sequence":"additional","affiliation":[{"name":"German Center for Infection Research, Partner Site Munich, 85764 Oberschlei\u00dfheim, Germany"},{"name":"Bundeswehr Institute of Microbiology, 80937 Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1370-4114","authenticated-orcid":false,"given":"Philipp","family":"Girl","sequence":"additional","affiliation":[{"name":"German Center for Infection Research, Partner Site Munich, 85764 Oberschlei\u00dfheim, Germany"},{"name":"Bundeswehr Institute of Microbiology, 80937 Munich, Germany"},{"name":"Chair of Bacteriology and Mycology, Department of Veterinary Sciences, Ludwig Maximilians University Munich (LMU Munich), 85764 Oberschlei\u00dfheim, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rosina","family":"Ehmann","sequence":"additional","affiliation":[{"name":"German Center for Infection Research, Partner Site Munich, 85764 Oberschlei\u00dfheim, Germany"},{"name":"Bundeswehr Institute of Microbiology, 80937 Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wolfgang","family":"Guggemos","sequence":"additional","affiliation":[{"name":"Munich Clinic Schwabing, Academic Teaching Hospital, Ludwig Maximilians University Munich (LMU Munich), 80804 Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Seilmaier","sequence":"additional","affiliation":[{"name":"Munich Clinic Schwabing, Academic Teaching Hospital, Ludwig Maximilians University Munich (LMU Munich), 80804 Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Clemens-Martin","family":"Wendtner","sequence":"additional","affiliation":[{"name":"Medical Clinic III, University Hospital, Ludwig Maximilians University Munich (LMU Munich), 80336 Munich, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8757-3896","authenticated-orcid":false,"given":"Asisa","family":"Volz","sequence":"additional","affiliation":[{"name":"Institute of Virology, University of Veterinary Medicine Hannover, 30559 Hannover, Germany"},{"name":"German Center for Infection Research, Partner Site Hannover-Braunschweig, 30559 Hannover, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6143-082X","authenticated-orcid":false,"given":"Gerd","family":"Sutter","sequence":"additional","affiliation":[{"name":"Division of Virology, Department of Veterinary Sciences, Ludwig Maximilians University Munich (LMU Munich), 85764 Oberschlei\u00dfheim, Germany"},{"name":"German Center for Infection Research, Partner Site Munich, 85764 Oberschlei\u00dfheim, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1149-1613","authenticated-orcid":false,"given":"Robert","family":"Fux","sequence":"additional","affiliation":[{"name":"Division of Virology, Department of Veterinary Sciences, Ludwig Maximilians University Munich (LMU Munich), 85764 Oberschlei\u00dfheim, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alina","family":"Tscherne","sequence":"additional","affiliation":[{"name":"Division of Virology, Department of Veterinary Sciences, Ludwig Maximilians University Munich (LMU Munich), 85764 Oberschlei\u00dfheim, Germany"},{"name":"German Center for Infection Research, Partner Site Munich, 85764 Oberschlei\u00dfheim, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,10,10]]},"reference":[{"key":"ref_1","first-page":"e2313","article-title":"COVID-19 vaccines and variants of concern: A review","volume":"32","year":"2021","journal-title":"Rev. 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