A high-throughput, fully automated competition assay to evaluate SARS-CoV-2 neutralizing responses and epitope specificity in clinical samples
Abstract Coronavirus disease-2019 (COVID-19) remains a critical global health concern. We developed a fully automated, high-throughput competition immunoassay to elucidate how epitope recognition on the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike receptor-binding domain (RBD)...
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Nature Portfolio
2025-04-01
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| Online Access: | https://doi.org/10.1038/s41598-025-94317-2 |
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| author | Yusuke Atarashi Jeeeun Kim Yasuhiro Irino Masayuki Amano Kiyoto Tsuchiya Kenji Maeda Mari Terada Noriko Iwamoto Shinya Shimada Hiroaki Mitsuya Masatoshi Yanagida Yuki Takamatsu |
| author_facet | Yusuke Atarashi Jeeeun Kim Yasuhiro Irino Masayuki Amano Kiyoto Tsuchiya Kenji Maeda Mari Terada Noriko Iwamoto Shinya Shimada Hiroaki Mitsuya Masatoshi Yanagida Yuki Takamatsu |
| author_sort | Yusuke Atarashi |
| collection | DOAJ |
| description | Abstract Coronavirus disease-2019 (COVID-19) remains a critical global health concern. We developed a fully automated, high-throughput competition immunoassay to elucidate how epitope recognition on the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike receptor-binding domain (RBD) correlates with neutralizing activity. Analysis of clinical samples from both SARS-CoV-2-infected and vaccinated individuals revealed that vaccination elicits significantly higher antibody titers across multiple S1 subunit epitopes compared to natural infection. Notably, median antibody levels against the receptor-binding motif (RBM) exceeded 50% in both cohorts, highlighting the RBM as a key target for antibody induction irrespective of immune origin. Furthermore, the strongest correlation with neutralizing activity was observed for antibodies directed against the broader S1 subunit, indicating that epitopes outside the RBM also contribute to neutralization. These findings underscore the importance of both RBM- and non-RBM-directed antibodies in effective immune defense against SARS-CoV-2. Our assay enables large-scale, reliable quantification of neutralizing antibodies and provides critical insights for developing improved diagnostic antigens and vaccine strategies aimed at eliciting robust, multi-epitope immune responses. |
| format | Article |
| id | doaj-art-1deb9dfa172c4ea088d48ecd837223e5 |
| institution | DOAJ |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-1deb9dfa172c4ea088d48ecd837223e52025-08-20T03:07:40ZengNature PortfolioScientific Reports2045-23222025-04-011511710.1038/s41598-025-94317-2A high-throughput, fully automated competition assay to evaluate SARS-CoV-2 neutralizing responses and epitope specificity in clinical samplesYusuke Atarashi0Jeeeun Kim1Yasuhiro Irino2Masayuki Amano3Kiyoto Tsuchiya4Kenji Maeda5Mari Terada6Noriko Iwamoto7Shinya Shimada8Hiroaki Mitsuya9Masatoshi Yanagida10Yuki Takamatsu11Central Research Laboratories, Sysmex CorporationCentral Research Laboratories, Sysmex CorporationCentral Research Laboratories, Sysmex CorporationDepartment of Clinical Retrovirology, Joint Research Center for Human Retrovirus Infection, Kumamoto UniversityAIDS Clinical Center, Center hospital of the National Center for Global Health and MedicineRefractory Viral Diseases, National Center for Global Health and Medicine Research InstituteDepartment of Disease Control Center, Center Hospital of the National Center for Global Health and MedicineDepartment of Disease Control Center, Center Hospital of the National Center for Global Health and MedicineJapan Community Healthcare Organization, Kumamoto General HospitalRefractory Viral Diseases, National Center for Global Health and Medicine Research InstituteCentral Research Laboratories, Sysmex CorporationRefractory Viral Diseases, National Center for Global Health and Medicine Research InstituteAbstract Coronavirus disease-2019 (COVID-19) remains a critical global health concern. We developed a fully automated, high-throughput competition immunoassay to elucidate how epitope recognition on the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike receptor-binding domain (RBD) correlates with neutralizing activity. Analysis of clinical samples from both SARS-CoV-2-infected and vaccinated individuals revealed that vaccination elicits significantly higher antibody titers across multiple S1 subunit epitopes compared to natural infection. Notably, median antibody levels against the receptor-binding motif (RBM) exceeded 50% in both cohorts, highlighting the RBM as a key target for antibody induction irrespective of immune origin. Furthermore, the strongest correlation with neutralizing activity was observed for antibodies directed against the broader S1 subunit, indicating that epitopes outside the RBM also contribute to neutralization. These findings underscore the importance of both RBM- and non-RBM-directed antibodies in effective immune defense against SARS-CoV-2. Our assay enables large-scale, reliable quantification of neutralizing antibodies and provides critical insights for developing improved diagnostic antigens and vaccine strategies aimed at eliciting robust, multi-epitope immune responses.https://doi.org/10.1038/s41598-025-94317-2SARS-CoV-2High throughput testingAntibody testNeutralizing activity |
| spellingShingle | Yusuke Atarashi Jeeeun Kim Yasuhiro Irino Masayuki Amano Kiyoto Tsuchiya Kenji Maeda Mari Terada Noriko Iwamoto Shinya Shimada Hiroaki Mitsuya Masatoshi Yanagida Yuki Takamatsu A high-throughput, fully automated competition assay to evaluate SARS-CoV-2 neutralizing responses and epitope specificity in clinical samples Scientific Reports SARS-CoV-2 High throughput testing Antibody test Neutralizing activity |
| title | A high-throughput, fully automated competition assay to evaluate SARS-CoV-2 neutralizing responses and epitope specificity in clinical samples |
| title_full | A high-throughput, fully automated competition assay to evaluate SARS-CoV-2 neutralizing responses and epitope specificity in clinical samples |
| title_fullStr | A high-throughput, fully automated competition assay to evaluate SARS-CoV-2 neutralizing responses and epitope specificity in clinical samples |
| title_full_unstemmed | A high-throughput, fully automated competition assay to evaluate SARS-CoV-2 neutralizing responses and epitope specificity in clinical samples |
| title_short | A high-throughput, fully automated competition assay to evaluate SARS-CoV-2 neutralizing responses and epitope specificity in clinical samples |
| title_sort | high throughput fully automated competition assay to evaluate sars cov 2 neutralizing responses and epitope specificity in clinical samples |
| topic | SARS-CoV-2 High throughput testing Antibody test Neutralizing activity |
| url | https://doi.org/10.1038/s41598-025-94317-2 |
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