Characterization and immunogenicity assessment of MERS-CoV pre-fusion spike trimeric oligomers as vaccine immunogen

Middle East respiratory syndrome coronavirus (MERS-CoV) is a lethal beta-coronavirus that emerged in 2012. The virus is part of the WHO blueprint priority list with a concerning fatality rate of 35%. Scientific efforts are ongoing for the development of vaccines, anti-viral and biotherapeutics, whic...

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Main Authors: Rahul Ahuja, Preeti Vishwakarma, Sneha Raj, Varun Kumar, Ritika Khatri, Bharat Lohiya, Shikha Saxena, Gurleen Kaur, Gagandeep Singh, Shailendra Asthana, Shubbir Ahmed, Sweety Samal
Format: Article
Language:English
Published: Taylor & Francis Group 2024-12-01
Series:Human Vaccines & Immunotherapeutics
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Online Access:https://www.tandfonline.com/doi/10.1080/21645515.2024.2351664
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author Rahul Ahuja
Preeti Vishwakarma
Sneha Raj
Varun Kumar
Ritika Khatri
Bharat Lohiya
Shikha Saxena
Gurleen Kaur
Gagandeep Singh
Shailendra Asthana
Shubbir Ahmed
Sweety Samal
author_facet Rahul Ahuja
Preeti Vishwakarma
Sneha Raj
Varun Kumar
Ritika Khatri
Bharat Lohiya
Shikha Saxena
Gurleen Kaur
Gagandeep Singh
Shailendra Asthana
Shubbir Ahmed
Sweety Samal
author_sort Rahul Ahuja
collection DOAJ
description Middle East respiratory syndrome coronavirus (MERS-CoV) is a lethal beta-coronavirus that emerged in 2012. The virus is part of the WHO blueprint priority list with a concerning fatality rate of 35%. Scientific efforts are ongoing for the development of vaccines, anti-viral and biotherapeutics, which are majorly directed toward the structural spike protein. However, the ongoing effort is challenging due to conformational instability of the spike protein and the evasion strategy posed by the MERS-CoV. In this study, we have expressed and purified the MERS-CoV pre-fusion spike protein in the Expi293F mammalian expression system. The purified protein was extensively characterized for its biochemical and biophysical properties. Thermal stability analysis showed a melting temperature of 58°C and the protein resisted major structural changes at elevated temperature as revealed by fluorescence spectroscopy and circular dichroism. Immunological assessment of the MERS-CoV spike immunogen in BALB/c mice with AddaVaxTM and Imject alum adjuvants showed elicitation of high titer antibody responses but a more balanced Th1/Th2 response with AddaVaxTM squalene like adjuvant. Together, our results suggest the formation of higher-order trimeric pre-fusion MERS-CoV spike proteins, which were able to induce robust immune responses. The comprehensive characterization of MERS-CoV spike protein warrants a better understanding of MERS spike protein and future vaccine development efforts.
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spelling doaj-art-4e44e7d9cd2b4a2bae31dfb3939ef7df2025-08-20T02:16:40ZengTaylor & Francis GroupHuman Vaccines & Immunotherapeutics2164-55152164-554X2024-12-0120110.1080/21645515.2024.2351664Characterization and immunogenicity assessment of MERS-CoV pre-fusion spike trimeric oligomers as vaccine immunogenRahul Ahuja0Preeti Vishwakarma1Sneha Raj2Varun Kumar3Ritika Khatri4Bharat Lohiya5Shikha Saxena6Gurleen Kaur7Gagandeep Singh8Shailendra Asthana9Shubbir Ahmed10Sweety Samal11Influenza and Respiratory Virus Laboratory, Translational Health Science and Technology Institute (THSTI), NCR-Biotech Science Cluster, Faridabad, Haryana, IndiaInfluenza and Respiratory Virus Laboratory, Translational Health Science and Technology Institute (THSTI), NCR-Biotech Science Cluster, Faridabad, Haryana, IndiaInfluenza and Respiratory Virus Laboratory, Translational Health Science and Technology Institute (THSTI), NCR-Biotech Science Cluster, Faridabad, Haryana, IndiaInfluenza and Respiratory Virus Laboratory, Translational Health Science and Technology Institute (THSTI), NCR-Biotech Science Cluster, Faridabad, Haryana, IndiaInfluenza and Respiratory Virus Laboratory, Translational Health Science and Technology Institute (THSTI), NCR-Biotech Science Cluster, Faridabad, Haryana, IndiaInfluenza and Respiratory Virus Laboratory, Translational Health Science and Technology Institute (THSTI), NCR-Biotech Science Cluster, Faridabad, Haryana, IndiaInfluenza and Respiratory Virus Laboratory, Translational Health Science and Technology Institute (THSTI), NCR-Biotech Science Cluster, Faridabad, Haryana, IndiaInfluenza and Respiratory Virus Laboratory, Translational Health Science and Technology Institute (THSTI), NCR-Biotech Science Cluster, Faridabad, Haryana, IndiaInfluenza and Respiratory Virus Laboratory, Translational Health Science and Technology Institute (THSTI), NCR-Biotech Science Cluster, Faridabad, Haryana, IndiaInfluenza and Respiratory Virus Laboratory, Translational Health Science and Technology Institute (THSTI), NCR-Biotech Science Cluster, Faridabad, Haryana, IndiaInfluenza and Respiratory Virus Laboratory, Translational Health Science and Technology Institute (THSTI), NCR-Biotech Science Cluster, Faridabad, Haryana, IndiaInfluenza and Respiratory Virus Laboratory, Translational Health Science and Technology Institute (THSTI), NCR-Biotech Science Cluster, Faridabad, Haryana, IndiaMiddle East respiratory syndrome coronavirus (MERS-CoV) is a lethal beta-coronavirus that emerged in 2012. The virus is part of the WHO blueprint priority list with a concerning fatality rate of 35%. Scientific efforts are ongoing for the development of vaccines, anti-viral and biotherapeutics, which are majorly directed toward the structural spike protein. However, the ongoing effort is challenging due to conformational instability of the spike protein and the evasion strategy posed by the MERS-CoV. In this study, we have expressed and purified the MERS-CoV pre-fusion spike protein in the Expi293F mammalian expression system. The purified protein was extensively characterized for its biochemical and biophysical properties. Thermal stability analysis showed a melting temperature of 58°C and the protein resisted major structural changes at elevated temperature as revealed by fluorescence spectroscopy and circular dichroism. Immunological assessment of the MERS-CoV spike immunogen in BALB/c mice with AddaVaxTM and Imject alum adjuvants showed elicitation of high titer antibody responses but a more balanced Th1/Th2 response with AddaVaxTM squalene like adjuvant. Together, our results suggest the formation of higher-order trimeric pre-fusion MERS-CoV spike proteins, which were able to induce robust immune responses. The comprehensive characterization of MERS-CoV spike protein warrants a better understanding of MERS spike protein and future vaccine development efforts.https://www.tandfonline.com/doi/10.1080/21645515.2024.2351664MERS-CoV vaccineprefusion spikeimmunogenicitythermostability, antibody response
spellingShingle Rahul Ahuja
Preeti Vishwakarma
Sneha Raj
Varun Kumar
Ritika Khatri
Bharat Lohiya
Shikha Saxena
Gurleen Kaur
Gagandeep Singh
Shailendra Asthana
Shubbir Ahmed
Sweety Samal
Characterization and immunogenicity assessment of MERS-CoV pre-fusion spike trimeric oligomers as vaccine immunogen
Human Vaccines & Immunotherapeutics
MERS-CoV vaccine
prefusion spike
immunogenicity
thermostability, antibody response
title Characterization and immunogenicity assessment of MERS-CoV pre-fusion spike trimeric oligomers as vaccine immunogen
title_full Characterization and immunogenicity assessment of MERS-CoV pre-fusion spike trimeric oligomers as vaccine immunogen
title_fullStr Characterization and immunogenicity assessment of MERS-CoV pre-fusion spike trimeric oligomers as vaccine immunogen
title_full_unstemmed Characterization and immunogenicity assessment of MERS-CoV pre-fusion spike trimeric oligomers as vaccine immunogen
title_short Characterization and immunogenicity assessment of MERS-CoV pre-fusion spike trimeric oligomers as vaccine immunogen
title_sort characterization and immunogenicity assessment of mers cov pre fusion spike trimeric oligomers as vaccine immunogen
topic MERS-CoV vaccine
prefusion spike
immunogenicity
thermostability, antibody response
url https://www.tandfonline.com/doi/10.1080/21645515.2024.2351664
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