| Summary: | 碩士 === 國立臺灣大學 === 生化科技學系 === 105 === In 2012, an outbreak in Saudi Arabia caused by the Middle East respiratory syndrome coronavirus (MERS-CoV) was reported by WHO. This novel coronavirus caused high fatality rates up to 36%, and was further spread to South Korea in 2015. Some researchers suggest that dromedary camels are the major reservoir host for MERS-CoV, and eventually cause human-to-human infections in health care settings. However, the actual role of dromedary camels in transmission of MERS-CoV is still unknown, and there is no specific antiviral drug against this virus. In this study, we focused on the spike protein (SP) on the virus surface and try to develop antibodies to inhibit the virus infection. The structure of spike protein contains two subunits, S1 and S2. The S1 subunit forms the globular head and binds to the host cell receptor DPP4 (dipeptidyl peptidase-4) with its receptor binding domain (RBD). Subsequently, the S2 subunit causes the conformation change to HR1/HR2, forming a six-helix bundle (6-HB) structure, and then insert its fusion peptide to the host membrane. Accordingly, S2 and HR1 is a good target to develop therapeutic antibodies against MERS-CoV. We firstly constructed and expressed S2 and HR1 domain in Bac-to-Bac expression system for the immunization of mice to produce antiserum (pAb). The mice were then sacrificed and their spleen cells collected for cell fusion with myeloma. The hybridoma cells producing specific monoclonal antibodies (mAb) were selected by enzyme-linked immunosorbent assay (ELISA). Both the pAb and mAb were successfully tested by neutralizing assays to confirm their capacity in inhibiting the invasion of MERS-CoV into host cells through its spike protein.
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