Generation and characterization of a monoclonal antibody against MERS-CoV targeting the spike protein using a synthetic peptide epitope-CpG-DNA-liposome complex

Abstract

Middle East respiratory syndrome coronavirus (MERS-CoV) uses the spike (S) glycoprotein to recognize and enter target cells. In this study, we selected two epitope peptide sequences within the receptor binding domain (RBD) of the MERS-CoV S protein. We used a complex consisting of the epitope peptide of the MERS-CoV S protein and CpG-DNA encapsulated in liposome complex to immunize mice, and produced the monoclonal antibodies 506-2G10G5 and 492-1G10E4E2. The western blotting data showed that both monoclonal antibodies detected the S protein and immunoprecipitated the native form of the S protein. Indirect immunofluorescence and confocal analysis suggested strong reactivity of the antibodies towards the S protein of MERS-CoV virus infected Vero cells. Furthermore, the 506-2G10G5 monoclonal antibody significantly reduced plaque formation in MERS-CoV infected Vero cells compared to normal mouse IgG and 492-1G10E4E2. Thus, we successfully produced a monoclonal antibody directed against the RBD domain of the S protein which could be used in the development of diagnostics and therapeutic applications in the future.

The Novel Coronavirus 2019 epidemic and kidneys

“…Monoclonal antibody directed against the RBD domain of the S protein of MERS-CoV has been found to have neutralizing activities in plaque assays in vitro. 29 A monoclonal antibody against COVID- 19 has not yet been developed. Trastuzumab, a monoclonal antibody against the IL-6 receptor, has achieved encouraging preliminary clinical results.…”

A pilot study of the antiviral activity of anionic and cationic polyamidoamine dendrimers against the Middle East respiratory syndrome coronavirus

“…The plaque reduction assay was performed as reported previously. 14 Most of the toxic effects of the dendrimers came from their size or surface charge, which interacted with the lipids of cell membranes.…”

CpG-DNA induces bacteria-reactive IgM enhancing phagocytic activity against Staphylococcus aureus infection

“…After 5 days, cell culture supernatants were obtained by centrifugation at 3,000 rpm for 10 min, and purified using DEAE Sepharose ™ Fast Flow (GE Healthcare Co.) and HiPrep ™ 16/60 Sephacryl ™ S-300 HR (Ge Healthcare Co.). The samples were loaded onto a SDS-PAGE, analyzed by Coomassie blue staining and Western blotting (28).…”