Ebola virus(EBOV) belongs to the Filoviridae family and causes severe illnesses such as hemorrhagic fever with a high mortality rate up to 90%. Now two antibody drugs termed Inmazeb and Ebanga have been approved for t...Ebola virus(EBOV) belongs to the Filoviridae family and causes severe illnesses such as hemorrhagic fever with a high mortality rate up to 90%. Now two antibody drugs termed Inmazeb and Ebanga have been approved for treating EBOV infection. However, clinical studies have demonstrated that the mortality rate of the patients who received these two antibody drugs remains above 30%. Therefore, novel therapeutics with better efficacy is still desired. The isolated human IgG1 constant domain 2(CH2 domain) has been proposed as a scaffold for the development of C-based single domain antibodies(C-sd Abs) as therapeutic candidates against viral infections and other diseases. Here, we screened and identified a novel C-sd Ab termed M24 that targets EBOV glycoprotein(GP) from a C-sd Ab phage display library. M24 neutralizes the pseudotype EBOV with IC;of 0.8 nmol/L(12 ng/mL) and has modest neutralizing activity against authentic EBOV.Epitope determination, including molecular docking and site mutation analysis, discloses that M24 binds to the internal fusion loop(IFL) within GP2, a transmembrane subunit of GP. Interestingly, we found that the binding of M24 to GP at pH5.5 has dramatically decreased compared to the binding at pH 7.5, which may lead to weak efficacy in the neutralization of authentic EBOV. Since no sd Ab against EBOV infection has been reported to date, our results not only give a proof of concept that sd Abs could be utilized for the development of potential therapeutic candidates against EBOV infection, but also provide useful information for the discovery and improvement of anti-EBOV agents.展开更多
基金This work was jointly supported by the Advanced Customer Cultivation Project of Wuhan National Biosafety Laboratory,Chinese Academy of Sciences(2021ACCP-MS01,2019ACCP-ZD03)the Natural Science Foundation of Hubei Province of China(2019CFA076)the National Natural Science Foundation of China(31870926)。
文摘Ebola virus(EBOV) belongs to the Filoviridae family and causes severe illnesses such as hemorrhagic fever with a high mortality rate up to 90%. Now two antibody drugs termed Inmazeb and Ebanga have been approved for treating EBOV infection. However, clinical studies have demonstrated that the mortality rate of the patients who received these two antibody drugs remains above 30%. Therefore, novel therapeutics with better efficacy is still desired. The isolated human IgG1 constant domain 2(CH2 domain) has been proposed as a scaffold for the development of C-based single domain antibodies(C-sd Abs) as therapeutic candidates against viral infections and other diseases. Here, we screened and identified a novel C-sd Ab termed M24 that targets EBOV glycoprotein(GP) from a C-sd Ab phage display library. M24 neutralizes the pseudotype EBOV with IC;of 0.8 nmol/L(12 ng/mL) and has modest neutralizing activity against authentic EBOV.Epitope determination, including molecular docking and site mutation analysis, discloses that M24 binds to the internal fusion loop(IFL) within GP2, a transmembrane subunit of GP. Interestingly, we found that the binding of M24 to GP at pH5.5 has dramatically decreased compared to the binding at pH 7.5, which may lead to weak efficacy in the neutralization of authentic EBOV. Since no sd Ab against EBOV infection has been reported to date, our results not only give a proof of concept that sd Abs could be utilized for the development of potential therapeutic candidates against EBOV infection, but also provide useful information for the discovery and improvement of anti-EBOV agents.
