Abstract
Filoviruses cause lethal hemorrhagic fever in humans. The filovirus nucleoprotein (NP) is expressed in high abundance in infected cells and is essential for virus replication. To generate anti-filovirus monoclonal antibodies (mAbs) against the NP, mice were immunized with peptides known as B-cell epitopes corresponding to different filovirus NPs, and hybridomas were screened using FLAG-tagged filovirus NP constructs. Numerous mAbs were identified, isotyped, and characterized. The anti-NP mAbs demonstrated different ranges of binding affinities to various filovirus NPs. Most of the clones specifically detected both recombinant and wild-type NPs from different filoviruses, including Ebola (EBOV), Sudan (SUDV), Bundibugyo (BDBV), Marburg (MARV), Tai Forest (TAFV), and Reston (RESTV) viruses in western blot analysis. The mAbs were also able to detect native NPs within the cytoplasm of infected cells by immunofluorescence confocal microscopy. Thus, this panel of mAbs represents an important set of tools that may be potentially useful for diagnosing filovirus infection, characterizing virus replication, and detecting NP–host protein interactions.
Highlights
Ebola virus (EBOV), an enveloped, negative-sense RNA virus, is a member of the Ebolavirus genus and the Filoviridae family
The remaining peptides, EBOV NP aa491–510, SUDV NP aa631–644, TAFV NP aa630–643, Reston virus (RESTV) NP aa630-643, and MARV NP aa635–652 corresponded to regions that showed little similarity among different filoviruses and were used to generate virus-specific antibodies [10]
The monoclonal antibodies against the filovirus NPs that were produced and characterized represent an important set of tools that may be used to develop novel diagnostic assays or to dissect the molecular biology of these viruses
Summary
Ebola virus (EBOV), an enveloped, negative-sense RNA virus, is a member of the Ebolavirus genus and the Filoviridae family. It is the causative agent of a severe and highly lethal hemorrhagic fever that has resulted in numerous outbreaks throughout Africa, including an unprecedented outbreak in. Other ebolaviruses, including Sudan (SUDV) and Bundibugyo (BDBV) viruses, as well as Marburg (MARV) virus, which belongs to the related Marburgvirus genus within the family Filoviridae, are highly virulent in humans and have caused numerous outbreaks [2]. Viruses 2019, 11, 259 several candidate vaccines and therapeutics, including monoclonal antibody therapeutics, have shown efficacy in clinical trials and are currently being deployed in the 2018 Democratic Republic of Congo. EBOV outbreak [4,5,6,7,8].
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