Development of a point-of-care immunoassay for Ebola Virus Disease through the detection of Ebola virus soluble glycoprotein

Abstract Ebola Virus Disease (EVD) is a deadly infectious disease that is caused by an infection with Ebola virus (EBOV). EVD is one of the deadliest viral diseases with a human case fatality rate of up to 90%. The largest outbreak to date was the 2013–2016 West African outbreak in which Zaire ebolavirus (ZEBOV) was responsible for over 11,000 deaths. Transmission of the virus can occur through contact with an infected animal or human. Early detection is imperative to prevent the spread of disease and reduce the risk of a potential epidemic by isolating known infected patients. The current gold standard for diagnosing EVD is polymerase chain reaction assays which requires complex laboratory equipment and highly trained personnel that is not easily accessible in resource-limited areas. There is a great need for an EBOV rapid diagnostic test that can be implemented at the point-of-care (POC). Multiple lateral flow immunoassays (LFIs) have been developed for this need and have received emergency use authorization. Many of these tests rely on the detection of Ebola virus VP40 antigen, however, soluble glycoprotein (sGP) has the potential to be a better biomarker for EVD rapid detection as it is abundant in the blood during an infection. The goal of this study was to produce an EBOV sGP LFI prototype. A library of seventeen monoclonal antibodies reactive to EBOV sGP were produced and characterized via enzyme-linked immunosorbent assay, surface plasmon resonance, and Western immunoblotting. From this mAb library, LFI prototypes were developed and assessed for the potential use as a rapid POC diagnostic for EVD. Supports by the National Institute of Allergy and Infectious Diseases under grant 1R41AI149940