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Abstract
In this work, we describe the selection and characterization of single-domain antibodies (sdAb) towards the E2/E3E2 envelope protein of the Western equine encephalitis virus (WEEV). Our purpose was to identify novel recognition elements which could be used for the detection, diagnosis, and perhaps treatment of western equine encephalitis (WEE). To achieve this goal, we prepared an immune phage display library derived from the peripheral blood lymphocytes of a llama that had been immunized with an equine vaccine that includes killed WEEV (West Nile Innovator + VEWT). This library was panned against recombinant envelope (E2/E3E2) protein from WEEV, and seven representative sdAb from the five identified sequence families were characterized. The specificity, affinity, and melting point of each sdAb was determined, and their ability to detect the recombinant protein in a MagPlex sandwich immunoassay was confirmed. Thus, these new binders represent novel recognition elements for the E2/E3E2 proteins of WEEV that are available to the research community for further investigation into their applicability for use in the diagnosis or treatment of WEE.
Highlights
Western equine encephalitis virus (WEEV), an alphavirus in the Togaviridae family, is an arbovirus transmitted to people and horses by mosquitoes and is the causative agent of western equine encephalitis (WEE)
WEEV, EEEV and the related Venezuelan equine encephalitis virus (VEEV) can spread to the central nervous system, causing symptoms ranging from mild febrile reactions to encephalitis, often resulting in permanent neurological damage that can lead to death
We describe the selection and characterization of seven new single-domain antibodies (sdAb) towards the E2/E3E2 envelope protein of WEEV
Summary
Western equine encephalitis virus (WEEV), an alphavirus in the Togaviridae family, is an arbovirus transmitted to people and horses by mosquitoes and is the causative agent of western equine encephalitis (WEE). WEEV originated when Eastern equine encephalitis virus (EEEV), a new world virus, and Sindbis virus, an old world virus, recombined [1,2,3]. WEEV, EEEV and the related Venezuelan equine encephalitis virus (VEEV) can spread to the central nervous system, causing symptoms ranging from mild febrile reactions to encephalitis, often resulting in permanent neurological damage that can lead to death. The disease is seen in South and Central American countries. WEEV causes serious disease in horses, with a case-fatality rate of 20–30% [5]. Human cases are relatively rare, in 1941 an outbreak in Canada caused more than 1000 human infections [6]
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