期刊
VACCINES
卷 8, 期 3, 页码 -出版社
MDPI
DOI: 10.3390/vaccines8030415
关键词
Trypanosoma evansi; Nanobody; enolase; diagnosis; homologous and heterologous detection assays
资金
- China Scholarship Council (CSC)
- UAntwerp-BOF research grant (DOCPRO1) [FFB190197]
- UGent-BOF `Startkrediet' [BOF.STG.2018.0009.01/01N01518]
- Foundation for Scientific Research/Fonds voorWetenschappelijk Onderzoek-Vlaanderen [G013518N]
Trypanosoma evansi is a widely spread parasite that causes the debilitating disease surra in several types of ungulates. This severely challenges livestock rearing and heavily weighs on the socio-economic development in the affected areas, which include countries on five continents. Active case finding requires a sensitive and specific diagnostic test. In this paper, we describe the application of an unbiased immunization strategy to identify potential biomarkers for Nanobody (Nb)-based detection of T. evansi infections. Alpaca immunization with soluble lysates from different T. evansi strains followed by panning against T. evansi secretome resulted in the selection of a single Nb (Nb11). By combining Nb11-mediated immuno-capturing with mass spectrometry, the T. evansi target antigen was identified as the glycolytic enzyme enolase. Four additional anti-enolase binders were subsequently generated by immunizing another alpaca with the recombinant target enzyme. Together with Nb11, these binders were evaluated for their potential use in a heterologous sandwich detection format. Three Nb pairs were identified as candidates for the further development of an antigen-based assay for Nb-mediated diagnosis of T. evansi infection.
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