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An Unbiased Immunization Strategy Results in the Identification of Enolase as a Potential Marker for Nanobody-Based Detection of Trypanosoma evansi

Authors
  • Li, Zeng1,
  • Pinto Torres, Joar Esteban1
  • Goossens, Julie1
  • Vertommen, Didier
  • Caljon, Guy
  • Sterckx, Yann G.-J.
  • Magez, Stefan1, 2, 3
  • 1 (J.G.)
  • 2 Department of Biochemistry and Microbiology, Ghent University, B-9000 Gent, Belgium
  • 3 Laboratory for Biomedical Research, Department of Molecular Biotechnology, Environment Technology and Food Technology, Ghent University Global Campus, Incheon 406-840, Korea
Type
Published Article
Journal
Vaccines
Publisher
MDPI
Publication Date
Jul 24, 2020
Volume
8
Issue
3
Identifiers
DOI: 10.3390/vaccines8030415
PMID: 32722150
PMCID: PMC7565430
Source
PubMed Central
Keywords
License
Green

Abstract

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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