Nanobodies, a promising tool for species-specific diagnosis of Taenia solium cysticercosis |
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Authors: | N. Deckers D. Saerens K. Kanobana K. Conrath B. Victor J. Vercruysse P. Dorny |
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Affiliation: | a Animal Health Department, Institute of Tropical Medicine, Nationalestraat 155, B-2000 Antwerp, Belgium b Laboratory of Parasitology, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium c Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, Belgium d Department of Molecular and Cellular Interactions, VIB, Brussels, Belgium e Central Veterinary Research Laboratory, Dubai, United Arab Emirates |
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Abstract: | Taenia solium cysticercosis is a major helminth zoonosis in developing countries. Pigs are the intermediate hosts mediating transmission of infection. Specific assays to diagnose living cysts in pigs are lacking. The monoclonal-based antigen detection ELISA is genus-specific and cross-reactions with Taenia hydatigena hamper the use of this test to screen pigs. We, therefore, aimed to introduce nanobodies, camelid-derived single-domain antibodies specific for T. solium cysticercosis, to develop unambiguous tests. Nanobodies were cloned following immunization of two dromedaries with T. solium antigen and eight T. solium-specific nanobodies were selected after phage display. Their binding characteristics and potential for the diagnosis of porcine cysticercosis were investigated. The nanobodies do not cross-react with T. hydatigena, Taenia saginata, Taenia crassiceps or Trichinella spiralis and were categorized into four epitope-binding groups. The target protein was identified as 14 kDa diagnostic glycoprotein (Ts14), but the nanobodies also reacted with other proteins of the same family. Nanobodies were tested in a sandwich ELISA with cyst fluid, and one particular nanobody detected its cognate serum antigens in a species-specific inhibition ELISA. Considering their beneficial production and stability properties, these highly specific nanobodies constitute a promising tool to diagnose cysticercosis after further improvement of the sensitivity and future assay validation. |
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Keywords: | Taenia solium Cysticercosis Pig Single-domain-antibodies 8 kDa protein family Epitope mapping |
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