Nanobodies, a promising tool for species-specific diagnosis of Taenia solium cysticercosis

被引:62
作者
Deckers, N. [1 ,2 ]
Saerens, D. [3 ]
Kanobana, K. [1 ]
Conrath, K. [3 ,4 ]
Victor, B. [1 ]
Wernery, U. [5 ]
Vercruysse, J. [2 ]
Muyldermans, S. [3 ]
Dorny, P. [1 ,2 ]
机构
[1] Inst Trop Med, Dept Anim Hlth, B-2000 Antwerp, Belgium
[2] Univ Ghent, Fac Vet Med, Parasitol Lab, Merelbeke, Belgium
[3] Vrije Univ Brussel, Cellular & Mol Immunol Lab, Brussels, Belgium
[4] VIB, Dept Mol & Cellular Interact, Brussels, Belgium
[5] Cent Vet Res Lab, Dubai, U Arab Emirates
基金
比利时弗兰德研究基金会;
关键词
Taenia solium; Cysticercosis; Pig; Single-domain-antibodies; 8 kDa protein family; Epitope mapping; SINGLE-DOMAIN ANTIBODIES; EXCRETORY-SECRETORY ANTIGENS; LIGAND-BINDING PROTEIN; MONOCLONAL-ANTIBODY; FRAGMENTS; PORCINE; NEUROCYSTICERCOSIS; METACESTODES; EPITOPES; SAMPLES;
D O I
10.1016/j.ijpara.2008.10.012
中图分类号
R38 [医学寄生虫学]; Q [生物科学];
学科分类号
07 ; 0710 ; 09 ; 100103 ;
摘要
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 monodonal-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. (C) 2008 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.
引用
收藏
页码:625 / 633
页数:9
相关论文
共 43 条
[1]   Gapped BLAST and PSI-BLAST: a new generation of protein database search programs [J].
Altschul, SF ;
Madden, TL ;
Schaffer, AA ;
Zhang, JH ;
Zhang, Z ;
Miller, W ;
Lipman, DJ .
NUCLEIC ACIDS RESEARCH, 1997, 25 (17) :3389-3402
[2]   Characterization of excretory/secretory endopeptidase and metallo-aminopeptidases from Taenia crassiceps metacestodes [J].
Baig, S ;
Damian, RT ;
Morales-Montor, J ;
Olecki, P ;
Talhouk, J ;
Hashmey, R ;
White, AC .
JOURNAL OF PARASITOLOGY, 2005, 91 (05) :983-987
[3]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[4]   A MONOCLONAL ANTIBODY-BASED ELISA FOR THE DETECTION OF CIRCULATING EXCRETORY SECRETORY ANTIGENS IN TAENIA-SAGINATA CYSTICERCOSIS [J].
BRANDT, JRA ;
GEERTS, S ;
DEDEKEN, R ;
KUMAR, V ;
CEULEMANS, F ;
BRIJS, L ;
FALLA, N .
INTERNATIONAL JOURNAL FOR PARASITOLOGY, 1992, 22 (04) :471-477
[5]   β-lactamase inhibitors derived from single-domain antibody fragments elicited in the Camelidae [J].
Conrath, KE ;
Lauwereys, M ;
Galleni, M ;
Matagne, A ;
Frère, JM ;
Kinne, J ;
Wyns, L ;
Muyldermans, S .
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 2001, 45 (10) :2807-2812
[6]   Camel single-domain antibodies as modular building units in bispecific and bivalent antibody constructs [J].
Conrath, KE ;
Lauwereys, M ;
Wyns, L ;
Muyldermans, S .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (10) :7346-7350
[7]   Molecular basis for the preferential cleft recognition by dromedary heavy-chain antibodies [J].
De Genst, E ;
Silence, K ;
Decanniere, K ;
Conrath, K ;
Loris, R ;
Kinne, R ;
Muyldermans, S ;
Wyns, L .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2006, 103 (12) :4586-4591
[8]   A SIMPLE AND RAPID TREATMENT (TRICHLOROACETIC-ACID PRECIPITATION) OF SERUM SAMPLES TO PREVENT NONSPECIFIC REACTIONS IN THE IMMUNOASSAY OF A PROTEOGLYCAN [J].
DEJONGE, N ;
FILLIE, YE ;
DEELDER, AM .
JOURNAL OF IMMUNOLOGICAL METHODS, 1987, 99 (02) :195-197
[9]   A Bayesian approach for estimating values for prevalence and diagnostic test characteristics of porcine cysticercosis [J].
Dorny, P ;
Phiri, IK ;
Vercruysse, J ;
Gabriel, S ;
Willingham, AL ;
Brandt, J ;
Victor, B ;
Speybroeck, N ;
Berkvens, D .
INTERNATIONAL JOURNAL FOR PARASITOLOGY, 2004, 34 (05) :569-576
[10]   Immunodiagnostic tools for human and porcine cysticercosis [J].
Dorny, P ;
Brandt, J ;
Zoli, A ;
Geerts, S .
ACTA TROPICA, 2003, 87 (01) :79-86