Structural basis for the high specificity of a Trypanosoma congolense immunoassay targeting glycosomal aldolase.
Animal African trypanosomosis (AAT) is a neglected tropical disease which imposes a heavy burden on the livestock industry in Sub-Saharan Africa. Its causative agents are Trypanosoma parasites, with T. congolense and T. vivax being responsible for the majority of the cases. Recently, we identified a...
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ftdoajarticles:oai:doaj.org/article:5f404419b70b4a13a2f7d237d7db0591 2023-05-15T15:11:47+02:00 Structural basis for the high specificity of a Trypanosoma congolense immunoassay targeting glycosomal aldolase. Joar Pinto Steven Odongo Felicity Lee Vaiva Gaspariunaite Serge Muyldermans Stefan Magez Yann G-J Sterckx 2017-09-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0005932 https://doaj.org/article/5f404419b70b4a13a2f7d237d7db0591 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC5617235?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0005932 https://doaj.org/article/5f404419b70b4a13a2f7d237d7db0591 PLoS Neglected Tropical Diseases, Vol 11, Iss 9, p e0005932 (2017) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2017 ftdoajarticles https://doi.org/10.1371/journal.pntd.0005932 2022-12-31T09:50:09Z Animal African trypanosomosis (AAT) is a neglected tropical disease which imposes a heavy burden on the livestock industry in Sub-Saharan Africa. Its causative agents are Trypanosoma parasites, with T. congolense and T. vivax being responsible for the majority of the cases. Recently, we identified a Nanobody (Nb474) that was employed to develop a homologous sandwich ELISA targeting T. congolense fructose-1,6-bisphosphate aldolase (TcoALD). Despite the high sequence identity between trypanosomatid aldolases, the Nb474-based immunoassay is highly specific for T. congolense detection. The results presented in this paper yield insights into the molecular principles underlying the assay's high specificity.The structure of the Nb474-TcoALD complex was determined via X-ray crystallography. Together with analytical gel filtration, the structure reveals that a single TcoALD tetramer contains four binding sites for Nb474. Through a comparison with the crystal structures of two other trypanosomatid aldolases, TcoALD residues Ala77 and Leu106 were identified as hot spots for specificity. Via ELISA and surface plasmon resonance (SPR), we demonstrate that mutation of these residues does not abolish TcoALD recognition by Nb474, but does lead to a lack of detection in the Nb474-based homologous sandwich immunoassay.The results show that the high specificity of the Nb474-based immunoassay is not determined by the initial recognition event between Nb474 and TcoALD, but rather by its homologous sandwich design. This (i) provides insights into the optimal set-up of the assay, (ii) may be of great significance for field applications as it could explain the potential detection escape of certain T. congolense strains, and (iii) may be of general interest to those developing similar assays. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 11 9 e0005932 |
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Directory of Open Access Journals: DOAJ Articles |
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English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Joar Pinto Steven Odongo Felicity Lee Vaiva Gaspariunaite Serge Muyldermans Stefan Magez Yann G-J Sterckx Structural basis for the high specificity of a Trypanosoma congolense immunoassay targeting glycosomal aldolase. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Animal African trypanosomosis (AAT) is a neglected tropical disease which imposes a heavy burden on the livestock industry in Sub-Saharan Africa. Its causative agents are Trypanosoma parasites, with T. congolense and T. vivax being responsible for the majority of the cases. Recently, we identified a Nanobody (Nb474) that was employed to develop a homologous sandwich ELISA targeting T. congolense fructose-1,6-bisphosphate aldolase (TcoALD). Despite the high sequence identity between trypanosomatid aldolases, the Nb474-based immunoassay is highly specific for T. congolense detection. The results presented in this paper yield insights into the molecular principles underlying the assay's high specificity.The structure of the Nb474-TcoALD complex was determined via X-ray crystallography. Together with analytical gel filtration, the structure reveals that a single TcoALD tetramer contains four binding sites for Nb474. Through a comparison with the crystal structures of two other trypanosomatid aldolases, TcoALD residues Ala77 and Leu106 were identified as hot spots for specificity. Via ELISA and surface plasmon resonance (SPR), we demonstrate that mutation of these residues does not abolish TcoALD recognition by Nb474, but does lead to a lack of detection in the Nb474-based homologous sandwich immunoassay.The results show that the high specificity of the Nb474-based immunoassay is not determined by the initial recognition event between Nb474 and TcoALD, but rather by its homologous sandwich design. This (i) provides insights into the optimal set-up of the assay, (ii) may be of great significance for field applications as it could explain the potential detection escape of certain T. congolense strains, and (iii) may be of general interest to those developing similar assays. |
format |
Article in Journal/Newspaper |
author |
Joar Pinto Steven Odongo Felicity Lee Vaiva Gaspariunaite Serge Muyldermans Stefan Magez Yann G-J Sterckx |
author_facet |
Joar Pinto Steven Odongo Felicity Lee Vaiva Gaspariunaite Serge Muyldermans Stefan Magez Yann G-J Sterckx |
author_sort |
Joar Pinto |
title |
Structural basis for the high specificity of a Trypanosoma congolense immunoassay targeting glycosomal aldolase. |
title_short |
Structural basis for the high specificity of a Trypanosoma congolense immunoassay targeting glycosomal aldolase. |
title_full |
Structural basis for the high specificity of a Trypanosoma congolense immunoassay targeting glycosomal aldolase. |
title_fullStr |
Structural basis for the high specificity of a Trypanosoma congolense immunoassay targeting glycosomal aldolase. |
title_full_unstemmed |
Structural basis for the high specificity of a Trypanosoma congolense immunoassay targeting glycosomal aldolase. |
title_sort |
structural basis for the high specificity of a trypanosoma congolense immunoassay targeting glycosomal aldolase. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2017 |
url |
https://doi.org/10.1371/journal.pntd.0005932 https://doaj.org/article/5f404419b70b4a13a2f7d237d7db0591 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 11, Iss 9, p e0005932 (2017) |
op_relation |
http://europepmc.org/articles/PMC5617235?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0005932 https://doaj.org/article/5f404419b70b4a13a2f7d237d7db0591 |
op_doi |
https://doi.org/10.1371/journal.pntd.0005932 |
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PLOS Neglected Tropical Diseases |
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11 |
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9 |
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e0005932 |
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