An Anti-proteome Nanobody Library Approach Yields a Specific Immunoassay for Trypanosoma congolense Diagnosis Targeting Glycosomal Aldolase

PLoS Neglected Tropical Diseases
Steven OdongoStefan Magez

Abstract

Infectious diseases pose a severe worldwide threat to human and livestock health. While early diagnosis could enable prompt preventive interventions, the majority of diseases are found in rural settings where basic laboratory facilities are scarce. Under such field conditions, point-of-care immunoassays provide an appropriate solution for rapid and reliable diagnosis. The limiting steps in the development of the assay are the identification of a suitable target antigen and the selection of appropriate high affinity capture and detection antibodies. To meet these challenges, we describe the development of a Nanobody (Nb)-based antigen detection assay generated from a Nb library directed against the soluble proteome of an infectious agent. In this study, Trypanosoma congolense was chosen as a model system. An alpaca was vaccinated with whole-parasite soluble proteome to generate a Nb library from which the most potent T. congolense specific Nb sandwich immunoassay (Nb474H-Nb474B) was selected. First, the Nb474-homologous sandwich ELISA (Nb474-ELISA) was shown to detect experimental infections with high Positive Predictive Value (98%), Sensitivity (87%) and Specificity (94%). Second, it was demonstrated under experimental conditio...Continue Reading

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Citations

Sep 16, 2017·PLoS Neglected Tropical Diseases·Joar PintoYann G-J Sterckx
Aug 12, 2020·The FEBS Journal·Serge Muyldermans
Jan 31, 2020·Acta Veterinaria Scandinavica·Edson KinimiGerald Misinzo
Jul 18, 2017·Frontiers in Immunology·Benoit StijlemansStefan Magez
Nov 26, 2020·Annual Review of Animal Biosciences·Serge Muyldermans
Jun 1, 2019·International Journal for Parasitology·Francisco J Morales-YanezSerge Muyldermans
Jun 11, 2021·Annals of Laboratory Medicine·Tahir S Pillay, Serge Muyldermans
Aug 28, 2021·Animals : an Open Access Journal From MDPI·Edson KinimiGerald Misinzo

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Datasets Mentioned

BETA
CCC93713.1
M19994.1
CAB55315.1

Methods Mentioned

BETA
PCR
enzyme-linked immunosorbent assay
ELISA
ELISAs
size exclusion chromatography

Software Mentioned

Proteome Discoverer
Clone Manager
BLASTN
Sequest HT
Gen5
SequestHT
GraphPad Prism

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