Advancing Tuberculosis Antigen Detection by Selecting Sandwich-Compatible Aptamers Against the Mycobacterium tuberculosis ESAT-6/CFP-10 Heterodimer Using Library-Immobilized Strategies.
- 1Interfibio Consolidated Research Group, Departament d'Enginyeria Química, Universitat Rovira i Virgili, 43007Tarragona, Spain.
- 2Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Flemingovo nam. 542/2, 160 00Prague 6, Czech Republic.
- 3Institució Catalana de Recerca i Estudis Avancats (ICREA), 08010Barcelona, Spain.
Abstract
Tuberculosis (TB) represents the world's leading infectious disease killer, surpassing COVID-19. Despite being preventable and curable, a substantial proportion of cases remain undiagnosed or unreported, sustaining transmission. The detection of antigens such as the Mycobacterium tuberculosis ESAT-6/CFP-10 heterodimer, which are secreted at different stages of infection, is a promising screening strategy. Traditional antigen tests relying on antibodies suffer from significant problems of specificity and sensitivity. Aptamers, selected in an in vitro technique termed SELEX (Systematic Evolution of Ligands by EXponential enrichment), are short, single-stranded oligonucleotides that fold into unique 3D structures, allowing them to bind to specific targets, including proteins, cells, or small molecules, with high affinity and specificity. However, conventional SELEX methods often yield suboptimal aptamers partly due to target immobilization that can promote nonspecific interactions and distort protein conformation. Here, we employed two library-immobilized strategies to generate aptamers against the M. tuberculosis ESAT-6/CFP-10 heterodimer. We first performed capture-SELEX to select aptamers against the unconstrained, conformationally native heterodimer target in solution. These selected aptamers were then exploited in a second SELEX, where aptamer-target complexes were "baited" by a target-specific antibody to isolate sandwich-compatible aptamers for downstream assay development. The selected aptamers exhibited high nanomolar affinity regardless of library and selection strategy employed, with bait-SELEX-derived candidates emerging after fewer selection rounds. These aptamers were successfully applied in antibody/aptamer hybrid sandwich assays, supporting their potential for TB antigen detection in nonsputum samples.
Related
- ESX-5 paralog deletions in Mycobacterium tuberculosis alter macrophage cytokine signaling and bacterial heavy metal response
- Fragment-based drug discovery strategy reveals an unprecedented allosteric binding site in Mycobacterium tuberculosis DNA gyrase
- X-ray crystallographic fragment screening reveals novel and conformationally dynamic ligand-binding sites in Mycobacterium tuberculosis FtsZ
- Mycobacterium tuberculosis gene expression dynamics in host environments with single-cell resolution
- Dynamic closure of the Mycobacterium tuberculosis InhA cofactor site reveals heterogeneous routes to isoniazid resistance
BibTeX
@article{acosta2026advancing,
title = {{Advancing Tuberculosis Antigen Detection by Selecting Sandwich-Compatible Aptamers Against the Mycobacterium tuberculosis ESAT-6/CFP-10 Heterodimer Using Library-Immobilized Strategies.}},
author = {Shaira Jane Acosta and M Carmen Bermudo Redondo and Lucie Mužíková Čechová and Marek Ondruš and Vasso Skouridou and Ciara K O'Sullivan},
journal = {ACS infectious diseases},
year = {2026},
month = oct,
doi = {10.1021/acsinfecdis.6c00529},
url = {https://europepmc.org/article/MED/42917103},
note = {CC BY},
}
RIS
TY - JOUR
TI - Advancing Tuberculosis Antigen Detection by Selecting Sandwich-Compatible Aptamers Against the Mycobacterium tuberculosis ESAT-6/CFP-10 Heterodimer Using Library-Immobilized Strategies.
AU - Shaira Jane Acosta
AU - M Carmen Bermudo Redondo
AU - Lucie Mužíková Čechová
AU - Marek Ondruš
AU - Vasso Skouridou
AU - Ciara K O'Sullivan
PY - 2026
DA - 2026/10/01
JO - ACS infectious diseases
DO - 10.1021/acsinfecdis.6c00529
UR - https://europepmc.org/article/MED/42917103
AB - Tuberculosis (TB) represents the world's leading infectious disease killer, surpassing COVID-19. Despite being preventable and curable, a substantial proportion of cases remain undiagnosed or unreported, sustaining transmission. The detection of antigens such as the Mycobacterium tuberculosis ESAT-6/CFP-10 heterodimer, which are secreted at different stages of infection, is a promising screening strategy. Traditional antigen tests relying on antibodies suffer from significant problems of specificity and sensitivity. Aptamers, selected in an in vitro technique termed SELEX (Systematic Evolution of Ligands by EXponential enrichment), are short, single-stranded oligonucleotides that fold into unique 3D structures, allowing them to bind to specific targets, including proteins, cells, or small molecules, with high affinity and specificity. However, conventional SELEX methods often yield suboptimal aptamers partly due to target immobilization that can promote nonspecific interactions and distort protein conformation. Here, we employed two library-immobilized strategies to generate aptamers against the M. tuberculosis ESAT-6/CFP-10 heterodimer. We first performed capture-SELEX to select aptamers against the unconstrained, conformationally native heterodimer target in solution. These selected aptamers were then exploited in a second SELEX, where aptamer-target complexes were "baited" by a target-specific antibody to isolate sandwich-compatible aptamers for downstream assay development. The selected aptamers exhibited high nanomolar affinity regardless of library and selection strategy employed, with bait-SELEX-derived candidates emerging after fewer selection rounds. These aptamers were successfully applied in antibody/aptamer hybrid sandwich assays, supporting their potential for TB antigen detection in nonsputum samples.
SN - 2373-8227
ER -