Widespread omission of aptamer-target binding verification in aptasensor development: Consequences for sensor performance and the need for a Kd gate
13 affiliations
- 1Centre for Research on Biomolecular Interactions, York University, Toronto, Ontario, M3J 1P3, Canada; Department of Chemistry, York University, Toronto, Ontario, M3J 1P3, Canada. Electronic address: [email protected]
- 2Centre for Research on Biomolecular Interactions, York University, Toronto, Ontario, M3J 1P3, Canada; Department of Chemistry, York University, Toronto, Ontario, M3J 1P3, Canada
- 3Freshman Research Initiative, The University of Texas, Austin, TX, 78712, USA; Department of Molecular Biosciences, The University of Texas, Austin, TX, 78712, USA
- 4Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, 5625 Renmin Street, Changchun, 130022, China; School of Applied Chemistry and Engineering, University of Science and Technology of China, No. 96 Jinzhai Road, Hefei, Anhui, 230026, China
- 5A.N. Bach Institute of Biochemistry, Research Center of Biotechnology, Russian Academy of Sciences, Leninsky Prospect 33, Moscow, 119071, Russia
- 6Department of Chemistry, Carleton University, Ottawa, Ontario, K1S 5B6, Canada
- 7Institute of Chemistry, São Paulo State University, IQ-UNESP, Araraquara, 14800-060, Brazil
- 8Department of Chemistry, University of Sherbrooke, Sherbrooke, Quebec, J1K 2R1, Canada
- 9Advanced Biomedical Instrumentation Center (ABIC), Hong Kong Science Park, Hong Kong Special Administrative Region of China
- 10State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing, 100085, China; University of Chinese Academy of Sciences, Beijing, 100049, China
- 11School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, China
- 12Laser Research Centre, Faculty of Health Science, University of Johannesburg, Doornfontein, P.O. Box 17011, Johannesburg, 2028, South Africa
- 13Interdisciplinary Nanoscience Center, Faculty of Natural Sciences, Aarhus University, Gustav Wieds Vej 1590-14, Aarhus, DK-8000, Denmark
Abstract
Aptamer-based biosensors (aptasensors) are widely explored for analytical and diagnostic applications because aptamers combine molecular recognition with scalable chemical synthesis. Yet aptasensors have achieved limited translational impact, motivating scrutiny of development practices. Critical reviews emphasize verifying target binding of the aptamer construct by determining its equilibrium dissociation constant ( K d , a standard measure of binding strength) prior to aptasensor development to avoid interface-contingent artifacts. We refer to this pre-development requirement as the K d gate: an auditable, construct-matched determination of K d in homogeneous, non-denaturing solution under sensing-relevant conditions. Here, we assess compliance with the K d gate by analyzing 209 aptasensor publications spanning six targets. Only 20 of 209 papers reported any solution-phase verification assay, and only one passed the K d gate; among the remaining numeric K d reports, the reported values were weakly constrained or format-contingent and therefore not auditable. As a result, reported sensor-performance claims often cannot be interpreted, transferred, or optimized on a fully rational basis. Although practical feasibility varies across constructs and targets, the observed pattern indicates that technical difficulty alone does not explain the persistent omission. We conclude that voluntary recommendations have not, in practice, functioned as an effective gate and that passing the K d gate should become a mandatory reporting requirement, supported by an auditable checklist for editors and reviewers.
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BibTeX
@article{krylov2026widespread,
title = {{Widespread omission of aptamer-target binding verification in aptasensor development: Consequences for sensor performance and the need for a Kd gate}},
author = {Sergey N. Krylov and Svetlana M. Krylova and An Thi Hoai Le and Philip E. Johnson and Ryan Hili and Derek J. Wilson and S. Brian Kim and Gwendolyn M. Stovall and Guobao Xu and Boris Borisovich Dzantiev and Maria C. DeRosa and Paulo R. Bueno and Philippe Dauphin‐Ducharme and Julian A. Tanner and William L. Whitehouse and Qiang Zhao and Jun‐Jie Zhu and Anine Crous and Elena E. Ferapontova},
journal = {Biosensors and Bioelectronics},
year = {2026},
month = may,
doi = {10.1016/j.bios.2026.118807},
url = {https://doi.org/10.1016/j.bios.2026.118807},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Widespread omission of aptamer-target binding verification in aptasensor development: Consequences for sensor performance and the need for a Kd gate
AU - Sergey N. Krylov
AU - Svetlana M. Krylova
AU - An Thi Hoai Le
AU - Philip E. Johnson
AU - Ryan Hili
AU - Derek J. Wilson
AU - S. Brian Kim
AU - Gwendolyn M. Stovall
AU - Guobao Xu
AU - Boris Borisovich Dzantiev
AU - Maria C. DeRosa
AU - Paulo R. Bueno
AU - Philippe Dauphin‐Ducharme
AU - Julian A. Tanner
AU - William L. Whitehouse
AU - Qiang Zhao
AU - Jun‐Jie Zhu
AU - Anine Crous
AU - Elena E. Ferapontova
PY - 2026
DA - 2026/05/15
JO - Biosensors and Bioelectronics
DO - 10.1016/j.bios.2026.118807
UR - https://doi.org/10.1016/j.bios.2026.118807
AB - Aptamer-based biosensors (aptasensors) are widely explored for analytical and diagnostic applications because aptamers combine molecular recognition with scalable chemical synthesis. Yet aptasensors have achieved limited translational impact, motivating scrutiny of development practices. Critical reviews emphasize verifying target binding of the aptamer construct by determining its equilibrium dissociation constant ( K d , a standard measure of binding strength) prior to aptasensor development to avoid interface-contingent artifacts. We refer to this pre-development requirement as the K d gate: an auditable, construct-matched determination of K d in homogeneous, non-denaturing solution under sensing-relevant conditions. Here, we assess compliance with the K d gate by analyzing 209 aptasensor publications spanning six targets. Only 20 of 209 papers reported any solution-phase verification assay, and only one passed the K d gate; among the remaining numeric K d reports, the reported values were weakly constrained or format-contingent and therefore not auditable. As a result, reported sensor-performance claims often cannot be interpreted, transferred, or optimized on a fully rational basis. Although practical feasibility varies across constructs and targets, the observed pattern indicates that technical difficulty alone does not explain the persistent omission. We conclude that voluntary recommendations have not, in practice, functioned as an effective gate and that passing the K d gate should become a mandatory reporting requirement, supported by an auditable checklist for editors and reviewers.
SN - 0956-5663
SN - 1873-4235
ER -