High-Affinity Anti-nivolumab DNA Aptamer: A Novel Capture Reagent for Its Bioanalysis by an Enzyme-Linked Aptamer Assay
Abstract
Bioanalysis of the immune checkpoint inhibitor nivolumab is crucial for pharmacokinetic studies. However, conventional ligand-binding assays (LBAs) often suffer from the lot-to-lot variability of capture antibodies. Here, we developed a high-affinity anti-nivolumab DNA aptamer as a robust alternative reagent. An integrated strategy combining Fast Protein Liquid Chromatography-based screening, next-generation sequencing, aptamer search algorithm, and computational modeling enabled rapid aptamer identification and optimization. The anti-nivolumab aptamer, mutant 2-3, discovered through in silico modification exhibited a dissociation constant (KD) of 33 nM. Using this aptamer, an enzyme-linked aptamer assay (ELAA) successfully quantified nivolumab in 400-fold diluted plasma samples over a range of 0.02–5.0 µg/mL, demonstrating good precision with a coefficient of variation less than 11.9%. This novel analytical method provides a sufficient quantitative range for the bioanalysis of nivolumab, thus offering a robust, specific, and cost-effective alternative for future pharmacokinetic studies.
Related
- Prussian blue electrocatalyst in bioanalysis: From oxidase biosensors to peroxidase kinetics assays
- Pharmaceutical Applications of Nucleic Acid Aptamers: Recent Advances and Future Perspectives in Therapeutics, Biosensing, Bioanalysis, and Process Analysis
- Looking Beyond KD for Aptamer Validation: Disentangling Specific Aptamer Binding from Electrostatic Association for Basic Protein Targets
- Preparation of electrochemical aptamer-based sensors: a direct aryl diazonium grafting approach
- Combining AlphaFold 3 with microscale thermophoresis for aptamer optimisation
BibTeX
@article{hojo2026highaffinity,
title = {{High-Affinity Anti-nivolumab DNA Aptamer: A Novel Capture Reagent for Its Bioanalysis by an Enzyme-Linked Aptamer Assay}},
author = {Taisei Hojo and Nao Kobayashi and Chiho Hirayama and Jiaxing Tong and Naruma Uchidate and Aogu Furusho and Kenji Kojima and Eiji Sugiyama and Hajime Mizuno and Hideki Hayashi and Kenichiro Todoroki},
journal = {Chemical and Pharmaceutical Bulletin},
year = {2026},
month = aug,
doi = {10.1248/cpb.c25-00788},
url = {https://www.jstage.jst.go.jp/article/cpb/74/8/74_c25-00788/_article},
note = {CC BY-NC},
}
RIS
TY - JOUR
TI - High-Affinity Anti-nivolumab DNA Aptamer: A Novel Capture Reagent for Its Bioanalysis by an Enzyme-Linked Aptamer Assay
AU - Taisei Hojo
AU - Nao Kobayashi
AU - Chiho Hirayama
AU - Jiaxing Tong
AU - Naruma Uchidate
AU - Aogu Furusho
AU - Kenji Kojima
AU - Eiji Sugiyama
AU - Hajime Mizuno
AU - Hideki Hayashi
AU - Kenichiro Todoroki
PY - 2026
DA - 2026/08/01
JO - Chemical and Pharmaceutical Bulletin
DO - 10.1248/cpb.c25-00788
UR - https://www.jstage.jst.go.jp/article/cpb/74/8/74_c25-00788/_article
AB - Bioanalysis of the immune checkpoint inhibitor nivolumab is crucial for pharmacokinetic studies. However, conventional ligand-binding assays (LBAs) often suffer from the lot-to-lot variability of capture antibodies. Here, we developed a high-affinity anti-nivolumab DNA aptamer as a robust alternative reagent. An integrated strategy combining Fast Protein Liquid Chromatography-based screening, next-generation sequencing, aptamer search algorithm, and computational modeling enabled rapid aptamer identification and optimization. The anti-nivolumab aptamer, mutant 2-3, discovered through in silico modification exhibited a dissociation constant (KD) of 33 nM. Using this aptamer, an enzyme-linked aptamer assay (ELAA) successfully quantified nivolumab in 400-fold diluted plasma samples over a range of 0.02–5.0 µg/mL, demonstrating good precision with a coefficient of variation less than 11.9%. This novel analytical method provides a sufficient quantitative range for the bioanalysis of nivolumab, thus offering a robust, specific, and cost-effective alternative for future pharmacokinetic studies.
SN - 1347-5223
SN - 0009-2363
ER -