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Updated: Apr 21, 2026

Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope

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Epitope-Directed Antibody Development and Enhancement for Broad-Spectrum Fentanyl-Class Substance Detection: From

Xiaoyi Chen^1,2, Xiang Chi^1,2, Yuxi Hu^1,2

¹Jiangsu Key Laboratory of Drug Design and Optimization, China Pharmaceutical University, Nanjing 210009, China.

Analytical Chemistry

|April 18, 2026

Summary

Developing new detection tools is vital for monitoring fentanyl and its analogues. This study created highly sensitive assays for 32 fentanyl analogues, aiding public health surveillance.

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Targeted Antibody Blocking by a Dual-Functional Conjugate of Antigenic Peptide and Fc-III Mimetics DCAF

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Published on: September 17, 2019

Area of Science:

Forensic Chemistry
Immunology
Drug Detection

Background:

Fentanyl analogue abuse is a major public health crisis.
Structural modifications of fentanyl analogues complicate detection and regulation.
Robust surveillance tools are needed to combat illicit fentanyl use.

Purpose of the Study:

To develop novel monoclonal antibodies (mAbs) targeting fentanyl analogues.
To create sensitive and specific detection assays for fentanyl analogues.
To establish a rational hapten design framework for antibody generation.

Main Methods:

Epitope-directed hapten design targeting fentanyl core structures.
Hybridoma technology for monoclonal antibody production.
Homology modeling and molecular docking for binding pattern analysis.
Development of indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and gold nanoparticle-based immunochromatographic assay (GICA).

Main Results:

Generated three mAbs with nanomolar affinity and tunable cross-reactivity.
Elucidated structural basis of antibody-hapten interactions.
Developed ic-ELISA detecting 32 fentanyl analogues (IC50: 0.072-101.7 ng/mL).
Developed GICA with a 5 ng/mL cutoff for rapid on-site detection.
Assays demonstrated high accuracy in human urine and hair samples (80.20-118.65% recovery).

Conclusions:

The developed mAbs and assays provide powerful tools for fentanyl analogue surveillance.
The study offers a rational hapten design strategy for generating antibodies with tailored properties.
This work supports enhanced public health strategies against fentanyl abuse.