Volume 137, Issue 30 e202506464
Forschungsartikel

Plasmonic DNA-Barcoded Virion Nano-Oscillators for Multiplexed Quantification of Small-Molecule Binding Kinetics to Membrane Proteins

Shuo-Hui Cao

Shuo-Hui Cao

Center for Bioelectronics and Biosensors, the Biodesign Institute, Arizona State University, Tempe, AZ, 85287 USA

MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, Fujian Provincial Key Laboratory of Plasma and Magnetic Resonance, Department of Electronic Science, Xiamen University, Xiamen, Fujian, 361005 China

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Zijian Wan

Zijian Wan

Center for Bioelectronics and Biosensors, the Biodesign Institute, Arizona State University, Tempe, AZ, 85287 USA

School of Electrical, Energy and Computer Engineering, Arizona State University, Tempe, AZ, 85287 USA

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Eric Johansen

Eric Johansen

Department of Pharmacology and Molecular Sciences, Johns Hopkins School of Medicine, Baltimore, MD, 21205 USA

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Guangzhong Ma

Guangzhong Ma

Center for Bioelectronics and Biosensors, the Biodesign Institute, Arizona State University, Tempe, AZ, 85287 USA

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Prashant Desai

Prashant Desai

Viral Oncology Program, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Baltimore, MD, 21231 USA

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Heng Zhu

Heng Zhu

Department of Pharmacology and Molecular Sciences, Johns Hopkins School of Medicine, Baltimore, MD, 21205 USA

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Shaopeng Wang

Corresponding Author

Shaopeng Wang

Center for Bioelectronics and Biosensors, the Biodesign Institute, Arizona State University, Tempe, AZ, 85287 USA

School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ, 85287 USA

E-mail: [email protected]

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First published: 19 May 2025

Abstract

A high-density nano-oscillator platform using self-assembled DNA-barcoded virion sensors is developed to address the critical need for high-throughput label-free measurement of small-molecule binding to membrane proteins. By integrating virion display technology with charge-sensitive plasmonic detection, our platform enables robust, label-free quantification of small-molecule binding kinetics to membrane proteins. Gold nanoparticle-virion conjugates are self-assembled onto a plasmonic sensor chip via a flexible molecular linker to form high-density nano-oscillators. Driven by an alternating electric field, the oscillation amplitudes of the nano-oscillators are precisely measured via widefield plasmonic imaging. This charge-sensitive mechanism can sensitively detect the binding of small-molecule ligands to the membrane proteins displayed on the virions at single-nanosensor resolution, overcoming the sensitivity limit of conventional mass-sensitive techniques. More importantly, the platform employs novel affinity-discriminated DNA barcodes for multistate decoding with exponential multiplexing capacity, enabling high-throughput screening of a library of membrane proteins. For a proof-of-concept demonstration, binding kinetics of five pairs of G-protein-coupled receptors and their corresponding small molecule ligands are measured on a single sensor chip, with all individual nano-oscillators identified by just two affinity-discriminated, quadra-state DNA decoders. This technology advances membrane protein research and drug screening capabilities, offering a practical solution for biomolecular interaction studies and biosensing applications.

Conflict of Interests

A US provisional patent application (63/649.551) has been filed by Skysong Innovations, LLC, on behalf of Arizona State University, based on an early draft of this article. Inventors are S.W. and S.C.

Data Availability Statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.

The full text of this article hosted at iucr.org is unavailable due to technical difficulties.