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Automated System for Single Molecule Fluorescence Measurements of Surface-immobilized Biomolecules

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AptaShield: A Universal Signal-Transduction System for Fast and High-Throughput Optical Molecular Biosensing.

Miguel António Dias Neves^1,2, Inês Mendes Pinto^1,2

¹Institute for Research and Innovation in Health (i3S), University of Porto, 4200-135 Porto, Portugal.

|April 15, 2024

Summary

A novel AptaShield biosensor enables universal, high-throughput detection of analytes in biological fluids. This solution-phase technology overcomes limitations of traditional assays, offering a versatile platform for point-of-care diagnostics.

Keywords:

Aptamers High-throughput Molecular profiling Optical detection Solution-phase biosensor Universal transducer

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Area of Science:

Biotechnology
Biosensing
Molecular Diagnostics

Background:

Biosensing commercialization is limited by reconfiguration difficulties and low throughput.
Immobilizing biomolecular probes requires tailored surface chemistry for optimal performance.
Existing methods struggle with universal analyte detection and high-throughput capabilities.

Purpose of the Study:

To develop a universal, high-throughput solution-phase biosensor overcoming current limitations.
To create an engineered aptamer (AptaShield) for universal immunoglobulin G (IgG) binding.
To enable facile biosensor reconfiguration for diverse analyte detection.

Main Methods:

Development of the AptaShield engineered aptamer for universal binding to rabbit IgG Fab' fragments.
Utilizing Förster resonance energy transfer (FRET) signal generation based on aptamer-IgG dynamic equilibrium.
Dissociation of the AptaShield-IgG complex upon analyte binding, causing a FRET signal decrease.

Main Results:

Demonstrated high selectivity, specificity, and reproducibility for analyte quantification.
Successful application in various biological fluids (urine, blood serum) with low sample volumes.
Achieved a universal signal transduction mechanism without aptamer modification for different rabbit antibodies.

Conclusions:

AptaShield offers a robust, high-throughput solution-phase biosensing technology.
The biosensor is suitable for point-of-care applications, outperforming traditional enzyme-linked immunosorbent assays (ELISA).
Provides a universal platform for molecular profiling, overcoming limitations in biosensor development.