Scalable Multiparametric Characterization of Aptamer-Target Interactions.

Area of Science:

• Biotechnology and Biosensing
• Molecular Biology and Aptamer Engineering

Background:

• Structure-switching aptamers are crucial for biosensing small molecules by converting conformational changes into signals.
• Understanding aptamer structural dynamics is key for rational design in biosensor development.
• Existing methods lack the scalability and spatiotemporal resolution for comprehensive aptamer dynamics analysis.

Purpose of the Study:

• To develop a scalable droplet microfluidic platform for high-resolution analysis of aptamer-target interactions.
• To enable multiparametric profiling of aptamer structural dynamics under physiologically relevant conditions.
• To bridge the gap between structural characterization and biosensor development for data-driven aptamer engineering.

Main Methods:

• Integration of Förster resonance energy transfer (FRET) with automated imaging in a droplet microfluidic system.
• Analysis of aptamer-target interactions in picoliter volumes across millisecond-to-hour timescales.
• Systematic investigation of serotonin aptamers with varying stem lengths to explore structure-function relationships.

Main Results:

• The platform provides high spatiotemporal resolution for characterizing aptamer structural transitions.
• Detailed analysis of aptamer-target interactions revealed structure-function relationships.
• Insights were translated into the selection of optimal aptamer candidates for specific applications.

Conclusions:

• The developed droplet microfluidic platform overcomes limitations in characterizing aptamer dynamics.
• This technology facilitates the rational design and engineering of structure-switching aptamers for biosensing.
• The platform lays the foundation for advancing translational biosensor development and diagnostics.