Recognition-Enhanced Metastably Shielded Aptamer for Digital Quantification of Small Molecules

Area of Science:

• Biotechnology
• Molecular Biology
• Analytical Chemistry

Background:

• Aptamers are valuable affinity reagents but struggle with low target binding affinity, especially for small molecules.
• Existing methods face limitations in precisely quantifying low-affinity small molecules using aptamers.

Purpose of the Study:

• To introduce and evaluate a novel aptamer probe design, the recognition-enhanced metastably shielded aptamer probe (RMSApt).
• To demonstrate the improved performance of RMSApt for the digital quantification of low-affinity small molecules.

Main Methods:

• Designed an allosteric aptamer probe (RMSApt) with a minor energy gap in the recognition switch process.
• Explored the probe's performance for digital quantification of targets with dissociation constants (K_d) from 10^-4 to 10^-9 M.

Main Results:

• The RMSApt design significantly enhances recognition efficiency for low-affinity targets.
• The probe strategy enables precise quantification across a broad range of small-molecule targets.
• Successfully demonstrated improved aptamer application for low-affinity small molecule detection.

Conclusions:

• RMSApt technology overcomes limitations in aptamer-based quantification of low-affinity small molecules.
• This advancement facilitates aptamer translation for critical applications in medical diagnosis, food safety, and environmental screening.
• The developed probe strategy broadens the utility of aptamers in detecting diverse small-molecule species.