A system for multiplexed selection of aptamers with exquisite specificity without counterselection

Area of Science:

• Biotechnology
• Molecular Biology
• Analytical Chemistry

Background:

• Aptamers are crucial for detecting small molecules due to their high specificity.
• Traditional aptamer selection relies on counterselection, which is inefficient and requires trial-and-error.
• Determining optimal counterselection parameters is challenging due to unknown library characteristics.

Purpose of the Study:

• To develop a high-throughput screening process for generating high-specificity aptamers.
• To enable parallel selection of aptamers against multiple targets without counterselection.
• To accelerate the development of aptamer reagents with exquisite specificity.

Main Methods:

• Utilized a modified benchtop sequencer for massively parallel aptamer screening.
• Implemented a novel screening process to select aptamers against multiple targets in parallel.
• Demonstrated the method by selecting aptamers for structurally similar kynurenine metabolites.

Main Results:

• Successfully generated aptamers with high affinity and exquisite specificity for three structurally similar kynurenine metabolites.
• The developed process eliminated the need for counterselection.
• Achieved parallel selection of highly specific aptamers against multiple targets in a single experiment.

Conclusions:

• The described high-throughput screening process is efficient for generating highly specific aptamers.
• This method significantly accelerates aptamer development for small-molecule detection.
• The platform is adaptable to various small-molecule analytes, enhancing aptamer reagent development.