A high-dimensional microfluidic approach for selection of aptamers with programmable binding affinities

Area of Science:

• Biotechnology
• Analytical Chemistry
• Molecular Biology

Background:

• Aptamers are valuable affinity reagents due to their stability and chemical modifiability.
• Traditional SELEX for aptamer generation struggles with quantitative control over binding affinity and requires multiple rounds to eliminate false positives.

Purpose of the Study:

• To introduce Pro-SELEX, a novel workflow for the rapid discovery of aptamers with precisely defined binding affinities.
• To overcome the limitations of conventional SELEX in controlling aptamer binding characteristics.

Main Methods:

• Pro-SELEX integrates efficient particle display, high-performance microfluidic sorting, and high-content bioinformatics.
• This approach allows for the investigation of individual aptamer candidate binding performance under varying selective pressures within a single selection round.

Main Results:

• Pro-SELEX successfully identified aptamers with dissociation constants spanning a 20-fold range of affinities in one round.
• Human myeloperoxidase was used as a target to demonstrate the efficacy of the Pro-SELEX workflow.

Conclusions:

• Pro-SELEX offers a significant advancement for aptamer discovery, enabling rapid generation of aptamers with desired binding affinities.
• This method provides a quantitative and efficient alternative to traditional SELEX for developing aptamer-based analytical reagents.