Particle display: a quantitative screening method for generating high-affinity aptamers

Area of Science:

• Biotechnology and Molecular Biology
• Biophysics
• Genomics and Proteomics

Background:

• Conventional aptamer selection methods can be time-consuming and may not always yield high-affinity aptamers.
• Discovering aptamers for challenging protein targets remains a significant hurdle in molecular biology.
• There is a need for more efficient and rapid technologies for aptamer discovery.

Purpose of the Study:

• To introduce and validate a novel aptamer discovery technology termed particle display.
• To demonstrate the superior efficiency and affinity of aptamers generated by particle display compared to conventional methods.
• To showcase the application of particle display for identifying high-affinity aptamers against diverse protein targets.

Main Methods:

• Transformation of solution-phase aptamer libraries into 'aptamer particles', with each particle displaying numerous copies of a single aptamer sequence.
• High-throughput screening and affinity measurement of individual aptamer particles using fluorescence-activated cell sorting (FACS).
• Mathematical analysis to optimize experimental parameters for particle display screening.

Main Results:

• Particle display reproducibly yields higher affinity aptamers in fewer selection rounds.
• Demonstrated enrichment performance exceeding theoretical maximums of conventional selection by orders of magnitude.
• Successfully obtained high-affinity DNA aptamers for four distinct protein targets within three rounds of selection.

Conclusions:

• Particle display is an extraordinarily efficient technology for rapid and economic generation of high-quality aptamers.
• This method overcomes limitations of conventional selection, enabling aptamer discovery for previously challenging targets.
• Particle display accelerates the exploration of the human proteome by providing a powerful tool for aptamer generation.