Multiparameter Particle Display (MPPD): A Quantitative Screening Method for the Discovery of Highly Specific Aptamers

Area of Science:

• Biochemistry
• Molecular Biology
• Biotechnology

Background:

• Aptamers are chemically synthesized affinity reagents with high reproducibility.
• Generating aptamers with both high affinity and specificity simultaneously is challenging with conventional methods.
• Current selection methods typically focus on either affinity or specificity, not both.

Purpose of the Study:

• To develop a high-throughput method for simultaneously screening aptamers for both affinity and specificity.
• To demonstrate the capability of fluorescence activated cell sorting (FACS) for multicolor aptamer selection.
• To generate high-affinity and high-specificity DNA aptamers for diverse protein targets.

Main Methods:

• Utilized fluorescence activated cell sorting (FACS) for multicolor sorting to screen aptamers.
• Achieved a screening throughput of 10^7 aptamers per hour.
• Generated DNA aptamers against three diverse protein targets in human serum.

Main Results:

• Developed DNA aptamers with picomolar to low nanomolar affinity in human serum.
• Demonstrated simultaneous screening for both affinity and specificity using FACS.
• The generated aptamers showed superior performance compared to monoclonal antibodies in ELISA.

Conclusions:

• FACS enables simultaneous selection of aptamers for both affinity and specificity at high throughput.
• This method overcomes limitations of conventional aptamer selection processes.
• The developed aptamers represent a promising alternative to antibodies for protein detection assays.