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Selection of optimised ligands by fluorescence-activated bead sorting.
Alexandra R Paul1, Mario Falsaperna1, Helen Lavender2
1School of Chemistry and Forensic Sciences, Division of Natural Sciences, University of Kent Canterbury CT2 7NH UK.
Chemical Science
|September 15, 2023
Summary
Researchers developed a new method to select modified aptamers with higher binding efficacy. This technique rapidly screens vast aptamer libraries, accelerating diagnostic and drug discovery for targets like EGFR.
Area of Science:
- Biochemistry and Molecular Biology
- Chemical Biology
- Nucleic Acid Chemistry
Background:
- Aptamer chemistry is primarily restricted to natural nucleotides.
- Nucleic acid modifications can improve aptamer affinity, but selection methods are lacking.
- Enzymatic amplification fails to convey sequence-specific modification information.
Purpose of the Study:
- To develop the first method for selecting specific nucleoside modifications that enhance aptamer binding efficacy.
- To demonstrate this novel strategy using the oncoprotein EGFR as a model target.
Main Methods:
- Utilized fluorescence-activated bead sorting (FABS) to select optimized aptamers.
- Screened a library of over 65,000 aptamer variations.
- Identified hits via tandem mass spectrometry and validated using EGFR binding assays and computational docking.
Main Results:
- Successfully selected optimized aptamers with specific nucleoside modifications.
- Demonstrated increased aptamer binding efficacy against the EGFR target.
- Provided proof of concept for a novel chemical optimization strategy for aptamers.
Conclusions:
- This study presents a groundbreaking method for selecting chemically optimized aptamers.
- The developed strategy accelerates the synthesis and screening of large aptamer libraries.
- This approach holds significant potential for advancing diagnostic and drug discovery applications.
