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High-affinity one-step aptamer selection using a non-fouling porous hydrogel
Naveen K Singh1,2, Yixun Wang1, Connie Wen1
1Department of Biomedical Engineering, The Penn State University, University Park, PA, USA.
Nature Biotechnology
|October 5, 2023
Summary
This study introduces a novel, instrument-free method for selecting high-affinity aptamers (chemical antibodies) using a specialized hydrogel. This technique simplifies aptamer enrichment, overcoming challenges like nonspecific binding and reducing selection cycles.
Area of Science:
- Biotechnology
- Molecular Biology
- Materials Science
Background:
- Aptamers, or chemical antibodies, are vital for applications like drug delivery and biosensing.
- Traditional aptamer selection is challenging due to multiple enrichment cycles and nonspecific binding issues.
Purpose of the Study:
- To develop a simple, fast, and instrument-free method for aptamer enrichment and selection.
- To overcome the limitations of conventional aptamer selection processes.
Main Methods:
- Utilized a diffusion-binding process within a 3D non-fouling porous hydrogel.
- Immobilized target proteins within the hydrogel to capture high-affinity aptamers.
- Separated aptamers based on differential binding affinities to the immobilized targets.
Main Results:
- Achieved one-step aptamer pool enrichment with strong binding to target proteins.
- Demonstrated consistent enrichment across five proteins with varying isoelectric points.
- Identified an anti-thrombin aptamer with binding affinity comparable to traditional multi-cycle methods.
Conclusions:
- The developed hydrogel-based method offers a significantly simplified and efficient approach to aptamer selection.
- This technique effectively enriches aptamer pools, yielding high-affinity candidates with reduced effort.
- The method shows broad applicability for various protein targets, advancing aptamer technology.
