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Engineering Robust Aptamers with High Affinity by Key Fragment Evolution and Terminal Fixation.
1College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China.
Analytical Chemistry
|November 2, 2022
Summary
Researchers developed a new strategy to stabilize aptamers, significantly enhancing their affinity and sensitivity for analytical applications. This method improves aptamer performance for detecting toxins and biomarkers like phosphatidylserine (PS).
Area of Science:
- Biotechnology
- Molecular Biology
- Analytical Chemistry
Background:
- Aptamers offer high specificity for analytical methods but often suffer from structural instability, limiting their affinity and sensitivity.
- Improving aptamer structural integrity is crucial for developing robust and highly sensitive biosensors.
- Current strategies for aptamer optimization require enhancement to meet real-world application demands.
Purpose of the Study:
- To develop a post-selection strategy for engineering robust aptamers with enhanced structural stability and high affinity.
- To demonstrate the efficacy of this strategy in improving aptamers for detecting specific targets, including toxins and biomarkers.
- To validate the performance of an engineered aptasensor for real-world sample analysis.
Main Methods:
- A post-selection strategy involving iterative embedding of key aptamer fragments to form chimeras.
- Fixing chimera termini via hybridization to reduce flexibility and enhance structural stability.
- Engineering and testing aptamers against okadaic acid, dinophysistoxin, and phosphatidylserine (PS).
- Developing and validating a polyA-nanotetrahedron-assisted electrochemical aptasensor for PS detection.
Main Results:
- Engineered aptamers exhibited significantly enhanced affinity, with improvements of 160.5-fold (anti-okadaic acid), 50.36-fold (anti-dinophysistoxin), and 39.28-fold (anti-PS) compared to parent aptamers.
- The engineered anti-PS aptamer demonstrated high sensitivity (limit of detection 1.741 nM), accuracy, and selectivity in a biosensor.
- The developed aptasensor successfully monitored PS in real biosynthesis samples, showcasing practical utility.
Conclusions:
- The proposed post-selection strategy is a facile and efficient method for generating robust aptamers with superior affinity.
- This approach enables the development of highly sensitive and selective aptasensors for diverse real-world applications.
- The engineered aptamers and aptasensor represent a significant advancement in analytical methodologies for toxin and biomarker detection.
