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Rational Truncation of Aptamer for Ultrasensitive Aptasensing of Chloramphenicol: Studies Using Bio-Layer
Richa Sharma1,2, Monali Mukherjee2,3, Praveena Bhatt2,3
1Department of Food Engineering, CSIR-Central Food Technological Research Institute (CFTRI), Mysore 570020, India.
Biosensors
|June 27, 2023
Summary
Researchers optimized a chloramphenicol aptamer for enhanced detection. Truncating the aptamer significantly improved its binding affinity and sensitivity for detecting chloramphenicol in honey samples.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Molecular Biology
Background:
- Aptamers offer selective small molecule detection.
- Previous chloramphenicol aptamers had low affinity due to steric hindrance, limiting assay sensitivity.
Purpose of the Study:
- To improve chloramphenicol aptamer binding affinity and sensitivity.
- To achieve stable, high-affinity aptamers through sequence truncation.
Main Methods:
- Systematic truncation of an 80-nucleotide chloramphenicol aptamer.
- Computational thermodynamic analysis of modified aptamer stability and folding.
- Bio-layer interferometry to evaluate binding affinities.
- Gold nanosphere aggregation assay for chloramphenicol detection in honey.
Main Results:
- Truncating 30 bases from the 3' end reduced the dissociation constant by 86.93%.
- The modified aptamer demonstrated a 32.87-fold lower detection limit (1.673 pg mL⁻¹).
- The optimized aptamer enabled sensitive chloramphenicol detection in honey via a colorimetric assay.
Conclusions:
- Aptamer truncation is an effective strategy to enhance binding affinity and analytical sensitivity.
- The modified aptamer is suitable for ultrasensitive chloramphenicol detection in real-world samples like honey.
- This work provides a foundation for developing improved aptamer-based sensors.
