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A Method for Selecting Structure-switching Aptamers Applied to a Colorimetric Gold Nanoparticle Assay
Published on: February 28, 2015
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Aptamer cleavage optimization, binding mechanism analysis, and dual-chemiluminescence sensor for microcystin
Zhongmei Peng1, Xuning Kang1, Zhenlin Fan1
1Engineering Research Center for Molecular Diagnosis, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, 650500, Yunnan, People's Republic of China.
Talanta
|March 17, 2025
Summary
Researchers developed a novel aptamer (M-Apt-6) for detecting multiple microcystins (MCs) in water. This aptamer enabled a sensitive chemiluminescence sensor for simultaneous MC detection.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Environmental Science
Background:
- Microcystins (MCs) are potent cyanotoxins posing significant risks to aquatic ecosystems and human health.
- Accurate and simultaneous detection of various MC congeners (e.g., MC-LR, MC-RR, MC-YR) is crucial for environmental monitoring and risk assessment.
- Existing detection methods may lack specificity, sensitivity, or the ability to detect multiple MC types concurrently.
Purpose of the Study:
- To develop a high-affinity and highly specific aptamer for simultaneous recognition of multiple microcystin conformations.
- To engineer a miniaturized and optimized aptamer sequence for efficient MC binding.
- To construct a sensitive chemiluminescence sensor for the simultaneous detection of MCs in environmental water samples.
Main Methods:
- Selection of a high-affinity aptamer (MCs-12) using Capture-Systematic Evolution of Ligands by Exponential Enrichment (SELEX) against immobilized microcystins.
- Optimization of the aptamer sequence through stepwise base removal to obtain a shorter, functional aptamer (M-Apt-6).
- Characterization of aptamer-microcystin interactions using circular dichroism, molecular docking, and molecular dynamics simulations.
- Development of a chemiluminescence sensor utilizing M-Apt-6, gold nanoparticles (AuNPs), and luminol-based reactions for MC detection.
Main Results:
- An optimized 27-base aptamer, M-Apt-6, was developed with high affinity and specificity for MC-LR, MC-RR, and MC-YR.
- The M-Apt-6 aptamer demonstrated specific binding to microcystins, forming complexes that induced AuNPs aggregation.
- The developed chemiluminescence sensor exhibited significant linear relationships between signal intensity and MC concentration.
- Detection limits of 0.22 ng/mL (luminol-AgNO3 system) and 0.01 ng/mL (luminol-H2O2 system) were achieved for MCs in Dianchi Lake water.
Conclusions:
- The M-Apt-6 aptamer is a promising candidate for the specific and simultaneous detection of multiple microcystins.
- The developed chemiluminescence sensor offers a sensitive and efficient method for monitoring microcystin contamination in aquatic environments.
- This approach provides a valuable tool for safeguarding water quality and public health from harmful algal blooms.
