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Selection of DNA aptamer against prostate specific antigen using a genetic algorithm and application to sensing
Nasa Savory1, Koichi Abe, Koji Sode
1Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan.
Biosensors & Bioelectronics
|August 10, 2010
Summary
Researchers improved DNA aptamers for prostate specific antigen (PSA) sensing using a genetic algorithm (GA) after SELEX. This new method yielded a highly effective aptamer for detecting PSA, significantly enhancing biosensor capabilities.
Area of Science:
- Biotechnology
- Molecular Biology
- Biosensor Development
Background:
- Aptasensors require high-affinity aptamers for target molecule detection.
- Systematic evolution of ligands by exponential enrichment (SELEX) can yield aptamers but sometimes lacks sufficient affinity.
- Improvements to aptamer selection methods are needed for enhanced biosensor performance.
Purpose of the Study:
- To develop an improved method for selecting high-affinity DNA aptamers against prostate specific antigen (PSA).
- To apply a genetic algorithm (GA) for in silico maturation of aptamers post-SELEX.
- To demonstrate the application of the optimized aptamer in a PSA biosensor.
Main Methods:
- Pre-selection of DNA aptamers against PSA using three rounds of SELEX.
- In silico maturation of pre-selected aptamers using a genetic algorithm (GA) involving replication, crossover, and mutation.
- In vitro synthesis and assay of oligonucleotide sequences to rank PSA-binding ability.
- Demonstration of PSA sensing using the optimized aptamer.
Main Results:
- The genetic algorithm (GA) post-SELEX screening significantly enhanced PSA-binding ability by 48-fold compared to SELEX-derived candidates.
- An aptamer with a dissociation constant (K(D)) in the tens of nanomolar range was identified.
- Successful sensing of PSA concentrations between 40 and 100 nM was achieved.
Conclusions:
- The combination of SELEX and GA provides an effective strategy for developing high-affinity aptamers.
- This study reports the first DNA aptamer against PSA and its successful application in biosensing.
- The developed aptasensor shows promise for sensitive PSA detection.
