The Analyst · 2017 · 80 citations · 15 references
PharmacyChemical BiologyMolecular BiologyDrug DiscoveryLead OptimizationMolecular OncologyAssay DevelopmentAssay ValidationLigand BindingPolymerase Chain ReactionAptamer Binding AssaysAptamer DiscoveryAptamer EngineeringCell-selexComplexityNucleic Acid Aptamer SelectionSelexSmall-molecule Aptamer Selection
The development of a simple, low-cost, time-saving and universally applicable method to monitor the progression of aptamer selection is particularly challenging. Herein, a combined strategy dependent on quantitative polymerase chain reaction amplification curve (AC) and melting curve analysis (MCA) is developed to monitor the convergence of the aptamer species during selection progress. As a parallel and complementary method to affinity tests and binding analyses, the AC-MCA method can be used to achieve the DNA complexity assay when affinity is undetectable. It is independent of the target properties and SELEX methods. Therefore, it has been demonstrated as a universal monitoring tool in different SELEX methods towards different targets (small molecules, proteins, bacteria and cancer cells). The AC-MCA method will facilitate current new aptamer discovery and aptamer-based wide application.
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