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Aptamer-Based Target Detection Facilitated by a 3-Stage G-Quadruplex Isothermal Exponential Amplification Reaction

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Thrombin-Binding Aptamer Quadruplex Formation: AFM and Voltammetric Characterization.

Victor Constantin Diculescu¹, Ana-Maria Chiorcea-Paquim, Ramon Eritja

¹Departamento de Quimica, Faculdade de Ciências e Tecnologia, Universidade de Coimbra, 3004-535 Coimbra, Portugal.

Journal of Nucleic Acids

|August 28, 2010

Summary

Thrombin-binding aptamers (TBAs) exhibit distinct adsorption and redox behaviors influenced by K+ ions. Guanine oxidation is altered when TBAs form quadruplex structures, impacting electron transfer.

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Area of Science:

Biophysical Chemistry
Nanotechnology
Electrochemistry

Background:

Thrombin-binding aptamers (TBAs) are short DNA sequences with potential therapeutic applications.
Understanding aptamer adsorption and redox behavior is crucial for biosensor development.

Purpose of the Study:

To investigate the adsorption and redox properties of thrombin-binding aptamer (TBA) and extended TBA (eTBA).
To correlate these properties with sequence composition, K+ ion presence, and molecular structure.

Main Methods:

Atomic force microscopy (AFM) for visualizing adsorption patterns.
Voltammetry to study electrochemical redox behavior.
Experiments conducted on highly oriented pyrolytic graphite (HOPG) and glassy carbon electrodes.

Main Results:

Adsorption and surface coverage vary with sequence base composition and K+ presence.
In the presence of K+, most TBA/eTBA molecules form quadruplexes, reducing adsorption.
Voltammetry revealed guanine oxidation, with K+ addition shifting peaks due to G-quartet formation.

Conclusions:

K+ ions induce quadruplex formation in TBA/eTBA, significantly altering their adsorption and electrochemical behavior.
The electron transfer from G-quartets is less facile than from single strands due to structural constraints.