This paper reports on a feasibility study of electrochemical in-vitro detection of prostate cancer biomarker PCA3 (prostate cancer antigen 3) in direct assay with specific RNA aptamer labelled with a redox group (ferrocene) and immobilized on a screen-printed gold electrode surface. The cyclic voltammograms and electrochemical impedance spectroscopy methods yield encouraging results on the detection of PCA3 in a range of concentrations from 1 μg/mL down to 0.1 ng/mL in buffer solutions. Both anodic and cathodic current values in cyclic voltammograms measurements and charge transfer resistance values in electrochemical impedance spectroscopy experiments correlate with the PCA3 concentration in the sample. Kinetics studies of the binding of the PCA3 to our aptamer demonstrated high specificity of the reaction with a characteristic affinity constant of approximately 4·10−10 molar. The results of this work provide a background for the future development of novel, highly sensitive and cost-effective diagnostic methodologies for prostate cancer detection.
This work is a continuation of our research into the development of simple, reliable, and cost-effective methods for the early diagnosis of prostate cancer (PCa). The proposed method is based on the electrochemical detection of the PCA3 biomarker of PCa (long non-coded RNA transcript expressed in urine) using a specific aptamer labeled with a redox group (methylene blue). The electrochemical measurements (cyclic voltammograms) obtained from electrodes functionalized with the aptamer were complemented in this work by another biosensing technique: total internal reflection ellipsometry (TIRE). In addition to proving the concept of the detection of PCA3 in low concentrations down to 90 pM, this study improved our understanding of the processes by which PCA3 binds to its specific aptamer. The high specificity of the binding of PCA3 to the aptamer was assessed by studying the binding kinetics, which yielded an affinity constant (KD) of 2.58 × 10−9 M. Additional XPS measurements confirmed the strong covalent binding of aptamers to gold and showed spectral features associated with PCA3 to aptamer binding.
Silica-based coatings prepared using sol-gel polymerizing technology have been shown to exhibit excellent chemical stability combined with reducing the corrosion of metal substrates, showing promising use in aerospace and marine applications to protect light alloys. Moreover, this technology is an eco-friendly technique route for producing surface coatings, showing high potential for replacing toxic pre-treatment coatings of traditional conversation chromate coatings. This study aims to investigate the enhancement in corrosion protection of a hybrid-organic-inorganic silica-based coating cured at 80 °C by increasing the hydrophobicity to work on the aluminium 2024-T3 alloy. This approach involving a novel silica-based hybrid coating was prepared by introducing 1H,1H,2H,2H-perfluorodecyltriethoxysilane (PFDTES) into the base hybrid formula created from tetraethylorthosilicatesilane (TEOS) and triethoxymethylsilane (MTMS) precursors; this formula was enhanced by introducing a Polydimethylsiloxane polymer (PDMS). The corrosion protection properties of these coatings were examined by being immersed in 3.5% NaCl with electrochemical impedance testing (EIS) and Potentiodynamic polarization scanning (PDPS). The chemical elements confirmation was performed using infrared spectroscopy (ATR-FTIR); all this was supported by analysing the surface morphology before and after the immersion by using scanning electron microscopy (SEM). The results of the electrochemical impedance testing analyses reveal the new open finite-length diffusion circuit element due to electrolyte media diffusion prevented by fluorinated groups. Additionally, it shows increases in corrosion protection arising from the increasing hydrophobicity of the fluorinated coating compared to other formulas cured under similar conditions and bare substrate. Additionally, the modified sol-gel exhibited improved resistance to cracking, while the increased hydrophobicity may also promote self-cleaning.
In the quest for the development of accurate, reliable, and cost-effective biosensing technology for early diagnostics of prostate cancer, we describe here an electrochemical biosensor combining a simple transducing method of differential pulse voltammetry (DPV) with an RNA-based aptamer labelled with a methylene blue redox group acting as a highly specific bioreceptor to the prostate cancer biomarker PCA3. A series of DPV measurements on screen-printed gold electrodes is functionalised with a redox-labelled aptamer in solutions (either buffer or synthetic urine) containing PCA3 in a wide range of concentrations from 0.1 picomolar (pM) to 10 nanomolar (nM). In these measurements, the current peak values correlate with the concentration of PCA3 and yield a low detection limit (LDL) of 0.1 pM. Furthermore, the binding kinetics study revealed the high affinity of the aptamer to the target PCA3 with the affinity constants KD of about 3.0 × 10−8 molar. In addition, the AFM study showed the increase in the molecular layer roughness caused by the binding of PCA3, which is a large RNA molecular fragment.
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