Optimization of Apta-Sensing Platform for Detection of Prostate Cancer Marker PCA3

• Published in: International journal of molecular sciences 2021-11, Vol.22 (23), p.12701
• Main Authors: Takita, Sarra, Nabok, Alexei, Lishchuk, Anna, Smith, David
• Format: Article
• Language: English
• Subjects: Antigens, Neoplasm - metabolism; Aptamers; Aptamers, Nucleotide - metabolism; Binding; Binding sites; Biomarkers; Biopsy; Biosensing Techniques - methods; Biosensors; cyclic voltammograms; Electrochemical analysis; Electrochemistry - methods; Electrodes; Ellipsometry; Gold; Humans; Low concentrations; Male; Methylene blue; PCA3; Prostate cancer; Prostatic Neoplasms - diagnosis; Prostatic Neoplasms - metabolism; spectroscopic ellipsometry; Spectrum analysis; Transcription; Urine; XPS
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms222312701

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 (K ) of 2.58 × 10 M. Additional XPS measurements confirmed the strong covalent binding of aptamers to gold and showed spectral features associated with PCA3 to aptamer binding.