Degree: Doctor

This CIQUP member does not yet have any projects linked with him.
Showing 5 latest publications. Total publications: 20
Show all publications
1. A Disposable Saliva Electrochemical MIP-Based Biosensor for Detection of the Stress Biomarker α-Amylase in Point-of-Care Applications, Rebelo, TSCR; Miranda, IM; Brandão, ATSC Sousa, LIG; Ribeiro, JA Silva, AF; Pereira, CM in Electrochem, 2021, Volume: 2, 
Article,  Indexed in: crossref  DOI: 10.3390/electrochem2030028 P-00V-S7N
Abstract <jats:p>The design and synthesis of artificial receptors based on molecular imprinting (MI) technology for the development of a new MIP-based biosensor for detection of the stress biomarker α-amylase in human saliva in point-of-care (PoC) applications is described in this work. The portable electrochemical devices for monitoring α-amylase consists of cost-effective and disposable gold screen-printed electrodes (AuSPEs). To build the electrochemical device, the template biomolecule was firstly immobilized directly over the working area of the gold chip previously activated with a self-assembled monolayer (SAM) of cysteamine (CA). Then, pyrrole (Py) monomer was selected as building block of a polymeric network prepared by CV electropolymerization. After the electropolymerization process, the enzyme was removed from the polymer film in order to build the specific recognition sites for the target enzyme. The MIP biosensor showed a very wide linear concentration range (between 3.0 × 10−4 to 0.60 mg mL−1 in buffer solution and between 3.0 × 10−4 to 3.0 × 10−2 mg mL−1 in human saliva) and low detection levels were achieved (LOD &lt; 3.0 × 10−4 mg mL−1) using square wave voltammetry (SWV) as the electroanalytical technique.</jats:p>

2. Electrochemical immunosensor for detection of CA 15-3 biomarker in point-of-care, Rebelo, TSCR Ribeiro, JA Sales, MGF; Pereira, CM in SENSING AND BIO-SENSING RESEARCH, 2021, ISSN: 2214-1804,  Volume: 33, 
Article,  Indexed in: crossref, wos  DOI: 10.1016/j.sbsr.2021.100445 P-00V-CE0
Abstract This work reports the development of a simple and rapid electrochemical immunosensor for the determination of breast cancer biomarker Cancer Antigen 15-3 (CA 15-3). Disposable and cost-effective chips, consisting of gold screen-printed electrodes (AuSPEs), were used to develop the portable electrochemical devices for monitoring the biomarker in point-of-care (PoC), under clinical context. The biosensor preparation consisted of two simple steps. First, a self-assembled monolayer (SAM) of mercaptosuccinic acid (MSA) was formed at the AuSPE surface. Then, the CA 15-3 antibody was covalently bound to the carboxylic groups standing at the electrode surface using EDC/NHS chemistry. The performance of the developed immunosensor was evaluated by assessing the sensor sensitivity, linear response interval, selectivity and detection limit (LOD). The developed immunosensor provided a wide linear concentration range (from 1.0 to 1000 U mL(-1)) and low detection levels were achieved (LOD of 0.95 U mL(-1)), enabling the sensitive detection of the cancer biomarker at clinically relevant levels, using square wave voltammetry (SWV) as electroanalytical technique. Moreover, selectivity studies performed against other cancer biomarkers (CA 125 and CA 19-9) revealed that the antibody has high selectivity for CA 15-3 antigen. The immunosensor was applied to the quantification of CA 15-3 in artificial serum samples with satisfactory results.

3. Electrochemistry-Assisted Surface Plasmon Resonance Biosensor for Detection of CA 15-3, Ribeiro, JA Sales, MGF; Pereira, CM in ANALYTICAL CHEMISTRY, 2021, ISSN: 0003-2700,  Volume: 93, 
Article,  Indexed in: crossref, wos  DOI: 10.1021/acs.analchem.0c05367 P-00V-WY0
Abstract In this work, we describe an innovative methodology based on combined surface plasmon resonance (SPR) and electrochemical responses (eSPR) in the same immunoassay for screening CA 15-3 cancer biomarker with high sensitivity (and selectivity), in a very simple, label-free, accurate, and fully automated manner. Detection was achieved by performing two simple steps. In the first step, direct SPR was used to monitor CA 15-3 interaction with surface immobilized antibody. Two linear response ranges were obtained and the detection limit achieved is poor (LOD of 21 U mL(-1)). However, in the second detection step, electrochemical measurements at the SPR gold surface were performed to measure the decrease of redox probe peak current upon antigen-antibody interaction, providing a suitable amplification strategy to lower detection levels of CA 15-3 (LOD of 0.0998 U mL(-1)), without the need of additional complex and/or expensive amplification steps to enhance the sensitivity. Moreover, selectivity studies were performed against other common cancer biomarkers and the results showed that the eSPR immunosensor is selective for the CA 15-3 protein. Finally, the clinical applicability of the developed eSPR biosensing methodology was successfully applied to detect CA 15-3 in human serum samples at clinically relevant levels due to the high sensitivity of electrochemical readout. The same concept may be further extended to other proteins of interest.

4. Electrochemistry-assisted surface plasmon resonance detection of miRNA-145 at femtomolar level, Ribeiro, JA Sales, MGF; Pereira, CM in SENSORS AND ACTUATORS B-CHEMICAL, 2020, ISSN: 0925-4005,  Volume: 316, 
Article,  Indexed in: crossref, scopus, wos  DOI: 10.1016/j.snb.2020.128129 P-00S-1H6
Abstract In this work, we combined electrochemical techniques with SPR (eSPR) for the label-free detection of cancer biomarker miRNA-145. Detection was performed in a simple two-step assay. In the first step, the gold sensor surface, previously functionalized with a self-assembled monolayer (SAM) of thiolated RNA probes is incubated with the sample containing the target RNA biomarker. In this step, hybridization of RNA fragments with complementary immobilized probes was monitored in real-time by SPR. In the second step, eSPR measurements were performed to improve the sensitivity of the hybridization assay. Potential-induced deposition of a redox probe at the sensor surface resulted in enhanced SPR response promoted by the electrochemical process, thereby allowing the detection of miRNA-145 at femtomolar level (LOD = 0.56 fM), without sample derivatization or post-hybridization treatment for signal amplification. Good linearity was achieved (R-2 = 0.984) over the concentration range from 1.0 fM and 10 nM. Furthermore, the developed eSPR biosensor showed high selectivity towards single-base and two-base mismatch sequences and detection of target miRNA-145 in synthetic human serum was successful achieved.

5. Electrochemical Characterization of Redox Probes at Gold Screen-Printed Electrodes: Efforts towards Signal Stability, Ribeiro, JA Silva, E; Moreira, PS; Pereira, CM in CHEMISTRYSELECT, 2020, ISSN: 2365-6549,  Volume: 5, 
Article,  Indexed in: crossref, scopus, wos  DOI: 10.1002/slct.202001411 P-00S-3D6
Abstract In this work, three universally used redox probes in amperometric biosensing devices, [Fe(CN)(6)](3-)/[Fe(CN)(6)](4-), Ru[(NH3)(6)](3+), and ferrocenedimethanol (FDM), were selected to evaluate the stability of electrochemical signals provide by the reporting systems. Studies were carried out at disposable gold screen-printed electrode (AuSPE) biosensing platforms, commonly used for screening chemical and biological relevant biomolecules. Firstly, electrochemical combined-surface plasmon resonance (eSPR) studies were performed to evaluated adsorption reversibility and/or formation of redox probe complexes at the bare gold surface when routinely used electrochemical techniques, namely cyclic voltammetry (CV) and square-wave voltammetry (SWV), are recorded. Then, the results obtained were compared with those obtained at the AuSPE under the same electrochemical conditions. Based on our findings, best experimental conditions, including the type of electrochemical technique used, are speculated for each reporting system in order to improve the analytical signal stability. Finally, a methodology based on SWV technique was applied to modified electrodes to provide a simple and easy tool to ensure diffusion controlled permeability of probes thorough the films to electrode surface.