Percorrer por autor "Carneiro, Pedro"
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- Alzheimer’s disease: Development of a sensitive label-free electrochemical immunosensor for detection of amyloid beta peptidePublication . Carneiro, Pedro; Loureiro, Joana; Delerue-Matos, Cristina; Morais, Simone; Pereira, Maria do CarmoIn this work, a highly sensitive label-free immunosensor for detection of the main biomarker of Alzheimer’s disease (AD), amyloid beta 1–42 (Aβ (1–42)), is presented. A gold electrode was modified with a mercaptopropionic acid (MPA) self-assembled monolayer, electrodeposited gold nanoparticles (AuNPs) and a monoclonal antibody mAb DE2B4 to recognize Aβ; all the relevant experimental variables were optimized. Antibodies were functionalized through chemical modification (thiolation) to promote the antibody immobilization on the AuNPs surface with proper orientation which enabled the direct detection of Aβ(1–42). Scanning electron microscopy, square-wave voltammetry and electrochemical impedance spectroscopy were used to characterize the construction of the biosensor. Using the proposed immunosensor, Aβ(1–42) was specifically detected within the linear range of 10–1000 pg mL−1 with a 5.2 pg mL−1 and 17.4 pg mL−1 detection and quantification limit, respectively; recovery values for the tested spiking levels ranged from 90.3 to 93.6%. The immunosensor enables rapid, accurate, precise, reproducible and highly sensitive detection (14.6%reduction mL pg−1) of Aβ with low-cost and opens the possibilities for diagnostic ex vivo applications and research-based in vivo studies.
- Alzheimer’s disease: Development of a sensitive label-freeelectrochemical immunosensor for detection of amyloid beta peptidePublication . Carneiro, Pedro; Loureiro, Joana; Delerue-Matos, Cristina; Morais, Simone; Pereira, Maria do CarmoIn this work, a highly sensitive label-free immunosensor for detection of the main biomarker of Alzheimer’s disease (AD), amyloid beta 1–42 (A (1–42)), is presented. A gold electrode was modified with a mercaptopropionic acid (MPA) self-assembled monolayer, electrodeposited gold nanoparticles (AuNPs) and a monoclonal antibody mAb DE2B4 to recognize A ; all the relevant experimental variables were optimized. Antibodies were functionalized through chemical modification (thiolation) to promote the antibody immobilization on the AuNPs surface with proper orientation which enabled the direct detection of A (1–42). Scanning electron microscopy, square-wave voltammetry and electrochemical impedance spectroscopy were used to characterize the construction of the biosensor. Using the pro-posed immunosensor, A (1–42) was specifically detected within the linear range of 10–1000 pg mL−1 with a 5.2 pg mL−1 and 17.4 pg mL−1 detection and quantification limit, respectively; recovery values for the tested spiking levels ranged from 90.3 to 93.6%. The immunosensor enables rapid, accurate, precise, reproducible and highly sensitive detection (14.6%reduction mL pg−1) of A with low-cost and opens the possibilities for diagnostic ex vivo applications and research-based in vivo studies.
- Nanomaterials towards Biosensing of Alzheimer’s Disease BiomarkersPublication . Carneiro, Pedro; Morais, Simone; Pereira, Maria CarmoAlzheimer's disease (AD) is an incurable and highly debilitating condition characterized by the progressive degeneration and/or death of nerve cells, which leads to manifestation of disabilities in cognitive functioning. In recent years, the development of biosensors for determination of AD's main biomarkers has made remarkable progress, particularly based on the tremendous advances in nanoscience and nanotechnology. The unique and outstanding properties of nanomaterials (such as graphene, carbon nanotubes, gold, silver and magnetic nanoparticles, polymers and quantum dots) have been contributing to enhance the electrochemical and optical behavior of transducers while offering a suitable matrix for the immobilization of biological recognition elements. Therefore, optical and electrochemical immuno- and DNA-biosensors with higher sensitivity, selectivity and longer stability have been reported. Nevertheless, strategies based on the detection of multiple analytes still need to be improved, as they will play a crucial role in minimizing misdiagnosis. This review aims to provide insights into the conjugation of nanomaterials with different transducers highlighting their crucial role in the construction of biosensors for detection of AD main biomarkers.
- Nanostructured label–free electrochemical immunosensor for detection of a Parkinson's disease biomarkerPublication . Carneiro, Pedro; Loureiro, Joana A.; Delerue-Matos, Cristina; Morais, Simone; Pereira, Maria do CarmoAggregation of α-synuclein has been recognized as a critical event in the pathogenesis of Parkinson's disease whose prevalence is increasing with great socio-economic challenges for future generations. Here, we developed a sensitive and specific electrochemical immunosensor for the detection and quantification of this biomarker, based on the voltammetric study of a redox indicator signal, which decreases upon the analyte recognition by the antibody due to the electronic resistance increase. The proposed immunosensor is based on a screen-printed carbon electrode modified in a layer-by-layer approach, which through extensive characterization led to the successful nanostructuration of the transducer, through the drop-cast of 3.0 μL of a 0.1 mg mL−1 single-walled carbon nanotubes suspension followed by electrodeposition of gold nanoparticles in a 3 mM HAuCl4 solution under a −0.2 V potential for 150 s. Monoclonal antibodies were immobilized on the gold nanoparticles surface through chemical modification at an optimal concentration of 200 μg mL−1. Using the proposed immunosensor, α-synuclein was detected in the range of 0.01–10 ng mL−1 with a 4.1 and 12.6 pg mL−1 limits of detection and quantification, respectively. Recovery values of 96.7, 106.2 and 102.9% were attained for the tested concentrations spiked in fetal bovine serum while also presenting excellent specificity and stability throughout one month. The nanostructured immunosensor provided a great interface for electronic transduction and biological recognition events, which enabled fast, sensitive and specific detection of α-synuclein while being based on a simple and inexpensive technology requiring small sample volumes, crucial characteristics for application in point-of-care testing.
- The simpler the better: Highly sensitive 17α-ethinylestradiol sensor based on an unmodified carbon paper transducerPublication . Torrinha, Álvaro; Carneiro, Pedro; Dias, Diana; Delerue-Matos, Cristina; Morais, SimoneThe remarkable features of a carbon fiber paper sensor (CP) were employed for detection of the estrogenic hormone 17α-ethinylestradiol (EE2), considered a contaminant of emerging concern due to its potential ecotoxicity and widespread in the aquatic ecosystems. In this work, an unpreceded CP pre-treatment study was conducted with the (Il)-hexacyanoferrate(III) ion pair, however a bare sensor without pre-treatment revealed higher efficiency on the oxidation of EE2 compared to a chemical and electrochemical pre-treated CP and a gold nanoparticles modified CP, being thus selected for EE2 determinations. The analytical conditions were thoroughly optimized in terms of electrolyte pH (pH 7), differential pulse voltammetry parameters (modulation time 0.003 s, amplitude 0.09 V, interval time 0.1 s and step potential 0.01 V), and analyte preconcentration potential (0.4 V) and time (180 s). The hormone can be determined by the CP in a wide linear range from 0.1 to 1000 nM, achieving a detection limit of 0.14 ± 0.005 nM and an outstanding sensitivity of 1636 ± 232 μA μM−1 cm−2 in the lowest linear zone (0.1–1 nM). The sensor was validated in river water and fish reaching good recoveries (91.2 ± 4.6 to 109.0 ± 7.1%), reproducibility and repeatability. Moreover, the sensor showed high selectivity to EE2 in the presence of several potential interfering compounds and frequently prescribed drugs, though it could not discriminate the similar hormone, 17β-estradiol, being the total concentration obtained in this case. CP-based sensors emerge as efficient electroanalytical tools, suggesting that modification of the surface may not always be beneficial in terms of sensitivity