Person: Pingarrón Carrazón, José Manuel
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First Name
José Manuel
Last Name
Pingarrón Carrazón
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Ciencias Químicas
Department
Area
Química Analítica
Identifiers
17 results
Search Results
Now showing 1 - 10 of 17
Item Development of an Electrochemical CCL5 Chemokine Immunoplatform for Rapid Diagnosis of Multiple Sclerosis(Biosensors, 2022) Guerrero Irigoyen, Sara; Sánchez Tirado, Esther; Agüí Chicharro, María Lourdes; González Cortés, Araceli; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José ManuelSerum level of CCL5 chemokine is considered an emerging biomarker for multiple sclerosis (MS). Due to the lack of specific assays for this disease, the development of a point-of-care test for rapid detection of MS could lead to avoiding diagnostics delays. In this paper, we report the first electrochemical immunoplatform for quantification of the CCL5 biomarker at the clinically required levels, able to discriminate between patients diagnosed with MS and healthy individuals. The immunosensing device involves protein capture from biological samples by complexation with biotinylated specific antibodies immobilized onto neutravidin-functionalized microparticles and sandwich assay with anti-CCL5 antibody and IgG labelled with horseradish peroxidase (HRP) for the enzyme-catalyzed amperometric detection of H2O2 using hydroquinone (HQ) as the redox mediator. The method shows excellent analytical performance for clinical application with a wide linear range of concentrations (0.1–300 ng·mL−1 CCL5, R2 = 0.998) and a low detection limit (40 pg·mL−1 CCL5). The biosensing platform was applied to the determination of the CCL5 endogenous content in 100-fold diluted sera both from healthy individuals and patients diagnosed with MS, with no further sample treatment in just two hours. The results were successfully compared with those obtained by the ELISA methodology.Item Electrochemical (Bio)Sensing Devices for Human-Microbiome-Related Biomarkers(Sensors, 2023) Sánchez Tirado, Esther; Agüí Chicharro, María Lourdes; González Cortés, Araceli; Campuzano Ruiz, Susana; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José ManuelThe study of the human microbiome is a multidisciplinary area ranging from the field of technology to that of personalized medicine. The possibility of using microbiota biomarkers to improve the diagnosis and monitoring of diseases (e.g., cancer), health conditions (e.g., obesity) or relevant processes (e.g., aging) has raised great expectations, also in the field of bioelectroanalytical chemistry. The well-known advantages of electrochemical biosensors—high sensitivity, fast response, and the possibility of miniaturization, together with the potential for new nanomaterials to improve their design and performance—position them as unique tools to provide a better understanding of the entities of the human microbiome and raise the prospect of huge and important developments in the coming years. This review article compiles recent applications of electrochemical (bio)sensors for monitoring microbial metabolites and disease biomarkers related to different types of human microbiome, with a special focus on the gastrointestinal microbiome. Examples of electrochemical devices applied to real samples are critically discussed, as well as challenges to be faced and where future developments are expected to go..Item Anti-double stranded DNA antibodies: Electrochemical isotyping in autoimmune and neurological diseases(Analytica Chimica Acta, 2023) Arévalo Pérez, Beatriz; Serafín González-Carrato, Verónica; Garranzo-Asensio, María; Montero-Calle, Ana; Barderas, Rodrigo; Yáñez-Sedeño Orive, Paloma; Campuzano Ruiz, Susana; Pingarrón Carrazón, José ManuelThis work reports the first amperometric biosensor for the simultaneous determination of the single or total content of the most relevant human immunoglobulin isotypes (hIgs) of anti-dsDNA antibodies, dsDNA-hIgG, dsDNA-hIgM, dsDNA-hIgA and dsDNA-three hIgs, which are considered relevant biomarkers in prevalent autoimmune diseases such as systemic lupus erythematosus (SLE) as well as of interest in neurodegenerative diseases such as Alzheimer’s disease (AD). The bioplatform involves the use of neutravidin-functionalized magnetic microparticles (NA-MBs) modified with a laboratory-prepared biotinylated human double-stranded DNA (b-dsDNA) for the efficient capture of specific autoantibodies that are enzymatically labeled with horseradish peroxidase (HRP) enzyme using specific secondary antibodies for each isotype or a mixture of secondary antibodies for the total content of the three isotypes. Transduction was performed by amperometry (− 0.20 V vs. the Ag pseudo-reference electrode) using the H2O2/hydroquinone (HQ) system after trapping the resulting magnetic bioconjugates on each of the four working electrodes of a disposable quadruple transduction platform (SP4CEs). The bioplatform demonstrated attractive operational characteristics for clinical application and was employed to determine the individual or total hIgs classes in serum from healthy individuals and from patients diagnosed with SLE and AD. The target concentrations in AD patients are provided for the first time in this work. In addition, the results for SLE patients and control individuals agree with those obtained by applying ELISA tests as well as with the clinical ranges reported by other authors, using individual detection methodologies restricted to centralized settings or clinical laboratories.Item Development of an Electrochemical CCL5 Chemokine Immunoplatform for Rapid Diagnosis of Multiple Sclerosis(Biosensors, 2022) Pingarrón Carrazón, José Manuel; Yáñez-Sedeño Orive, Paloma; Guerrero Irigoyen, Sara; González Cortés, Araceli; Agüí Chicharro, María Lourdes; Sánchez Tirado, EstherSerum level of CCL5 chemokine is considered an emerging biomarker for multiple sclerosis (MS). Due to the lack of specific assays for this disease, the development of a point-of-care test for rapid detection of MS could lead to avoiding diagnostics delays. In this paper, we report the first electrochemical immunoplatform for quantification of the CCL5 biomarker at the clinically required levels, able to discriminate between patients diagnosed with MS and healthy individuals. The immunosensing device involves protein capture from biological samples by complexation with biotinylated specific antibodies immobilized onto neutravidin-functionalized microparticles and sandwich assay with anti-CCL5 antibody and IgG labelled with horseradish peroxidase (HRP) for the enzyme-catalyzed amperometric detection of H2O2 using hydroquinone (HQ) as the redox mediator. The method shows excellent analytical performance for clinical application with a wide linear range of concentrations (0.1–300 ng·mL−1 CCL5, R2 = 0.998) and a low detection limit (40 pg·mL−1 CCL5). The biosensing platform was applied to the determination of the CCL5 endogenous content in 100-fold diluted sera both from healthy individuals and patients diagnosed with MS, with no further sample treatment in just two hours. The results were successfully compared with those obtained by the ELISA methodology.Item An electrochemical immunosensor for adiponectin using reduced graphene oxide–carboxymethylcellulose hybrid as electrode scaffold(Sensors and Actuators B: Chemical, 2015) Arenas, C.B.; Sánchez Tirado, Esther; Ojeda, I.; Gómez-Suárez, C.A.; González Cortés, Araceli; Villalonga Santana, Reynaldo; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José ManuelReduced graphene oxide–carboxymethylcellulose hybrid (CMC–rGO) was used for the development of a novel electrochemical immunosensor for the determination of adiponectin (APN) cytokine. The hybrid material was synthesized by covalent binding of oxidized CMC to GO layers followed by chemical reduction with sodium borohydride. A sandwich-type immunoassay was employed involving the commercial metal-complexes based polymer Mix & Go™ for the stable and oriented immobilization of anti-APN capture antibody. Biotinylated-anti-APN and HRP-Strept were used for the assay configuration. The APN quantification was performed by amperometry at −200 mV using the hydrogen peroxide/hydroquinone enzyme substrate/mediator system. A sigmoidal calibration plot for APN in the 0.1–50 μg/mL range, with a linear portion in the 0.5–10.0 μg/mL APN concentration range was obtained. The calculated limit of detection (3sb/m) was 61 ng/mL APN. The usefulness of the immunosensor was evaluated by analyzing human serum from hypercholesterolemia or diabetes patientsItem Amperometric immunosensor for the determination of ceruloplasmin in human serum and urine based on covalent binding to carbon nanotubes-modified screen-printed electrodes(Talanta, 2013) Garcinuño, Belit; Ojeda Fernández, Irene; Moreno Guzmán, María; González Cortés, Araceli; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José ManuelA novel electrochemical immunosensor for the determination of ceruloplasmin (Cp) in human serum and urine is reported. The immunosensor configuration involves an indirect competitive immunoassay implying covalent immobilization of Cp on activated carboxylic groups at carbon nanotubes-modified screen-printed electrodes (CNTs/SPE). After Cp immobilization and reaction between the target analyte and anti-ceruloplasmin antibodies in solution, the remaining non-conjugated antibody is attached on the Cp-CNTs modified electrode. Monitoring of Cp is performed by means of a secondary antibody labeled with peroxidase (HRP-anti-IgG) and measurement of the amperometric current resulting from the addition of hydrogen peroxide in the presence of hydroquinone as the redox mediator. The experimental variables affecting the analytical performance of the immunosensor were optimized. Calibration curves for Cp provided a linear range between 0.07 and 250 μg/mL (r=0.997). The limit of detection achieved was 21 ng/mL. These analytical characteristics allow the immunosensor to be successfully used for the determination of Cp in spiked human serum and urine at various concentration levelsItem Carbon Nanohorns as a Scaffold for the Construction of Disposable Electrochemical Immunosensing Platforms. Application to the Determination of Fibrinogen in Human Plasma and Urine(Analytical Chemistry, 2014) Ojeda Fernández, Irene; Garcinuño, Belit; Moreno Guzmán, María; González Cortés, Araceli; Yudasaka, Masako; Iijima, Sumio; Langa, Fernando; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José ManuelWe describe in this work a novel electrochemical immunosensor design making use of carbon nanohorns (CNHs) as a scaffold for the preparation of disposable immunosensing platforms for the determination of fibrinogen (Fib). The approach involved the immobilization of Fib onto activated CNHs deposited on screen-printed carbon electrodes (SPCEs) and the implementation of an indirect competitive assay using anti-Fib labeled with horseradish peroxidase (HRP) and hydroquinone (HQ) as the redox mediator. Both CNHs and the Fib-CNHs covalent assembly were characterized by microscopic and electrochemical techniques. The different variables affecting the analytical performance of the amperometric immunosensing strategy were optimized. The calibration plot for Fib allowed a range of linearity between 0.1 and 100 μg/mL (r 2 = 0.994) and a detection limit of 58 ng/ mL to be achieved. The Fib-CNHs/SPCEs exhibited an excellent storage stability of at least 42 days. The developed immunosensor provides, in general, an analytical performance better than that reported for other Fib immunosensors and commercial ELISA kits. This simple and relatively low cost immunosensor configuration permitted the sensitive and selective determination of Fib in human plasma and urineItem Grafted-double walled carbon nanotubes as electrochemical platforms for immobilization of antibodies using a metallic-complex chelating polymer: Application to the determination of adiponectin cytokine in serum(Biosensors and Bioelectronics, 2015) Ojeda Fernández, Irene; Barrejón, Myriam; Arellano, Luis ; González Cortés, Araceli; Yáñez-Sedeño Orive, Paloma; Langa, Fernando; Pingarrón Carrazón, José ManuelAn electrochemical immunosensor for adiponectin (APN) using screen printed carbon electrodes (SPCEs) modified with functionalized double-walled carbon nanotubes (DWCNTs) as platforms for immobilization of the specific antibodies is reported. DWCNTs were functionalized by treatment with 4-aminobenzoic acid (HOOC-Phe) in the presence of isoamylnitrite resulting in the formation of 4-carboxyphenyl-DWCNTs. The oriented binding of specific antibodies toward adiponectin was accomplished by using the metallic-complex chelating polymer Mix&Go™. The HOOC-Phe-DWCNTs-modified SPCEs were characterized by cyclic voltammetry and compared with HOOC-Phe-SWCNTs/SPCE. The different variables affecting the performance of the developed immunosensor were optimized. Under the selected conditions, a calibration plot for APN was constructed showing a range of linearity extending between 0.05 and 10.0 μg/mL which is adequate for the determination of the cytokine in real samples. A detection limit of 14.5 ng/mL was achieved. The so prepared immunosensor exhibited a good reproducibility for the APN measurements, excellent storage stability and selectivity, and a much shorter assay time than the available ELISA kits. The usefulness of the immunosensor for the analysis of real samples was demonstrated by analyzing human serum from female or male healthy patientsItem Electrochemical biosensor for the simultaneous determination of rheumatoid factor and anti-cyclic citrullinated peptide antibodies in human serum(Analyst, 2020) Guerrero Irigoyen, Sara; Sánchez Tirado, Esther; Martínez-García, Gonzalo; González Cortés, Araceli; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José ManuelThis paper reports a dual electrochemical biosensor involving carboxylated- or neutravidin-functionalized magnetic microbeads and dual screen-printed carbon electrodes for the simultaneous determination of rheumatoid factor (RF) and anti-cyclic citrullinated peptide (CCPA) autoantibodies used as biomarkers for the detection of rheumatoid arthritis autoimmune disease. Sandwich-type biosensors involving Fc fragments of IgG Fc(IgG) and biotinylated cyclic cytrullinated peptide (CCP-biotin) to form CCP-biotin-Neutr-MBs for the specific immobilization of RF and CCPA, respectively, as well as conjugation with HRP-IgM and HRP-IgG for RF and CCPA, respectively, were prepared. Amperometric detection was performed at −0.20 V vs. Ag pseudo-reference electrode using the H2O2/hydroquinone (HQ) system upon capturing the bioconjugates onto the corresponding working electrode (WE1 or WE2) of SPCdEs. The dual biosensor exhibits high sensitivity for RF and CCPA with LOD values of 0.8 and 2.5 IU mL−1, respectively. The simultaneous determination can be completed in about two hours using a simple protocol and a sample volume (25 μL) four times smaller than that required by the ELISA method. The dual electrochemical biosensor was used for the determination of both target biomarkers in human serum.Item Electrochemical immunosensor for rapid and sensitive determination of estradiol(Analytica Chimica Acta, 2012) Ojeda Fernández, Irene; Lopez-Montero, Judith; Moreno Guzmán, María; Janegitz, Bruno; González Cortés, Araceli; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José ManuelThis work describes the preparation of an electrochemical immunosensor for estradiol based on the surface modification of a screen printed carbon electrode with grafted p-aminobenzoic acid followed by covalent binding of streptavidin (Strept) and immobilization of biotinylated anti-estradiol (antiestradiol-Biotin). The hormone determination was performed by applying a competitive immunoassay with peroxidase-labelled estradiol (HRP–estradiol) and measurement of the amperometric response at −200 mV using hydroquinone (HQ) as redox mediator. The calibration curve for estradiol exhibited a linear range between 1 and 250 pg mL−1 (r = 0.990) and a detection limit of 0.77 pg mL−1 was achieved. Cross-reactivity studies with other hormones related with estradiol at physiological concentration levels revealed the practical specificity of the developed method for estradiol. A good reproducibility, with RSD = 5.9% (n = 8) was also observed. The operating stability of a single bioelectrode modified with anti-estradiol-Biotin-Strept was nine days when it was stored at 8 ◦C under humid conditions between measurements. The developed immunosensor was applied to the analysis of certified serum and spiked urine samples with good results