Person:
Serafín González-Carrato, Verónica

First Name

Verónica

Last Name

Serafín González-Carrato

URI

https://hdl.handle.net/20.500.14352/76204

Affiliation

Universidad Complutense de Madrid

Faculty / Institute

Ciencias Químicas

Department

Química Analítica

Area

Química Analítica

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Now showing 1 - 10 of 23

Angiogenesis inhibitor or aggressiveness marker? The function of endostatin in cancer through electrochemical biosensing
(Bioelectrochemistry, 2024) Tejerina Miranda, Sandra; Pedrero Muñoz, María; Blázquez García, Marina; Serafín González-Carrato, Verónica; Montero Calle, Ana; Garranzo Asensio, María; Reviejo García, Ángel Julio; Pingarrón Carrazón, José Manuel; Barderas Manchado, Rodrigo; Campuzano Ruiz, Susana
This work reports the first electrochemical bioplatform developed for the determination of human endostatin (HE), a biomarker with recognized antiangiogenic potential whose elevated circulating levels have also been associated with the development of aggressive cancers. The developed electroanalytical biotool combines the benefits of using magnetic microparticles for the implementation of sandwich immunoassays and amperometric transduction on disposable carbon electrodes. A limit of detection (LOD) of 34.1 pg mL−1 for HE standards and a selectivity suitable for its foray into the clinical oncology area, are demonstrated. The determination of HE in clinical samples such as lysates and secretomes of colorectal cancer (CRC) cells, plasma, and tissue samples from patients with CRC in different stages, has been faced with satisfactory results showing the ability for discriminating the metastatic capabilities of cells and for identifying and staging CRC patients. The developed bioplatform allows precise quantitative determinations, requiring minimal pre-treatments and sample amounts in only 75 min. In addition, due to the instrumentation and the type of substrates used in the detection step, the biotool is compatible with implementation in multiplexed and/or point-of-need devices, features in which this bioplatform is advantageous with respect to the enzyme linked immunosorbent assay (ELISA) or immunoblotting technologies.
Beyond Sensitive and Selective Electrochemical Biosensors: Towards Continuous, Real-Time, Antibiofouling and Calibration-Free Devices
(Sensors, 2020) Campuzano Ruiz, Susana; Pedrero Muñoz, María; Gamella Carballo, Maria; Serafín González-Carrato, Verónica; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José Manuel
Nowadays, electrochemical biosensors are reliable analytical tools to determine a broad range of molecular analytes because of their simplicity, affordable cost, and compatibility with multiplexed and point-of-care strategies. There is an increasing demand to improve their sensitivity and selectivity, but also to provide electrochemical biosensors with important attributes such as near real-time and continuous monitoring in complex or denaturing media, or in vivo with minimal intervention to make them even more attractive and suitable for getting into the real world. Modification of biosensors surfaces with antibiofouling reagents, smart coupling with nanomaterials, and the advances experienced by folded-based biosensors have endowed bioelectroanalytical platforms with one or more of such attributes. With this background in mind, this review aims to give an updated and general overview of these technologies as well as to discuss the remarkable achievements arising from the development of electrochemical biosensors free of reagents, washing, or calibration steps, and/or with antifouling properties and the ability to perform continuous, real-time, and even in vivo operation in nearly autonomous way. The challenges to be faced and the next features that these devices may offer to continue impacting in fields closely related with essential aspects of people’s safety and health are also commented upon.
Simultaneous amperometric immunosensing of the metastasis-related biomarkers IL-13Rα2 and CDH-17 by using grafted screen-printed electrodes and a composite prepared from quantum dots and carbon nanotubes for signal amplification
(Microchimica Acta, 2019) Serafín González-Carrato, Verónica; Valverde De La Fuente, Alejandro; Garranzo-Asensio, María; Barderas, Rodrigo; Campuzano Ruiz, Susana; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José Manuel
This paper describes a dual electrochemical immunoassay for the simultaneous determination of IL-13Rα2 and CDH-17, two biomarkers of emerging relevance in metastatic processes. The sandwich assay uses a screen-printed dual carbon electrode that was electrochemically grafted with p-aminobenzoic acid to allow the covalent immobilization of capture antibodies. A hybrid composed of graphene quantum dots (GQDs) and multiwalled carbon nanotubes (MWCNTs) act as nanocarriers for the detection antibodies and horseradish peroxidase. The use of this hybrid material considerably improves the assay (in comparison to the use of MWCNTs) due to the peroxidase mimicking activity of the GQDs. The method works at a low working potential (0.20 V vs. Ag pseudo-reference electrode) and thus is not readily interfered by unknown electroactive species. The dual immunoassay allows for the selective determination of both biomarkers with LOD values of 1.4 (IL-13sRα2) and 0.03 ng mL-1 (CDH-17). The simultaneous determination of IL-13Rα2 and CDH-17 was accomplished in lysates from breast and colorectal cancer cells with different metastatic potential, and in paraffin-embedded tumor tissues extracts from patients diagnosed with colorectal cancer at different stages. The applicability to discriminate the metastatic potential even in intact cells through the detection of both extracellular receptors has been demonstrated also. The assay can be performed within 3 h, requires small sample amounts (0.5 μg), and has a simple protocol. Graphical abstract Dual amperometric immunosensing of the metastasis-related biomarkers IL-13Rα2 and CDH-17 in human colorectal cancer cells and tissues by using grafted screen-printed electrodes and composites of quantum dots and carbon nanotubes as nanocarriers.
Biosensing and Delivery of Nucleic Acids Involving Selected Well-Known and Rising Star Functional Nanomaterials
(Nanomaterials, 2019) Campuzano Ruiz, Susana; Gamella Carballo, Maria; Serafín González-Carrato, Verónica; Pedrero Muñoz, María; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José Manuel
In the last fifteen years, the nucleic acid biosensors and delivery area has seen a breakthrough due to the interrelation between the recognition of nucleic acid’s high specificity, the great sensitivity of electrochemical and optical transduction and the unprecedented opportunities imparted by nanotechnology. Advances in this area have demonstrated that the assembly of nanoscaled materials allows the performance enhancement, particularly in terms of sensitivity and response time, of functional nucleic acids’ biosensing and delivery to a level suitable for the construction of point-of-care diagnostic tools. Consequently, this has propelled detection methods using nanomaterials to the vanguard of the biosensing and delivery research fields. This review overviews the striking advancement in functional nanomaterials’ assisted biosensing and delivery of nucleic acids. We highlight the advantages demonstrated by selected well-known and rising star functional nanomaterials (metallic, magnetic and Janus nanomaterials) focusing on the literature produced in the past five years.
Electrochemical biosensor for creatinine based on the immobilization of creatininase, creatinase and sarcosine oxidase onto a ferrocene/horseradish peroxidase/gold nanoparticles/multi-walled carbon nanotubes/Teflon composite electrode
(Electrochimica Acta, 2013) Serafín González-Carrato, Verónica; Hernández, Paloma; Agüí Chicharro, María Lourdes; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José Manuel
A composite electrode consisted of gold nanoparticles (AuNPs), multi-walled carbon nanotubes (MWCNTs) and Teflon, to which peroxidase (HRP) and ferrocene (Fc) were incorporated as auxiliary enzyme and redox mediator, respectively, was constructed. The enzymes creatininase, creatinase and sarcosine oxidase were then co-immobilized onto the surface of the resulting HRP/Fc/AuNPs/MWCNTs/Teflon electrode for the preparation of a creatinine biosensor. Amperometry in stirred solutions using a detection potential of 0.0 V vs Ag/AgCl allowed a linear calibration plot to be obtained in the 0.003–1.0 mM creatinine concentration range with a limit of detection of 0.1 M (S/N = 3). The apparent Michaelis-Menten constant for creatininase was KMap= 4.1 ± 0.4 mM. The developed biosensor was validated with good results by determining creatinine in human serum and correlating with the spectrophotometric Jaffe’s method.
Multiplexed determination of human growth hormone and prolactin at a label free electrochemical immunosensor using dual carbon nanotube–screen printed electrodes modified with gold and PEDOT nanoparticles
(Analyst, 2014) Serafín González-Carrato, Verónica; Martínez-García, Gonzalo; Agüí Chicharro, María Lourdes; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José Manuel
A label-free dual electrochemical immunosensor was constructed for the multiplexed determination of human growth (hGH) and prolactin (PRL) hormones. The immunosensor used an electrochemical platform composed of carbon nanotube–screen printed carbon electrodes (CNT/SPCEs) modified with poly(ethylene-dioxythiophene) (PEDOT) and gold nanoparticles, on which the corresponding hGH and PRL antibodies were immobilized. The affinity reactions were monitored by measuring the decrease in the differential pulse voltammetric oxidation response of the redox probe dopamine. The experimental variables involved in the preparation of both AuNP/PEDOT/CNT/SPC modified electrodes and the dual immunosensor were optimized. The immunosensor exhibited an improved analytical performance for hGH and PRL with respect to other electrochemical immunosensor designs, showing wide ranges of linearity and low detection limits of 4.4 and 0.22 pg mL−1, respectively. An excellent selectivity against other hormones and in the presence of ascorbic and uric acids was found. The usefulness of the dual immunosensor for the simultaneous analysis of hGH and PRL was demonstrated by analyzing human serum and saliva samples spiked with the hormones at different concentration levels.
Comparative evaluation of the performance of electrochemical immunosensors using magnetic microparticles and nanoparticles. Application to the determination of tyrosine kinase receptor AXL
(Microchimica Acta, 2017) Serafín González-Carrato, Verónica; Torrente Rodríguez, Rebeca Magnolia; Batlle, Montserrat; García de Frutos, Pablo; Campuzano Ruiz, Susana; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José Manuel
Electrochemical sandwich immunoassay strategies involving the use of carboxyl-functionalized magnetic microbeads (cMBs) and magnetic nanoparticles (cMNPs) have been evaluated and compared. The proteolytically cleaved soluble Tyrosine kinase receptor sAXL was used as the target analyte. Antibodies against AXL were covalently immobilized on cMBs or cMNPs. Immunobinding of AXL was detected by means of a secondary biotinylated antibody and a streptavidinhorseradish peroxidase conjugate. The electrochemical transduction was accomplished by capturing the cMBs or cMNPs bearing the immunoconjugates onto screen-printed carbon electrodes (SPCEs) by using a small magnet. The amperometric response was measured at −0.20 V (vs the silver pseudoreference electrode of the SPCE) upon the addition of H2O2 in the presence of hydroquinone as the redox mediator. The calibration plots for AXL extended up to 7.5 ng mL−1 when cMBs were used for the preparation of the immunosensor and up to 40 ng mL−1 in the case of using cMNPs. The respective slope values were 158 (cMBs) and 43 nA mL ng−1 (cMNPs), while the achieved LODs were 74 (cMBs) and 75 pg mL−1 (cMNPs). Although the immunosensors prepared with cMBs provided a shorter range of linearity, they exhibited a 3.7-times larger sensitivity than those constructed with cMNPs. The successful application of the new strategies was demonstrated for the determination of the endogenous content of sAXL in real human serum samples (a cut-off value of 71 ng mL−1 have been established for patients with risk of heart failure). The immunosensors constructed using cMBs or cMNPs can be advanta geously compared, in terms of sensitivity and fabrication time, with the only immunosensor for AXL previously reported. In addition, these new immunosensors took approximately half time than ELISA to perform the assay.
Magnetic Janus Particles for Static and Dynamic (Bio)Sensing
(Magnetochemistry, 2019) Campuzano Ruiz, Susana; Gamella Carballo, Maria; Serafín González-Carrato, Verónica; Pedrero Muñoz, María; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José Manuel
Magnetic Janus particles bring together the ability of Janus particles to perform two different functions at the same time in a single particle with magnetic properties enabling their remote manipulation, which allows headed movement and orientation. This article reviews the preparation procedures and applications in the (bio)sensing field of static and self-propelled magnetic Janus particles. The main progress in the fabrication procedures and the applicability of these particles are critically discussed, also giving some clues on challenges to be dealt with and future prospects. The promising characteristics of magnetic Janus particles in the (bio)sensing field, providing increased kinetics and sensitivity and decreased times of analysis derived from the use of external magnetic fields in their manipulation, allows foreseeing their great and exciting potential in the medical and environmental remediation fields.
Opportunities, Challenges, and Prospects in Electrochemical Biosensing of Circulating Tumor DNA and its Specific Features
(Sensors, 2019) Campuzano Ruiz, Susana; Serafín González-Carrato, Verónica; Gamella Carballo, Maria; Pedrero Muñoz, María; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José Manuel
Nowadays, analyzing circulating tumor DNA (ctDNA), a very small part of circulating free DNA (cfDNA) carried by blood, is considered to be an interesting alternative to conventional single-site tumor tissue biopsies, both to assess tumor burden and provide a more comprehensive snapshot of the time-related and spatial heterogeneity of cancer genetic/epigenetic scenery. The determination of ctDNA and/or mapping its characteristic features, including tumor-specific mutations, chromosomal aberrations, microsatellite alterations, and epigenetic changes, are minimally invasive, powerful and credible biomarkers for early diagnosis, follow-up, prediction of therapy response/resistance, relapse monitoring, and tracking the rise of new mutant subclones, leading to improved cancer outcomes This review provides an outline of advances published in the last five years in electrochemical biosensing of ctDNA and surrogate markers. It emphasizes those strategies that have been successfully applied to real clinical samples. It highlights the unique opportunities they offer to shift the focus of cancer patient management methods from actual decision making, based on clinic-pathological features, to biomarker-driven treatment strategies, based on genotypes and customized targeted therapies. Also highlighted are the unmet hurdles and future key points to guide these devices in the development of liquid biopsy cornerstone tools in routine clinical practice for the diagnosis, prognosis, and therapy response monitoring in cancer patients.
Fast and sensitive diagnosis of autoimmune disorders through amperometric biosensing of serum anti-dsDNA autoantibodies
(Biosensors & Bioelectronics, 2020) López Ruiz, María Beatriz; Arévalo Pérez, Beatriz; Serafín González-Carrato, Verónica; Sánchez-Paniagua López, Marta; Montero Calle, Ana; Barderas Manchado, Rodrigo; Campuzano Ruiz, Susana; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José Manuel
This work reports the first amperometric biosensor involving the use of neutravidin-functionalized magnetic microbeads (NA-MBs) modified with a biotinylated-anti-dsDNA (b-dsDNA) as efficient magnetic microcarriers to selectively capture anti-dsDNA autoantibodies (IgG, IgA and IgM AAbs) present in the sera of patients with rheumatoid arthritis (RA). Subsequently, the attached anti-dsDNA AAbs are detected with a mixture of conventional HRP-labeled secondary antibodies (HRP-anti-human IgG/IgM/IgA mixture). The biorecognition event is monitored by amperometric transduction using the hydroquinone (HQ)/H2O2 system upon capturing the modified MBs on the surface of screen-printed carbon electrodes (SPCEs). The developed bioplatform exhibits a linear calibration plot ranging from 1 to 200 IU mL−1 with a LOD of 0.3 IU mL−1 for anti-dsDNA AAbs standards. In addition, the biosensor allows performing the determination of the anti-dsDNA AAbs levels directly in 100-times diluted serum samples from patients diagnosed with RA and in just 75 min. The obtained results are in agreement with those provided by an ELISA kit and allow discrimination between positive and negative samples.