Person:
Ruiz Valdepeñas Montiel, Víctor

First Name

Víctor

Last Name

Ruiz Valdepeñas Montiel

URI

https://hdl.handle.net/20.500.14352/81630

Affiliation

Universidad Complutense de Madrid

Faculty / Institute

Ciencias Químicas

Department

Química en Ciencias Farmacéuticas

Identifiers

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Now showing 1 - 10 of 28

Amperometric Immunosensing Scaffolds for Rapid, Simple, Non-Invasive and Accurate Determination of Protein Biomarkers of Well-Accepted and Emerging Clinical Importance
(Proceedings, 2017) Pedrero Muñoz, María; Muñoz San Martín, Cristina; Torrente Rodríguez, Rebeca Magnolia; Ruiz Valdepeñas Montiel, Víctor; Vargas Orgaz, Eva; Manuel de Villena Rueda, Francisco Javier; Barderas Manchado, Rodrigo; Campuzano Ruiz, Susana; Pingarrón Carrazón, José Manuel
Toward Liquid Biopsy: Determination of the Humoral Immune Response in Cancer Patients Using HaloTag Fusion Protein-Modified Electrochemical Bioplatforms
(Analytical Chemistry, 2016) Garranzo Asensio, María; Guzmán Aránguez, Ana Isabel; Povés Francés, Carmen; Fernández Aceñero, Mª Jesús; Torrente Rodríguez, Rebeca Magnolia; Ruiz Valdepeñas Montiel, Víctor; Domínguez Muñóz, Gemma; San Frutos Llorente, Luis; Rodríguez Salas, Nuria; Villalba Díaz, Mayte; Pingarrón Carrazón, José Manuel; Campuzano Ruiz, Susana; Barderas Manchado, Rodrigo
Autoantibodies raised against tumor-associated antigens have shown high promise as clinical biomarkers for reliable diagnosis, prognosis, and therapy monitoring of cancer. An electrochemical disposable biosensor for the specific and sensitive determination of p53-specific autoantibodies has been developed for the first time in this work. This biosensor involves the use of magnetic microcarriers (MBs) modified with covalently immobilized HaloTag fusion p53 protein as solid supports for the selective capture of specific autoantibodies. After magnetic capture of the modified MBs onto screen-printed carbon working electrodes, the amperometric signal using the system hydroquinone/H2O2 was related to the levels of p53-autoantibodies in the sample. The biosensor was applied for the analysis of sera from 24 patients with high-risk of developing colorectal cancer and 6 from patients already diagnosed with colorectal (4) and ovarian (2) cancer. The developed biosensor was able to determine p53 autoantibodies with a sensitivity higher than that of a commercial standard ELISA using a just-in-time produced protein in a simpler protocol with less sample volume and easily miniaturized and cost-effective instrumentation.
p53 and p63 Proteoforms Derived from Alternative Splicing Possess Differential Seroreactivity in Colorectal Cancer with Distinct Diagnostic Ability from the Canonical Proteins
(Cancers, 2023) Montero Calle, Ana; Garranzo Asensio, María; Torrente Rodríguez, Rebeca Magnolia; Ruiz Valdepeñas Montiel, Víctor; Poves, Carmen; Dziakova, Jana; Sanz, Rodrigo; Díaz Del Arco, Cristina; Pingarrón Carrazón, José Manuel; Fernández Aceñero, María Jesús; Campuzano Ruiz, Susana; Barderas Manchado, Rodrigo
Colorectal cancer (CRC) is the third most common cancer and the second most frequent cause of cancer-related death worldwide. The detection in plasma samples of autoantibodies against specific tumor-associated antigens has been demonstrated to be useful for the early diagnosis of CRC by liquid biopsy. However, new studies related to the humoral immune response in cancer are needed to enable blood-based diagnosis of the disease. Here, our aim was to characterize the humoral immune response associated with the different p53 and p63 proteoforms derived from alternative splicing and previously described as aberrantly expressed in CRC. Thus, here we investigated the diagnostic ability of the twelve p53 proteoforms and the eight p63 proteoforms described to date, and their specific N-terminal and C-terminal end peptides, by means of luminescence HaloTag beads immunoassays. Full-length proteoforms or specific peptides were cloned as HaloTag fusion proteins and their seroreactivity analyzed using plasma from CRC patients at stages I-IV (n = 31), individuals with premalignant lesions (n = 31), and healthy individuals (n = 48). p53γ, Δ40p53β, Δ40p53γ, Δ133p53γ, Δ160p53γ, TAp63α, TAp63δ, ΔNp63α, and ΔNp63δ, together with the specific C-terminal end α and δ p63 peptides, were found to be more seroreactive against plasma from CRC patients and/or individuals with premalignant lesions than from healthy individuals. In addition, ROC (receiver operating characteristic) curves revealed a high diagnostic ability of those p53 and p63 proteoforms to detect CRC and premalignant individuals (AUC higher than 85%). Finally, electrochemical biosensing platforms were employed in POC-like devices to investigate their usefulness for CRC detection using selected p53 and p63 proteoforms. Our results demonstrate not only the potential of these biosensors for the simultaneous analysis of proteoforms’ seroreactivity, but also their convenience and versatility for the clinical detection of CRC by liquid biopsy. In conclusion, we here show that p53 and p63 proteoforms possess differential seroreactivity in CRC patients in comparison to controls, distinctive from canonical proteins, which should improve the diagnostic panels for obtaining a blood-based biomarker signature for CRC detection.
Simultaneous Determination of the Main Peanut Allergens in Foods Using Disposable Amperometric Magnetic Beads-Based Immunosensing Platforms
(Chemosensors, 2016) Ruiz Valdepeñas Montiel, Víctor; Torrente Rodríguez, Rebeca Magnolia; Campuzano Ruiz, Susana; Pellicanò, Alessandro; Reviejo García, Ángel Julio; Cosio, Maria; Pingarrón Carrazón, José Manuel
In this work, a novel magnetic beads (MBs)-based immunosensing approach for the rapid and simultaneous determination of the main peanut allergenic proteins (Ara h 1 and Ara h 2) is reported. It involves the use of sandwich-type immunoassays using selective capture and detector antibodies and carboxylic acid-modified magnetic beads (HOOC-MBs). Amperometric detection at −0.20 V was performed using dual screen-printed carbon electrodes (SPdCEs) and the H2O2/hydroquinone (HQ) system. This methodology exhibits high sensitivity and selectivity for the target proteins providing detection limits of 18.0 and 0.07 ng/mL for Ara h 1 and Ara h 2, respectively, with an assay time of only 2 h. The usefulness of the approach was evaluated by detecting the endogenous content of both allergenic proteins in different food extracts as well as trace amounts of peanut allergen (0.0001% or 1.0 mg/kg) in wheat flour spiked samples. The developed platform provides better Low detection limits (LODs) in shorter assay times than those claimed for the allergen specific commercial ELISA kits using the same immunoreagents and quantitative information on individual food allergen levels. Moreover, the flexibility of the methodology makes it readily translate to the detection of other food-allergens.
Automated Bioanalyzer Based on Amperometric Enzymatic Biosensors for the Determination of Ethanol in Low-Alcohol Beers
(Beverages, 2017) Vargas Orgaz, Eva; Conzuelo, Felipe; Ruiz, M.; Campuzano Ruiz, Susana; Ruiz Valdepeñas Montiel, Víctor; González de Rivera, Guillermo; López-Colino, Fernando; Reviejo García, Ángel Julio; Pingarrón Carrazón, José Manuel
In this work, a new automated bioanalyzer based on the use of enzymatic biosensors as amperometric detectors is reported. This automatic bioanalyzer is configurable both as continuous flow and flow injection analysis systems and enables both on-line and off-line monitoring of ethanol in low-alcohol beer to be performed. The attractive analytical and operational characteristics demonstrated by the automated bioanalyzer make it a promising, simple, rapid, and reliable tool for quality control of this beverage in the beer industry, either during the manufacturing process or in the final product. Moreover its applicability to the analysis of the ethanol content in different non-alcoholic beers working at different modes was successfully demonstrated.
Multiplexed monitoring of a novel autoantibody diagnostic signature of colorectal cancer using HaloTag technology-based electrochemical immunosensing platform
(Theranostics, 2020) Garranzo Asensio, María; Guzmán Aránguez, Ana Isabel; Povedano, Eloy; Ruiz Valdepeñas Montiel, Víctor; Povés Francés, Carmen; Fernández Aceñero, María Jesús; Montero Calle, Ana; Solís Fernández, Guillermo; Fernández Díez, Servando; Camps, Jordi; Arenas, Meritxell; Rodríguez Tomás, Elisabeth; Joven, Jorge; Sánchez Martínez, Maricruz; Rodrígez, Nuria; Domínguez Muñóz, Gemma; Yáñez Sedeño, Paloma; Pingarrón Carrazón, José Manuel; Campuzano Ruiz, Susana; Barderas Manchado, Rodrigo
Background and Purpose: The humoral immune response in cancer patients can be used for early detection of the disease. Autoantibodies raised against tumor-associated antigens (TAAs) are promising clinical biomarkers for reliable cancer diagnosis, prognosis, and therapy monitoring. In this study, an electrochemical disposable multiplexed immunosensing platform able to integrate difficult- and easy-to-express colorectal cancer (CRC) TAAs is reported for the sensitive determination of eight CRC-specific autoantibodies. Methods: The electrochemical immunosensing approach involves the use of magnetic microcarriers (MBs) as solid supports modified with covalently immobilized HaloTag fusion proteins for the selective capture of specific autoantibodies. After magnetic capture of the modified MBs onto screen-printed carbon working electrodes, the amperometric responses measured using the hydroquinone (HQ)/H2O2 system were related to the levels of autoantibodies in plasma. Results: The biosensing platform was applied to the analysis of autoantibodies against 8 TAAs described for the first time in this work in plasma samples from healthy asymptomatic individuals (n=3), and patients with high-risk of developing CRC (n=3), and from patients already diagnosed with colorectal (n=3), lung (n=2) or breast (n=2) cancer. The developed bioplatform demonstrated an improved discrimination between CRC patients and controls (asymptomatic healthy individuals and breast and lung cancer patients) compared to an ELISA-like luminescence test. Conclusions: The proposed methodology uses a just-in-time produced protein in a simpler protocol, with low sample volume, and involves cost-effective instrumentation, which could be used in a high-throughput manner for reliable population screening to facilitate the detection of early CRC patients at affordable cost.
Magnetic Beads-Based Sensor with Tailored Sensitivity for Rapid and Single-Step Amperometric Determination of miRNAs
(International Journal of Molecular Sciences, 2017) Vargas Orgaz, Eva; Torrente Rodríguez, Rebeca Magnolia; Ruiz Valdepeñas Montiel, Víctor; Povedano Muñumel, Eloy; Pedrero Muñoz, María; Montoya, Juan; Campuzano Ruiz, Susana; Pingarrón Carrazón, José Manuel
This work describes a sensitive amperometric magneto-biosensor for single-step and rapid determination of microRNAs (miRNAs). The developed strategy involves the use of direct hybridization of the target miRNA (miRNA-21) with a specific biotinylated DNA probe immobilized on streptavidin-modified magnetic beads (MBs), and labeling of the resulting heteroduplexes with a specific DNA–RNA antibody and the bacterial protein A (ProtA) conjugated with an horseradish peroxidase (HRP) homopolymer (Poly-HRP40) as an enzymatic label for signal amplification. Amperometric detection is performed upon magnetic capture of the modified MBs onto the working electrode surface of disposable screen-printed carbon electrodes (SPCEs) using the H2O2/hydroquinone (HQ) system. The magnitude of the cathodic signal obtained at −0.20 V (vs. the Ag pseudo-reference electrode) demonstrated linear dependence with the concentration of the synthetic target miRNA over the 1.0 to 100 pM range. The method provided a detection limit (LOD) of 10 attomoles (in a 25 μL sample) without any target miRNA amplification in just 30 min (once the DNA capture probe-MBs were prepared). This approach shows improved sensitivity compared with that of biosensors constructed with the same anti-DNA–RNA Ab as capture instead of a detector antibody and further labeling with a Strep-HRP conjugate instead of the Poly-HRP40 homopolymer. The developed strategy involves a single step working protocol, as well as the possibility to tailor the sensitivity by enlarging the length of the DNA/miRNA heteroduplexes using additional probes and/or performing the labelling with ProtA conjugated with homopolymers prepared with different numbers of HRP molecules. The practical usefulness was demonstrated by determination of the endogenous levels of the mature target miRNA in 250 ng raw total RNA (RNAt) extracted from human mammary epithelial normal (MCF-10A) and cancer (MCF-7) cells and tumor tissues.
First PCR-free electrochemical bioplatform for the detection of mustard Sin a 1 protein as a potential “hidden” food allergen
(Bioelectrochemistry, 2023) Gamella Carballo, María; Laza, Anabel; Parrón Ballesteros, Jorge; Bueno Díaz, Cristina; Ruiz Valdepeñas Montiel, Víctor; Pedrero Muñoz, María; Bertolino, Franco ; Pingarrón Carrazón, José Manuel; Villalba Díaz, María Teresa; Campuzano Ruiz, Susana
A disposable electrochemical PCR-free biosensor for the selective detection of a fragment encoding the protein Sin a 1, a 2S albumin considered a diagnostic marker for sensitization to mustard, is reported. The methodology is based on the formation of DNA/RNA heterohybrids by sandwich hybridization of a specific fragment of the Sin a 1 allergen coding sequence with appropriately designed RNA probes. Labeling with commercial antibodies specific to the heteroduplexes and secondary antibodies conjugated with horseradish peroxidase (HRP) was carried out onto the surface of magnetic beads (MBs). Amperometric transduction was undertaken on screen-printed electrodes using H2O2 as enzyme substrate and hydroquinone (HQ) a redox mediator. The electrochemical biosensor allows the simple and fast detection (75 min) of Sin a 1 reaching a limit of detection of 3 pM. The bioplatform was successfully applied to the analysis of the targeted Sin a 1 gene specific region using just 50 ng of non-fragmented denatured genomic DNA extracted from yellow mustard seeds.
Disposable electrochemical immunoplatform to shed light on the role of the multifunctional glycoprotein TIM-1 in cancer cells invasion
(Talanta, 2024) Quinchia, Jennifer; Blázquez-García, Marina; Torrente Rodríguez, Rebeca Magnolia; Ruiz Valdepeñas Montiel, Víctor; Serafín González-Carrato, Verónica; Rejas-González, Raquel; Montero-Calle, Ana; Orozco, Jahir; Pingarrón Carrazón, José Manuel; Barderas Manchado, Rodrigo; Campuzano Ruiz, Susana
Detecting overexpression of cancer biomarkers is an excellent tool for diagnostic/prognostic and follow-up of patients with cancer or their response to treatment. This work illustrates the relevance of interrogating the levels of T-cell immunoglobulin and mucin domain 1 (TIM-1) protein as a diagnostic/prognostic biomarker of high-prevalence breast and lung cancers by using an amperometric disposable magnetic microparticles-assisted immunoplatform. The developed method integrates the inherent advantages of carboxylic acid-functionalized magnetic beads (HOOC-MBs) as pre-concentrator support and the amperometric transduction at screen-printed carbon electrodes (SPCEs). The immunoplatform involves a sandwich-type immunoassay assembled on HOOC-MBs through the specific capture/labeling of TIM-1 using capture antibodies and horseradish peroxidase (HRP)-conjugated biotinylated detection antibodies as biorecognition elements. The magnetic immunoconjugates were confined onto the working electrode (WE) surface of the SPCEs for amperometric detection using the hydroquinone/hydrogen peroxide/HRP (HQ/H2O2/HRP) redox system. The method allows the selective detection of TIM-1 protein over the 87–7500 pg mL−1 concentration range in only 45 min, with a limit of detection of 26 pg mL−1. The developed bioplatform was successfully applied to the analysis of breast and lung cancer cell extracts, providing the first quantitative results of the target glycoprotein in these types of samples.
Improving Cancer Outcomes through Electrochemical Biosensing of Early Diagnosis/Prognosis Biomarkers in Human Biopsies
(Proceedings, 2017) Pingarrón Carrazón, José Manuel; Campuzano Ruiz, Susana; Torrente Rodríguez, Rebeca Magnolia; Ruiz Valdepeñas Montiel, Víctor; Vargas Orgaz, Eva; Barderas Manchado, Rodrigo