Celiac disease detection using a transglutaminase electrochemical immunosensor fabricated on nanohybrid screen-printed carbon electrodes

Celiac disease is a gluten-induced autoimmune enteropathy characterized by the presence of tissue tranglutaminase (tTG) autoantibodies. A disposable electrochemical immunosensor (EI) for the detection of IgA and IgG type anti-tTG autoantibodies in real patient’s samples is presented. Screen-printed carbon electrodes (SPCE) nanostructurized with carbon nanotubes and gold nanoparticles were used as the transducer surface. This transducer exhibits the excellent characteristics of carbon–metal nanoparticle hybrid conjugation and led to the amplification of the immunological interaction. The immunosensing strategy consisted of the immobilization of tTG on the nanostructured electrode surface followed by the electrochemical detection of the autoantibodies present in the samples using an alkaline phosphatase (AP) labelled anti-human IgA or IgG antibody. The analytical signal was based on the anodic redissolution of enzymatically generated silver by cyclic voltammetry. The results obtained were corroborated with a commercial ELISA kit indicating that the electrochemical immunosensor is a trustful analytical screening tool

Voltammetric immunosensor for the simultaneous analysis of the breast cancer biomarkers CA 15-3 and HER2-ECD

Cancer Antigen 15-3 (CA 15-3) and the extracellular domain of the human epidermal growth factor receptor 2 (HER2-ECD) are independent breast cancer biomarkers. The combination of their profiles (presence and concentration) could provide an important contribution to diagnostics and patient follow-up. Therefore, a disposable electrochemical immunosensor for the simultaneous detection of CA 15-3 and HER2-ECD was developed in this work. The immunosensor was constructed on a customized dual screen-printed carbon electrode. The carbon working electrodes' surfaces were first modified with in situ electrodeposited gold nanoparticles and then individually coated with either a monoclonal anti-human CA 15-3 or a monoclonal anti-human HER2-ECD antibody. After incubation with the biomarkers and monoclonal biotin-labelled detection antibodies, the antigen-antibody interactions were detected by linear sweep voltammetric analysis of enzymatically (alkaline phosphatase) generated metallic silver. The immunosensor’s limits of detection for the selected biomarkers were 5.0 U mL−1 for CA 15-3 and 2.9 ng mL−1 for HER2-ECD. These values could allow the use of the sensor in the non-invasive control of these biomarkers in breast cancer patients.This work received financial support from the European Union (FEDER funds through COMPETE) and National Funds (FCT, Fundação para a Ciência e a Tecnologia) through projects PTDC/SAU-ENB/114786/2009 and UID/QUI/50006/2013. Estefanía Costa Rama thanks the Government of Principado de Asturias for the award of a Severo Ochoa predoctoral grant (BP11-097).info:eu-repo/semantics/publishedVersio

An electrochemical deamidated gliadin antibody immunosensor for celiac disease clinical diagnosis

The first electrochemical immunosensor (EI) for the detection of antibodies against deamidated gliadin peptides (DGP) is described here. A disposable nanohybrid screen-printed carbon electrode modified with DGP was employed as the transducer's sensing surface. Real serumsampleswere successfully assayed and the results were corroborated with an ELISA kit. The presented EI is a promising analytical tool for celiac disease diagnosis

Competitive electrochemical immunosensor for the detection of unfolded p53 protein in blood as biomarker for Alzheimer’s disease

Alzheimer's disease is one of the most common causes of dementia nowadays, and its prevalence increases over time. Because of this and the difficulty of its diagnosis, accurate methods for the analysis of specific biomarkers for an early diagnosis of this disease are much needed. Recently, the levels of unfolded isoform of the multifunctional protein p53 in plasma have been proved to increase selectively in Alzheimer's Disease patients in comparison with healthy subjects, thus entering the list of biomarkers that can be used for the diagnosis of this illness. We present here the development of an electrochemical immunosensor based on nanostructured screen-printed carbon electrodes for the quantification of unfolded p53 in plasma samples. The sensor shows a suitable linear range (from 2 to 50 nM) for its application in real blood samples and a very low limit of detection (0.05 nM). The concentration of unfolded p53 has been accurately detected in plasma of elderly people in healthy conditions, subjects with mild cognitive impairment (MCI) and Alzheimer's Disease (AD) subjects, obtaining results with no significant differences to those provided by an ELISA assay. These results support the possibility of measuring unfolded p53 levels with a cheap, simple and miniaturized device with a promising future for point-of-care applications in the early diagnosis of Alzheimer's dementia.This work has been supported by the FC-15-GRUPIN14-021 project from the Asturias Regional Government and the CTQ2014-58826-R and MINECO-18-CTQ2017-86994-R projects from the Spanish Ministry of Economy and Competitiveness (MINECO). O. Amor-Gutiérrez thanks Vicerrectorado de Investigación from University of Oviedo for the award of a grant “Ayudas para la realización de tesis doctorales” (PAPI-18-PF-13) through Plan de Apoyo y Promoción de la Investigación.info:eu-repo/semantics/publishedVersio

Celiac disease diagnosis and gluten-free food analytical control

Celiac disease (CD) is an autoimmune enteropathy, characterized by an inappropriate T-cell-mediated immune response to the ingestion of certain dietary cereal proteins in genetically susceptible individuals. This disorder presents environmental, genetic, and immunological components. CD presents a prevalence of up to 1% in populations of European ancestry, yet a high percentage of cases remain underdiagnosed. The diagnosis and treatment should be made early since untreated disease causes growth retardation and atypical symptoms, like infertility or neurological disorders. The diagnostic criteria for CD, which requires endoscopy with small bowel biopsy, have been changing over the last few decades, especially due to the advent of serological tests with higher sensitivity and specificity. The use of serological markers can be very useful to rule out clinical suspicious cases and also to help monitor the patients, after adherence to a gluten-free diet. Since the current treatment consists of a life-long glutenfree diet, which leads to significant clinical and histological improvement, the standardization of an assay to assess in an unequivocal way gluten in gluten-free foodstuff is of major importance

Bioelectroanalysis in a Drop: Construction of a Glucose Biosensor

This lab experiment describes a complete method to fabricate an enzymatic glucose electroanalytical biosensor by students. Using miniaturized and disposable screen-printed electrodes (SPEs), students learn how to use them as transducers and understand the importance SPEs have acquired in sensor development during the last years. Students can also revise concepts related to enzymatic assays, with glucose oxidase and horseradish peroxidase involved in subsequent reactions. Moreover, they learn the trends that current analytical chemistry follows presently such as miniaturization, portability, and low cost. At the same time, this experiment serves to teach basic analytical concepts (accuracy, precision, sensitivity, and selectivity) in a practical way. The high clinical interest of glucose, due to a large number of diabetes patients around the world, and the application of the sensor to analysis of real food samples make this experiment very attractive to students. The questions set out along this experiment help students to acquire skills for solving analytical problems from the very beginning

Mercury determination in urine samples by gold nanostructured screen-printed carbon electrodes after vortex-assisted ionic liquid dispersive liquid–liquid microextraction

A novel approach is presented to determine mercury in urine samples, employing vortex-assisted ionic liquid dispersive liquid–liquid microextraction and microvolume back-extraction to prepare samples, and screen-printed electrodes modified with gold nanoparticles for voltammetric analysis. Mercury was extracted directly from non-digested urine samples in a water-immiscible ionic liquid, being back-extracted into an acidic aqueous solution. Subsequently, it was determined using gold nanoparticle-modified screen-printed electrodes. Under optimized microextraction conditions, standard addition calibration was applied to urine samples containing 5, 10 and 15 μg L−1 of mercury. Standard addition calibration curves using standards between 0 and 20 μg L−1 gave a high level of linearity with correlation coefficients ranging from 0.990 to 0.999 (N = 5). The limit of detection was empirical and statistically evaluated, obtaining values that ranged from 0.5 to 1.5 μg L−1, and from 1.1 to 1.3 μg L−1, respectively, which are significantly lower than the threshold level established by the World Health Organization for normal mercury content in urine (i.e., 10–20 μg L−1). A certified reference material (REC-8848/Level II) was analyzed to assess method accuracy finding 87% and 3 μg L−1 as the recovery (trueness) and standard deviation values, respectively. Finally, the method was used to analyze spiked urine samples, obtaining good agreement between spiked and found concentrations (recovery ranged from 97 to 100%).The authors would like to thank the Spanish Ministry of Science and Innovation (projects n. CTQ2011-23968 and CTQ2011-24560), Generalitat Valenciana (Spain) (projects n. ACOMP/2013/072, GVA/2014/096 and PROMETEO/2013/038) and University of Alicante (Spain) (project n. GRE12-45) for the financial support. E. Fernández also thanks Ministry of Education (FPU13/03125) for her FPU grant

High Performance Tunable Catalysts Prepared by Using 3D Printing

Honeycomb monoliths are the preferred supports in many industrial heterogeneous catalysis reactions, but current extrusion synthesis only allows obtaining parallel channels. Here, we demonstrate that 3D printing opens new design possibilities that outperform conventional catalysts. High performance carbon integral monoliths have been prepared with a complex network of interconnected channels and have been tested for carbon dioxide hydrogenation to methane after loading a Ni/CeO2 active phase. CO2 methanation rate is enhanced by 25% at 300 ◦C because the novel design forces turbulent flow into the channels network. The methodology and monoliths developed can be applied to other heterogeneous catalysis reactions, and open new synthesis options based on 3D printing to manufacture tailored heterogeneous catalysts