Direct Covalent Immobilization of new Nitrogen-doped Carbon Nanodots by Electrografting for Sensing Applications

This paper reports a facile strategy to covalently immobilize nanosized carbon dots (CD) onto carbon conductive surfaces for sensing applications. The carbon nanodots designed with surface amine groups (N-CD) can be electrografted onto carbon electrodes and, thus, easily covalently immobilized on these conductive surfaces. They have been synthetized by a carbonization method microwave-assisted using preselected low cost and biocompatible precursors, such as D-fructose as primary carbon source and urea as N-donor reagent to obtain peripheral enriched nitrogen CD. The synthetized nanomaterial has been characterized by different techniques, that confirm the presence of size-regular amorphous structures with blue fluorescence when are irradiated with UV light. The highly stable immobilization of N-CD onto the electrode surfaces by electrografting provides hybrid electrodes with greater relative surface area and improved electron transfer properties, demonstrating to be a great promise for electrochemical sensing. Because of its good electrical conductivity, electrical properties, abundant edges sites and high catalytic activity, N-CD immobilized on carbon electrodes efficient amplify the electrochemiluminiscence (ECL) signal from the luminophore [Ru(bpy)3]2+ in a taurine sensor. A linear concentration range from 126 to 1000 μM, a sensitivity of 7.40ⅹ10-4 μM-1 and a detection limit of 37.8 μM were determined for the taurine sensorThis work has been supported by the Spanish Ministerio de Ciencia, Innovación y Universidades through projects CTQ2017-84309-C2-1-R and RED2018-102412-T, and Comunidad Autónoma de Madrid (S2018/NMT-4349 TRANSNANOAVANSENS-CM Program and 2017-T1/BIO-5435 Atracción de Talento Project). The authors thank Professor Hector Abruña the critical review of this wor

Bifunctional carbon nanodots for highly sensitive HER2 determination based on electrochemiluminescence

Early detection of breast cancer increases the chances of achieving adequate and successful treatment as soon as possible. In this work, a promising disposable electrochemiluminescent immunosensor has been developed for simple, efficient detection of the HER2 protein, a breast cancer biomarker. Nitrogen-rich carbon nanodots were synthesized with two functions: to provide functional groups for covalent immobilization of HER2 antibodies and to act as co-reactants in the electrochemiluminescent process. The proposed immunosensor responded linearly to HER2 concentration over a wide range, showing a detection limit of 20.4 pg mL−1. The reliability of this biosensor was confirmed by analyzing HER2 in the presence of another tumor biomarker (CEA), as well as various proteins and sugars. In addition, this proposed strategy presented good stability and applicability in the analysis of human serum samples, showing great potential for applications in the early diagnosis of breast cancerThis work has been supported by the Spanish Ministry of Science, Innovation and Universities (CTQ2017-84309-C2-1-R; RED2018- 102412-T) and by the Autonomous Community of Madrid (Talent Attraction Project 2017-T1/BIO-5435 and the P2018/NMT4349 TRANSNANOAVANSENS Program

Evaluation of the oxygen reduction reaction electrocatalytic activity of postsynthetically modified covalent organic frameworks

The pyrolysis of organic precursors to produce heteroatomic-doped carbonaceous materials has emerged as a powerful tool to construct metal-free heterogeneous electrocatalysts due to their low cost and their environmental friendliness. However, the lack of control in the atomic positions or the location of the chemical functionalities makes it difficult to establish structure-property relationships. Herein, we report an easy strategy to compare the electrocatalytic oxygen reduction reaction (ORR) performance of metal-free and nonpyrolyzed materials by postsynthetic modification of covalent organic frameworks (COFs) via click-chemistry. This method facilitates the evaluation of different active centers using materials with the same morphology and prevents active site agglomeration by covalently anchoring these moieties inside of a porous and crystalline framework. In this study we developed a series of diimide-based materials (XDI0.17-COFs) with a loading of 7.65 × 10-4 mol of active site/mg of host COF. The bulk COFs have been delaminated to perform electrode modification by drop-casting. The electrocatalytic response toward the ORR has been studied in alkaline media obtaining the best results for the NDI0.17-COF with an onset potential of 0.77 V (vs reversible hydrogen electrode, RHE) and a limiting current of 4.2 mA/cm2 by a preferred pathway toward water electroreduction. Finally, an adequate combination of density functional theory with the thermochemical Gibbs free energy formalism has been used to theoretically rationalize the ORR mechanism in these metal-free and nonpyrolyzed materials. We have obtained theoretical ORR overpotentials for each COF system agreeing with the experimental observation, which correlate with the ability of the NDI, BzDI, and PDI molecular blocks to accommodate electrons. Our work provides a guideline on how to study the electrocatalytic performance of different organic moieties in metal-free and non-pyrolyzed COFs avoiding their de novo synthesis by using the click postsynthetic methodologyTED2021-129886B-C43, PID2019-106268GB-C32, RED2018-102412-T, PID2020-116728RB-I00, PID2020-113142RB-C21, PLEC2021-007906, 2018/NMT-4349TRANSNANOAVANSENS, S2018/NMT-4367, Y2020/NMT646

MoS2-Carbon Nanodots as a New Electrochemiluminescence Platform for Breast Cancer Biomarker Detection

In this work, we present the combination of two different types of nanomaterials, 2D molybdenum disulfide nanosheets (MoS2-NS) and zero-dimensional carbon nanodots (CDs), for the development of a new electrochemiluminescence (ECL) platform for the early detection and quantification of the biomarker human epidermal growth factor receptor 2 (HER2), whose overexpression is associated with breast cancer. MoS2-NS are used as an immobilization platform for the thiolated aptamer, which can recognize the HER2 epitope peptide with high affinity, and CDs act as coreactants of the anodic oxidation of the luminophore [Ru(bpy)3]2+. The HER2 biomarker is detected by changes in the ECL signal of the [Ru(bpy)3]2+/CD system, with a low detection limit of 1.84 fg/mL and a wide linear range. The proposed method has been successfully applied to detect the HER2 biomarker in human serum samplesThis work has been financially supported by the Spanish Ministry of Economy and Competitiveness (PID2020-116728RB-I00, PID2020-116661RB-I00, CTQ2015-71955-REDT (ELECTROBIONET)), Community of Madrid (TRANSNANOAVANSENS, S2018/NMT-4349, and FotoArt (P2018/NMT4367), project S2018/NMT-4291 TEC2SPACE), MINECO (project CSIC13-4E-1794) and EU (FEDER, FSE). IMDEA Nanociencia receives support from the “Severo Ochoa” Programme for Centres of Excellence in R&D (MINECO, Grant CEX2020-001039-S)

Oxygen reduction using a metal-free naphthalene diimide-based covalent organic framework electrocatalyst

A novel naphthalene diimide-based covalent organic framework (NDI-COF) has been synthesized and successfully exfoliated into COF nanosheets (CONs). Electrochemical measurements reveal that the naphthalene diimide units incorporated into NDI-CONs act as efficient electrocatalyst for oxygen reduction in alkaline media, showing its potential for the development of metal-free fuel cellsFinancial support from the Spanish Government (projects MAT2016-77608-C3-1-P, MAT2016-77608-C3-2-P, CTQ2017-84309-C2-1-R, MAT2017-85089-C2-1-R, FJCI-2017-33536 and RYC-2015-17730), the UCM (INV.GR.00.1819.10759) and the Madrid Regional Government (TRANSNANOAVANSENS-CM (S2018/NMT-4349)) is acknowledge

Azure A embedded in carbon dots as NADH electrocatalyst: Development of a glutamate electrochemical biosensor

Carbon nanodots modified with azure A (AA-CDs) have been synthesized and applied as redox mediator of bioelectrocatalytic reactions. A deep characterization of AA-CDs nanomaterial has been carried out, proving the covalent attachment of azure A molecules into the carbon dots nanostructure. Disposable screen-printed carbon electrodes (SPCE) have been modified with AA-CDs, through the action of chitosan polymer (Chit-AA-CDs/SPCE). The Chit-AA-CDs/SPCE electrocatalytic activity towards the oxidation of NADH has been proved, obtaining excellent results regarding the low oxidation potential achieved (−0.15 V vs. Ag) and low detection and quantification limits (LOD and LOQ) for NADH, 16 and 53 µM, respectively. The developed electrochemical platform has been applied for the construction of a glutamate biosensor by immobilizing L-glutamic dehydrogenase (GLDH/Chit-AA-CDs/SPCE). The morphology of GLDH/Chit-AA-CDs/SPCE platform was analysed by AFM at each different step of the electrode modification process. The resulting biosensing platform is capable of detect NADH enzymatically generated by GLDH in the presence of glutamate and NAD+. Good analytical parameters were obtained for glutamate analysis using GLDH/Chit-AA-CDs/SPCE, as LOD and LOQ of 3.3 and 11 µM, respectively. The biosensor has been successfully applied to the analysis of food and biological samplesThis work has been supported by the Spanish Ministerio de Ciencia e Innovacion (PID2020–116728RB-I00) and Comunidad Autonoma de Madrid (SI3/PJI/2021–00341, P2018/NMT-4349 TRANSNANOAVANSENS Program

Synergistic enhancement of electrochemiluminescence through hybridization of α-Ge nanolayers and gold nanoparticles for highly sensitive detection of tyramine

This work presents a novel approach for detecting biogenic amine tyramine using a sensitive and disposable electrochemiluminescent sensor. The sensor is fabricated by modifying a screen-printed carbon electrode surface with two nanomaterials, α-Ge nanolayers and AuNP, which synergistically enhance the electrochemiluminescence response. The sensor was characterized using various techniques such as SEM-EDX, EIS, Raman, and AFM. The principle of the biosensor relays on the fact that tyramine molecule acts as an analyte and co-reactant, which interacts with the luminophore [Ru(bpy)3] 2+ on the sensor surface. The proposed sensor shows a linear response to tyramine concentration, with a detection limit of 2.28 µM. The sensor successfully detected tyramine in avocado samples, demonstrating its potential for practical applicationsThis work has been supported the Comunidad Autonoma ´ de Madrid (2021-5A/BIO-20943 Talent Attraction Project, SI3/PJI/2021-00341 and S2018/NMT-4349 TRANSNANOAVANSENS-CM Program) and by the Spanish Ministerio de Ciencia e Innovacion ´ (PID2020-116728RBI00, PDC2021-120782-C21, PID2019-106268GB-C32 and TED2021- 129738B-I00). This work has also been supported by the Spanish MINECO (PID2019-106268GB-C32, CEX2018-000805-M and PDC2021- 120782-C21). We acknowledge the support from the “(MAD2D-CM)- UAM” project funded by Comunidad de Madrid, by the Recovery, Transformation and Resilience Plan, and by NextGenerationEU from the European Unio

Carbon nanodots modified-electrode for peroxide-free cholesterol biosensing and biofuel cell design

The determination of cholesterol is greatly important because high concentrations of this biomarker are associated to heart disease. Moreover, cholesterol can be used as a fuel in enzymatic fuel cells operating under physiological conditions. Here, we present a cholesterol biosensor and a peroxide-free biofuel cell based on the electrocatalytic oxidation of the NADH generated during the enzymatic reaction of cholesterol dehydrogenase (ChDH) as an alternative to the H2O2 biosensing strategies used with cholesterol oxidase-bioelectrodes. Azure A functionalized-carbon nanodots were used as NADH oxidation electrocatalysts and for ChDH covalent immobilization. The biosensor responded linearly to cholesterol concentrations up to 1.7 mM with good sensitivity (4.50 mA cm−2 M−1) and at a low potential. The ChDH bioelectrode was combined with an O2-reducing bilirubin oxidase cathode to produce electrical energy using cholesterol as fuel and O2 as oxidant. Furthermore, the resulting enzymatic fuel cell was tested in human serum naturally containing free cholesterolA.L.DL. and M.P. thank MCIU/AEI/FEDER, EU for funding project RTI2018–095090-B-I00. M.B. acknowledges funding from the European Union’s Horizon 2020 Research and Innovation Program under the Marie Skłodowska-Curie grant agreement No. 713366. This work was also supported by Talent Attraction Project from CAM (SI3/PJI/ 2021–00341 and 2021–5A/BIO-20943), Spanish Ministerio de Ciencia e Innovacion (PID2020–116728RB-I00) and TRANSNANOAVANSENSCAM Program (S2018/NMT-4349

Bifunctional Au@Pt/Au nanoparticles as electrochemiluminescence signaling probes for SARS-CoV-2 detection

A novel immunosensor based on electrochemiluminescence resonance energy transfer (ECL-RET) for the sensitive determination of N protein of the SARS-CoV-2 coronavirus is described. For this purpose, bifunctional core@shell nanoparticles composed of a Pt-coated Au core and finally decorated with small Au inlays (Au@Pt/Au NPs) have been synthesized to act as ECL acceptor, using [Ru (bpy)3]2+ as ECL donor. These nanoparticles are efficient signaling probes in the immunosensor developed. The proposed ECL-RET immunosensor has a wide linear response to the concentration of N protein of the SARS-CoV-2 coronavirus with a detection limit of 1.27 pg/mL. Moreover, it has a high stability and shows no response to other proteins related to different virus. The immunosensor has achieved the quantification of N protein of the SARS-CoV-2 coronavirus in saliva samples. Results are consistent with those provided by a commercial colorimetric ELISA kit. Therefore, the developed immunosensor provides a feasible and reliable tool for early and effective detection of the virus to protect the populationThe authors wish to express their sincere thanks to the Spanish Ministerio de Ciencia e Innovacion (MICINN) (PID2020-116728RB-I00 and PID2020-115204RB-I00) and the Comunidad Autonoma ´ de Madrid (S2018/NMT-4349 TRANSNANOAVANSENS-CM Program, SI3/PJI/ 2021–00341 and 2021-5A/BIO-20943 Talent Attraction Project) for the financial support. C. Toyos-Rodríguez acknowledges the MICINN for the award of a FPI Grant (PRE2018-084953). A. de la Escosura-Muniz ˜ also thanks the MICINN for the research founding by a “Ramon ´ y Cajal” contract (RyC-2016-20299

Pathogen sensing device based on 2D MoS2/graphene heterostructure

In this work we propose a new methodology for selective and sensitive pathogen detection based on a 2D layered heterostructured biosensing platform. As a proof of concept, we have chosen SARS-CoV-2 virus because the availability of new methods to detect this virus is still a great deal of interest. The prepared platform is based on the covalent immobilization of molybdenum disulphide functionalized with a diazonium salt (f-MoS2) onto graphene screen-printed electrodes (GPH SPE) by electrografting of the diazonium salt. This chemistry-based method generates an improved heterostructured biosensing platform for aptamer immobilization and aptasensor development. Electrochemical impedance spectroscopy (EIS) is used to obtain the signal response of the device, proving the ability of the sensor platform to detect the virus. SARS-CoV-2 spike RBD recombinant protein (SARS-CoV-2 S1 protein) has been detected and quantified with a low detection limit of 2.10 fg/mL. The selectivity of the developed biosensor has been confirmed after detecting the S1 protein even in presence of other interfering proteins. Moreover, the ability of the device to detect SARS-CoV-2 S1 protein has been also tested in nasopharyngeal swab samplesThis work has been financially supported by the Spanish Ministry of Economy and Competitiveness (PID2020-116728RB-I00, PID2020- 116661RB-I00, CTQ2015-71955-REDT (ELECTROBIONET)) and Community of Madrid (TRANSNANOAVANSENS, S2018/NMT-4349, and PhotoArt P2018/NMT-4367). E. Enebral thank the financial support of “Nanotecnología para detección del SARS-CoV-2 y sus variantes. NANOCOV” project. IMDEA Nanociencia receives support from the “Severo Ochoa” Programme for Centres of Excellence in R&D (MINECO, Grant CEX2020-001039-S). We also thank the Spanish Ministry of Universities for supporting Laura Gutiérrez-Galvez with the Formación del Profesorado Universitario (FPU) grant (FPU19/06309