Development of Photonic Multi-Sensing Systems Based on Molecular Gates Biorecognition and Plasmonic Sensors: The PHOTONGATE Project

[EN] This paper presents the concept of a novel adaptable sensing solution currently being developed under the EU Commission-founded PHOTONGATE project. This concept will allow for the quantification of multiple analytes of the same or different nature (chemicals, metals, bacteria, etc.) in a single test with levels of sensitivity and selectivity at/or over those offered by current solutions. PHOTONGATE relies on two core technologies: a biochemical technology (molecular gates), which will confer the specificity and, therefore, the capability to be adaptable to the analyte of interest, and which, combined with porous substrates, will increase the sensitivity, and a photonic technology based on localized surface plasmonic resonance (LSPR) structures that serve as transducers for light interaction. Both technologies are in the micron range, facilitating the integration of multiple sensors within a small area (mm2). The concept will be developed for its application in health diagnosis and food safety sectors. It is thought of as an easy-to-use modular concept, which will consist of the sensing module, mainly of a microfluidics cartridge that will house the photonic sensor, and a platform for fluidic handling, optical interrogation, and signal processing. The platform will include a new optical concept, which is fully European Union Made, avoiding optical fibers and expensive optical components.The micro-nanofabrication capabilities required in the PHOTONGATE project- 101093042 are funded by the Pluri-Regional FEDER funding Plan 2014-2020 European Commission. This research project has received funding from the European Union¿s HORIZON-CL4-2022 research and innovation programme under grant agreement ID 101093042, PHOTONGATE projectNieves-Paniagua, Ó.; Ortiz De Zárate-Díaz, D.; Aznar, E.; Caballos-Gómez, MI.; Garrido-García, EM.; Martínez-Máñez, R.; Dortu, F.... (2023). Development of Photonic Multi-Sensing Systems Based on Molecular Gates Biorecognition and Plasmonic Sensors: The PHOTONGATE Project. Sensors. 23(20):1-13. https://doi.org/10.3390/s23208548113232

Aptamer-capped nanoporous anodic alumina for SARS-CoV-2 spike protein detection

The COVID-19 pandemic, which began in 2019, has highlighted the importance of testing and tracking infected individuals as a means of mitigating the spread of the virus. In this context, the development of sensitive and rapid methods for the detection of SARS-CoV-2, the virus responsible for COVID-19, is crucial. Herein, a biosensor based on oligonucleotide-gated nanomaterials for the specific detection of SARS-CoV-2 spike protein is presented. The sensing system consists of a nanoporous anodic alumina disk loaded with the fluorescent indicator rhodamine B and capped with a DNA aptamer that selectively binds the SARS-CoV-2 spike protein. The system is initially evaluated using pseudotype virus systems based on vesicular stomatitis virus carrying different SARS-CoV-2 S-proteins on their surface. When the pseudotype virus is present, the cap of the solid is selectively removed, triggering the release of the dye from the pore voids to the medium. The nanodevice demonstrated its ability to detect pseudotype virus concentrations as low as 7.5·103 PFU mL. In addition, the nanodevice is tested on nasopharyngeal samples from individuals suspected of having COVID-19.This study was supported by the Spanish Government (projects PID2021-126304OB-C41, and PID2021-122875OB-100 (MCUI/AEI/FEDER, UE)), the Generalitat Valenciana (project no.2 RD 180/2020, CIPROM/2021/007), Supera COVID-19 Fund (DIACOVID project), the Universitat Politècnica de València−Instituto de Investigación Sanitaria La Fe (IIS-LaFe) (SARS-COV-2-SEEKER and VISION-COV projects), and by the European Commission –NextGenerationEU, through CSIC's Global Health Platform (PTI Salud Global) to Ron Geller. The project leading to this application has received funding from the European Union's Horizon EUROPE research and innovation programme under grant agreement No 101093042. Isabel Caballos thanks the Instituto de Salud Carlos III for her predoctoral fellowship (IFI21/00008). Alba López-Palacios thanks the Ministerio de Universidades for her predoctoral grant (FPU20/05297). Ron Geller holds a Ramon y Cajal fellowship from the Spanish Ministerio de Economía y Competitividad (RYC-2015-17517).Peer reviewe

Aptamer-Capped Nanoporous Anodic Alumina for SARS-CoV-2 Spike Protein Detection

[EN] The COVID-19 pandemic, which began in 2019, has highlighted the importance of testing and tracking infected individuals as a means of mitigating the spread of the virus. In this context, the development of sensitive and rapid methods for the detection of SARS-CoV-2, the virus responsible for COVID-19, is crucial. Here, a biosensor based on oligonucleotide-gated nanomaterials for the specific detection of SARS-CoV-2 spike protein is presented. The sensing system consists of a nanoporous anodic alumina disk loaded with the fluorescent indicator rhodamine B and capped with a DNA aptamer that selectively binds the SARS-CoV-2 spike protein. The system is initially evaluated using pseudotype virus systems based on vesicular stomatitis virus carrying different SARS-CoV-2 S-proteins on their surface. When the pseudotype virus is present, the cap of the solid is selectively removed, triggering the release of the dye from the pore voids to the medium. The nanodevice demonstrated its ability to detect pseudotype virus concentrations as low as 7.5 center dot 10(3) PFU mL. In addition, the nanodevice is tested on nasopharyngeal samples from individuals suspected of having COVID-19.This study was supported by the Spanish Government (projects PID2021-126304OB-C41, and PID2021-122875OB-100 (MCUI/AEI/FEDER, UE)), the Generalitat Valenciana (project no.2 RD 180/2020, CIPROM/2021/007), Supera COVID-19 Fund (DIACOVID project), the Universitat Politecnica de Valencia-Instituto de Investigacion Sanitaria La Fe (IIS-LaFe) (SARS-COV-2-SEEKER and VISION-COV projects), and by the European Commission -NextGenerationEU, through CSIC's Global Health Platform (PTI Salud Global) to Ron Geller. The project leading to this application has received funding from the European Union's Horizon EUROPE research and innovation programme under grant agreement No 101093042. Isabel Caballos thanks the Instituto de Salud Carlos III for her predoctoral fellowship (IFI21/00008). Alba Lopez-Palacios thanks the Ministerio de Universidades for her predoctoral grant (FPU20/05297). Ron Geller holds a Ramon y Cajal fellowship from the Spanish Ministerio de Economia y Competitividad (RYC-2015-17517). Figure 1 done with BioRender.com. The use of samples from human subjects was approved by the Medicaments Research Ethics Commmittee, CEIm of Hospital Universitari i Politecnic La Fe (no. 2021-012-1). Informed written consent was obtained from all participants or next of kin prior to the research.Caballos-Gómez, MI.; Aranda, MN.; López-Palacios, A.; Pla, L.; Santiago Felipe, S.; Hernández-Montoto, A.; Tormo-Mas, MÁ.... (2023). Aptamer-Capped Nanoporous Anodic Alumina for SARS-CoV-2 Spike Protein Detection. Advanced Materials Technologies. 8(11):1-10. https://doi.org/10.1002/admt.20220191311081

Human Papilloma Virus DNA Detection in Clinical Samples Using Fluorogenic Probes Based on Oligonucleotide Gated Nanoporous Anodic Alumina Films

[EN] In this work, fluorogenic probes based on oligonucleotide capped nanoporous anodic alumina films are developed for specific and sensitive detection of human papilloma virus (HPV) DNA. The probe consists of anodic alumina nanoporous films loaded with the fluorophore rhodamine B (RhB) and capped with oligonucleotides bearing specific base sequences complementary to genetic material of different high-risk (hr) HPV types. Synthesis protocol is optimized for scale up production of sensors with high reproducibility. The sensors' surfaces are characterized by scanning electron microscopy (HR-FESEM) and atomic force microscopy (AFM) and their atomic composition is determined by energy dispersive X-ray spectroscopy (EDXS). Oligonucleotide molecules onto nanoporous films block the pores and avoid diffusion of RhB to the liquid phase. Pore opening is produced when specific DNA of HPV is present in the medium, resulting in RhB delivery, that is detected by fluorescence measurements. The sensing assay is optimized for reliable fluorescence signal reading. Nine different sensors are synthesized for specific detection of 14 different hr-HPV types in clinical samples with very high sensitivity (100%) and high selectivity (93-100%), allowing rapid screening of virus infections with very high negative predictive values (100%).The authors gratefully acknowledge financial support projects for PID2021-126304OB-C41 and PDI2021-122875OB-100 funded by MCIN/AEI /10.13039/501100011033 / FEDER, UE, project DTS18/00090 from Instituto de Salud Carlos III and FEDER and Generalitat Valenciana (Project PROMETEO CIPROM/2021/007). This work was also supported by CIBER -Consorcio Centro de Investigacion Biomedica en Red- (CB06/01/2012), Instituto de Salud Carlos III, Ministerio de Ciencia e Innovacion. The authors thank UPV electron microscopy service for technical support.Hernández-Montoto, A.; Aranda, MN.; Caballos-Gómez, MI.; López-Palacios, A.; Tormo-Mas, MÁ.; Peman, J.; Prieto Rodríguez, M.... (2023). Human Papilloma Virus DNA Detection in Clinical Samples Using Fluorogenic Probes Based on Oligonucleotide Gated Nanoporous Anodic Alumina Films. Advanced Healthcare Materials (Online). 12(22). https://doi.org/10.1002/adhm.202203326122