1. Early stress detection in forest trees using a nanobody-functionalized electrochemical biosensor for ascorbate peroxidaseClaudia D'Ercole, Rossella Svigelj, Tanja Mrak, Ario De Marco, 2025, original scientific article Abstract: Forest environments are exposed to multiple stressful factors of both abiotic and biotic nature such as heavy metal contamination, drought, or pest infestations which may lead to their massive decline. We designed a comprehensive approach for isolating, producing and functionalizing reagents suitable for the affordable detection of forest plant stress biomarkers with the aim to provide quantitative data to assess plant stress fluctuation and, possibly, to design mitigation strategies. We first optimized a panning protocol to recover nanobodies targeting shared sequences that could cross-react with both Pisum sativum and Populus nigra ascorbate peroxidase (APX). After their production as recombinant constructs and their extensive biophysical and biochemical characterization, such reagents were exploited as the immunocapture element of an electrochemical biosensor conceived as a potential point-of-care device. Such biosensor could detect both pea and poplar APX in leaf extracts and could be used to clearly discriminate between control and heavy metal-stressed poplar plants based on their APX activity, even before the appearance of any phenotypic symptom. The combination of fast and inexpensive reagent production with the development of portable diagnostics opens the opportunity for large-scale, on-site surveys of forest trees.
Keywords: plant stress, scavengers, diagnostics, nanobodies, biosensors
Published in RUNG: 11.04.2025; Views: 301; Downloads: 2
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6. Fluorescent covalent organic frameworks : promising bioimaging materialsChimatahalli Santhakumar Karthik, Tina Škorjanc, Dinesh Shetty, 2024, original scientific article Abstract: Fluorescent covalent organic frameworks (COFs) have emerged as promising candidates for imaging living cells due to their unique properties and adjustable fluorescence. In this mini-review, we provide an overview of recent advancements in fluorescent COFs for bioimaging applications. We discuss the strategies used to design COFs with desirable properties such as high photostability, excellent biocompatibility, and pH sensitivity. Additionally, we explore the various ways in which fluorescent COFs are utilized in bioimaging, including cellular imaging, targeting specific organelles, and tracking biomolecules. We delve into their applications in sensing intracellular pH, reactive oxygen species (ROS), and specific biomarkers. Furthermore, we examine how functionalization techniques enhance the targeting and imaging capabilities of fluorescent COFs. Finally, we discuss the challenges and prospects in the field of fluorescent COFs for bioimaging in living cells, urging further research in this exciting area.
Keywords: covalent organic frameworks, fluorescent materials, imaging, bioimaging, biosensors
Published in RUNG: 05.03.2024; Views: 2435; Downloads: 8
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8. Biological applications of porous polymers and covalent organic frameworks : lecture at the Otto-von-Guericke Universität Magdeburg, Tuesday, 27th June 2023, Magdeburg, GermanyTina Škorjanc, 2023, unpublished conference contribution Abstract: Porous organic polymers (POPs) and covalent organic frameworks (COFs) have gained significant attention in the scientific community for a wide array of applications because of their attractive physical and chemical properties. Porosity of these materials provides ample surface area for interaction with targets, while crystallinity allows for highly specific structural tuning. In this seminar, I will present two strategies of utilizing these features of newly prepared materials in biosensing. Firstly, a cationic POP was synthesized, deposited onto interdigitated electrode arrays via a nontraditional electrophoresis technique, and utilized for electrochemical sensing of bacterial cells. As the principle of detection relied on electrostatic interactions between the cationic POP and the anionic bacterial surface, the sensor operated for both Gram-positive and Gram-negative bacteria. Secondly, a small-molecule nitroimidazole target for hypoxia, a low oxygen environment present in tumors, was post-synthetically conjugated to the pores of a fluorescent COF. This material served as a useful hypoxia imaging agent in cancerous cells. The seminar will conclude with some future perspectives on POPs and COFs in biological applications followed by Q & A.
Keywords: Porous organic polymers, covalent organic frameworks, biosensors, hypoxia
Published in RUNG: 13.07.2023; Views: 2450; Downloads: 0
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10. Native llama nanobody library panning performed by phage and yeast display provides binders suitable for C-reactive protein detectionSandra Oloketuyi, Robert Alvin Bernedo-Navarro, Andreas Christmann, Justyna Borkowska, Giulia Cazzaniga, Horst Wilhelm Schuchmann, Joanna Niedziółka-Jönsson, Katarzyna Szot-Karpińska, Harald Kolmar, Ario De Marco, 2021, original scientific article Keywords: phage display, yeast display, nanobodies, electrochemical impedance biosensors
Published in RUNG: 06.12.2021; Views: 3157; Downloads: 46
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