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- Duznovic, I., et al.
(författare)
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Ultrasensitive and Selective Protein Recognition with Nanobody-Functionalized Synthetic Nanopores
- 2021
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Ingår i: Small. - : Wiley. - 1613-6810 .- 1613-6829. ; 17:33
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Tidskriftsartikel (refereegranskat)abstract
- The development of flexible and reconfigurable sensors that can be readily tailored toward different molecular analytes constitutes a key goal and formidable challenge in biosensing. In this regard, synthetic nanopores have emerged as potent physical transducers to convert molecular interactions into electrical signals. Yet, systematic strategies to functionalize their surfaces with receptor proteins for the selective detection of molecular analytes remain scarce. Addressing these limitations, a general strategy is presented to immobilize nanobodies in a directional fashion onto the surface of track-etched nanopores exploiting copper-free click reactions and site-specific protein conjugation systems. The functional immobilization of three different nanobodies is demonstrated in ligand binding experiments with green fluorescent protein, mCherry, and α-amylase (α-Amy) serving as molecular analytes. Ligand binding is resolved using a combination of optical and electrical recordings displaying quantitative dose–response curves. Furthermore, a change in surface charge density is identified as the predominant molecular factor that underlies quantitative dose–responses for the three different protein analytes in nanoconfined geometries. The devised strategy should pave the way for the systematic functionalization of nanopore surfaces with biological receptors and their ability to detect a variety of analytes for diagnostic purposes. © 2021 The Authors. Small published by Wiley-VCH GmbH
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| 2. |
- Muller, L. K., et al.
(författare)
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Ultrasensitive and Selective Copper(II) Detection: Introducing a Bioinspired and Robust Sensor
- 2020
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Ingår i: Chemistry-a European Journal. - : Wiley. - 0947-6539 .- 1521-3765. ; 26:39, s. 8511-8517
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Tidskriftsartikel (refereegranskat)abstract
- A nanopore-based Cu-II-sensing system is reported that allows for an ultrasensitive and selective detection of Cu(II)with the possibility for a broad range of applications, for example in medical diagnostics. A fluorescent ATCUN-like peptide 5/6-FAM-Dap-beta-Ala-His is employed to selectively bind Cu(II)ions in the presence of Ni(II)and Zn(II)and was crafted into ion track-etched nanopores. Upon Cu(II)binding the fluorescence of the peptide sensor is quenched, permitting the detection of Cu(II)in solution. The ion transport characteristics of peptide-modified nanopore are shown to be extremely sensitive and selective towards Cu(II)allowing to sense femtomolar Cu(II)concentrations in human urine mimics. Washing with EDTA fully restores the Cu-II-binding properties of the sensor, enabling multiple repetitive measurements. The robustness of the system clearly has the potential to be further developed into an easy-to-use, lab-on-chip Cu-II-sensing device, which will be of great importance for bedside diagnosis and monitor of Cu(II)levels in patients with copper-dysfunctional homeostasis.
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