Duznovic, Ivana ; Gräwe, Alexander ; Weber, Wadim ; Müller, Lena K. ; Ali, Mubarak ; Ensinger, Wolfgang ; Tietze, Alesia ; Stein, Viktor (2023)
Ultrasensitive and Selective Protein Recognition with Nanobody‐Functionalized Synthetic Nanopores.
In: Small : nano micro, 2021, 17 (33)
doi: 10.26083/tuprints-00020980
Artikel, Zweitveröffentlichung, Verlagsversion
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Kurzbeschreibung (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.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2023 |
| Autor(en): | Duznovic, Ivana ; Gräwe, Alexander ; Weber, Wadim ; Müller, Lena K. ; Ali, Mubarak ; Ensinger, Wolfgang ; Tietze, Alesia ; Stein, Viktor |
| Art des Eintrags: | Zweitveröffentlichung |
| Titel: | Ultrasensitive and Selective Protein Recognition with Nanobody‐Functionalized Synthetic Nanopores |
| Sprache: | Englisch |
| Publikationsjahr: | 22 Dezember 2023 |
| Ort: | Darmstadt |
| Publikationsdatum der Erstveröffentlichung: | 2021 |
| Ort der Erstveröffentlichung: | Weinheim |
| Verlag: | Wiley-VCH |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Small : nano micro |
| Jahrgang/Volume einer Zeitschrift: | 17 |
| (Heft-)Nummer: | 33 |
| Kollation: | 9 Seiten |
| DOI: | 10.26083/tuprints-00020980 |
| URL / URN: | https://tuprints.ulb.tu-darmstadt.de/20980 |
| Zugehörige Links: | |
| Herkunft: | Zweitveröffentlichung DeepGreen |
| Kurzbeschreibung (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. |
| Freie Schlagworte: | biosensors, current rectification, molecular diagnostics, nanobodies, nanofluidic devices, synthetic nanopores, track‐etched membrane |
| ID-Nummer: | 2101066 |
| Status: | Verlagsversion |
| URN: | urn:nbn:de:tuda-tuprints-209801 |
| Sachgruppe der Dewey Dezimalklassifikatin (DDC): | 500 Naturwissenschaften und Mathematik > 540 Chemie 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie |
| Fachbereich(e)/-gebiet(e): | 10 Fachbereich Biologie 10 Fachbereich Biologie > Protein Engineering of Ion Conducting Nanopores 11 Fachbereich Material- und Geowissenschaften 11 Fachbereich Material- und Geowissenschaften > Materialwissenschaft 11 Fachbereich Material- und Geowissenschaften > Materialwissenschaft > Fachgebiet Materialanalytik Interdisziplinäre Forschungsprojekte Interdisziplinäre Forschungsprojekte > Centre for Synthetic Biology 07 Fachbereich Chemie 07 Fachbereich Chemie > Clemens-Schöpf-Institut > Fachgebiet Biochemie |
| Hinterlegungsdatum: | 22 Dez 2023 13:18 |
| Letzte Änderung: | 28 Dez 2023 07:33 |
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| Suche nach Titel in: | TUfind oder in Google |
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- Ultrasensitive and Selective Protein Recognition with Nanobody‐Functionalized Synthetic Nanopores. (deposited 22 Dez 2023 13:18) [Gegenwärtig angezeigt]
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