Oloketuyi, Sandra ; Bernedo, Robert ; Christmann, Andreas ; Borkowska, Justyna ; Cazzaniga, Giulia ; Schuchmann, Horst Wilhelm ; Niedziółka-Jönsson, Joanna ; Szot-Karpińska, Katarzyna ; Kolmar, Harald ; Marco, Ario de (2022)
Native llama Nanobody Library Panning Performed by Phage and Yeast Display Provides Binders Suitable for C-Reactive Protein Detection.
In: Biosensors, 11 (12)
doi: 10.3390/bios11120496
Artikel, Bibliographie
Dies ist die neueste Version dieses Eintrags.
Kurzbeschreibung (Abstract)
C-reactive protein (CRP) is an inflammation biomarker that should be quantified accurately during infections and healing processes. Nanobodies are good candidates to replace conventional antibodies in immunodiagnostics due to their inexpensive production, simple engineering, and the possibility to obtain higher binder density on capture surfaces. Starting from the same pre-immune library, we compared the selection output resulting from two independent panning strategies, one exclusively exploiting the phage display and another in which a first round of phage display was followed by a second round of yeast display. There was a partial output convergence between the two methods, since two clones were identified using both panning protocols but the first provided several further different sequences, whereas the second favored the recovery of many copies of few clones. The isolated anti-CRP nanobodies had affinity in the low nanomolar range and were suitable for ELISA and immunoprecipitation. One of them was fused to SpyTag and exploited in combination with SpyCatcher as the immunocapture element to quantify CRP using electrochemical impedance spectroscopy. The sensitivity of the biosensor was calculated as low as 0.21 μg/mL.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2022 |
| Autor(en): | Oloketuyi, Sandra ; Bernedo, Robert ; Christmann, Andreas ; Borkowska, Justyna ; Cazzaniga, Giulia ; Schuchmann, Horst Wilhelm ; Niedziółka-Jönsson, Joanna ; Szot-Karpińska, Katarzyna ; Kolmar, Harald ; Marco, Ario de |
| Art des Eintrags: | Bibliographie |
| Titel: | Native llama Nanobody Library Panning Performed by Phage and Yeast Display Provides Binders Suitable for C-Reactive Protein Detection |
| Sprache: | Englisch |
| Publikationsjahr: | 2022 |
| Verlag: | MDPI |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Biosensors |
| Jahrgang/Volume einer Zeitschrift: | 11 |
| (Heft-)Nummer: | 12 |
| Kollation: | 14 Seiten |
| DOI: | 10.3390/bios11120496 |
| Zugehörige Links: | |
| Kurzbeschreibung (Abstract): | C-reactive protein (CRP) is an inflammation biomarker that should be quantified accurately during infections and healing processes. Nanobodies are good candidates to replace conventional antibodies in immunodiagnostics due to their inexpensive production, simple engineering, and the possibility to obtain higher binder density on capture surfaces. Starting from the same pre-immune library, we compared the selection output resulting from two independent panning strategies, one exclusively exploiting the phage display and another in which a first round of phage display was followed by a second round of yeast display. There was a partial output convergence between the two methods, since two clones were identified using both panning protocols but the first provided several further different sequences, whereas the second favored the recovery of many copies of few clones. The isolated anti-CRP nanobodies had affinity in the low nanomolar range and were suitable for ELISA and immunoprecipitation. One of them was fused to SpyTag and exploited in combination with SpyCatcher as the immunocapture element to quantify CRP using electrochemical impedance spectroscopy. The sensitivity of the biosensor was calculated as low as 0.21 μg/mL. |
| Freie Schlagworte: | phage display, yeast display, nanobodies, biopanning, CRP |
| Sachgruppe der Dewey Dezimalklassifikatin (DDC): | 500 Naturwissenschaften und Mathematik > 540 Chemie 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie |
| Fachbereich(e)/-gebiet(e): | 07 Fachbereich Chemie 07 Fachbereich Chemie > Clemens-Schöpf-Institut > Fachgebiet Biochemie 07 Fachbereich Chemie > Clemens-Schöpf-Institut |
| Hinterlegungsdatum: | 02 Aug 2024 12:40 |
| Letzte Änderung: | 02 Aug 2024 12:40 |
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| Suche nach Titel in: | TUfind oder in Google |
Verfügbare Versionen dieses Eintrags
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Native llama Nanobody Library Panning Performed by Phage and Yeast Display Provides Binders Suitable for C-Reactive Protein Detection. (deposited 29 Apr 2022 09:08)
- Native llama Nanobody Library Panning Performed by Phage and Yeast Display Provides Binders Suitable for C-Reactive Protein Detection. (deposited 02 Aug 2024 12:40) [Gegenwärtig angezeigt]
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