Bogen, Jan P. ; Storka, Juliana ; Yanakieva, Desislava ; Fiebig, David ; Grzeschik, Julius ; Hock, Björn ; Kolmar, Harald (2020)
Isolation of common light chain antibodies from immunized chickens using yeast biopanning and fluorescence‐activated cell sorting.
In: Biotechnology Journal : Systems and Synthetic Biology, Nanobiotech, Medicine, 16 (3)
doi: 10.1002/biot.202000240
Artikel, Bibliographie
Dies ist die neueste Version dieses Eintrags.
Kurzbeschreibung (Abstract)
The phylogenetic distance between chickens and humans accounts for a strong immune response and a broader epitope coverage compared to rodent immunization approaches. Here the authors report the isolation of common light chain (cLC)‐based chicken monoclonal antibodies from an anti‐epidermal growth factor receptor (EGFR) immune library utilizing yeast surface display in combination with yeast biopanning and fluorescence‐activated cell sorting (FACS). For the selection of high‐affinity antibodies, a yeast cell library presenting cLC‐comprising fragment antigen binding (Fab) fragments is panned against hEGFR‐overexpressing A431 cells. The resulting cell–cell‐complexes are sorted by FACS resulting in gradual enrichment of EGFR‐binding Fabs in three sorting rounds. The isolated antibodies share the same light chain and show high specificity for EGFR, resulting in selective binding to A431 cells with notable EC50 values. All identified antibodies show very good aggregation propensity profiles and thermostabilities. Additionally, epitope binning demonstrates that these cLC antibodies cover a broad epitope space. Isolation of antibodies from immunized chickens by yeast cell biopanning makes an addition to the repertoire of methods for antibody library screening, paving the way for the generation of cLC‐based bispecific antibodies against native mammalian receptors.
| Typ des Eintrags: | Artikel | ||||
|---|---|---|---|---|---|
| Erschienen: | 2020 | ||||
| Autor(en): | Bogen, Jan P. ; Storka, Juliana ; Yanakieva, Desislava ; Fiebig, David ; Grzeschik, Julius ; Hock, Björn ; Kolmar, Harald | ||||
| Art des Eintrags: | Bibliographie | ||||
| Titel: | Isolation of common light chain antibodies from immunized chickens using yeast biopanning and fluorescence‐activated cell sorting | ||||
| Sprache: | Englisch | ||||
| Publikationsjahr: | 2020 | ||||
| Ort: | Weinheim | ||||
| Verlag: | Wiley-VCH | ||||
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Biotechnology Journal : Systems and Synthetic Biology, Nanobiotech, Medicine | ||||
| Jahrgang/Volume einer Zeitschrift: | 16 | ||||
| (Heft-)Nummer: | 3 | ||||
| Kollation: | 10 Seiten | ||||
| DOI: | 10.1002/biot.202000240 | ||||
| Zugehörige Links: | |||||
| Kurzbeschreibung (Abstract): | The phylogenetic distance between chickens and humans accounts for a strong immune response and a broader epitope coverage compared to rodent immunization approaches. Here the authors report the isolation of common light chain (cLC)‐based chicken monoclonal antibodies from an anti‐epidermal growth factor receptor (EGFR) immune library utilizing yeast surface display in combination with yeast biopanning and fluorescence‐activated cell sorting (FACS). For the selection of high‐affinity antibodies, a yeast cell library presenting cLC‐comprising fragment antigen binding (Fab) fragments is panned against hEGFR‐overexpressing A431 cells. The resulting cell–cell‐complexes are sorted by FACS resulting in gradual enrichment of EGFR‐binding Fabs in three sorting rounds. The isolated antibodies share the same light chain and show high specificity for EGFR, resulting in selective binding to A431 cells with notable EC50 values. All identified antibodies show very good aggregation propensity profiles and thermostabilities. Additionally, epitope binning demonstrates that these cLC antibodies cover a broad epitope space. Isolation of antibodies from immunized chickens by yeast cell biopanning makes an addition to the repertoire of methods for antibody library screening, paving the way for the generation of cLC‐based bispecific antibodies against native mammalian receptors. |
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| Alternatives oder übersetztes Abstract: |
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| Freie Schlagworte: | antibody discovery, cell panning, common light chain, fluorescence‐activated cell sorting, yeast display | ||||
| ID-Nummer: | Artikel-ID: 2000240 | ||||
| Sachgruppe der Dewey Dezimalklassifikatin (DDC): | 500 Naturwissenschaften und Mathematik > 540 Chemie 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie |
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| Fachbereich(e)/-gebiet(e): | 07 Fachbereich Chemie 07 Fachbereich Chemie > Clemens-Schöpf-Institut > Fachgebiet Biochemie 07 Fachbereich Chemie > Clemens-Schöpf-Institut |
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| Hinterlegungsdatum: | 31 Jan 2024 06:59 | ||||
| Letzte Änderung: | 31 Jan 2024 06:59 | ||||
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| Suche nach Titel in: | TUfind oder in Google | ||||
Verfügbare Versionen dieses Eintrags
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Isolation of Common Light Chain Antibodies from Immunized Chickens Using Yeast Biopanning and Fluorescence‐Activated Cell Sorting. (deposited 30 Jan 2024 13:26)
- Isolation of common light chain antibodies from immunized chickens using yeast biopanning and fluorescence‐activated cell sorting. (deposited 31 Jan 2024 06:59) [Gegenwärtig angezeigt]
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