Elter, Adrian ; Bogen, Jan P. ; Hinz, Steffen C. ; Fiebig, David ; Macarrón Palacios, Arturo ; Grzeschik, Julius ; Hock, Björn ; Kolmar, Harald (2024)
Humanization of Chicken‐Derived scFv Using Yeast Surface Display and NGS Data Mining.
In: Biotechnology Journal : Systems & Synthetic Biology, Nanobiotech, Medicine, 2021, 16 (3)
doi: 10.26083/tuprints-00017792
Artikel, Zweitveröffentlichung, Verlagsversion
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Kurzbeschreibung (Abstract)
Generation of high‐affinity monoclonal antibodies by immunization of chickens is a valuable strategy, particularly for obtaining antibodies directed against epitopes that are conserved in mammals. A generic procedure is established for the humanization of chicken‐derived antibodies. To this end, high‐affinity binders of the epidermal growth factor receptor extracellular domain are isolated from immunized chickens using yeast surface display. Complementarity determining regions (CDRs) of two high‐affinity binders are grafted onto a human acceptor framework. Simultaneously, Vernier zone residues, responsible for spatial CDR arrangement, are partially randomized. A yeast surface display library comprising ≈300 000 variants is screened for high‐affinity binders in the scFv and Fab formats. Next‐generation sequencing discloses humanized antibody variants with restored affinity and improved protein characteristics compared to the parental chicken antibodies. Furthermore, the sequencing data give new insights into the importance of antibody format, used during the humanization process. Starting from the antibody repertoire of immunized chickens, this work features an effective and fast high‐throughput approach for the generation of multiple humanized antibodies with potential therapeutic relevance.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2024 |
| Autor(en): | Elter, Adrian ; Bogen, Jan P. ; Hinz, Steffen C. ; Fiebig, David ; Macarrón Palacios, Arturo ; Grzeschik, Julius ; Hock, Björn ; Kolmar, Harald |
| Art des Eintrags: | Zweitveröffentlichung |
| Titel: | Humanization of Chicken‐Derived scFv Using Yeast Surface Display and NGS Data Mining |
| Sprache: | Englisch |
| Publikationsjahr: | 12 Februar 2024 |
| Ort: | Darmstadt |
| Publikationsdatum der Erstveröffentlichung: | 2021 |
| Ort der Erstveröffentlichung: | Weinheim |
| Verlag: | Wiley-VCH |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Biotechnology Journal : Systems & Synthetic Biology, Nanobiotech, Medicine |
| Jahrgang/Volume einer Zeitschrift: | 16 |
| (Heft-)Nummer: | 3 |
| Kollation: | 9 Seiten |
| DOI: | 10.26083/tuprints-00017792 |
| URL / URN: | https://tuprints.ulb.tu-darmstadt.de/17792 |
| Zugehörige Links: | |
| Herkunft: | Zweitveröffentlichung DeepGreen |
| Kurzbeschreibung (Abstract): | Generation of high‐affinity monoclonal antibodies by immunization of chickens is a valuable strategy, particularly for obtaining antibodies directed against epitopes that are conserved in mammals. A generic procedure is established for the humanization of chicken‐derived antibodies. To this end, high‐affinity binders of the epidermal growth factor receptor extracellular domain are isolated from immunized chickens using yeast surface display. Complementarity determining regions (CDRs) of two high‐affinity binders are grafted onto a human acceptor framework. Simultaneously, Vernier zone residues, responsible for spatial CDR arrangement, are partially randomized. A yeast surface display library comprising ≈300 000 variants is screened for high‐affinity binders in the scFv and Fab formats. Next‐generation sequencing discloses humanized antibody variants with restored affinity and improved protein characteristics compared to the parental chicken antibodies. Furthermore, the sequencing data give new insights into the importance of antibody format, used during the humanization process. Starting from the antibody repertoire of immunized chickens, this work features an effective and fast high‐throughput approach for the generation of multiple humanized antibodies with potential therapeutic relevance. |
| Freie Schlagworte: | chicken antibody, fluorescence‐activated cell sorting, humanization, next‐generation sequencing, yeast surface display |
| ID-Nummer: | Artikel-ID: 2000231 |
| Status: | Verlagsversion |
| URN: | urn:nbn:de:tuda-tuprints-177923 |
| 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: | 12 Feb 2024 13:42 |
| Letzte Änderung: | 13 Feb 2024 07:37 |
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| Suche nach Titel in: | TUfind oder in Google |
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- Humanization of Chicken‐Derived scFv Using Yeast Surface Display and NGS Data Mining. (deposited 12 Feb 2024 13:42) [Gegenwärtig angezeigt]
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