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Humanization of chicken‐derived scFv using yeast surface display and NGS data mining

Elter, Adrian ; Bogen, Jan P. ; Hinz, Steffen C. ; Fiebig, David ; Macarrón Palacios, Arturo ; Grzeschik, Julius ; Hock, Björn ; Kolmar, Harald (2021)
Humanization of chicken‐derived scFv using yeast surface display and NGS data mining.
In: Biotechnology Journal : Systems & Synthetic Biology, Nanobiotech, Medicine, 16 (3)
doi: 10.1002/biot.202000231
Article, Bibliographie

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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.

Item Type: Article
Erschienen: 2021
Creators: Elter, Adrian ; Bogen, Jan P. ; Hinz, Steffen C. ; Fiebig, David ; Macarrón Palacios, Arturo ; Grzeschik, Julius ; Hock, Björn ; Kolmar, Harald
Type of entry: Bibliographie
Title: Humanization of chicken‐derived scFv using yeast surface display and NGS data mining
Language: English
Date: 2021
Place of Publication: Weinheim
Publisher: Wiley-VCH
Journal or Publication Title: Biotechnology Journal : Systems & Synthetic Biology, Nanobiotech, Medicine
Volume of the journal: 16
Issue Number: 3
Collation: 9 Seiten
DOI: 10.1002/biot.202000231
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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.

Uncontrolled Keywords: chicken antibody, fluorescence‐activated cell sorting, humanization, next‐generation sequencing, yeast surface display
Identification Number: Artikel-ID: 2000231
Classification DDC: 500 Science and mathematics > 540 Chemistry
500 Science and mathematics > 570 Life sciences, biology
Divisions: 07 Department of Chemistry
07 Department of Chemistry > Clemens-Schöpf-Institut > Fachgebiet Biochemie
07 Department of Chemistry > Clemens-Schöpf-Institut
Date Deposited: 13 Feb 2024 07:37
Last Modified: 13 Feb 2024 07:38
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