Humanization of Chicken-Derived Antibodies by Yeast Surface Display.
| Autor: | Bogen JP; Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Darmstadt, Germany.; Ferring Darmstadt Laboratory, Biologics Technology and Development, Darmstadt, Germany., Elter A; Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Darmstadt, Germany.; Merck Lab @ Technical University of Darmstadt, Darmstadt, Germany., Grzeschik J; Ferring Darmstadt Laboratory, Biologics Technology and Development, Darmstadt, Germany., Hock B; Ferring International Center S.A., Saint-Prex, Switzerland., Kolmar H; Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Darmstadt, Germany. Kolmar@Biochemie-TUD.de. |
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| Jazyk: | angličtina |
| Zdroj: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2022; Vol. 2491, pp. 335-360. |
| DOI: | 10.1007/978-1-0716-2285-8_18 |
| Abstrakt: | Chicken-derived antibodies emerged as a promising tool for diagnostic and therapeutic usage. Due to the phylogenetic distance between birds and mammals, chicken immunization campaigns with human antigens result in a chicken antibody (IgY) repertoire targeting epitopes not addressed by rodent-derived antibodies. However, this phylogenetic distance accounts for a low homology of IgY molecules to human antibodies, resulting in potential immunogenicity and thus excluding IgYs from therapeutic applications. Herein, we describe a straightforward method to efficiently humanize chicken-derived antibodies by a CDR-grafting-based approach, including a simultaneous randomization of key residues (Vernier residues). Utilizing yeast surface display (YSD) and fluorescence-activated cell sorting (FACS), yeast cells displaying functional humanized scFvs and Fab variants are isolated, and subsequent next-generation sequencing (NGS) enables the identification of humanized antibody variants with restored affinity and beneficial protein characteristics. (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.) |
| Databáze: | MEDLINE |
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