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. 2023:2681:231-248.
doi: 10.1007/978-1-0716-3279-6_13.

Affinity Maturation of the Natural Ligand (B7-H6) for Natural Cytotoxicity Receptor NKp30 by Yeast Surface Display

Stefan Zielonka  1   2 , Simon Krah  1 , Paul Arras  1 , Britta Lipinski  1   2 , Jasmin Zimmermann  1   2 , Ammelie Svea Boje  3 , Katja Klausz  3 , Matthias Peipp  3 , Lukas Pekar  4
Affiliations

Affinity Maturation of the Natural Ligand (B7-H6) for Natural Cytotoxicity Receptor NKp30 by Yeast Surface Display

Stefan Zielonka et al. Methods Mol Biol. 2023.

Abstract

In recent years, the development of bispecific antibodies (bsAbs) has experienced tremendous progress for disease treatment, and consequently, a plethora of bsAbs is currently scrutinized in clinical trials. Besides antibody scaffolds, multifunctional molecules referred to as immunoligands have been developed. These molecules typically harbor a natural ligand entity for the engagement of a specific receptor, while binding to the additional antigen is facilitated by an antibody-derived paratope. Immunoligands can be exploited to conditionally activate immune cells, e.g., natural killer (NK) cells, in the presence of tumor cells, ultimately causing target-dependent tumor cell lysis. However, many ligands naturally show only moderate affinities toward their cognate receptor, potentially hampering killing capacities of immunoligands. Herein, we provide protocols for yeast surface display-based affinity maturation of B7-H6, the natural ligand of NK cell-activating receptor NKp30.

Keywords: Affinity maturation; B7-H6; Bispecifics; Fluorescence-activated cell sorting; Immunoligand; NKp30; Protein engineering; Yeast surface display.

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