Generation of a symmetrical trispecific NK cell engager based on a two-in-one antibody

Julia Harwardt; Stefania C Carrara; Jan P Bogen; Katrin Schoenfeld; Julius Grzeschik; Björn Hock; Harald Kolmar

doi:10.3389/fimmu.2023.1170042

Generation of a symmetrical trispecific NK cell engager based on a two-in-one antibody

Front Immunol. 2023 Apr 4:14:1170042. doi: 10.3389/fimmu.2023.1170042. eCollection 2023.

Authors

Julia Harwardt¹, Stefania C Carrara^{1

2}, Jan P Bogen^{1

2}, Katrin Schoenfeld¹, Julius Grzeschik³, Björn Hock¹, Harald Kolmar^{1

4}

Affiliations

¹ Institute for Organic Chemistry and Biochemistry, Technical University of Darmstadt, Darmstadt, Germany.
² Biologics Technology and Development, Ferring Darmstadt Laboratory, Darmstadt, Germany.
³ Biologics Technology and Development, Ferring Biologics Innovation Centre, Epalinges, Switzerland.
⁴ Centre for Synthetic Biology, Technical University of Darmstadt, Darmstadt, Germany.

Abstract

To construct a trispecific IgG-like antibody at least three different binding moieties need to be combined, which results in a complex architecture and challenging production of these molecules. Here we report for the first time the construction of trispecific natural killer cell engagers based on a previously reported two-in-one antibody combined with a novel anti-CD16a common light chain module identified by yeast surface display (YSD) screening of chicken-derived immune libraries. The resulting antibodies simultaneously target epidermal growth factor receptor (EGFR), programmed death-ligand 1 (PD-L1) and CD16a with two Fab fragments, resulting in specific cellular binding properties on EGFR/PD-L1 double positive tumor cells and a potent ADCC effect. This study paves the way for further development of multispecific therapeutic antibodies derived from avian immunization with desired target combinations, valencies, molecular symmetries and architectures.

Keywords: NK cell engager; common light chain; trispecific antibody; two-in-one antibody; yeast surface display.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antibodies / metabolism
B7-H1 Antigen* / metabolism
ErbB Receptors / metabolism
Killer Cells, Natural*

Substances

B7-H1 Antigen
Antibodies
ErbB Receptors

Grants and funding

This work was supported by the Ferring Darmstadt Laboratories at Technical University of Darmstadt and by the department of GPRD at Ferring Holding S.A., Saint-Prex. The funders had no role in study design, data collection and analysis, interpretation of data, decision to publish, or preparation of the manuscript.