mAbs (Dec 2022)

Beyond bispecificity: Controlled Fab arm exchange for the generation of antibodies with multiple specificities

  • Desislava Yanakieva,
  • Lukas Pekar,
  • Andreas Evers,
  • Markus Fleischer,
  • Stephan Keller,
  • Dirk Mueller-Pompalla,
  • Lars Toleikis,
  • Harald Kolmar,
  • Stefan Zielonka,
  • Simon Krah

DOI
https://doi.org/10.1080/19420862.2021.2018960
Journal volume & issue
Vol. 14, no. 1

Abstract

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Controlled Fab arm exchange (cFAE) has proven to be a generic and versatile technology for the efficient generation of IgG-like bispecific antibodies (DuoBodies or DBs), with several in clinical development and one product, amivantamab, approved by the Food and Drug Administration. In this study, we expand the cFAE-toolbox by incorporating VHH-modules at the C-termini of DB-IgGs, termed DB-VHHs. This approach enables the combinatorial generation of tri- and tetraspecific molecules with flexible valencies in a straightforward fashion. Using cFAE, a variety of multispecific molecules was produced and assessed for manufacturability and physicochemical characteristics. In addition, we were able to generate DB-VHHs that efficiently triggered natural killer cell mediated lysis of tumor cells, demonstrating the utility of this format for potential therapeutic applications.

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