Heterodimeric antibody Fc-containing proteins and methods for production thereof

Inventors

Labrijn, Aran Frank • Meesters, Joyce I. • Van Den Bremer, Ewald T. J. • Neijssen, Joost J. • van Berkel, Patrick • De Goeij, Bart • Vink, Tom • van de Winkel, Jan • Schuurman, Janine • Parren, Paul

Assignees

Genmab AS

Abstract

Novel heterodimeric antibody-Fc-containing proteins, such as bispecific antibodies, and novel methods for producing such proteins.

Core Innovation

The invention relates to a heterodimeric antibody comprising a first and a second heavy chain, each comprising a first or second Fc region with CH3 regions of human IgG4. The first Fc region comprises a first CH3 region of human IgG4, while the second Fc region comprises a second CH3 region of human IgG4 comprising amino acid substitutions at positions 405 and 409 according to the EU Index, and the sequences of the first and second CH3 regions are different.

In certain embodiments, both heavy chains comprise a hinge region comprising a Cys-Pro-Pro-Cys (SEQ ID NO:4) sequence, and the antibody is human IgG4 isotype except for the specified substitutions and hinge region. The heavy-chain Fc CH3-region asymmetry with amino acid substitutions at positions 405 and 409 is used to strengthen heterodimeric CH3–CH3 interaction relative to homodimeric interactions and to achieve directional heterodimeric Fc formation.

The disclosed antibodies are described as stable heterodimeric Fc-containing proteins, including bispecific antibodies, produced through in vitro Fab-arm (half-molecule) exchange or by co-expressing nucleic acid constructs encoding the first and second heavy chains in a host cell and obtaining the antibody from cell culture. In further embodiments, the first and second heavy chains bind different epitopes, and heavy-chain/light-chain pairing remains intact while undesired heavy/light mispairing is avoided.

Claims Coverage

The consolidated claim coverage includes four independent claim families and centers on a heterodimeric antibody with human IgG4 CH3-region differences defined by EU Index substitutions, hinge-region constraints, and production or formation conditions. The main inventive features total five core components.

Overall, the claim coverage centers on heterodimeric Fc-containing antibodies using human IgG4 CH3-region asymmetry with amino acid substitutions at EU Index positions 405 and 409, combined with hinge-region disulfide-bond isomerization to obtain the heterodimeric antibody, with optional constraints including hinge-region Cys-Pro-Pro-Cys (SEQ ID NO: 4), full-length light chains, human IgG4 isotype framework limitations, and different epitope binding.

Stated Advantages

Documented Applications