Design of orthogonal constant domain interfaces to aid proper heavy/light chain pairing of bispecific antibodies.

ABSTRACT: The correct pairing of cognate heavy and light chains is critical to the efficient manufacturing of IgG-like bispecific antibodies (bsAbs) from a single host cell. We present a general solution for the elimination of heavy chain (HC):light chain (LC) mispairs in bsAbs with κ LCs via the use of two orthogonal constant domain (C_H1:Cκ) interfaces comprising computationally designed amino acid substitutions. Substitutions were designed by Rosetta to introduce novel hydrogen bond (H-bond) networks at the C_H1:Cκ interface, followed by Rosetta energy calculations to identify designs with enhanced pairing specificity and interface stability. Our final design, featuring a total of 11 amino acid substitutions across two Fab constant regions, was tested on a set of six IgG-like

SUBMITTER: Barlow KA 

PROVIDER: S-EPMC11934185 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature