Novel multispecific heterodimeric antibody format allowing modular assembly of variable domain fragments

• Published in: mAbs Vol. 9; no. 1; pp. 68 - 84
• Main Authors: Egan, Timothy J., Diem, Dania, Weldon, Richard, Neumann, Tessa, Meyer, Sebastian, Urech, David M.
• Format: Report
• Language: English
• Published: Taylor & Francis 02.01.2017
• Subjects: Antibody; antibody fragment; framework; heterodimer; humanization; multispecific; tetraspecific
• ISSN: 1942-0862; 1942-0870
• DOI: 10.1080/19420862.2016.1248012

Multispecific antibody formats provide a promising platform for the development of novel therapeutic concepts that could facilitate the generation of safer, more effective pharmaceuticals. However, the production and use of such antibody-based multispecifics is often made complicated by: 1) the instability of the antibody fragments of which they consist, 2) undesired inter-subunit associations, and 3) the need to include recombinant heterodimerization domains that confer distribution-impairing bulk or enhance immunogenicity. In this paper, we describe a broadly-applicable method for the stabilization of human or humanized antibody Fv fragments that entails replacing framework region IV of a V κ 1/V H 3-consensus Fv framework with the corresponding germ-line sequence of a λ-type V L chain. We then used this stable Fv framework to generate a novel heterodimeric multispecific antibody format that assembles by cognate V L /V H associations between 2 split variable domains in the core of the complex. This format, termed multispecific antibody-based therapeutics by cognate heterodimerization (MATCH), can be applied to produce homogeneous and highly stable antibody-derived molecules that simultaneously bind 4 distinct antigens. The heterodimeric design of the MATCH format allows efficient in-format screening of binding domain combinations that result in maximal cooperative activity.