title: Novel multispecific heterodimeric antibody format allowing modular assembly of variable domain fragments.
abstract: 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 Vκ1/VH3-consensus Fv framework with the corresponding germ-line sequence of a λ-type VLchain. We then used this stable Fv framework to generate a novel heterodimeric multispecific antibody format that assembles by cognate VL/VHassociations 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.
authors: a Numab AG, Wadenswil , Switzerland.|b Cartilage Engineering & Regeneration Lab, Department of Health , Science & Technology, The Swiss Federal Institute of Technology (ETH) , Zurich , Switzerland.
authors: Timothy J
authors: TJ
authors: Egan
authors: a Numab AG, Wadenswil , Switzerland.
authors: Dania
authors: D
authors: Diem
authors: a Numab AG, Wadenswil , Switzerland.
authors: Richard
authors: R
authors: Weldon
authors: a Numab AG, Wadenswil , Switzerland.
authors: Tessa
authors: T
authors: Neumann
authors: a Numab AG, Wadenswil , Switzerland.
authors: Sebastian
authors: S
authors: Meyer
authors: a Numab AG, Wadenswil , Switzerland.
authors: David M
authors: DM
authors: Urech
mesh_terms: D000906:Antibodies / Q000737:chemistry* / Q000276:immunology; D000918:Antibody Specificity; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000737:chemistry* / Q000276:immunology; D011487:Protein Conformation; D055550:Protein Stability; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology
chemical_list: D000906:Antibodies; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins
keywords: Antibody; antibody fragment; framework; heterodimer; humanization; multispecific; tetraspecific