title: Overcoming a species-specificity barrier in development of an inhibitory antibody targeting a modulator of tumor stroma.
abstract: The secreted disulfide catalyst Quiescin sulfhydryl oxidase-1 (QSOX1) affects extracellular matrix organization and is overexpressed in various adenocarcinomas and associated stroma. Inhibition of extracellular human QSOX1 by a monoclonal antibody decreased tumor cell migration in a cell co-culture model and hence may have therapeutic potential. However, the species specificity of the QSOX1 monoclonal antibody has been a setback in assessing its utility as an anti-metastatic agent in vivo, a common problem in the antibody therapy industry. We therefore used structurally guided engineering to expand the antibody species specificity, improving its affinity toward mouse QSOX1 by at least four orders of magnitude. A crystal structure of the re-engineered variant, complexed with its mouse antigen, revealed that the antibody accomplishes dual-species targeting through altered contacts between its heavy and light chains, plus replacement of bulky aromatics by flexible side chains and versatile water-bridged polar interactions. In parallel, we produced a surrogate antibody targeting mouse QSOX1 that exhibits a new QSOX1 inhibition mode. This set of three QSOX1 inhibitory antibodies is compatible with various mouse models for pre-clinical trials and biotechnological applications. In this study we provide insights into structural blocks to cross-reactivity and set up guideposts for successful antibody design and re-engineering.
authors.affiliation: Department of Structural Biology, Weizmann Institute of Science, Rehovot 7610001, Israel.
authors.forename: Iris
authors.initials: I
authors.lastname: Grossman
authors.affiliation: Department of Structural Biology, Weizmann Institute of Science, Rehovot 7610001, Israel.
authors.forename: Tal
authors.initials: T
authors.lastname: Ilani
authors.affiliation: Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 7610001, Israel.
authors.forename: Sarel Jacob
authors.initials: SJ
authors.lastname: Fleishman
authors.affiliation: Department of Structural Biology, Weizmann Institute of Science, Rehovot 7610001, Israel deborah.fass@weizmann.ac.il.
authors.forename: Deborah
authors.initials: D
authors.lastname: Fass
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000970:Antineoplastic Agents / Q000737:chemistry* / Q000378:metabolism; D002478:Cells, Cultured; D055808:Drug Discovery; D006801:Humans; D007797:Laminin; D051379:Mice; D008958:Models, Molecular; D050862:Oxidoreductases Acting on Sulfur Group Donors / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D013045:Species Specificity
chemical_list: D000911:Antibodies, Monoclonal; D000970:Antineoplastic Agents; D007797:Laminin; D050862:Oxidoreductases Acting on Sulfur Group Donors; C117183:QSOX1 protein, human
keywords: anti-metastatic agents; dual-specificity; laminin; monoclonal antibodies; structure-guided engineering
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