title: Targeted Reconstitution of Cytokine Activity upon Antigen Binding using Split Cytokine Antibody Fusion Proteins.
abstract: The targeted assembly of antibody products upon antigen binding represents a novel strategy for the reconstitution of potent therapeutic activity at the site of disease, sparing healthy tissues. We demonstrate that interleukin-12, a heterodimeric pro-inflammatory cytokine consisting of the disulfide-linked p40 and p35 subunits, can be reconstituted by sequential reassembly of fusion proteins based on antibody fragments and interleukin-12 subunit mutants. Analysis of the immunostimulatory properties of interleukin-12 and its derivatives surprisingly revealed that the mutated p35 subunit partially retained the activity of the parental cytokine, whereas the p40 subunit alone was not able to stimulate T cells or natural killer cells. The concept of stepwise antibody-based reassembly of split cytokines could be useful for the development of other anticancer therapeutics with improved safety and tolerability.
authors.affiliation: From the Department of Chemistry and Applied Biosciences, Swiss Federal Institute of Technology (ETH Zürich), Vladimir-Prelog-Weg 1-5/10, CH-8093 Zürich, Switzerland dario.venetz@pharma.ethz.ch.
authors.forename: Dario
authors.initials: D
authors.lastname: Venetz
authors.affiliation: From the Department of Chemistry and Applied Biosciences, Swiss Federal Institute of Technology (ETH Zürich), Vladimir-Prelog-Weg 1-5/10, CH-8093 Zürich, Switzerland.
authors.forename: Danil
authors.initials: D
authors.lastname: Koovely
authors.affiliation: From the Department of Chemistry and Applied Biosciences, Swiss Federal Institute of Technology (ETH Zürich), Vladimir-Prelog-Weg 1-5/10, CH-8093 Zürich, Switzerland.
authors.forename: Bruce
authors.initials: B
authors.lastname: Weder
authors.affiliation: From the Department of Chemistry and Applied Biosciences, Swiss Federal Institute of Technology (ETH Zürich), Vladimir-Prelog-Weg 1-5/10, CH-8093 Zürich, Switzerland dario.neri@pharma.ethz.ch.
authors.forename: Dario
authors.initials: D
authors.lastname: Neri
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D006801:Humans; D053765:Interleukin-12 Subunit p35 / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D053762:Interleukin-12 Subunit p40 / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology
chemical_list: D000906:Antibodies; C508858:IL12A protein, human; C500879:IL12B protein, human; C508859:Il12a protein, mouse; D053765:Interleukin-12 Subunit p35; D053762:Interleukin-12 Subunit p40; D011993:Recombinant Fusion Proteins
keywords: activity reconstitution; activity-on-demand; antibody; cancer; cytokine; immunotherapy; interleukin-12; pharmacology; split protein; tumor targeting