Cytokine-based therapies and methods

A method for developing a multi-specific binding molecule that redirects an active form of a cytokine to a target cell or a target tissue of interest, the method comprising:

• (a) providing a panel that comprises a plurality of cytokine-binding domains that bind to the cytokine and a plurality of target-binding domains that bind to a target molecule, wherein the target molecule is a marker on the target cell in the target tissue;
• (b) screening the panel using an assay that measures the ability of the cytokine, when complexed with at least one cytokine-binding domain, to bind to and/or agonize its cognate receptor compared to the ability of an unbound cytokine to bind to and/or agonize its cognate receptor;
• (c) screening the plurality of target-binding domains for binding to the target molecule;
• (d) selecting at least one cytokine-binding domain among the panel that does not block or only partially blocks the ability of the cytokine to bind to and/or agonize its cognate receptor;
• (e) generating a panel of multi-specific binding molecules, wherein the panel of multispecific binding molecules comprise the at least one cytokine-binding domain selected in (d) and at least one target-binding domain selected in (c);
• and
• (f) screening the panel of multi-specific binding molecules in an in vitro cell-based assay that measures the ability of the cytokine to bind to and agonize its cognate receptor in the presence of varying amounts of the multi-specific binding molecules.