Patent ID: 8987417
Filing Date: 2015-03-24
Classification: A61K,A61P,C07K,G01N

Abstract:
1. A method of producing a covalently dimerized bivalent binding agent comprising: (a) providing a first fusion protein comprising a first binding domain fused to a first coiled-coil, a first cysteine residue between the first binding domain and the first coiled-coil, and a first cleavage site between the first cysteine residue and the first coiled-coil; (b) providing a second fusion protein comprising a second binding domain fused to a second coiled-coil capable of dimerizing non-covalently with the first coiled-coil, a second cysteine residue between the second binding domain and the second coiled-coil, and a second cleavage site between the second cysteine residue and the second coiled-coil; (c) mixing the first fusion protein with the second fusion protein, the first and second coiled-coils non-covalently dimerizing to bring the first and second cysteine residues into proximity to form a disulphide bond between the first and second cysteine residues; and, (d) excising the first and second coiled-coils by cleaving at the cleavage sites to produce the covalently dimerized bivalent binding agent having a disulphide bond between the first and second binding domains.