	US 11,965,030 B2
	Multispecific binding proteins with mutant fab domains
Marta Amaral, Frankfurt am Main (DE); Christian Beil, Frankfurt am Main (DE); Ingo Focken, Frankfurt am Main (DE); Gerhard Hessler, Frankfurt am Main (DE); Soraya Hoelper, Frankfurt am Main (DE); Jennifer Jung, Frankfurt am Main (DE); Christian Lange, Frankfurt am Main (DE); Wulf-Dirk Leuschner, Frankfurt am Main (DE); Ercole Rao, Frankfurt am Main (DE); Garima Tiwari, Frankfurt am Main (DE); and Sandra Weil, Frankfurt am Main (DE)
Assigned to SANOFI, Paris (FR)
Filed by SANOFI, Paris (FR)
Filed on Dec. 23, 2019, as Appl. No. 16/725,228.
Claims priority of application No. 18306843 (EP), filed on Dec. 24, 2018; and application No. 19305812 (EP), filed on Jun. 21, 2019.
Prior Publication US 2020/0262926 A1, Aug. 20, 2020
Int. Cl. C07K 16/28 (2006.01)

CPC C07K 16/2878 (2013.01) [C07K 16/2827 (2013.01); C07K 2317/31 (2013.01); C07K 2317/522 (2013.01); C07K 2317/524 (2013.01); C07K 2317/526 (2013.01); C07K 2317/53 (2013.01); C07K 2317/55 (2013.01); C07K 2317/94 (2013.01)]

15 Claims

1. A multispecific antigen-binding protein comprising at least two VL regions respectively paired with at least two VH regions to form at least two antigen-binding sites and at least two CH1 regions respectively paired with two CL regions,

wherein at least one CH1/CL pair comprises a CH1 region comprising K221E and K228D mutations at Kabat positions 221 and 228 and CL region comprising D122K and E123K mutations at Kabat positions 122 and 123 to facilitate pairing, and

wherein at least one VH/VL pair comprises opposite charged mutations to facilitate pairing, said opposite charged mutations comprising (1) a mutated residue in the VH region at Kabat position 39 selected from E, D, K, R, or H, and (2) a mutated residue in the VL region at Kabat position 38 selected from E, D, K, R, or H, and

wherein the mutated residue in the VH region has an opposite charge from the mutated residue in the VL region.