MULTISPECIFIC MUTATED ANTIBODY FAB FRAGMENTS

摘要

The invention relates to multispecific antibody constructs comprising Fab fragments having mutations at the interface of the CH1 and CL domains, said mutations preventing heavy/light chain mispairing.

主权项

1. A multispecific antigens-binding fragment comprising at least two Fab fragments with different CH1 and CL domains, wherein said Fab fragments are selected from the group consisting of:
a. a wild-type Fab fragment consisting of wild-type CH1 and CL domains of an immunoglobulin, and the VH and VL domains of an antibody recognizing an epitope of interest; b. a mutated Fab fragment consisting of:
i. the VH and VL domains of an antibody recognizing an epitope of interest;ii. a CH1 domain which is derived from the CH1 domain of an immunoglobulin by substitution of the threonine residue at position 192 of said CH1 domain with a glutamic acid residue; andiii. a CL domain which is derived from the CL domain of an immunoglobulin by substitution of the asparagine residue at position 137 of said CL domain with a lysine residue and substitution of the serine residue at position 114 of said CL domain with an alanine residue; c. a mutated Fab fragment consisting of:
i. the VH and VL domains of an antibody recognizing an epitope of interest;ii. a CH1 domain which is derived from the CH1 domain of an immunoglobulin by substitution of the leucine residue at position 143 of said CH1 domain with a glutamine residue and substitution of the serine residue at position 188 of said CH1 domain with a valine residue; andiii. a CL domain which is derived from the CL domain of an immunoglobulin by substitution of the valine residue at position 133 of said CL domain with a threonine residue and substitution of the serine residue at position 176 of said CL domain with an valine residue. d. a mutated Fab fragment consisting of:
i. the VH and VL domains of an antibody recognizing an epitope of interest;ii. a CH1 domain which is derived from the CH1 domain of an IgG immunoglobulin by substitution of the leucine residue at position 124 of said CH1 domain with an alanine residue and substitution of the leucine residue at position 143 of said CH1 domain with a glutamic acid residue;iii. a CL domain which is derived from the CL domain of an IgG immunoglobulin by substitution of the valine residue at position 133 of said CL domain with a tryptophane residue; e. a mutated Fab fragment consisting of:
i. the VH and VL domains of an antibody recognizing an epitope of interest;ii. a CH1 domain which is derived from the CH1 domain of an immunoglobulin by substitution of the valine residue at position 190 of said CH1 domain with an alanine residue; andiii. a CL domain which is derived from the CL domain of an immunoglobulin by substitution of the leucine residue at position 135 of said CL domain with a tryptophane residue, and substitution of the asparagine residue at position 137 of said CL domain with an alanine residue;each Fab fragment recognizing a different epitope of interest, and said Fab fragments being tandemly arranged in any order, the C-terminal end of the CH1 domain of a first Fab fragment being linked to the N-terminal end of the VH domain of the following Fab fragment through a polypeptide linker.