Hetero-Dimeric Immunoglobulins

Abstract

The present invention relates to engineered hetero-dimeric immunoglobulins or fragments thereof and methods of making the same.

1 . A hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof comprising:
 (a) a first engineered immunoglobulin chain comprising an engineered domain with a protein-protein interface, wherein at least one amino acid residue of the protein-protein interface of the parent domain of said first engineered immunoglobulin chain is substituted with an amino acid residue at the equivalent 3D structural position from a protein-protein interface of a donor domain of a first member of the naturally occurring immunoglobulin super-family; and   (b) a second engineered immunoglobulin chain comprising an engineered domain with a protein-protein interface, wherein at least one amino acid residue of the protein-protein interface of the parent domain of said second engineered immunoglobulin chain is substituted with an amino acid residue at the equivalent 3D structural position from a protein-protein interface of a donor domain of a second member of the naturally occurring immunoglobulin super-family,   wherein the engineered domain of the first engineered immunoglobulin chain and the engineered domain of the second engineered immunoglobulin chain are not identical and,   wherein the donor domain of the first member of the naturally occurring immunoglobulin super-family and the donor domain of the second member of the naturally occurring immunoglobulin super-family form a naturally occurring hetero-dimer or a naturally occurring homo-dimer and,   wherein the parent domain of the first and/or the second engineered immunoglobulin chain is a CH3 domain, wherein the amino acid residue which is substituted in the protein-protein interface of the parent domain of the first engineered immunoglobulin chain comprises the amino acid residue at position 88, and optionally a further amino acid residue at a position selected from the group consisting of 3, 5, 7, 20, 22, 26, 27, 79, 81, 84, 84.2, 85.1, 86, and 90 and, wherein the amino acid residue which is substituted in the protein-protein interface of the parent domain of the second engineered immunoglobulin chain comprises the amino acid residue at position 85.1 and/or 86 and optionally a further amino acid residue at a position selected from the group consisting of 3, 5, 7, 20, 22, 26, 27, 79, 81, 84, 84.2, 88, and 90 or at a position selected from the group consisting of 3, 5, 7, 20, 22, 26, 27, 79, 81, 84, 84.2, 85.1, 88, and 90 or at a position selected from the group consisting of 3, 5, 7, 20, 22, 26, 27, 79, 81, 84, 84.2, 86, 88, and 90,   wherein the amino acid residue substituted at position 88 in the parent domain of the first engineered immunoglobulin chain is interacting with the amino acid residue substituted at position 85.1 and/or 86 in the parent domain of the second engineered immunoglobulin chain,   wherein the amino acid position of each group member is indicated according to the IMGT® numbering.   
     
     
         2 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the parent domain of the first and/or the second engineered immunoglobulin chain is a naturally occurring CH3 domain. 
     
     
         3 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the amino acid sequence of the engineered domains of the first engineered immunoglobulin chain and/or the second engineered immunoglobulin chain do not contain an insertion of one or more amino acid residues compared to the amino acid sequence of the parent domains of the first engineered immunoglobulin chain and/or the second engineered immunoglobulin chain. 
     
     
         4 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the donor domain of the first member of the naturally occurring immunoglobulin super-family and the donor domain of the second member of the naturally occurring immunoglobulin super-family form a naturally occurring hetero-dimer. 
     
     
         5 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the amino acid residue which is substituted in the protein-protein interface of the parent domain of the first engineered immunoglobulin chain at position 88 is not a charged amino acid or is not 88I. 
     
     
         6 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the amino acid residue which is substituted in the protein-protein interface of the parent domain of the first engineered immunoglobulin chain at position 88 is 88W and conservative amino acid substitutions thereof, wherein the amino acid position is indicated according to the IMGT® numbering. 
     
     
         7 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the amino acid residue which is substituted at position 85 and/or 86 in the protein-protein interface of the parent domain of the second engineered immunoglobulin chain is selected from the group consisting of 85.1A, 85.1S, 85.1C and 85.1N and conservative amino acid substitutions thereof and/or is selected from the group consisting of 86S and 86V and conservative amino acid substitutions thereof, wherein the amino acid position is indicated according to the IMGT® numbering. 
     
     
         8 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the amino acid residue which is substituted in the protein-protein interface of the parent domain of the first engineered immunoglobulin chain at position 88 is selected from the group consisting of 88Q, 88L, 88V, 88R, 88E, 88I, 88T, 88Y, 88K and 88 W and/or wherein the further amino acid residue substituted in the protein-protein interface of the parent domain of the first engineered immunoglobulin chain is selected from the group consisting of 20V, 20T, 20A, 20N, 20Q, 20E, 20S, 20K, 20W, 22A, 22G, 22T, 22L, 22I, 22V, 26R, 26Q, 26T, 26K, 26V, 26S, 26N, 26E, 85.1T, 85.1M, 85.1A, 85.1S, 85.1R, 85.1H, 85.1K, 85.1F, 85.1C, 85.1N, 85.1W, 86S, 86I, 86T, 86H, 86Q, 86V, 86W, 86Y and 86F, wherein the amino acid position of each group member is indicated according to the IMGT® numbering. 
     
     
         9 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the amino acid residue which is substituted in the protein-protein interface of the parent domain of the first engineered immunoglobulin chain at position 88 is selected from the group consisting of 88Q, 88L, 88V, 88R, 88E, 88I, 88T, 88Y, and 88W and/or wherein the further amino acid residue substituted in the protein-protein interface of the parent domain of the first engineered immunoglobulin chain is selected from the group consisting of 20V, 20T, 20A, 20N, 20Q, 20E, 20S, 20K, 22A, 22G, 22T, 22L, 22I, 22V, 26Q, 26T, 26K, 26V, 26S, 26N, 26E, 85.1T, 85.1M, 85.1A, 85.1S, 85.1R, 85.1H, 85.1K, 85.1F, 85.1C, 85.1N, 86S, 86I, 86T, 86H, 86Q, 86V, and 86F, wherein the amino acid position of each group member is indicated according to the IMGT® numbering. 
     
     
         10 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the further amino acid residue which is substituted in the protein-protein interface of the parent domain of the first engineered immunoglobulin chain is selected from the group consisting of 3E, 3K, 5A, 5T, 7F, 7M, 20K, 20N, 20T, 22A, 22L, 22V, 26E, 26T, 27E, 27K, 79F, 79Y, 81A, 81G, 81D, 84L, 84M, 84.2E, 84.2S, 85.1A, 85.1C, 85.1M, 85.1N, 85.1S, 86F, 86S, 86V, 90K, 90N, and 90R and conservative amino acid substitutions thereof, wherein the amino acid position of each group member is indicated according to the IMGT® numbering. 
     
     
         11 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the amino acid residue which is substituted in the protein-protein interface of the parent domain of the first engineered immunoglobulin chain comprises the amino acid residues at position 88 and at position 20, and optionally a further amino acid residue at a position selected from the group consisting of 3, 5, 7, 22, 26, 27, 79, 81, 84, 84.2, 85.1, 86, and 90, wherein the amino acid position of each group member is indicated according to the IMGT® numbering. 
     
     
         12 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the amino acid residue which is substituted in the protein-protein interface of the parent domain of the second engineered immunoglobulin chain comprises the amino acid residues at position 85.1 and/or 86 and at position 26 and optionally an amino acid residue at a further position, wherein the amino acid position is indicated according to the IMGT® numbering. 
     
     
         13 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the further amino acid residue which is substituted in the protein-protein interface of the parent domain of the first engineered immunoglobulin chain is at a position selected from the group consisting of 20, 22, 26, 79, 85.1, 86, and 90, wherein the amino acid position of each group member is indicated according to the IMGT® numbering. 
     
     
         14 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the further amino acid residue which is substituted in the protein-protein interface of the parent domain of the first engineered immunoglobulin chain is selected from the group consisting of 20K, 22V, 26T, 79Y, 85.1S, 86V, and 90N and conservative amino acid substitutions thereof, wherein the amino acid position of each group member is indicated according to the IMGT® numbering. 
     
     
         15 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the further amino acid residue which is substituted in the protein-protein interface of the parent domain of the second engineered immunoglobulin chain is at position 84 or is 84L and conservative amino acid substitutions thereof, and optionally a further amino acid at a position selected from the group consisting of 3, 5, 7, 20, 22, 26, 81, 84.2, 88, and 90 or at a position selected from the group consisting of 3, 5, 7, 20, 22, 26, 81, 84.2, 85.1, 88, and 90 or at a position selected from the group consisting of 3, 5, 7, 20, 22, 26, 81, 84.2, 86, 88, and 90, wherein the amino acid position of each group member is indicated according to the IMGT® numbering. 
     
     
         16 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the further amino acid residue which is substituted in the protein-protein interface of the parent domain of the second engineered immunoglobulin chain is at a position selected from the group consisting of 3, 5, 7, 20, 22, 26, 81, 84, 84.2, 88, and 90 or at a position selected from the group consisting of 3, 5, 7, 20, 22, 26, 81, 84, 84.2, 85.1, 88, and 90 or at a position selected from the group consisting of 3, 5, 7, 20, 22, 26, 81, 84, 84.2, 86, 88, and 90, wherein the amino acid position of each group member is indicated according to the IMGT® numbering. 
     
     
         17 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the amino acid residue which is substituted in the protein-protein interface of the parent domain of the second engineered immunoglobulin chain comprises the amino acid residues at position 85.1 and 86 or comprises 85.1C and conservative amino acid substitutions thereof and 86S and conservative amino acid substitutions thereof, and wherein the further amino acid residue which is substituted in the protein-protein interface of the parent domain of the second engineered immunoglobulin chain is selected from the group consisting of 3E, 5A, 7F, 20T, 22V, 26T, 81D, 84L, 84.2E, 88R and 90R and conservative amino acid substitutions thereof, wherein the amino acid position of each group member is indicated according to the IMGT® numbering. 
     
     
         18 . (canceled) 
     
     
         19 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the further amino acid residue which is substituted in the protein-protein interface of the parent domain of the first engineered immunoglobulin chain is at a position selected from the group consisting of 3, 5, 20, 22, 26, 27, 81, 84, 85.1, and 86, wherein the amino acid position of each group member is indicated according to the IMGT® numbering. 
     
     
         20 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the further amino acid residue which is substituted in the protein-protein interface of the parent domain of the first engineered immunoglobulin chain is selected from the group consisting of 3K, 5T, 20T, 22L, 26E, 27K, 81G, 84M, 85.1M, 86F and conservative amino acid substitutions thereof, wherein the amino acid position of each group member is indicated according to the IMGT® numbering. 
     
     
         21 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the further amino acid residue which is substituted in the protein-protein interface of the parent domain of the second engineered immunoglobulin chain is at a position selected from the group consisting of 7, 20, 22, 27, 79, 81, 84.2, 88, and 90 or at a position selected from the group consisting of 7, 20, 22, 27, 79, 81, 84.2, 85.1, 88, and 90 or at a position selected from the group consisting of 7, 20, 22, 27, 79, 81, 84.2, 86, 88, and 90, wherein the amino acid position of each group member is indicated according to the IMGT® numbering. 
     
     
         22 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the further amino acid residue which is substituted in the protein-protein interface of the parent domain of the second engineered immunoglobulin chain is at position 79 or is 79F and conservative amino acid substitutions thereof, and optionally a further amino acid residue at a position selected from the group consisting of 7, 20, 22, 27, 81, 84.2, 88, and 90 or at a position selected from the group consisting of 7, 20, 22, 27, 81, 84.2, 85.1, 88, and 90 or at a position selected from the group consisting of 7, 20, 22, 27, 81, 84.2, 86, 88, and 90, wherein the amino acid position of each group member is indicated according to the IMGT® numbering. 
     
     
         23 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the amino acid residue which is substituted in the protein-protein interface of the parent domain of the second engineered immunoglobulin chain comprises the amino acid residues at position 85.1 and 86 or comprises 85.1N and conservative amino acid substitutions thereof and 86V and conservative amino acid substitutions thereof, and wherein the further amino acid residue which is substituted in the protein-protein interface of the parent domain of the second engineered immunoglobulin chain is selected from the group consisting of 7M, 20N, 22A, 27E, 79F, 81A, 84.2S, 85.1N, 86V, 88L, and 90K and conservative amino acid substitutions thereof, wherein the amino acid position of each group member is indicated according to the IMGT® numbering. 
     
     
         24 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the amino acid residue which is substituted in the protein-protein interface of the parent domain of the first engineered immunoglobulin chain is 88I and conservative amino acid substitutions thereof and the optional further amino acid residue which is substituted in the protein-protein interface of the parent domain of the first engineered immunoglobulin chain is 81W and conservative amino acid substitutions thereof, wherein the amino acid position of each group member is indicated according to the IMGT® numbering. 
     
     
         25 - 55 . (canceled) 
     
     
         56 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the amino acid residues of the protein-protein interface of the parent domain of the first and/or second engineered immunoglobulin chain which are substituted are not adjacent. 
     
     
         57 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein at least two amino acid residues of the protein-protein interface of the parent domain of the first and/or second engineered immunoglobulin chain are substituted. 
     
     
         58 - 61 . (canceled) 
     
     
         62 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the first engineered immunoglobulin chain and/or the second engineered immunoglobulin chain comprise a further engineered domain with a protein-protein interface, wherein at least one amino acid residue of the protein-protein interface of the parent domain of said further engineered domain of said first engineered immunoglobulin chain and/or said second engineered immunoglobulin chain is substituted with an amino acid residue at the equivalent 3D structural position from a protein-protein interface of a donor domain of a third member of the naturally occurring immunoglobulin super-family, and
 wherein the protein-protein interface of the further engineered domain of the first engineered immunoglobulin chain and/or the second engineered immunoglobulin chain is interacting with the protein-protein interface of an engineered domain of a third engineered immunoglobulin chain by hetero-dimerization or by homo-dimerization, wherein at least one amino acid residue of the protein-protein interface of the parent domain of said engineered domain of said third engineered immunoglobulin chain is substituted with an amino acid residue at the equivalent 3D structural position from a protein-protein interface of a donor domain of a fourth member of the naturally occurring immunoglobulin super-family, and   wherein the donor domain of the third member of the naturally occurring immunoglobulin super-family and the donor domain of the fourth member of the naturally occurring immunoglobulin super-family form a naturally occurring hetero-dimer or a naturally occurring homo-dimer and,   wherein the protein-protein interface of the further engineered domain of the first engineered immunoglobulin chain and/or the second engineered immunoglobulin chain and of the engineered domain of the third engineered immunoglobulin chain is different from the protein-protein interface of the engineered domain of the first engineered immunoglobulin chain and of the engineered domain of the second engineered immunoglobulin chain.   
     
     
         63 - 74 . (canceled) 
     
     
         75 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the first and second engineered immunoglobulin chains comprise an Fc region. 
     
     
         76 . (canceled) 
     
     
         77 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein the hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof is a full-length antibody. 
     
     
         78 - 79 . (canceled) 
     
     
         80 . The hetero-dimeric immunoglobulin or hetero-dimeric fragment thereof of  claim 1 , wherein at least one additional polypeptide is fused to the first and/or second engineered immunoglobulin chain. 
     
     
         81 - 211 . (canceled)