Novel multivalent immunoglobulins

Abstract

The present invention provides a multivalent immunoglobulin or part thereof binding specifically to at least two cell surface molecules of a single cell with at least one modification in at least one structural loop region of said immunoglobulin determining binding to an epitope of said cell surface molecules wherein the unmodified immunoglobulin does not significantly bind to said epitope, its use and methods for producing it.

1 . A multivalent immunoglobulin or part thereof binding specifically to at least two cell surface molecules of a single cell with at least one modification in at least one structural loop region of said immunoglobulin determining binding to an epitope of said cell surface molecules wherein the unmodified immunoglobulin does not significantly bind to said epitope. 
   
   
       2 . Immunoglobulin according to  claim 1  wherein the modified structural loop region is within the constant domain of said immunoglobulin, preferably within CH1, CH2, CH3, CH4, Igk-C, Igl-C, or a part thereof. 
   
   
       3 . Immunoglobulin according to  claim 2 , characterised in that said modified structural loop region comprises at least 6 amino acid modifications. 
   
   
       4 . Immunoglobulin according to  claim 2 , characterised in that the modified structural loop region is within a constant domain selected from the group consisting of a CH1, a CH2, a CH3 and a CH4 domain of human or murine origin and comprises at least one modification within amino acids 7 to 21, amino acids 25 to 39, amino acids 41 to 81, amino acids 83 to 85, amino acids 89 to 103, or amino acids 106 to 117. 
   
   
       5 . Immunoglobulin according to  claim 2 , characterised in that the immunoglobulin comprises Igk-C or Igl-C modified structural loop regions that are of human origin and comprise at least one modification within amino acids 8 to 18, amino acids 27 to 35, amino acids 42 to 78, amino acids 83 to 85, amino acids 92 to 100, amino acids 108 to 117, or amino acids 123 to 126. 
   
   
       6 . Immunoglobulin according to  claim 2 , characterised in that the modified structural loop regions of Igk-C or Igl-C are of murine origin and comprise at least one modification within amino acids 8 to 20, amino acids 26 to 36, amino acids 43 to 79, amino acids 83 to 85, amino acids 90 to 101, amino acids 108 to 116, or amino acids 122 to 125. 
   
   
       7 . Immunoglobulin according to  claim 1 , characterised in that the immunoglobulin comprises a constant domain of camelid origin. 
   
   
       8 . Immunoglobulin according to  claim 7 , characterised in that the immunoglobulin comprises at least one modified constant domain selected from the group consisting of a CH1, CH2 and CH3 domain. 
   
   
       9 . Immunoglobulin according to  claim 8 , characterised in that the modified constant domain comprises at least one modification within amino acids 8 to 20, amino acids 24 to 39, amino acids 42 to 78, amino acids 82 to 85, amino acids 91 to 103, or amino acids 108 to 117. 
   
   
       10 . Immunoglobulin according to  claim 1 , characterised in that the immunoglobulin comprises a variable domain selected from the group consisting of VH, Vkappa, Vlambda, VHH, and combinations thereof. 
   
   
       11 . Immunoglobulin according to  claim 1 , characterised in that the immunoglobulin comprises a modified structural loop region of a VH, a Vkappa, a Vlambda or a VHH domain comprising at least one modification within amino acids 7 to 21, amino acids 25 to 39, amino acids 41 to 81, amino acids 83 to 85, amino acids 89 to 103, or amino acids 106 to 117, where the numbering of the amino acid position of the domains is that of the IMGT. 
   
   
       12 . Immunoglobulin according to  claim 1 , characterised in that the immunoglobulin comprises a modified structural loop region of a VH, Vkappa, or Vlambda domain of human origin and comprises at least one modification within amino acids 8 to 20, amino acids 44 to 50, amino acids 67 to 76 and amino acids 89 to 101, most preferably amino acid positions 12 to 17, amino acid positions 45 to 50, amino acid positions 69 to 75, and amino acid positions 93 to 98, where the numbering of the amino acid position of the domains is that of the IMGT. 
   
   
       13 . Immunoglobulin according to  claim 1 , characterised in that the immunoglobulin comprises a modified structural loop region of a VH domain of murine origin and comprises at least one modification within amino acids 6 to 20, amino acids 44 to 52, amino acids 67 to 76, and amino acids 92 to 101, where the numbering of the amino acid position of the domains is that of the IMGT. 
   
   
       14 . Immunoglobulin according to  claim 1 , characterised in that the immunoglobulin comprises a modified structural loop region of a VHH domain of camelid origin and comprises at least one modification within amino acids 7 to 18, amino acids 43 to 55, amino acids 68 to 75, and amino acids 91 to 101, where the numbering of the amino acid position of the domains is that of the IMGT. 
   
   
       15 . Immunoglobulin according to  claim 1 , characterized in that the immunoglobulin is further combined with one or more additional modified immunoglobulins or with unmodified immunoglobulins, or parts thereof, to obtain a combination immunoglobulin. 
   
   
       16 . Immunoglobulin according to  claim 1 , characterised in that the modification is a deletion, a substitution, an insertion or a combination thereof. 
   
   
       17 . Nucleic acid encoding an immunoglobulin according to  claim 1  or part thereof. 
   
   
       18 . Method for engineering a multivalent immunoglobulin according to  claim 1 , comprising the steps of:
 (a) providing a nucleic acid encoding an immunoglobulin comprising at least one structural loop region,   (b) modifying at least one nucleotide residue of said structural loop region,   (c) transferring said modified nucleic acid in an expression system,   (d) expressing said multivalent immunoglobulin,   (e) contacting the expressed multivalent immunoglobulin with an epitope, and   (f) determining whether said multivalent immunoglobulin binds to said epitope.   
   
   
       19 . (canceled)