Functional heavy chain antibodies, fragments thereof, library thereof and methods of production thereof

Abstract

The present invention relates to functional heavy chain antibodies, functional single domain heavy chain antibodies, functional VH domains, or functional fragments thereof comprising an amino acid which is neither a charged amino acid nor a C at position 45, and comprising an amino acid at position 103 independently chosen from the group consisting of R, G, K, S, Q, L, and P, and optionally an amino acid at position 108 independently chosen from the group consisting of Q, L and R, said positions determined according to the Kabat numbering.

1 - 87 . (canceled)  
     
     
         88 . A functional heavy chain antibody, a functional single domain heavy chain antibody, a functional VH domain, or a functional fragment thereof, comprising 
 an amino acid which is neither a charged amino acid nor a C at position 45, and    an amino acid at position 103 selected from the group consisting of R, G, K, S, Q, L, and P,    said positions determined according to the Kabat numbering.    
     
     
         89 . The functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof of  claim 88 , further comprising an amino acid at position 108 selected from the group consisting of Q, L and R.  
     
     
         90 . The functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof of  claim 88 , further comprising an amino acid sequence, starting at position 103, selected from the group consisting of RGQGTQ (SEQ ID NO: 13), RGKGTQ (SEQ ID NO: 14), LGQGTQVTVSS (SEQ ID NO: 16), QGQGTGVTVSS (SEQ ID NO:17), PGQGTQVTVSS (SEQ ID NO:18), and SSQGTQVTSS (SEQ ID NO:19):  
     
     
         91 . The functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof of  claim 88 , further comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:1, 2, 3, 4, 5, 6, 7, 8, 9, and 10.  
     
     
         92 . The functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof of  claim 88 , further comprising an amino acid sequence selected from the group consisting of SEQ ID NOs:20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64,  65 , 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, and 79.  
     
     
         93 . A functional heavy chain antibody, a functional single domain heavy chain antibody, a functional VH domain, or a functional fragment thereof, comprising the amino acid sequence VXXXXXXGLXW, wherein the V of the amino acid sequence is at position 37 according to the Kabat numbering, whereby X is any amino acid.  
     
     
         94 . The functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof of  claim 88 , which is a functional heavy chain antibody.  
     
     
         95 . The functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof of  claim 88 , which is a functional single domain heavy chain antibody.  
     
     
         96 . The functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof of  claim 88 , which is a functional VH domain.  
     
     
         97 . The functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof of  claim 88 , which is a functional fragment thereof.  
     
     
         98 . The functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof of  claim 88 , which is an artificial mutant.  
     
     
         99 . The functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof of  claim 93 , which is an artificial mutant.  
     
     
         100 . The artificial mutant of  claim 98 , comprising at least one derivatized amino acid.  
     
     
         101 . The artificial mutant of  claim 98 , derived from a protein from a Camelidae.  
     
     
         102 . The artificial mutant of  claim 98 , derived from a protein from a human.  
     
     
         103 . The artificial mutant of  claim 98 , derived from a protein from a mouse.  
     
     
         104 . The artificial mutant of  claim 98 , derived from a protein from a vertebrate species other than a Camelidae, human or mouse.  
     
     
         105 . A polypeptide comprising the functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof of  claim 88 .  
     
     
         106 . A method to functionalize a heavy chain antibody, a single domain heavy chain antibody, a VH domain, or a fragment thereof, the method comprising replacing the amino acid at position 103 with an amino acid independently chosen from the group consisting of R, G, K, S and P, said position determined by the Kabat numbering.  
     
     
         107 . A functional heavy chain antibody, a single domain heavy chain antibody, a VH domain, or a fragment thereof, produced by the method of  claim 106 .  
     
     
         108 . A method to humanize and functionalize a heavy chain antibody, a single domain heavy chain antibody, a VH domain, or a fragment thereof, said method comprising replacing the amino acid at position 45 with L, and replacing the amino acid at position 103 with an amino acid selected from the group consisting of R, G; K, S and P, said positions determined by the Kabat numbering, wherein the resulting product is the functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof of  claim 88 .  
     
     
         109 . A library comprising more than one functional mutant of  claim 98 .  
     
     
         110 . A library comprising more than one functional mutant of  claim 99 .  
     
     
         111 . A nucleic acid comprising a DNA sequence encoding the functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof of  claim 88 .  
     
     
         112 . A nucleic acid comprising a DNA sequence encoding the functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof of  claim 93 .  
     
     
         113 . A host cell comprising the nucleic acid of  claim 111 , wherein the host cell is capable of expressing the functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof.  
     
     
         114 . A host cell comprising the nucleic acid of  claim 112 , wherein the host cell is capable of expressing the functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof.  
     
     
         115 . The functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof of  claim 88 , in a pharmaceutically suitable excipient.  
     
     
         116 . The functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof of  claim 93 , in a pharmaceutically suitable excipient.  
     
     
         117 . A method of treating a mammal, the method comprising administering to the mammal the functional heavy chain antibody, functional single domain heavy chain antibody, functional VH domain, or functional fragment thereof of  claim 115 .  
     
     
         118 . The method of  claim 117 , wherein the mammal is a human with a disease selected from the group consisting of asthma, rhinoconjunctivitis, allergic disorders, acute allograft rejection, Crohn's disease and ulcerative colitis.  
     
     
         119 . A nucleic acid comprising a sequence selected from the group consisting of SEQ ID NOs:80, 81, 82, 83, 84, 85, 86, 87, and 88.