Antibodies binding to phosphorylcholine (pc) and/or pc conjugates

Abstract

The present invention relates to an antibody or antibody fragment capable of binding to phosphorylcholine and/or a phosphorylcholine conjugate, wherein the antibody or antibody fragment comprises a variable heavy chain (VH) domain and/or a variable light chain (VL) domain, and wherein—(a) the VH domain comprises an amino acid sequence that includes one, two or three complementarity determining regions (CDRs) selected from the group consisting of:a CDR1 sequence comprising an amino acid sequence having at least 20%, 40%, 60%, 80% or 100% sequence identity to the sequence of SEQ ID NO: 7;a CDR2 sequence comprising an amino acid sequence having at least 5%, 11%, 17%, 23%, 29%, 35%, 41%, 47%, 52%, 58%, 64%, 70%, 76%, 82%, 88%, 94%, or 100% sequence identity to the sequence of SEQ ID NO: 8; anda CDR3 sequence comprising an amino acid sequence having at least 11%, 22%, 33%, 44%, 55%, 66%, 77%, 88% or 100% sequence identity to the sequence of SEQ ID NO: 9 or 10; and/or(b) the VL domain comprises an amino acid sequence that includes one, two or three complementarity determining regions (CDRs) selected from the group consisting of:a CDR4 sequence comprising an amino acid sequence having at least 7.5%, 15%, 23%, 30%, 38%, 46%, 53%, 61%, 69%, 76%, 84%, 92% or 100% sequence identity to the sequence of SEQ ID NO: 11;a CDR5 sequence comprising an amino acid sequence having at least 14%, 28%, 42%, 57%, 71%, 85% or 100% sequence identity to the sequence of SEQ ID NO: 12;a CDR6 sequence comprising an amino acid sequence having at least 9%, 18%, 27%, 36%, 45%, 54%, 63%, 72%, 81%, 90% or 100% sequence identity to the sequence of SEQ ID NO: 13.

1 . An antibody or antibody fragment capable of binding to phosphorylcholine and/or a phosphorylcholine conjugate,
 wherein the antibody or antibody fragment comprises a variable heavy chain (VH) domain and/or a variable light chain (VL) domain, and wherein—   (a) the VH domain comprises an amino acid sequence that includes one, two or three complementarity determining regions (CDRs) selected from the group consisting of:
 a CDR1 sequence comprising an amino acid sequence having at least 20%, 40%, 60%, 80% or 100% sequence identity to the sequence of SEQ ID NO: 7; 
 a CDR2 sequence comprising an amino acid sequence having at least 5%, 11%, 17%, 23%, 29%, 35%, 41%, 47%, 52%, 58%, 64%, 70%, 76%, 82%, 88%, 94%, or 100% sequence identity to the sequence of SEQ ID NO: 8; and 
 a CDR3 sequence comprising an amino acid sequence having at least 11%, 22%, 33%, 44%, 55%, 66%, 77%, 88% or 100% sequence identity to the sequence of SEQ ID NO: 9 or 10; and/or 
   (b) the VL domain comprises an amino acid sequence that includes one, two or three complementarity determining regions (CDRs) selected from the group consisting of:
 a CDR4 sequence comprising an amino acid sequence having at least 7.5%, 15%, 23%, 30%, 38%, 46%, 53%, 61%, 69%, 76%, 84%, 92% or 100% sequence identity to the sequence of SEQ ID NO: 11; 
 a CDR5 sequence comprising an amino acid sequence having at least 14%, 28%, 42%, 57%, 71%, 85% or 100% sequence identity to the sequence of SEQ ID NO: 12; 
 a CDR6 sequence comprising an amino acid sequence having at least 9%, 18%, 27%, 36%, 45%, 54%, 63%, 72%, 81%, 90% or 100% sequence identity to the sequence of SEQ ID NO: 13. 
   
     
     
         2 . The antibody or antibody fragment of  claim 1  wherein the VH domain comprises an amino acid sequence that includes a CDR1 sequence, a CDR2 and a CDR3 sequence as defined by  claim 1 , and/or the VL domain comprises an amino acid sequence that includes a CDR4 sequence, a CDR5 and a CDR6 sequence as defined by  claim 1 . 
     
     
         3 . The antibody or antibody fragment of  claim 1  or  2  wherein—
 the VH domain comprises an amino acid sequence that includes the CDR1, CDR2 and CDR3 sequences present in an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 3 or 5 or an amino acid sequence having at least 80%, 85%, 90%, or 95% sequence identity to an amino acid sequence of any of SEQ ID NOs: 1, 3 or 5; and/or 
 the VL domain comprises an amino acid sequence that includes the CDR4, CDR5 and CDR6 sequences present in an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 4 or 6 or an amino acid sequence having at least 80%, 85%, 90%, or 95% sequence identity to an amino acid sequence of any of SEQ ID NOs: 2, 4 or 6. 
 
     
     
         4 . The antibody or antibody fragment of any preceding claim wherein—
 the VH domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 3, or 5 or an amino acid sequence having at least 80%, 85%, 90%, or 95% sequence identity to an amino acid sequence of any of SEQ ID NOs: 1, 3, or 5; and 
 the VL domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 4, or 6 or an amino acid sequence having at least 80%, 85%, 90%, or 95% sequence identity to an amino acid sequence of any of SEQ ID NOs: 2, 4, or 6. 
 
     
     
         5 . The antibody or antibody fragment according to any preceding claim, wherein—
 the VH domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95% or 100% sequence identity SEQ ID NO:1; and 
 the VL domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95% or 100% sequence identity SEQ ID NO: 2. 
 
     
     
         6 . The antibody or antibody fragment according to any of  claims 1  to  4 , wherein—
 the VH domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95% or 100% sequence identity SEQ ID NO:3; and 
 the VL domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95% or 100% sequence identity SEQ ID NO: 4. 
 
     
     
         7 . The antibody or antibody fragment according to any of  claims 1  to  4 , wherein—
 the VH domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95% or 100% sequence identity SEQ ID NO:5; and 
 the VL domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95% or 100% sequence identity SEQ ID NO: 6. 
 
     
     
         8 . The antibody or antibody fragment according to any preceding claim, wherein the VH domain, the VL domain, or preferably both of the VH and VL domains, comprise an amino acid sequence having 100% sequence identity to the, or one or more (such as all) of each, stated SEQ ID NO. 
     
     
         9 . The antibody or antibody fragment according to any preceding claim, wherein the VH domain, the VL domain, or both of the VH and VL domains, comprise an amino acid sequence having less than 100%, but at least 80%, 85%, 90%, 95%, sequence identity to the, or one or more (such as all) of each, stated SEQ ID NO. 
     
     
         10 . The antibody or antibody fragment according to any of preceding claim,
 wherein the VH domain, the VL domain, or both of the VH and VL domains, comprise an amino acid sequence having less than 100%, but at least 80%, 85%, 90%, 95%, sequence identity to the, or one or more (such as all) of each, stated SEQ ID NO, and wherein   the ability of the antibody or antibody fragment to bind to phosphorylcholine and/or a phosphorylcholine conjugate is equivalent to (that is, at least 80%, 85%, 90% or 95%, of), or greater than, the ability of a corresponding antibody or antibody fragment, wherein the VH domain and the VL domain of the corresponding antibody or antibody fragment each comprise an antigen-binding sequence comprising an amino acid sequence having 100% sequence identity to the, or each, stated SEQ ID NO.   
     
     
         11 . The antibody or antibody fragment according to any preceding claim wherein the VH domain and the VL domain are present in a linear polypeptide sequence. 
     
     
         12 . The antibody or antibody fragment according to any preceding claim wherein the VH domain and the VL domain are each present in a separate polypeptide sequence, and preferably wherein the separate polypeptide sequence are directly or indirectly bound together (such as by one or more disulphide bonds between the separate polypeptide sequence). 
     
     
         13 . The antibody according to any of  claim 1 - 10  or  12  wherein the antibody is a monoclonal antibody. 
     
     
         14 . The antibody fragment according to any of  claims 1  to  12  wherein the antibody fragment is a single chain antibody, Fv, scFv, Fab, F(ab′) 2 , Fab′, Fd, dAb, CDR, or scFv-Fc fragment, a nanobody, and diabody, or any such fragment that has been stabilized such as by PEGylation. 
     
     
         15 . The antibody or antibody fragment according to any preceding claim which is a human or humanized antibody or antibody fragment, such as a human or humanized monoclonal antibody. 
     
     
         16 . The antibody or antibody fragment according to any preceding claim which is capable of binding to a phosphorylcholine conjugate. 
     
     
         17 . The antibody or antibody fragment according to  claim 16  wherein the phosphorylcholine conjugate is a phosphorylcholine moiety linked to a carrier, optionally via a spacer, and preferably the antibody or antibody fragment binds specifically to the phosphorylcholine moiety in the phosphorylcholine conjugate. 
     
     
         18 . A pharmaceutical composition comprising or consisting essentially of an antibody or an antibody fragment according to any of the preceding claims and a pharmaceutically acceptable carrier or excipient, optionally wherein the only antibodies or antibody fragments present in the composition are those defined by any of the preceding claims. 
     
     
         19 . An antibody or antibody fragment according to any of  claims 1  to  17 , or a pharmaceutical composition according to  claim 18  for use in medicine. 
     
     
         20 . An antibody or antibody fragment according to any of  claims 1  to  17 , or a pharmaceutical composition according to  claim 18  for use in the prevention, prophylaxis and/or treatment of mammals, including humans, against atherosclerosis, an atherosclerotic related disease or ischemic cardiovascular disease. 
     
     
         21 . An antibody or antibody fragment according to any of  claims 1  to  17 , or a pharmaceutical composition according to  claim 18  for use in the prophylaxis, prevention and/or treatment of Alzheimer's disease. 
     
     
         22 . An antibody or antibody fragment according to any of  claims 1  to  17 , or a pharmaceutical composition according to  claim 18  for use in the immunization or prophylaxis against, or the prevention or treatment of, metabolic diseases in mammals, including humans. 
     
     
         23 . A method for prevention, prophylaxis and/or treatment of a mammal, including a human, against atherosclerosis, an atherosclerotic related disease, or ischemic cardiovascular disease, the method comprising the step of administering to the mammal an antibody or antibody fragment according to any of  claims 1  to  17 , or a pharmaceutical composition according to  claim 18 . 
     
     
         24 . A method for immunization and prophylaxis, prevention and/or treatment of a subject against Alzheimer's disease, the method comprising the step of administering to the subject an antibody or antibody fragment according to any of  claims 1  to  17 , or a pharmaceutical composition according to  claim 18 . 
     
     
         25 . A method for the immunization or prophylaxis against, or the treatment of, metabolic diseases in a mammal, such as a human, the method comprising the step of administering to the mammal an antibody or antibody fragment according to any of  claims 1  to  17 , or a pharmaceutical composition according to  claim 18 . 
     
     
         26 . The antibody or antibody fragment according to any of  claims 1  to  17 , or a pharmaceutical composition according to  claim 18  for use according to  claim 22 , or the method according to  claim 25 , wherein the metabolic disease is a condition selected from the group consisting of metabolic syndrome, insulin resistance, glucose intolerance, hyperglycemia, type I diabetes, type II diabetes, hyperlipidemia, hypertriglyceridemia, hypercholesterolemia, dyslipidemia, and polycystic ovary syndrome (PCOS). 
     
     
         27 . A nucleic acid sequences encoding an antibody or an antibody fragment according to any of  claims 1  to  17 . 
     
     
         28 . A vector or plasmid comprising the nucleic acid sequence of  claim 27 . 
     
     
         29 . A host cell comprising the nucleic acid sequence of  claim 27  and/or a vector or plasmid according to  claim 28 . 
     
     
         30 . The host cell of  claim 29  wherein the cell is a prokaryotic cell, such as an  Escherichia coli  cell, or a eukaryotic cell, such as animal, plant, or fungal cell. 
     
     
         31 . The host cell of  claim 29  or  30  which expresses the nucleic acid sequence of  claim 27  and thereby produces an antibody or an antibody fragment according to any of  claims 1  to  17 . 
     
     
         32 . A method of producing an antibody or an antibody fragment according to any of  claims 1  to  17  comprising culturing a host cell according to  claim 31 , and recovering therefrom an antibody or an antibody fragment according to any of  claims 1  to  17 . 
     
     
         33 . A method of preparing a variant of the antibody or antibody fragments of according to any of  claims 1  to  17 , which variant retains the ability to bind to phosphorylcholine and/or a phosphorylcholine conjugate, the method comprising—
 (i) providing a nucleic acid according to  claim 27  encoding a parent antibody or antibody fragment; 
 (ii) introducing one or more nucleotide mutations into the amino acid coding regions of the nucleic acid sequence, optionally within the regions encoding the VH and/or VL domain(s), such that the mutated nucleic acid encodes a variant antibody or antibody fragment having a different amino acid sequence compared to the parent antibody or antibody fragment; 
 (iii) expressing the variant antibody or antibody fragment that is encoded by the mutated nucleic acid sequence; and 
 (iv) comparing the ability of the variant antibody or antibody fragment and the parent antibody or antibody fragment to bind to phosphorylcholine and/or a phosphorylcholine conjugate. 
 
     
     
         34 . The method of  claim 33  wherein the step of assessing the ability of the variant antibody or antibody fragment to bind to phosphorylcholine and/or a phosphorylcholine conjugate further comprises selecting those variants that have substantially equal or enhanced ability to bind to phosphorylcholine and/or a phosphorylcholine conjugate compared to the parent. 
     
     
         35 . The method of  claim 33  or  34  further comprising recovering a nucleic acid molecule that comprises the mutated nucleic acid sequence that encodes the variant antibody or antibody fragment, and optionally transforming a host cell with a composition comprising the recovered nucleic acid molecule and further optionally expressing the variant antibody or antibody fragment from the host cell, and yet further optionally recovering the thus-expressed variant antibody or antibody fragment from the host cell. 
     
     
         36 . The method of  claim 35  that comprises the step of recovering the thus-expressed variant antibody or antibody fragment from the host cell, and further comprises the step of formulating the recovered variant antibody or antibody fragment into a pharmaceutically acceptable composition. 
     
     
         37 . A variant antibody or antibody fragment obtained or obtainable by the method of  claim 33 - 35 , or a pharmaceutically acceptable obtained or obtainable by the method of  claim 36  for use in medicine and/or for use in accordance with  claims 20 ,  12  and/or  22  or for use in a method according to any of  claims 24 ,  25  and/or  26 .