Human BNP immunospecific antibodies

Abstract

The present invention relates to antibodies that immunospecifically bind to human brain natriuretic peptide or a human brain natriuretic peptide fragment with a high binding affinity, methods for producing and selecting said antibodies, immunoassays for human brain natriuretic peptide or a human brain natriuretic peptide fragment that employ said antibodies and therapeutic compositions containing said antibodies.

1 . An isolated antibody which immunospecifically binds to an epitope comprising amino acid residues 5 through 13 of human brain natriuretic peptide (“hBNP”) with at least about a two fold improvement in its equilibrium dissociation constant (K D ) when compared with an antibody produced by hybridoma cell line 106.3, said cell line having A.T.C.C. Accession No. HB-12044.  
     
     
         2 . The antibody of  claim 1 , wherein said isolated antibody exhibits at least about a five fold improvement in its K D  when compared with an antibody produced by hybridoma cell line 106.3.  
     
     
         3 . The antibody of  claim 1 , wherein said isolated antibody exhibits at least about a ten fold improvement in its K D  when compared with an antibody produced by hybridoma cell line 106.3.  
     
     
         4 . The antibody of  claim 1 , wherein said isolated antibody exhibits at least about a fifteen fold improvement in its K D  when compared with an antibody produced by hybridoma cell line 106.3.  
     
     
         5 . The antibody of  claim 1 , wherein said isolated antibody exhibits at least about a twenty fold improvement in its K D  when compared with an antibody produced by hybridoma cell line 106.3.  
     
     
         6 . The antibody of  claim 1 , wherein said isolated antibody exhibits at least about a twenty-five fold improvement in its K D  when compared with an antibody produced by hybridoma cell line 106.3.  
     
     
         7 . The antibody of  claim 1 , wherein said isolated antibody is a monoclonal antibody, a multispecific antibody, a human antibody, a fully humanized antibody, a partially humanized antibody, an animal antibody, a recombinant antibody, a chimeric antibody, a single-chain Fv, a single chain antibody, a single domain antibody, a Fab fragment, a F(ab′) 2  fragment, a disulfide-linked Fv, an anti-idiotypic antibody, or a functionally active epitope-binding fragment thereof.  
     
     
         8 . An isolated antibody which immunospecifically binds to hBNP, wherein said antibody has an association rate (k a ) of between about 5.0×10 4  and about 1.0×10 8  M −1 s −1 .  
     
     
         9 . The antibody of  claim 8 , wherein said antibody has an association rate of between about 3.3×10 4  and about 1.0×10 9  M −1 s −1 .  
     
     
         10 . The antibody of  claim 8 , wherein said antibody has an association rate of between about 2.5×10 4  and about 1.0×10 8  M −1 s −1 .  
     
     
         11 . The antibody of  claim 8 , wherein said antibody has an association rate of between about 2.4×10 4  and about 1.35×10 7  M −1 s −1 .  
     
     
         12 . The antibody of  claim 8 , wherein said antibody is a monoclonal antibody, a multispecific antibody, a human antibody, a fully humanized antibody, a partially humanized antibody, an animal antibody, a recombinant antibody, a chimeric antibody, a single-chain Fv, a single chain antibody, a single domain antibody, a Fab fragment, a F(ab′) 2  fragment, a disulfide-linked Fv, an anti-idiotypic antibody, or a functionally active epitope-binding fragment thereof.  
     
     
         13 . The antibody of  claim 8 , wherein said antibody immunospecifically binds to an epitope comprising amino acid residues 5 through 13 of hBNP.  
     
     
         14 . An isolated antibody which immunospecifically binds to hBNP, wherein said antibody has a dissociation rate (k d ) of between about 1.0×10 −3  and about 1.0×10 −6 ·s −1 .  
     
     
         15 . The antibody of  claim 14 , wherein said antibody has a dissociation rate of between about 1.0×10 −3  and about 1.0×10 −5 ·s −1 .  
     
     
         16 . The antibody of  claim 14 , wherein said antibody has a dissociation rate of between about 1.0×10 −3  and about 1.0×10 −4 ·s −1 .  
     
     
         17 . The antibody of  claim 14 , wherein said antibody is a monoclonal antibody, a multispecific antibody, a human antibody, a fully humanized antibody, a partially humanized antibody, an animal antibody, a recombinant antibody, a chimeric antibody, a single-chain Fv, a single chain antibody, a single domain antibody, a Fab fragment, a F(ab′) 2  fragment, a disulfide-linked Fv, an anti-idiotypic antibody, or a functionally active epitope-binding fragment thereof.  
     
     
         18 . The antibody of  claim 14 , wherein said antibody immunospecifically binds to an epitope comprising amino acid residues 5 through 13 of hBNP.  
     
     
         19 . An isolated antibody which immunospecifically binds to hBNP wherein said antibody has an equilibrium dissociation constant (K D ) of between about 2×10 −11  M and about 1×10 −15  M.  
     
     
         20 . The antibody of  claim 19 , wherein said antibody has an equilibrium dissociation constant of between about 3.0×10 −11  M and about 1.0×10 −14  M.  
     
     
         21 . The antibody of  claim 19 , wherein said antibody has an equilibrium dissociation constant of between about 4.0×10 −11  M and about 8.0×10 −13  M.  
     
     
         22 . The antibody of  claim 19 , wherein said antibody has an equilibrium dissociation constant of between about 4.2×10 −11  M and about 7.4×10 −13  M.  
     
     
         23 . The antibody of  claim 19 , wherein said antibody is a monoclonal antibody, a multispecific antibody, a human antibody, a fully humanized antibody, a partially humanized antibody, an animal antibody, a recombinant antibody, a chimeric antibody, a single-chain Fv, a single chain antibody, a single domain antibody, a Fab fragment, a F(ab′) 2  fragment, a disulfide-linked Fv, an anti-idiotypic antibody, or a functionally active epitope-binding fragment thereof.  
     
     
         24 . The antibody of  claim 19 , wherein said antibody immunospecifically binds to an epitope comprising amino acid residues 5 through 13 of hBNP.  
     
     
         25 . Chinese hamster ovary (“CHO”) cell line AM1 having A.T.C.C. Accession No. PTA-6987.  
     
     
         26 . An antibody made from DNA extracted from the CHO cell line AM1 having A.T.C.C. Accession No. PTA-6987.  
     
     
         27 . A chimeric antibody or a hBNP-epitope binding fragment thereof produced by CHO cell line AM1, wherein said cell line has A.T.C.C. Accession No. PTA-6987.  
     
     
         28 . An isolated antibody which immunospecifically binds to hBNP, wherein said antibody has a variable heavy domain and a variable light domain, the variable heavy domain comprising a heavy chain complementarity determining region (“CDR”) 1, a heavy chain CDR 2 and a heavy chain CDR 3, the variable light domain comprising a light chain CDR 1, a light chain CDR 2 and a light chain CDR 3, wherein 
 (a) Heavy Chain CDR 1 has an amino acid sequence of: Gly-Tyr-Thr-Phe-Thr-His-Tyr-Gly-Ile-Asn (SEQ ID NO:6);    (b) Heavy Chain CDR 2 has an amino acid sequence having a formula of:                          (SEQ ID NO:12)                   Trp-Ile-Asn-Thr-His-Thr-Gly-Glu-Xaa 1 -Xaa 2 -Tyr-Ala-               Asp-Asp-Phe-Lys-Gly                                 wherein Xaa 1  is selected from the group consisting of proline and alanine;    wherein Xaa 2  is selected from the group consisting of isoleucine and tyrosine;    (c) Heavy Chain CDR 3 has an amino acid sequence of: Ser-His-Arg-Phe-Gly-Leu-Asp-Tyr (SEQ ID NO:8);    (d) Light Chain CDR 1 has an amino acid sequence having a formula of:                          (SEQ ID NO:13)                           Lys-Ala-Xaa 3 -Xaa 4 -Xaa 5 -Val-Asp-Tyr-Asn-Gly-Asp-                       Ser-Tyr-Leu-Asn                                     wherein Xaa 3  is selected from the group consisting of: serine, alanine, asparagine, glutamine, tyrosine, threonine and arginine;    wherein Xaa 4  is selected from the group consisting of: glutamine, tyrosine, tryptophan, alanine and phenylalanine;    wherein Xaa 5  is selected from the group consisting of: serine, glycine, proline, alanine and aspartic acid;    (e) Light Chain CDR 2 has an amino acid sequence having the formula of:                          (SEQ ID NO:14)                           Ala-Ala-Ser-Xaa 6 -Xaa 7 -Xaa 8 -Ser                                       wherein Xaa 6  is selected from the group consisting of: asparagine and cysteine;    wherein Xaa 7  is selected from the group consisting of: leucine, glycine and alanine;    wherein Xaa 8  is selected from the group consisting of glutamic acid, tryptophan and proline; and    (f) Light Chain CDR 3 has an amino acid sequence of: Gln-Gln-Ser-Asn-Glu-Asp-Pro-Phe-Thr (SEQ ID NO: 11),    wherein the heavy chain CDR 2 has an amino acid sequence other than Trp-Ile-Asn-Thr-His-Thr-Gly-Glu-Pro-Ile-Tyr-Ala-Asp-Asp-Phe-Lys-Gly (SEQ ID NO:7) when the light chain CDR 1 has the amino acid sequence of Lys-Ala-Ser-Gln-Ser-Val-Asp-Tyr-Asn-Gly-Asp-Ser-Tyr-Leu-Asn (SEQ ID NO:9) and the light chain CDR 2 has the amino acid sequence of Ala-Ala-Ser-Asn-Leu-Glu-Ser (SEQ ID NO:10), the light chain CDR 1 has an amino acid sequence other than Lys-Ala-Ser-Gln-Ser-Val-Asp-Tyr-Asn-Gly-Asp-Ser-Tyr-Leu-Asn (SEQ ID NO:9) when the heavy chain CDR 2 has the amino acid sequence Trp-Ile-Asn-Thr-His-Thr-Gly-Glu-Pro-Ile-Tyr-Ala-Asp-Asp-Phe-Lys-Gly (SEQ ID NO:7) and the light chain CDR 2 has the amino acid sequence Ala-Ala-Ser-Asn-Leu-Glu-Ser (SEQ ID NO:19), or the light chain CDR 2 has an amino acid sequence other than Ala-Ala-Ser-Asn-Leu-Glu-Ser (SEQ ID NO:10) when the heavy chain CDR 2 has the amino acid sequence of Trp-Ile-Asn-Tnr-His-Thr-Gly-Glu-Pro-Ile-Tyr-Ala-Asp-Asp-Phe-Lys-Gly (SEQ ID NO:7) and the light chain CDR 1 has the amino acid sequence of Lys-Ala-Ser-Gln-Ser-Val-Asp-Tyr-Asn-Gly-Asp-Ser-Tyr-Leu-Asn (SEQ ID NO:9).    
     
     
         29 . The antibody of  claim 28 , wherein: 
 Xaa 1  is alanine.    Xaa 2  is tyrosine.    Xaa 3  is serine.    Xaa 4  is glutamine.    Xaa 5  is serine.    Xaa 6  is asparagine.    Xaa 7  is leucine.    Xaa 8  is glutamic acid.    
     
     
         30 . The antibody of  claim 28 , wherein: 
 Xaa 1  is proline.    Xaa 2  is isoleucine.    Xaa 3  is glutamine.    Xaa 4  is phenylalanine.    Xaa 5  is alanine.    Xaa 6  is asparagine.    Xaa 7  is leucine.    Xaa 8  is glutamic acid.    
     
     
         31 . The antibody of  claim 28 , wherein: 
 Xaa 1  is proline.    Xaa 2  is isoleucine.    Xaa 3  is tyrosine.    Xaa 4  is alanine.    Xaa 5  is serine.    Xaa 6  is asparagine.    Xaa 7  is leucine.    Xaa 8  is glutamic acid.    
     
     
         32 . The antibody of  claim 28 , wherein: 
 Xaa 1  is proline.    Xaa 2  is isoleucine.    Xaa 3  is glutamine.    Xaa 4  is tryptophan.    Xaa 5  is glycine.    Xaa 6  is asparagine.    Xaa 7  is leucine.    Xaa 8  is glutamic acid.    
     
     
         33 . The antibody of  claim 28 , wherein: 
 Xaa 1  is proline.    Xaa 2  is isoleucine.    Xaa 3  is threonine.    Xaa 4  is tryptophan.    Xaa 5  is aspartic acid.    Xaa 6  is asparagine.    Xaa 7  is leucine.    Xaa 8  is glutamic acid.    
     
     
         34 . The antibody of  claim 28 , wherein: 
 Xaa 1  is proline.    Xaa 2  is isoleucine.    Xaa 3  is arginine.    Xaa 4  is tryptophan.    Xaa 5  is proline.    Xaa 6  is asparagine.    Xaa 7  is leucine.    Xaa 8  is glutamic acid.    
     
     
         35 . The antibody of  claim 28 , wherein: 
 Xaa 1  is proline.    Xaa 2  is isoleucine.    Xaa 3  is alanine.    Xaa 4  is tyrosine.    Xaa 5  is glycine.    Xaa 6  is asparagine.    Xaa 7  is leucine.    Xaa 8  is glutamic acid.    
     
     
         36 . The antibody of  claim 28 , wherein: 
 Xaa 1  is proline.    Xaa 2  is isoleucine.    Xaa 3  is asparagine.    Xaa 4  is tryptophan.    Xaa 5  is proline.    Xaa 6  is asparagine.    Xaa 7  is leucine.    Xaa 8  is glutamic acid.    
     
     
         37 . The antibody of  claim 28 , wherein: 
 Xaa 1  is proline.    Xaa 2  is isoleucine.    Xaa 3  is serine.    Xaa 4  is glutamine.    Xaa 5  is serine.    Xaa 6  is cysteine.    Xaa 7  is glycine.    Xaa 8  is tryptophan.    
     
     
         38 . The antibody of  claim 28 , wherein: 
 Xaa 1  is proline.    Xaa 2  is isoleucine.    Xaa 3  is serine.    Xaa 4  is glutamine.    Xaa 5  is serine.    Xaa 6  is cysteine.    Xaa 7  is alanine.    Xaa 8  is proline.    
     
     
         39 . The antibody of  claim 28 , wherein said antibody has an equilibrium dissociation constant of between about 2.0×10 −11  M and about 1.0×10 −15  M.  
     
     
         40 . The antibody of  claim 28 , wherein said antibody has an equilibrium dissociation constant of between about 3.0×10 −11  M and about 1.0×10 −14  M.  
     
     
         41 . The antibody of  claim 28 , wherein said antibody has an equilibrium dissociation constant of between about 4.0×10 −11  M and about 8.0×10 −13  M.  
     
     
         42 . The antibody of  claim 28 , wherein said antibody has an equilibrium dissociation constant of between about 4.2×10 −11  M and about 7.4×10 −13  M.  
     
     
         43 . The antibody of  claim 28 , wherein said antibody further comprises an association rate (k a ) of between about 5.0×10 4  and about 1.0×10 8  M −1 s −1 .  
     
     
         44 . The antibody of  claim 28 , wherein said antibody further comprises a dissociation rate (k d ) of between about 1.0×10 −3  and about 1.0×10 −6  s −1 .  
     
     
         45 . The antibody of  claim 28 , wherein said antibody is a monoclonal antibody, a multispecific antibody, a human antibody, a fully humanized antibody, a partially humanized antibody, an animal antibody, a recombinant antibody, a chimeric antibody, a single-chain Fv, a single chain antibody, a single domain antibody, a Fab fragment, a F(ab′) 2  fragment, a disulfide-linked Fv, an anti-idiotypic antibody, or a functionally active epitope-binding fragment thereof.  
     
     
         46 . The antibody of  claim 28 , wherein said antibody immunospecifically binds to an epitope comprising amino acid residues 5 through 13 of hBNP.  
     
     
         47 . An immunoassay for hBNP or hBNP fragment, wherein said immunoassay comprises an antibody of  claim 1 .  
     
     
         48 . The immunoassay of  claim 47 , wherein said immunoassay comprises a single antibody that immunospecifically binds to hBNP or hBNP fragment.  
     
     
         49 . The immunoassay of  claim 47 , wherein said immunoassay further comprising an additional specific binding partner for hBNP or hBNP fragment.  
     
     
         50 . A pharmaceutical composition comprising a therapeutically effective amount of an antibody of  claim 1.