Antibodies recognizing genetic variants of NT-proBNP
The present disclosure relates to an antibody that specifically binds a mutated NT-proBNP having i) a mutation substituting arginine at position 46 with histidine or ii) a mutation substituting glutamic acid at position 43 with aspartic acid. Moreover, the present disclosure relates to a mutated NT-proBNP or fragment thereof. Further, envisaged by the present disclosure are kits containing the antibody of the present disclosure, or the mutated NT-proBNP of the present disclosure. The present disclosure also concerns a method for diagnosing heart failure.
1. An antibody that specifically binds a mutated NT-proBNP (N-terminal of the prohormone brain natriuretic peptide), wherein said antibody is selected from
(a) an antibody that specifically binds a mutated NT-proBNP comprising a mutation substituting arginine at position 46 with histidine, and wherein the light chain variable domain of the antibody comprises a CDR1 having the sequence LLDDAY (SEQ ID NO: 16), a CDR2 having the sequence KDS, and a CDR3 having the sequence LSVDSSEYSV (SEQ ID NO: 17), and the heavy chain variable domain comprises a CDR1 having the sequence GFSLIGEY (SEQ ID NO: 18), a CDR2 having the sequence MASGGTI (SEQ ID NO: 19), and a CDR3 having the sequence VRSSVSPGDDRDV (SEQ ID NO: 20), and
(b) an antibody that specifically binds a mutated NT-proBNP comprising a mutation substituting glutamic acid at position 43 with aspartic acid, and wherein the light chain variable domain of the antibody comprises a CDR1 having the sequence SSNVGYGNY (SEQ ID NO: 21), a CDR2 having the sequence SAT, and a CDR3 having the sequence VSYDSSSKFGV (SEQ ID NO: 22), and the heavy chain variable domain comprises a CDR1 having the sequence GFSVTNSG (SEQ ID NO: 23), a CDR2 having the sequence INNDGVA (SEQ ID NO: 24), and a CDR3 having the sequence GTRDLPSDVRYGNMYINY (SEQ ID NO: 25).
2. The antibody of claim 1 , wherein said antibody is a monoclonal antibody.
3. The antibody of claim 2 , wherein the antibody is a monoclonal sheep antibody.
4. The antibody of claim 1 , wherein said NT-proBNP is mutated human NT-proBNP, and/or wherein
(i) said mutated NT-proBNP comprising a mutation substituting arginine at position 46 with histidine comprises an amino acid sequence as shown in SEQ ID NO: 1, or
(ii) said mutated NT-proBNP comprising a mutation substituting glutamic acid at position 43 with aspartic acid comprises an amino acid sequence as shown in SEQ ID NO: 2.
5. The antibody of claim 1 , wherein said antibody does not bind to wild-type NT-proBNP.
6. The antibody of claim 1 , wherein said antibody is linked to a detectable label.
7. The antibody of claim 6 , wherein said detectable label is an enzyme, biotin, a radioactive label, a fluorescent label, a chemiluminescent label, an electrochemiluminescent label, a gold label, or a magnetic label.
8. The antibody of claim 6 wherein said detectable label is an electrochemiluminescent label comprising ruthenium.
9. An antigen-binding fragment of the antibody of claim 1 , wherein the antigen-binding fragment is selected from the group consisting of a Fab fragment, a Fab′ fragment, a Facb fragment, a F(ab′) 2 fragment, a scFv fragment, and a Fv fragment.
10. The antigen-binding fragment of claim 9 , wherein said antigen-binding fragment is linked to a detectable label.
11. The antigen-binding fragment of claim 10 , wherein said detectable label is an enzyme, biotin, a radioactive label, a fluorescent label, a chemiluminescent label, an electrochemiluminescent label, a gold label, or a magnetic label.
12. A kit or composition comprising
a) i) an antibody that specifically binds a mutated NT-proBNP comprising a mutation substituting arginine at position 46 with histidine, or an antigen-binding fragment thereof, wherein the light chain variable domain of the antibody comprises a CDR1 having the sequence LLDDAY (SEQ ID NO: 16), a CDR2 having the sequence KDS, and a CDR3 having the sequence LSVDSSEYSV (SEQ ID NO: 17), and the heavy chain variable domain comprises a CDR1 having the sequence GFSLIGEY (SEQ ID NO: 18), a CDR2 having the sequence MASGGTI (SEQ ID NO: 19), and a CDR3 having the sequence VRSSVSPGDDRDV (SEQ ID NO: 20), and
ii) an antibody that specifically binds a mutated NT-proBNP comprising a mutation substituting glutamic acid at position 43 with aspartic acid, or an antigen-binding fragment thereof, wherein the light chain variable domain of the antibody comprises a CDR1 having the sequence SSNVGYGNY (SEQ ID NO: 21), a CDR2 having the sequence SAT, and a CDR3 having the sequence VSYDSSSKFGV (SEQ ID NO: 22), and the heavy chain variable domain comprises a CDR1 having the sequence GFSVTNSG (SEQ ID NO: 23), a CDR2 having the sequence INNDGVA (SEQ ID NO: 24), and a CDR3 having the sequence GTRDLPSDVRYGNMYINY (SEQ ID NO: 25) or
b) i) an antibody that specifically binds a mutated NT-proBNP comprising a mutation substituting arginine at position 46 with histidine, or an antigen-binding fragment thereof, wherein the light chain variable domain of the antibody comprises a CDR1 having the sequence LLDDAY (SEQ ID NO: 16), a CDR2 having the sequence KDS, and a CDR3 having the sequence LSVDSSEYSV (SEQ ID NO: 17), and the heavy chain variable domain comprises a CDR1 having the sequence GFSLIGEY (SEQ ID NO: 18), a CDR2 having the sequence MASGGTI (SEQ ID NO: 19), and a CDR3 having the sequence VRSSVSPGDDRDV (SEQ ID NO: 20), and
ii) an antibody that specifically binds to wild-type NT-proBNP, wherein the antibody specifically binds to a region of the wild-type NT-proBNP comprising amino acid residues 42 to 46 of the wild-type NT-proBNP, or an antigen-binding fragment thereof or
c) i) an antibody that specifically binds a mutated NT-proBNP comprising a mutation substituting glutamic acid at position 43 with aspartic acid, or an antigen-binding fragment thereof, wherein the light chain variable domain of the antibody comprises a CDR1 having the sequence SSNVGYGNY (SEQ ID NO: 21), a CDR2 having the sequence SAT, and a CDR3 having the sequence VSYDSSSKFGV (SEQ ID NO: 22), and the heavy chain variable domain comprises a CDR1 having the sequence GFSVTNSG (SEQ ID NO: 23), a CDR2 having the sequence INNDGVA (SEQ ID NO: 24), and a CDR3 having the sequence GTRDLPSDVRYGNMYINY (SEQ ID NO: 25), and
ii) an antibody that specifically binds to wild-type NT-proBNP, wherein the antibody specifically binds to a region of the wild-type NT-proBNP comprising amino acid residues 42 to 46 of the wild-type NT-proBNP, or an antigen-binding fragment thereof, or
d) i) an antibody that specifically binds a mutated NT-proBNP comprising a mutation substituting arginine at position 46 with histidine, or an antigen-binding fragment thereof, wherein the light chain variable domain of the antibody comprises a CDR1 having the sequence LLDDAY (SEQ ID NO: 16), a CDR2 having the sequence KDS, and a CDR3 having the sequence LSVDSSEYSV (SEQ ID NO: 17), and the heavy chain variable domain comprises a CDR1 having the sequence GFSLIGEY (SEQ ID NO: 18), a CDR2 having the sequence MASGGTI (SEQ ID NO: 19), and a CDR3 having the sequence VRSSVSPGDDRDV (SEQ ID NO: 20), and
ii) an antibody that specifically binds a mutated NT-proBNP comprising a mutation substituting glutamic acid at position 43 with aspartic acid, or an antigen-binding fragment thereof, wherein the light chain variable domain of the antibody comprises a CDR1 having the sequence SSNVGYGNY (SEQ ID NO: 21), a CDR2 having the sequence SAT, and a CDR3 having the sequence VSYDSSSKFGV (SEQ ID NO: 22), and the heavy chain variable domain comprises a CDR1 having the sequence GFSVTNSG (SEQ ID NO: 23), a CDR2 having the sequence INNDGVA (SEQ ID NO: 24), and a CDR3 having the sequence GTRDLPSDVRYGNMYINY (SEQ ID NO: 25), and
iii) an antibody that specifically binds to wild-type NT-proBNP, wherein the antibody specifically binds to a region of the wild-type NT-proBNP comprising amino acid residues 42 to 46 of the wild-type NT-proBNP, or an antigen-binding fragment thereof.
13. The kit or composition of claim 12 further comprising an antibody which binds to an epitope comprising amino acids 27 to 31 of NT-proBNP having a sequence shown in SEQ ID NO: 3.