Nucleic acid encoding hepatocyte growth factor (HGF) binding proteins
View Patent ↗The present invention provides a family of binding proteins that bind and neutralize the activity of hepatocyte growth factor (HGF), in particular human HGF. The binding proteins can be used as diagnostic and/or therapeutic agents. With regard to their therapeutic activity, the binding proteins can be used to treat certain HGF responsive disorders, for example, certain HGF responsive tumors.
1. An isolated nucleic acid comprising a nucleotide sequence encoding an immunoglobulin heavy chain variable region comprising the structure CDR H1 -CDR H2 -CDR H3 , wherein
(i) CDR H1 comprises the amino acid sequence of SEQ ID NO: 15,
(ii) CDR H2 comprises the amino acid sequence of SEQ ID NO: 204, and
(iii) CDR H3 comprises the amino acid sequence of SEQ ID NO: 17; and
an immunoglobulin light chain variable region comprising the structure CDR L1 -CDR L2 -CDR L3 , wherein
(i) CDR L1 comprises the amino acid sequence of SEQ ID NO: 18,
(ii) CDR L2 comprises the amino acid sequence of SEQ ID NO: 19, and
(iii) CDR L3 comprises the amino acid sequence of SEQ ID NO: 20;
wherein the immunoglobulin heavy chain variable region and the immunoglobulin light chain variable region together bind human hepatocyte growth factor (HGF).
2. The isolated nucleic acid of claim 1 , wherein the complementarity determining sequences (CDRs) are interposed between human or humanized immunoglobulin framework regions.
3. The isolated nucleic acid of claim 1 , wherein the nucleotide sequence encodes an immunoglobulin heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 183.
4. The isolated nucleic acid of claim 1 , wherein the nucleotide sequence encodes an immunoglobulin light chain variable region comprising the amino acid sequence of SEQ ID NO: 193.
5. The isolated nucleic acid of claim 1 , wherein the nucleotide sequence encodes an immunoglobulin heavy chain comprising the amino acid sequence of SEQ ID NO: 187.
6. The isolated nucleic acid of claim 1 , wherein the nucleotide sequence encodes an immunoglobulin light chain comprising the amino acid sequence of SEQ ID NO: 197.
7. An expression vector comprising the nucleic acid of claim 1 .
8. A host cell comprising the expression vector of claim 7 .
9. A host cell comprising a first expression vector comprising a nucleic acid sequence encoding an immunoglobulin heavy chain variable region comprising the structure CDR H1 -CDR H2 -CDR H3 , wherein
(i) CDR H1 comprises the amino acid sequence of SEQ ID NO: 15,
(ii) CDR H2 comprises the amino acid sequence of SEQ ID NO: 204, and
(iii) CDR H3 comprises the amino acid sequence of SEQ ID NO: 17; and
a second expression vector comprising a nucleic acid sequence encoding an immunoglobulin light chain variable region comprising the structure CDR L1 -CDR L2 -CDR L3 , wherein
(i) CDR L1 comprises the amino acid sequence of SEQ ID NO: 18,
(ii) CDR L2 comprises the amino acid sequence of SEQ ID NO: 19, and
(iii) CDR L3 comprises the amino acid sequence of SEQ ID NO: 20;
wherein the immunoglobulin heavy chain variable region and the immunoglobulin light chain variable region together bind human hepatocyte growth factor (HGF).
10. A method of producing a monoclonal antibody that binds human hepatocyte growth factor (HGF) or an antigen binding fragment of the antibody, the method comprising:
(a) growing the host cell of claim 8 under conditions that permit the expression of a polypeptide comprising the immunoglobulin heavy chain variable region and a polypeptide comprising the immunoglobulin light chain variable region, thereby producing the antibody or antigen binding fragment of the antibody; and
(b) purifying the antibody or antigen binding fragment of the antibody.
11. A method of producing a monoclonal antibody that binds human hepatocyte growth factor (HGF) or an antigen binding fragment of the antibody, the method comprising:
(a) growing the host cell of claim 9 under conditions that permit the expression of a polypeptide comprising the immunoglobulin heavy chain variable region and a polypeptide comprising the immunoglobulin light chain variable region, thereby producing the antibody or the antigen-binding fragment of the antibody; and
(b) purifying the antibody or the antigen-binding fragment of the antibody.
12. An isolated nucleic acid comprising a nucleotide sequence encoding an immunoglobulin heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 183, and encoding an immunoglobulin light chain variable region comprising the amino acid sequence of SEQ ID NO: 193, wherein the immunoglobulin heavy chain variable region and the immunoglobulin light chain variable region together bind human hepatocyte growth factor (HGF).
13. An expression vector comprising the nucleic acid of claim 12 .
14. A host cell comprising the expression vector of claim 13 .
15. A host cell comprising:
a first expression vector comprising a nucleic acid sequence encoding an immunoglobulin heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 183; and
a second expression vector comprising a nucleic acid sequence encoding an immunoglobulin light chain variable region comprising amino acid sequence of SEQ ID NO: 193;
wherein the immunoglobulin heavy chain variable region and the immunoglobulin light chain variable region together bind human hepatocyte growth factor (HGF).
16. A method of producing a monoclonal antibody that binds human hepatocyte growth factor (HGF) or an antigen binding fragment of the antibody, the method comprising:
(a) growing the host cell of claim 14 under conditions that permit the expression of a polypeptide comprising the immunoglobulin heavy chain variable region and a polypeptide comprising the immunoglobulin light chain variable region, thereby producing the antibody or antigen binding fragment of the antibody; and
(b) purifying the antibody or antigen binding fragment of the antibody.
17. A method of producing a monoclonal antibody that binds human hepatocyte growth factor (HGF) or an antigen binding fragment of the antibody, the method comprising:
(a) growing the host cell of claim 15 under conditions that permit the expression of a polypeptide comprising the immunoglobulin heavy chain variable region and a polypeptide comprising the immunoglobulin light chain variable region, thereby producing the antibody or the antigen-binding fragment of the antibody; and
(b) purifying the antibody or the antigen-binding fragment of the antibody.
18. An isolated nucleic acid comprising a nucleotide sequence encoding an immunoglobulin heavy chain comprising the amino acid sequence of SEQ ID NO: 187, and encoding an immunoglobulin light chain comprising the amino acid sequence of SEQ ID NO: 197, wherein the immunoglobulin heavy chain variable region and the immunoglobulin light chain variable region together bind human hepatocyte growth factor (HGF).
19. An expression vector comprising the nucleic acid of claim 18 .
20. A host cell comprising the expression vector of claim 19 .
21. A host cell comprising:
a first expression vector comprising a nucleic acid sequence encoding an immunoglobulin heavy chain comprising the amino acid sequence of SEQ ID NO: 187; and
a second expression vector comprising a nucleic acid sequence encoding an immunoglobulin light chain comprising the amino acid sequence of SEQ ID NO: 197;
wherein the immunoglobulin heavy chain and the immunoglobulin light chain together bind human hepatocyte growth factor (HGF).
22. A method of producing a monoclonal antibody that binds human hepatocyte growth factor (HGF) or an antigen binding fragment of the antibody, the method comprising:
(a) growing the host cell of claim 20 under conditions that permit the expression of a polypeptide comprising the immunoglobulin heavy chain and a polypeptide comprising the immunoglobulin light chain, thereby producing the antibody or antigen binding fragment of the antibody; and
(b) purifying the antibody or antigen binding fragment of the antibody.
23. A method of producing a monoclonal antibody that binds human hepatocyte growth factor (HGF) or an antigen binding fragment of the antibody, the method comprising:
(a) growing the host cell of claim 21 under conditions that permit the expression of a polypeptide comprising the immunoglobulin heavy chain and a polypeptide comprising the immunoglobulin light chain, thereby producing the antibody or the antigen-binding fragment of the antibody; and
(b) purifying the antibody or the antigen-binding fragment of the antibody.