High specificity monoclonal antibody against a protein or a polypeptide having oxidative modification
The present invention provides an antibody, which reacts with a FGA or FGA partial peptide in which a part of prolines in the molecule thereof are hydroxylated, and which does not react with an unmodified FGA or FGA partial peptide.
1. An isolated antibody that specifically binds to hydroxylated fibrinogen comprising at least one hydroxylated proline, wherein the isolated antibody has a higher affinity for the hydroxylated fibrinogen than for fibrinogen having no hydroxylated prolines,
wherein when the hydroxylated fibrinogen is attached to a solid-phase in a reaction system for a competitive reaction between hydroxylated fibrinogen not attached to the solid-phase (antigen 1) and fibrinogen haying no hydroxylated prolines (antigen 2), the antibody satisfies measurement conditions in which 50% inhibition activity of an antibody bound by the antigen 1 is at least 10-fold when compared to 50% inhibition activity of an antibody bound by the antigen 2, and
wherein the hydroxylated fibrinogen attached to the solid-phase and the antigen 1 are represented by the amino acid sequence TFP(OH)GFFSPMLGEFVSETESR (SEQ ID NO: 3) or ESSSHEIP(OH)GIAEFPSR (SEQ ID NO: 4) (P(OH) represents hydroxylated proline), and wherein the amino acid sequence of the antigen 2 is represented by TFPGFFSPMLGEFVSETESR (SEQ ID NO: 1) or ESSSHHPGIAEFPSR (SEQ ID NO: 2).
2. The isolated antibody according to claim 1 , wherein the antibody is a monoclonal antibody.
3. A detection reagent for a protein or polypeptide comprising an amino acid sequence in which a part of prolines in the molecule thereof are hydroxylated, the reagent comprising the isolated antibody according to claim 1 .
4. The reagent according to claim 3 , wherein the protein is fibrinogen.
5. The reagent according to claim 3 , wherein the protein or polypeptide comprising an amino acid sequence in which a part of prolines in the molecule thereof are hydroxylated comprises an amino acid sequence in which a part of prolines in the amino acid sequence of (a) or (b) below are hydroxylated:
(a)
TFPGFFSPMLGEFVSETESR
(SEQ ID NO: 1)
(b)
ESSSHHPGIAEFPSR.
(SEQ ID NO: 2)
6. The reagent according to claim 3 , wherein the protein or polypeptide comprising an amino acid sequence in which a part of prolines in the molecule thereof are hydroxylated comprises the amino acid sequence of (c) or (d) below:
(c)
TFP(OH)GFFSPMLGEFVSETESR
(SEQ ID NO: 3)
(d)
ESSSHHP(OH)GIAEFPSR
(SEQ ID NO: 4)
(P(OH) represents hydroxylated proline).
7. A detection or diagnostic agent for cancer, comprising the isolated antibody according to claim 1 or a reagent for a protein or polypeptide comprising an amino acid sequence in which a part of prolines in the molecule thereof are hydroxylated, the reagent comprising the isolated antibody according to claim 1 .
8. The detection or diagnostic agent according to claim 7 , wherein the cancer is pancreatic cancer.
9. A monoclonal antibody to hydroxylated fibrinogen comprising at least one hydroxylated praline, wherein the antibody has a higher affinity for the hydroxylated fibrinogen than for fibrinogen having no hydroxylated prolines, which is produced by a hybridoma whose accession number is FERM BP-11163.
10. The monoclonal antibody according to claim 9 , wherein the hydroxylated fibrinogen is represented by TFP(OH)GFFSPMLGEFVSETESR (SEQ ID NO: 3) or ESSSHHP(OH)GIAEFPSR (SEQ ID NO: 4) (P(OH) represents hydroxylated proline).
11. A detection or diagnostic agent for cancer, comprising the isolated antibody according to claim 9 or a reagent for a protein or polypeptide comprising an amino acid sequence in which a part of prolines in the molecule thereof are hydroxylated, the reagent comprising the antibody according to claim 9 .
12. The monoclonal antibody according to claim 9 , wherein, in order to inhibit an immune reaction between the monoclonal antibody and the hydroxylated fibrinogen, wherein hydroxylated fibrinogen is attached to a solid-phase in a reaction system for a competitive reaction between hydroxylated fibrinogen not attached to the solid-phase (antigen 1) and fibrinogen having no hydroxylated prolines (antigen 2), the monoclonal antibody satisfies measurement conditions in which 50% inhibition activity to the immune reaction caused by the antigen 1 is at least 10-fold when compared to 50% inhibition activity to the immune reaction caused by the antigen 2.
13. The monoclonal antibody according to claim 12 , wherein the amino acid sequence of the antigen 2 is represented by TFPGFFSPMLGEFVSETESR (SEQ ID NO: 1) or ESSSHHPGIAEFPSR (SEQ ID NO: 2).
14. The monoclonal antibody according to claim 12 , wherein the hydroxylated fibrinogen attached to the solid-phase and the antigen 1 comprise the amino acid sequence TFP(OH)GFFSPMLGEFVSETESR (SEQ ID NO: 3) or ESSSHHEP(OH)GIAEFPSR (SEQ ID NO: 4) (P(OH) represents hydroxylated proline).
15. A detection reagent for a protein or polypeptide comprising an amino acid sequence in which a part of prolines in the molecule thereof are hydroxylated, the reagent comprising the monoclonal antibody according to claim 9 .
16. The reagent according to claim 15 , wherein the protein is fibrinogen.
17. The reagent according to claim 15 , wherein the protein or polypeptide comprising an amino acid sequence in which a part of prolines in the molecule thereof are hydroxylated comprises the amino acid sequence of (c) or (d) below:
(c)
TFP(OH)GFFSPMLGEFVSETESR
(SEQ ID NO: 3)
(d)
ESSSHHP(OH)GIAEFPSR
(SEQ ID NO: 4)
(P(OH) represents hydroxylated proline).
18. An isolated hybridoma cell which produces the monoclonal antibody according to claim 9 .
19. An isolated antibody that specifically binds to hydroxylated fibrinogen comprising at least one hydroxylated proline, wherein the isolated antibody has a higher affinity for the hydroxylated fibrinogen than for fibrinogen having no hydroxylated prolines,
wherein when the hydroxylated fibrinogen is attached to a solid-phase in a reaction system for a competitive reaction between hydroxylated fibrinogen not attached to the solid-phase (antigen 1) and fibrinogen having no hydroxylated prolines (antigen 2), the antibody satisfies measurement conditions in which 50% inhibition activity of an antibody bound by the antigen 1 is at least 10-fold when compared to 50% inhibition activity of an antibody bound by the antigen 2, and wherein the hydroxylated fibrinogen attached to the solid-phase and the antigen 1 are represented by the amino acid sequence TFP(OH)GFFSPMLGEFVSETESR (SEQ ID NO: 3) or ESSSHHP(OH)GIAEFPSR (SEQ ID NO: 4) (P(OH) represents hydroxylated proline), and wherein the amino acid sequence of the antigen 2 is represented by TFPGFFSPMLGEFVSETESR (SEQ ID NO: 1) or ESSSHHPGIAEFPSR (SEQ ID NO: 2), and
wherein the isolated antibody specifically binds to a site to which a monoclonal antibody which is produced by the hybridoma whose accession number is PERM BP-11163 specifically binds.
20. An isolated hybridoma cell which produces an antibody to hydroxylated fibrinogen comprising at least one hydroxylated proline, wherein the antibody has a higher affinity for the hydroxylated fibrinogen than for fibrinogen having no hydroxylated prolines, wherein the accession number of the cell line is FERM BP-11163.
21. A monoclonal antibody or an antigen binding fragment thereof produced by hybridoma having accession number PERM BP-11163.
22. A monoclonal antibody or an antigen binding fragment thereof which binds to the same epitope as the monoclonal antibody produced by hybridoma having accession number FERM BP-11163.