Antibodies directed against prothrombin fragment F
View Patent ↗The invention relates to antibodies directed against F 1+2 , to the preparation and use thereof, especially in therapy and diagnosis. The antibodies bind to an epitope on the N-terminal half of the F 2 fragment of prothrombin.
1. An isolated monoclonal antibody which binds specifically to a peptide comprising an epitope having the amino acid sequence Ser-Pro-Pro-Leu-Glu-Gln-Cys (amino acids 9-15 of SEQ ID NO:2), wherein the antibody specifically binds said sequence.
2. The isolated antibody as claimed in claim 1 wherein the antibody is associated with at least one of a solid phase and a component of a signal-generating system.
3. An isolated antibody which is produced by the hybridoma cell line DSM ACC2607.
4. A composition comprising the antibody as claimed in claim 1 and a pharmaceutically acceptable, sterile injection medium.
5. A reagent comprising the antibody as claimed in claim 1 .
6. A test kit comprising the reagent as claimed in claim 5 .
7. An isolated cell which produces the antibody as claimed in claim 1 .
8. The isolated cell as claimed in claim 7 , wherein the cell is an animal, plant, prokaryotic, or human cell.
9. A hybridoma cell line which produces the isolated antibody as claimed in claim 1 .
10. The hybridoma cell line DSM ACC2607, which produces the antibody of claim 3 .
11. An isolated monoclonal antibody which binds specifically to an epitope consisting of the amino acid sequence Ser-Pro-Pro-Leu-Glu-Gln-Cys (amino acids 9-15 of SEQ ID NO:2).
12. A method of diagnosing a hypercoagulability state or thrombotic event, comprising contacting a sample with a first antibody, wherein the first antibody specifically binds to an epitope that includes the four carboxy-terminal amino acids of the prothrombin fragments F 1+2 and F 2 ; removing substances from said sample that are not bound by the first antibody; contacting the resulting sample with a second antibody as claimed in claim 1 ; allowing said second antibody to bind to F 1+2 and F 2 ; removing unbound second antibody from said sample; and measuring the concentration, amount, or presence of F 1+2 and F 2 in said sample via a label associated with the second antibody.
13. A method of detecting prothrombin fragments F 1+2 and F 2 in a sample, comprising contacting a sample with a first antibody, wherein the first antibody specifically binds to an epitope that includes the four carboxy-terminal amino acids of the F 2 and F 1+2 fragments; removing substances from said sample that are not bound by the first antibody; contacting the resulting sample with a second antibody as claimed in claim 1 ; allowing said second antibody to bind to F 1+2 and F 2 ; removing unbound second antibody from said sample; and detecting or measuring the signal produced by a label associated with the second antibody in said sample.
14. A method for the quantitative or qualitative detection of at least one sample analyte chosen from prothrombin fragments F 1+2 and F 2 in a sample comprising performing a competitive binding assay comprising (a) contacting the sample with an analyte-specific binding partner and a reagent analyte, a component of a signal-generating system being associated, directly or indirectly, with the analyte-specific binding partner or the reagent analyte, and (b) detecting binding, quantitatively or qualitatively, of the reagent analyte to the analyte-specific binding partner by detecting signal from the signal-generating system, wherein the analyte-specific binding partner comprises at least one antibody as claimed in claim 1 , and wherein the reagent analyte comprises at least one peptide consisting of 5 to 25 amino acids, which includes amino acids 11 to 15 of SEQ ID NO: 2.
15. A method for the quantitative or qualitative detection of at least one sample analyte chosen from prothrombin fragments F 1+2 and F 2 in a sample comprising performing a competitive binding assay comprising (a) contacting the sample with an analyte-specific binding partner and a reagent analyte, and (b) detecting binding, quantitatively or qualitatively, of the reagent analyte to the analyte-specific binding partner, wherein the analyte-specific binding partner comprises at least one antibody as claimed in claim 1 , wherein the reagent analyte comprises at least one peptide consisting of 5 to 25 amino acids, which includes amino acids 11 to 15 of SEQ ID NO: 2, and wherein (i) the reagent analyte is directly or indirectly associated with a component of a signal-generating system and the analyte-specific binding partner is directly or indirectly solid-phase associated, or (ii) the analyte-specific binding partner is directly or indirectly associated with a component of a signal-generating system and the reagent analyte is directly or indirectly solid-phase associated.
16. A method for the quantitative or qualitative detection of at least one sample analyte chosen from prothrombin fragments F 1+2 and F 2 in a sample comprising performing a competitive binding assay comprising (a) contacting the sample with an analyte-specific binding partner and a reagent analyte, (b) allowing the reagent analyte and the sample analyte to compete for binding to said analyte-specific binding partner, wherein binding of analyte-specific binding partner to reagent analyte results in solid-phase association of a component of a signal-generating system directly or indirectly associated with the analyte-specific binding partner or the reagent analyte; and (c) detecting or measuring signal from the signal-generating system qualitatively or quantitatively, wherein the analyte-specific binding partner comprises at least one antibody as claimed in claim 1 , and wherein the reagent analyte comprises at least one peptide consisting of 5 to 25 amino acids, which includes amino acids 11 to 15 of SEQ ID NO: 2.