Novel methods and antibodies for treating coagulapathy
A technology for coagulation disorders and antibodies, applied in the fields of antibodies, gene therapy, chemical instruments and methods, etc., can solve problems such as lack of high blood clots
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[0267] The following embodiments are provided to aid understanding of the present invention, however, the present invention is not limited to the following embodiments.
[0268] In one embodiment, the invention relates to inhibitors (such as but not limited to antibodies, Fab or other fragments, peptides or aptamers) that bind to protein S and inhibit the interaction of protein S with APCs.
[0269] In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to protein S and inhibits the interaction of protein S with APC.
[0270] In one embodiment, the invention relates to an antibody or antigen-binding fragment thereof that binds to protein S and inhibits the interaction of protein S with APCs without interfering with known non-coagulation functions of protein S.
[0271] In one embodiment, the present invention relates to the use of an antibody or antigen-binding fragment thereof that binds to protein S and inhibits the interaction...
Embodiment 1
[0472] Example 1: Polyclonal Antibodies Against Protein S Improve APTT in Human Hemophiliac Plasma
[0473] In FVIII-deficient human plasma, polyclonal anti-Protein S antibody concentration-dependently shortened clotting time in the presence of APC ( figure 1 ). Congenitally FVIII-deficient human plasma (George King Biomedical Inc.) was treated with 0.3 μg / ml APC (Innovative Research) and indicated levels of polyclonal anti-protein S (DAKO #A0384) together with APTT reagent (APTT-SP, IL) before recalcification. Incubate at 37°C for 300 sec. The time to fibrin clot formation was measured using an ACL9000 (ILS). Average EC 50 It was 37.1 μg / ml (SD=2.4, n=3 experiments), corresponding to about 250 nM.
Embodiment 2
[0474] Example 2: Procoagulant effect of anti-Protein S antibody compared to FVIII in hemophilia A plasma
[0475] The maximum effect of anti-Protein S antibody in FVIII-deficient plasma was compared with the clotting times of normal human plasma and human plasma with 1%, 5% and 10% FVIII (internal), respectively ( figure 2 ). The data indicated that the complete response with anti-protein S was similar to the clotting time of plasma with 5-10% FVIII. The effect of complete neutralization of protein S was confirmed by establishing the clotting time of protein S-deficient plasma (HaemochromDiagnostica) with excess neutralizing FVIII antibody (internal), similar to protein S and FVIII doubly deficient plasma.
[0476]Plasma was mixed with APC (0.3 μg / ml) and antiprotein S (DAKO, #A0384) or FVIII in various combinations together with APTT reagent (APTT-SP, IL) and incubated at 37°C before recalcification 300sec. The time to fibrin clot formation was measured using an ACL9000 ...
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