Complement-mediated thrombophilic assay
View Patent ↗A method for assessing the presence of an acquired thrombophilia disorder in a patient exhibiting hypercoagulation is disclosed.
1. A method for assessing the presence of an acquired thrombophilia disorder in a patient exhibiting hypercoagulation, the method comprising:
a) incubating primary healthy blood cells in a first incubation condition, the first incubation condition comprising the presence of serum obtained from the patient;
b) incubating primary healthy blood cells in a second incubation condition, the second incubation condition comprising the presence of serum obtained from the patient, wherein the serum further comprises a complement inhibitor; and
c) detecting the presence of tissue factor in the first incubation condition and in the second incubation condition, wherein the presence of tissue factor in the first incubation condition and not in the second incubation condition is indicative of an acquired thrombophilia disorder in the patient.
2. The method of claim 1 , wherein the detecting step comprises contacting the incubation conditions with an agent capable of binding tissue factor.
3. The method of claim 2 , wherein the agent is an anti-tissue factor antibody or functional fragment thereof.
4. The method of claim 1 , wherein the detecting step comprises detecting the presence of tissue factor in a first cell supernatant and in a second cell supernatant, wherein the first cell supernatant is obtained from the first incubation condition and the second cell supernatant is obtained from the second incubation.
5. The method of claim 4 , wherein detecting the presence of tissue factor in a first cell supernatant and in a second cell supernatant comprises measuring a first rate of coagulation using the first cell supernatant and a second rate of coagulation using the second cell supernatant, wherein an acquired thrombophilia disorder is indicated in the patient if the first rate of coagulation is accelerated compared to the second rate of coagulation.
6. The method of claim 1 , wherein the complement inhibitor is a C3 inhibitor.
7. The method of claim 6 , wherein the C3 inhibitor is selected from the group consisting of: compstatin, a compstatin analog, a compstatin peptidomimetic, a compstatin derivative, and combinations thereof.
8. The method of claim 1 , wherein the complement inhibitor is a C5aR inhibitor.
9. The method of claim 8 , wherein the C5aR inhibitor is selected from the group consisting of: an anti-C5 antibody, an anti-C5 aptamer, an anti-C5aR antibody and a C5aR antagonist.
10. The method of claim 9 , wherein the C5aR antagonist is selected from the group consisting of: PMX-53, a PMX-53 analog, and combinations thereof.
11. The method of claim 1 , wherein the acquired thrombophilia disorder is one of antiphospholipid syndrome and systemic lupus erythymatosus.