Methods and kits for detection of thromboxane A2 metabolites
View Patent ↗Methods, compositions and kits are provided for measuring aspirin's anti-thrombotic effectiveness on a subject. Included are a novel assay for quickly and specifically measuring TxA2 metabolite levels in urine and correlating the levels with aspirin dose in a subject. The methods, compositions and kits utilize a novel anti TxA2 metabolite antibody.
1. A method for identifying aspirin response in a subject comprising:
administering to the subject a first dose of aspirin;
collecting a urine sample from the subject;
determining an amount of two thromboxane A 2 metabolites in the urine sample, wherein the determining the amount of thromboxane A 2 metabolites comprises using a monoclonal antibody capable of binding to 11-dehydro thromboxane B 2 and 11-dehydro-2,3-dinor Thromboxane B 2 ;
normalizing the amount of the two thromboxane A 2 metabolites to an amount of a standard protein in the subject's urine to obtain a first thromboxane A 2 metabolite/standard protein ratio; and
comparing the first thromboxane A 2 metabolite/standard protein ratio to a threshold ratio value wherein a thromboxane A 2 metabolite/standard protein value below the threshold value indicates aspirin responsiveness in the subject.
2. The method of claim 1 further comprising:
administering to the subject a second dose of aspirin different from the first dose of aspirin, the subject having a first thromboxane A 2 metabolite/standard protein ratio above the threshold value, and wherein the first dose and second dose of aspirin are at least 50 mg apart;
collecting a second urine sample from the subject;
determining an amount of the two thromboxane A 2 metabolites in the urine sample;
normalizing the amount of the two thromboxane A 2 metabolites to an amount of a standard protein in the subject's urine to obtain a second thromboxane A 2 metabolite/standard protein ratio; and
comparing the first and second thromboxane A 2 metabolite/standard protein ratios;
wherein the subject having substantially the same first and second thromboxane A 2 metabolite/standard protein ratio is identified as not responsive to aspirin.
3. The method of claim 2 wherein the first dose and second dose of aspirin are at least 100 mg apart.
4. The method of claim 1 wherein the monoclonal antibody recognizes an epitope common to both 11-dehydro Thromboxane B 2 and 11-dehydro-2,3-dinor Thromboxane B 2 .
5. The method of claim 1 , wherein the subject is suffering from or suspected of suffering from thrombotic disease.
6. The method of claim 1 , wherein the subject is suffering from or suspected of suffering from cardiovascular disease.
7. The method of claim 1 , wherein the subject is suffering from or suspected of suffering from cerebrovascular ischemia.
8. A method for identifying aspirin response in a subject comprising:
administering to the subject a first dose of aspirin;
collecting a blood plasma sample from the subject;
determining an amount of two thromboxane A 2 metabolites in the sample, wherein the determining the amount of thromboxane A 2 metabolites comprises using a monoclonal antibody capable of binding to 11-dehydro thromboxane B 2 and 11-dehydro-2,3-dinor Thromboxane B 2 ;
normalizing the amount of the two thromboxane A 2 metabolites to an amount of a standard protein in the subject's plasma to obtain a first thromboxane A 2 metabolite/standard protein ratio; and
comparing the first thromboxane A 2 metabolite/standard protein ratio to a threshold ratio value wherein a thromboxane A 2 metabolite/standard protein value below the threshold value indicates aspirin responsiveness in the subject.
9. The method of claim 8 further comprising:
administering to the subject a second dose of aspirin different from the first dose of aspirin, the subject having a first thromboxane A 2 metabolite/standard protein ratio above the threshold value, and wherein the first dose and second dose of aspirin are at least 50 mg apart;
collecting a second plasma sample from the subject;
determining an amount of the two thromboxane A 2 metabolites in the plasma sample;
normalizing the amount of the two thromboxane A 2 metabolites to an amount of a standard protein in the subject's plasma to obtain a second thromboxane A 2 metabolite/standard protein ratio; and
comparing the first and second thromboxane A 2 metabolite/standard protein ratios;
wherein the subject having substantially the same first and second thromboxane A 2 metabolite/standard protein ratio is identified as not responsive to aspirin.
10. The method of claim 9 wherein the first dose and second dose of aspirin are at least 100 mg apart.
11. The method of claim 8 wherein the monoclonal antibody recognizes an epitope common to both 11-dehydro Thromboxane B 2 and 11-dehydro-2,3-dinor Thromboxane B 2 .
12. The method of claim 8 , wherein the subject is suffering from or suspected of suffering from cardiovascular disease.