Controlled platelet activation to monitor therapy of ADP antagonists

• US 9,341,637 B2
• Filed: 11/10/2010
• Issued: 05/17/2016
• Est. Priority Date: 07/08/2003
• Status: Active Grant

• Current Assignee: Instrumentation Lab Co (Instrumentation Lab)
• Original Assignee: Accumetrics, Inc. (Accriva Diagnostics, Inc.)
• Inventors: Coller, Barry S., Durbin, Dennis
• Primary Examiner(s): DRISCOLL, LORA E BARNHART
• Application Number:
  US12/943,413
• Publication Number:
  US 20110081657A1
• Time in Patent Office: 2,015 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: G01N 33/86 involving blood coagulating time or factors, or their receptors

Abstract

A method is provided of determining whether an individual has reduced ability to form platelet thrombi due to inhibition of platelet activation initiation, signal transduction and/or GPIIb/IIIa blockade. A blood sample is obtained from the individual being assessed. The blood sample is mixed in combination with 1) an anticoagulant; 2) sufficient buffer to maintain the pH and salt concentration of the anticoagulated blood within a range suitable for platelet aggregation; 3) a platelet GPIIb/IIIa receptor ligand immobilized on a solid surface; 4) one or more agents to enhance a signal transduction pathway and 5) a receptor activator. The combination is incubated under conditions for agglutinating particles. Platelet-mediated agglutination is assessed in the agitated mixture. The absence of agglutination indicates that the individual has a reduced ability to form platelet thrombi.

Claims

1. A method for determining a patient'"'"'s response to a P2Y12 antagonist, comprising the steps of:

providing a platelet-containing blood sample from a patient an individual treated with a P2Y12 antagonist;

contacting the platelet-containing blood sample with adenosine diphosphate (ADP), prostaglandin E1 (PGE1) and particles comprising a GPIIb/IIIa receptor ligand, under conditions suitable for platelet aggregation as measured by agglutination of the particles,

measuring the level of particle agglutination;

determining the level of platelet aggregation as measured by the level of particle agglutination;

wherein a reduced level of agglutination, as compared to a control level, indicates an effective response to said P2Y12 antagonist treatment.

2. The method of claim 1, wherein said P2Y12 antagonist is a thienopyridine.

3. The method of claim 2, wherein said thienopyridine is clopidogrel.

4. The method of claim 2, wherein said thienopyridine is ticlopidine.

5. The method of claim 1, wherein platelet-containing blood sample is from a patient treated with a P2Y12 antagonist and aspirin.

6. The method of claim 1, wherein the platelet-containing blood sample is contacted with PGE1 at a concentration of 20 nM to 25 nM.

7. The method of claim 1, wherein said platelet-containing blood sample is a whole blood sample.

8. The method of claim 1, wherein said platelet-containing blood sample is a plasma sample.

9. The method of claim 8, wherein said plasma sample is a platelet rich plasma sample.

10. The method of claim 1, wherein the level of platelet aggregation is determined in an aqueous medium comprising the blood sample.

11. The method of claim 1, wherein the blood sample volume is from 30 μ

L to 5000 μ

L.