Coagulation assay reagents containing lanthanides

• US 6,902,904 B2
• Filed: 08/27/2001
• Issued: 06/07/2005
• Est. Priority Date: 08/27/2001
• Status: Expired due to Fees

• Current Assignee: PharmaNetics, Inc.
• Original Assignee: PharmaNetics, Inc.
• Inventors: Mahan, Donald E., Cutsforth, Gwyn A.
• Primary Examiner(s): GITOMER, RALPH J
• Application Number:
  US09/938,728
• Publication Number:
  US 20030044871A1
• Time in Patent Office: 1,380 Days
• Field of Search: 435/13, 422/58, 422/61, 422/7, 422/73, 436/82, 436/172
• US Class Current: 435/13
• CPC Class Codes: C12Q 1/56 involving blood clotting factors, e.g. involving thrombin, thromboplastin, fibrinogen
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/86 involving blood coagulating time or factors, or their receptors

Abstract

A method, kit, system and reagent for performing coagulation assays with higher sensitivity and greater dynamic range is provided which involves the use of one or more metal compounds that interact with calcium binding sites in the blood coagulation cascade, particularly lanthanide compounds, manganese compounds and magnesium compounds. A Protein C reagent, kit, and assay method is also provided using the same type of metal compounds to provide greater detection sensitivity and dynamic range between samples.

Claims

1. A dry coagulation assay reagent comprising magnetic particles distributed substantially homogeneously therethrough wherein said reagent is selected from the group consisting of:

(1) one member selected from the group consisting of prothrombin time reagents and (ii) prothrombin time reagents and calcium salts;

(2) partial thromboplastin time reagents with calcium chloride;

(3) partial thromboplastin time reagents with calcium chloride and clot formation activators;

(4) thrombin or a snake venom with thrombotic activity;

(5) fibrin, or plasminogen and fibrin;

(6) plasminogen activator assay reagents containing (i) plasminogen and (ii) either fibrin, a snake venom with thrombotic activity, or thrombin;

(7) plasminogen assay reagents containing (i) a plasminogen activator and (ii) either fibrin, a snake venom with thrombotic activity, or thrombin;

(8) natural and synthetic plasminogen activators;

(9) alpha-2-antiplasmin assay reagents containing fibrin and plasmin; and

(10) combinations thereof,

wherein the reagent further comprises one or more lanthanide metal compounds that interact with calcium binding sites in a blood coagulation cascade.

2. The reagent of claim 1, wherein said one or more lanthanide metal compounds are selected from the group consisting of compounds of europium, lanthanum, gadolinium and terbium.

3. The reagent of claim 1, wherein said one or more lanthanide metal compounds are selected from the group consisting of salts of organic acids, halides, C2-C6 alkyl esters of organic acids, hexafluorophosphates, nitrates, oxalates, perchlorates, tetrafluoroborates, trifluoromethanesulfonates, substituted and unsubstituted C4-C8 alkanedionates, acetylacetonates, and sulfates.

4. The reagent of claim 1, wherein said reagent is thromboplastin or thromboplastin and a calcium salt.

5. The reagent of claim 1, wherein said reagent is a partial thromboplastin time reagent with calcium chloride.

6. The reagent of claim 1, wherein said reagent is a partial thromboplastin reagent with calcium chloride and a clot formation activator.

7. The reagent of claim 1, wherein said reagent is thrombin or a snake venom with thrombotic activity.

8. The reagent of claim 1, wherein said reagent is fibrin, plasminogen and fibrin, a snake venom with thrombin-like activity, or thrombin.

9. The reagent of claim 8, comprising either said snake venom with thrombotic activity or a combination of said snake venom with thrombotic activity and thrombin.

10. The reagent of claim 1, wherein said reagent is a plasminogen assay reagent containing (i) a plasminogen activator and (ii) either fibrin, a snake venom with thrombotic activity, or thrombin.

11. The reagent of claim 10, comprising either said snake venom with thrombotic activity or a combination of said snake venom with thrombotic activity and thrombin.

12. The reagent of claim 1, wherein said reagent is one member selected from the group consisting of streptokinase and urokinase.

13. The reagent of claim 1, wherein said reagent comprises one member selected from the group consisting of natural and synthetic tissue plasminogen activators.

14. The reagent of claim 1, wherein said reagent is an alpha-2-antiplasmin assay reagent containing fibrin and plasmin.

15. The reagent of claim 1, wherein said reagent comprises:

(4) thrombin or a snake venom with thrombotic activity; and

(8) one member selected from the group consisting of natural and synthetic plasminogen activators.

16. The reagent of claim 1, wherein said reagent comprises at least one member selected from the group consisting of streptokinase, tissue plasminogen activator and urokinase.

17. The reagent of claim 1, wherein said reagent is thrombomodulin or recombinant thrombomodulin.

18. The reagent of claim 1, wherein said one or more metals is at least one member selected from the group consisting of cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, ytterbium, and lutetium.

19. The reagent of claim 18, wherein said reagent is thrombomodulin or recombinant thrombomodulin.

20. A reagent for performing a Protein C assay, comprising:

an initiator of an intrinsic blood coagulation pathway or Factor X;

a Protein C activator;

calcium ions; and

one or more lanthanide metal compounds that interact with calcium binding sites in a blood coagulation cascade.

21. The reagent of claim 20, wherein said Protein C activator is a thrombomodulin.

22. The reagent of claim 20, wherein said one or more metal compounds are one or more members selected from the group consisting of compounds of terbium, gadolinium, europium and lanthanum.

23. The reagent of claim 20, further comprising one or more bulkers, stabilizers or both.