COMPOSITIONS, KIT AND ONE-STEP METHOD FOR MONITORING COMPOUNDS HAVING ANTI-FACTOR Xa AND/OR ANTI FACTOR IIa ACTIVITIES

Abstract

Provided are compositions and methods for accurate determination of the concentration of anticoagulant in a sample such as a blood or plasma sample. The compositions can contain a Factor X compound and Factor V compound, and additional components as well. Methods for performing the assay include one-step methods in which a sample is added to a coagulation assay composition, and time is monitored from the point of adding the sample and coagulation assay composition to an endpoint, such as clot formation.

1 . A blood coagulation assay composition, comprising: 
 (a) a partially or substantially pure Factor X compound;    (b) Factor V compound; and    (c) a multivalent cation,    wherein the components of the composition are substantially unreacted with one another.    
   
   
       2 . The composition of  claim 1 , wherein the composition is substantially free of liquid or gaseous H 2 O.  
   
   
       3 . The composition of  claim 1 , wherein the composition is substantially free of products of a clotting reaction.  
   
   
       4 . The composition of  claim 1 , further characterized by providing a linear anticoagulant curve when anticoagulant concentration is plotted versus clotting time.  
   
   
       5 . The composition of  claim 1 , further comprising a calcium salt.  
   
   
       6 . The composition of  claim 1 , further comprising lipid or detergent.  
   
   
       7 . The composition of  claim 1 , wherein Factor V compound is a component of a plasma fraction.  
   
   
       8 . The composition of  claim 1 , further comprising fibrinogen.  
   
   
       9 . The composition of  claim 1 , further comprising a plasma fraction containing Factor V compound, and further containing fibrinogen and lipid or detergent added thereto.  
   
   
       10 . The composition of  claim 1 , further comprising a plasma fraction containing Factor V compound, and further containing prothrombin complex, anticoagulant, lipid or detergent, and normal plasma added thereto.  
   
   
       11 . The composition of  claim 1 , wherein the composition has been lyophilized.  
   
   
       12 . The composition of  claim 1 , wherein the composition provides a linear correlation between clotting time and heparin concentration range, wherein the heparin concentration range is selected from the group consisting of 0 units/mL to about 20 units/mL, 0 units/mL to about 15 units/mL, 0 units/mL to about 12 units/mL, 0 units/mL to about 10 units/mL, 0 units/mL to about 7.5 units/mL, and 0 units/mL to about 5 units/mL.  
   
   
       13 . The composition of  claim 1 , wherein the Factor X compound is Factor X a .  
   
   
       14 . The composition of  claim 1 , wherein the Factor V compound is Factor V.  
   
   
       15 . The composition of  claim 1 , further comprising benzamidine.  
   
   
       16 . The composition of  claim 1 , wherein Factor X compound is present in excess relative to the amount of sample to be added to the composition.  
   
   
       17 . The composition of  claim 1 , wherein Factor V compound is present in excess relative to the amount of sample to be added to the composition.  
   
   
       18 . A blood coagulation assay composition, comprising: 
 (a) a Factor X compound; and    (b) Factor V compound,    wherein said Factor X compound and said Factor V compound are substantially not admixed.    
   
   
       19 . A combination comprising the composition of  claim 1  located in a single chamber vessel.  
   
   
       20 . The combination of  claim 19 , wherein the vessel is sealed with a vapor-impermeable seal.  
   
   
       21 . The combination of  claim 19 , wherein a steel ball is located within the vessel.  
   
   
       22 . A kit, comprising the composition of  claim 19 , further including a reference for quantitating the results of a coagulation assay.  
   
   
       23 . A method for preparing the blood coagulation assay composition of  claim 1 , comprising: 
 (a) placing Factor X compound into a vessel; and    (b) placing Factor V compound into the vessel, whereby said Factor X compound and said Factor V compound are substantially not admixed, thereby forming the composition of  claim 1 .    
   
   
       24 . A method of assaying coagulation of a sample, comprising: 
 (a) mixing a sample with the coagulation assay composition of  claim 1;  and    (b) determining the time period beginning upon performing step (a) and ending upon clot formation.    
   
   
       25 . A method for preparing a blood coagulation assay composition, comprising: 
 (a) placing Factor X compound into a vessel; and    (b) placing Factor V compound into the vessel;    whereby said Factor X compound and said Factor V compound are substantially not admixed.    
   
   
       26 . A method of assaying coagulation of a sample, comprising: 
 (a) mixing a sample with a coagulation assay composition, wherein Factor X compound and Factor V compound in said coagulation assay composition are substantially not admixed prior to said mixing of the sample with the coagulation assay composition; and    (b) determining the time period beginning upon performing step (a) and ending upon clot formation.