Method for separating target molecules or particles from fibrinogen-containing samples including blood components

Abstract

A method for separating target molecules or particles from a fibrinogen containing sample comprises: (a) trapping the said target molecules or particles in a fibrin network by converting at least partially the fibrinogen contained in the sample into fibrin; (b) retracting the said fibrin network to form a fibrin clot; (c) separating the said fibrin clot from the surrounding sample medium.

1 . A sample collection device for separating target molecules or particles from a sample comprising: (i) an identification code; (ii) a container for containing the sample; (iii) a dry reagents formulation in the container, the device being operable to form a fibrin clot that traps in a separable manner the target molecules or particles upon the exposure of the sample to thrombin within the device in the presence of fibrinogen that is native to the sample and/or artificially added to the sample, wherein the dry reagents formulation comprises a chelating agent and said thrombin present at an amount between 0.01 to 10 IU per 1 mg of fibrinogen, and wherein the fibrin clot is reduced to a size less than 1/10 th  of the initial sample volume. 
     
     
         2 . The device according to  claim 1 , wherein the volume of the sample container is between 0.1 and 20 ml. 
     
     
         3 . The device according to  claim 1 , wherein the concentration of fibrinogen within the sample is between 0.1 to 10 mg/ml. 
     
     
         4 . The device of  claim 1 , wherein said fibrinogen is native to the sample. 
     
     
         5 . The device according to  claim 1 , wherein said fibrinogen is initially included within the device, prior to adding the sample into the device. 
     
     
         6 . The device according to  claim 1 , wherein said thrombin is included within the device in a lyophilized format. 
     
     
         7 . The device according to  claim 1 , which further includes additives selected from the group consisting of calcium, activated platelet cells, activated platelet cell lysate, and factor XIII. 
     
     
         8 . The device according to  claim 1 , which further includes magnetic particles. 
     
     
         9 . The device according to  claim 8 , wherein the magnetic particles are coated with a fibrinogen/fibrin binding moiety. 
     
     
         10 . The device according to  claim 9 , which further comprises molecules having: (I) fibrin/fibrinogen-binding moiety and (II) a substance-capturing moiety directed against said target molecules or particles. 
     
     
         11 . The device according to  claim 1 , wherein the fibrinogen in the sample is recombinant. 
     
     
         12 . The device according to  claim 11 , wherein the fibrinogen in the sample is a fibrinogen fusion protein with a capturing moiety domain directed against said target molecules or particles. 
     
     
         13 . The device according to  claim 1 , wherein said target molecules or particles comprise bacteria, virus, yeast, proteins, peptides and/or nucleic acids. 
     
     
         14 . The device according to  claim 1 , wherein the dry reagents formulation further comprises fibrinogen. 
     
     
         15 . A sample collection device for separating target molecules or particles from a sample comprising: (i) an identification code; (ii) a container for containing the sample; (iii) a dry reagents formulation in the container, the device being operable to form a fibrin clot that traps in a separable manner the target molecules or particles upon the exposure of the sample to thrombin within the device in the presence of fibrinogen that is native to the sample and/or artificially added to the sample, wherein the dry reagents formulation comprises a chelating agent selected from the group consisting of GDTA, EDTA, and citrate, and said thrombin present at an amount between 0.01 to 10 IU per 1 mg of fibrinogen, and wherein the fibrin clot is reduced to a size less than 1/10 th  of the initial sample volume.