System and method for isolating alpha 2m molecules

Abstract

A method includes: depositing whole blood into at least one separator tube; subjecting the at least one separator tube to a first centrifugal force to cause a combination of the first centrifugal force and separator gel within each separator tube of the at least one separator tube to separate plasma of the whole blood from red and white blood cells of the whole blood within the at least one separator tube, wherein the plasma includes α2M molecules; transferring one or more portions of the plasma from within the at least one separator tube and into at least one isolator; and subjecting the at least one isolator to a second centrifugal force to cause a combination of the second centrifugal force and a filter within each isolator of the at least one isolator to isolate the α2M molecules from other components of the plasma within the at least one isolator.

1 . A method for isolating Alpha-2 Macroglobulin (α2M) molecules from whole blood comprising:
 depositing whole blood into at least one separator tube, wherein each separator tube of the at least one separator tube contains an amount of separator gel; 
 subjecting the at least one separator tube to a first centrifugal force in a first centrifuging stage for a first predetermined period of time to cause a combination of the first centrifugal force and the separator gel within each separator tube of the at least one separator tube to separate plasma of the whole blood within the at least one separator tube from red blood cells and white blood cells of the whole blood within the at least one separator tube, wherein the plasma includes the α2M molecules; 
 transferring one or more portions of the plasma from within the at least one separator tube and into at least one isolator, wherein each isolator of the at least one isolator comprises a filter; and 
 subjecting the at least one isolator to a second centrifugal force in a second centrifuging stage for a second predetermined period of time to cause a combination of the second centrifugal force and the filter within each isolator of the at least one isolator to isolate the α2M molecules from other components of the plasma within the at least one isolator. 
 
     
     
         2 . The method of  claim 1 , further comprising:
 retrieving at least a portion of the α2M molecules from within the at least one isolator; and   injecting at least the portion of the α2M molecules into a musculoskeletal structure of a patient from which the whole blood was drawn.   
     
     
         3 . The method of  claim 1 , wherein:
 each separator tube of the at least one separator tube comprises an elongate transparent tube that defines an opening at one end that is sealed with a cap;   the cap is formed from a flexible material that allows a hollow needle of a syringe to penetrate therethrough while sealing around the hollow needle, and that re-seals after the hollow needle is withdrawn; and   depositing the whole blood into at least one separator tube comprises:
 inserting a hollow needle of the syringe through the cap of each separator tube of the at least one separator tube; and 
 providing at least a portion of the whole blood into each separator tube of the at least one separator tube through the hollow needle. 
   
     
     
         4 . The method of  claim 3 , comprising:
 using a syringe comprising the hollow needle to draw the whole blood from a patient into which the α2M molecules are to be injected; and   partially pre-filling the syringe with an anticoagulant before using the syringe to draw the whole blood from the patient.   
     
     
         5 . The method of  claim 4 , wherein the anticoagulant comprises a citrate dextrose solution (ACD-A). 
     
     
         6 . The method of  claim 3 , wherein:
 each separator tube of the at least one separator tube comprises a vacuum separator tube that is pre-provided with a vacuum therein when in an unused condition.   
     
     
         7 . The method of  claim 1 , wherein:
 subjecting the at least one separator tube to the first centrifugal force in the first centrifuging stage comprises placing the at least one separator tube within a first holder of a centrifuge, and operating the centrifuge to exert the first centrifugal force on the at least one separator tube; and   subjecting the at least one isolator to the second centrifugal force in the second centrifuging stage comprises placing the at least one isolator within a second holder of the centrifuge, and operating the centrifuge to exert the second centrifugal force on the at least one isolator;   
     
     
         8 . The method of  claim 7 , wherein:
 the first holder comprises either a first removable holder configured to be inserted into a bucket of the centrifuge, or a first exchangeable rotor of the centrifuge; and   the second holder comprises either a second removable holder configured to be inserted into a bucket of the centrifuge, or a second exchangeable rotor of the centrifuge.   
     
     
         9 . The method of  claim 7 , wherein transferring the one or more portions of the plasma from within the at least one separator tube and into the at least one isolator comprises:
 using a transfer syringe to withdraw the one or more portions of the plasma from within the at least one separator tube; and   using the transfer syringe to deposit the one or more portions of the plasma into the at least one isolator.   
     
     
         10 . The method of  claim 9 , wherein:
 the at least one separator tube, the centrifuge, the transfer syringe and the at least one isolator, together, form a kit for isolating the α2M molecules from the whole blood; and   multiple versions of the kit are defined by at least one of a quantity of separator tubes included in the at least one separator tube and a quantity of isolators included in the at least one isolator.   
     
     
         11 . The method of  claim 10 , wherein the multiple versions of the kit include a kit comprising 4, 8 or 16 separator tubes. 
     
     
         12 . The method of  claim 10 , the kit further includes a dummy isolator of a shape and size similar to the at least one separator tube to provide a counterbalance to the weight of a single separator tube of the at least one separator tube after the single separator tube is filled with plasma to enable balancing of the centrifuge. 
     
     
         13 . The method of  claim 1 , wherein transferring the one or more portions of the plasma from within the at least one separator tube and into the at least one isolator further comprises:
 coupling a transfer syringe to a syringe port of a transfer device;   coupling each separator tube of the at least one separator tube, one at a time, to a separator tube port of the transfer device, wherein the separator tube port comprises at least one hollow needle configured to simultaneously couple the separator tube port to the syringe port and to a filtered air port of the transfer device;   while each separator tube of the at least one separator tube is coupled to the separator tube port, operating a plunger of the transfer syringe to withdraw at least one portion of the one or more portions of the plasma from the separator tube and into the transfer syringe through the transfer device, and to simultaneously cause air to be drawn through an air filter at the filtered air port and into the separator tube through the transfer device; and   following transfer of the one or more portions of the plasma from each separator tube of the at least one separator tube, using the transfer syringe to inject the one or more portions of the plasma into the at least one isolator.   
     
     
         14 . The method of  claim 13 , wherein the at least one hollow needle comprises:
 a first hollow needle coupling the separator tube port to the syringe port; and   a second hollow needle coupling the separator tube port to the filtered air port.   
     
     
         15 . The method of  claim 1 , wherein transferring the one or more portions of the plasma from within the at least one separator tube and into the at least one isolator further comprises:
 coupling a transfer syringe to a syringe port of a three-way valve;   coupling each separator tube of the at least one separator tube, one at a time, to a separator tube port of the three-way valve;   while each separator tube of the at least one separator tube is coupled to the separator tube port, performing operations comprising:
 with the three-way valve operated to couple a filtered air port of the three-way valve to the syringe port, operating a plunger of the transfer syringe to draw air through an air filter at the filtered air port, through the three-way valve, and into the transfer syringe; and 
 with the three-way valve operated to couple the separator tube port to the syringe port, operating the plunger of the transfer syringe to inject the filtered air into the separator tube through the three-way valve, and to withdraw at least one of the one or more portions of the plasma from the separator tube and into the transfer syringe through the three-way valve; and 
   following transfer of the one or more portions of the plasma from each separator tube of the at least one separator tube, using the transfer syringe to inject the one or more portions of the plasma into the at least one isolator.   
     
     
         16 . The method of  claim 1 , wherein:
 each isolator of the at least one isolator comprises a first cylinder defined by a first cylindrical wall and a second cylinder defined by a second cylindrical wall;   a first end of the first cylindrical wall of the first cylinder defines an opening that is configured to be closable with a septum cap, and a second end of the first cylindrical wall is closed with the filter;   a first end of the second cylindrical wall of the second cylinder is closed where the second cylindrical wall narrows to form a conically-shaped end portion, and the second end of the second cylindrical wall of the second cylinder defines an opening that is configured to be coupled to the second end of the first cylinder in a manner that causes a first interior space of the first cylinder and a second interior space of the second cylinder to be separated by the filter;   at least a portion of the septum cap is formed from a flexible material that allows a hollow needle of a syringe to penetrate therethrough while sealing around the hollow needle, and that re-seals after the hollow needle is withdrawn;   transferring the one or more portions of the plasma from within the at least one separator tube and into the at least one isolator comprises:
 using a transfer syringe to withdraw the one or more portions of the plasma from within the at least one separator tube; 
 inserting a needle of the transfer syringe through the septum cap of each isolator of the at least one isolator; and 
 injecting the one or more portions of the plasma into the first interior space from within the transfer syringe; and 
   isolating the α2M molecules from other components of the plasma within the at least one isolator comprises isolating the α2M molecules from other components within the first interior space of each isolator of the at least one isolator from the other components of the plasma within the second interior space of each isolator of the at least one isolator.   
     
     
         17 . The method of  claim 16 , wherein the septum cap further comprises a third cylindrical wall configured to serve as an extension to the first cylindrical wall to increase a volume of the first interior space when the first end of the first cylindrical wall is closed with the septum cap. 
     
     
         18 . The method of  claim 16 , wherein the filter of each isolator of the at least one isolator has a molecular weight cut off ranging from 100 kD to 500 kD.