Patent Publication Number: US-2009223080-A1

Title: Apparatus and methods for making, storing, and administering freeze-dried materials such as freeze-dried plasma

Description:
RELATED APPLICATIONS 
     This application is a continuation-in-part of co-pending U.S. patent application Ser. No. 12/283,885, filed 16 Sep. 2008, entitled “Apparatus and Methods for Making, Storing, and Administering Freeze-Dried Materials Such as Freeze-Dried Plasma”, which is a continuation-in-part of co-pending U.S. patent application Ser. No. 12/228,745, filed 15 Aug. 2008, entitled “Apparatus and Methods for Making, Storing, and Administering Freeze-Dried Materials Such as Freeze-Dried Plasma”, which is a continuation-in-part of co-pending U.S. patent application Ser. No. 12/077,397, filed 19 Mar. 2008, entitled “Apparatus and Methods for Making, Storing, and Administering Freeze-Dried Materials Such as Freeze-Dried Plasma”, which is a continuation-in-part of co-pending U.S. patent application Ser. No. 11/881,493, filed 27 Jul. 2007, which is a continuation-in-part of co-pending U.S. patent application Ser. No. 11/725,352, filed 19 Mar. 2007, all of which are incorporated herein by reference. 
    
    
     FIELD OF THE INVENTION 
     The present invention relates to methods, systems, and apparatuses for manufacturing, storing and administering freeze-dried materials, such as single donor units of freeze-dried human plasma. 
     BACKGROUND OF THE INVENTION 
     First aid is critical for the survival of a person that has suffered a serious injury, such as a trauma victim. For instance, initial treatment of a severely wounded person in combat situations can often mean the difference between life and death. While it is necessary to treat the wounds and stop the bleeding of the person, it is also important to ensure that the person&#39;s body is capable of properly functioning. Thus, it is necessary to take steps to ensure that the person&#39;s body is properly hydrated after losing fluids due to the injury. The present invention addresses these issues. 
     Previously, fluids were replenished within the patient by delivering saline intravenously. While effective, research has indicated that delivery of plasma to the patient is even more effective in replenishing fluid to the patient than the use of saline. However, delivery and storage of the plasma is critical to prevent contamination of the plasma. An ideal way of delivering the plasma is to deliver the plasma in a freeze dried form and reconstituting the plasma when it is administered to a person. 
     SUMMARY OF THE INVENTION 
     The invention provides methods, systems, and apparatuses for manufacturing, storing and administering freeze-dried biological materials, such as plasma derived from e.g., single unit blood plasma, or pre-treated single unit blood plasma, or single donor units of blood plasma, and converted into freeze-dried human plasma. 
     According to one aspect of the invention, a freeze-dried material, e.g., freeze-dried human plasma, is stored in a first chamber of a container along with a reconstituting liquid for the freeze-dried material, e.g., de-gassed water. The reconstituting liquid is stored in a second chamber of the container. A sealing wall within the container forms a barrier between the first chamber and the second chamber preventing contact between the freeze-dried material and the reconstituting liquid. At least one valve assembly in the sealing wall can be manipulated to selectively open at least one region of the sealing wall to establish fluid flow communication between the first and second chambers. This allows the freeze dried material to be reconstituted within the container. The reconstituted freeze-dried material can also be administered directly from the same container to a recipient. 
     In one arrangement, the valve assembly includes a pressure sensitive valve, e.g., a flap valve. The valve is operative between a normally closed condition, normally resisting fluid flow communication between the first and second chambers, and an opened condition, establishing fluid flow condition communication between the first and second chambers. The pressure sensitive valve can be placed in its open condition in response to establishing a pressure differential across the valve, e.g., by preferentially squeezing a chamber of the container. 
     In one arrangement, the valve assembly includes a normally closed septum. The septum is operative in a normally closed condition, maintaining closure between the first and second chambers, and an opened condition establishing fluid flow communication between the first and second chambers in response to at least a partially tearing of the septum. The septum can, e.g., include a tear member coupled to a pulling member to at least partially tear open the septum. 
     The pressure sensitive valve and the septum can be arranged serially to provide a redundant valve assembly. In this arrangement, the normally closed septum is operative in a normally closed condition, maintaining closure between the first and second chambers, independent of the valve and an opened condition establishing fluid flow communication between the first and second chambers in response to at least a partially tearing of the septum and a pressure differential applied across the valve. 
     In one arrangement, an outer skirt is provided that overlays an exterior wall of the container in a region of the sealing wall. The outer skirt can include a tear member coupled to a pulling member to tear open the outer skirt for removal. 
     Another embodiment of the invention provides a method that provides a flexible container as above generally described, with first and second chambers. The first chamber holds a freeze-dried material, such as freeze-dried human plasma, in a dry state. The second chamber holds a reconstituting liquid for the freeze-dried material. An interior sealing wall within the container is sized and configured to form a barrier between the first chamber and the second chamber preventing contact between the freeze-dried material and the reconstituting liquid. At least one valve assembly in the sealing wall is operative by manipulation to open at least one region of the sealing wall to establish fluid flow communication between the first and second chambers. According to this aspect of the invention, the valve assembly is manipulated to open the region, and the reconstituting liquid is expressed from the second chamber through the valve assembly into the first chamber into contact with the freeze-dried material. 
     In one arrangement, an outer skirt overlays an exterior wall of the container in a region of the sealing wall and blocking manipulation of the valve assembly. In this arrangement, the outer skirt is removed to expose the valve assembly to manipulation prior to manipulating the valve assembly to open the region in the sealing wall. 
     In another arrangement, the reconstituted freeze-dried plasma is administered directly from the container to a recipient. 
     According to another aspect of the invention, a freeze-dried material comprising freeze-dried human plasma is prepared and stored, transported, reconstituted, and administered using a container as just generally described in any of the foregoing paragraphs. In one arrangement, liquid human plasma is loaded in molds. The molds are cooled until they reach approximately −45° C. The plasma is dried so the moisture content is below 5% w/w, thereby forming the freeze-dried human material that can be stored, transported, reconstituted, and administered using a container. In another arrangement, liquid human plasma is freeze-dried in situ within the container. 
     According to another aspect of the invention, a freeze-dried material, e.g., freeze-dried human plasma, is stored in a first container, and a reconstituting liquid for the freeze-dried material, e.g., de-gassed water is stored in a separate second container. A transfer set can be manipulated to couple the two containers together, to establish fluid flow communication between the first and second containers. This allows the freeze dried material to be reconstituted within one of the containers. The reconstituted freeze-dried material can also be administered directly from the same container to a recipient. 
     According to another aspect of the invention, a system is provided that comprises a vessel including first and second end components each comprising a rigid or semi-rigid material defining, respectively, first and second frames providing structural strength. A transparent gas impermeable material peripherally is sealed to the first frame, and a gas permeable material is peripherally sealed to the second frame. A flexible side wall component is peripherally sealed to side edges of the first and second frames. The first end component, the second end component, and the side wall component peripherally define an interior space. At least one port component on the side wall provides fluid communication with the interior space. 
     The system makes it possible for a material such as fresh human plasma to be freeze-dried, transported, stored, reconstituted, and administered in a single, multifunctional vessel. 
     Another aspect of the invention provides a method that makes use of the technical features of the multifunctional vessel just described. The method includes introducing a liquid material, such as fresh human plasma, through a first port component on the vessel. The method includes freeze-drying the liquid material in situ within the interior space of the vessel, during which time the gas permeable material of the second end component provides gas transport to accommodate sublimation of water vapor. The method also includes introducing a reconstituting liquid for mixing with the freeze-dried material within the interior space through a second port component of the vessel, to reconstitute the freeze-dried material. The method further includes conveying the reconstituted freeze-dried material from the interior space through a third port component of the vessel. 
     As defined, the single, multifunctional vessel accommodates freeze-drying a material within the vessel; the transport and storage of the freeze-dried material within the vessel; and the reconstitution and administration of the material from the vessel. 
     In one embodiment, the method further includes, after freeze drying, introducing an oxygen-free inert gas into the interior space through the gas permeable material of the second end component. The oxygen-free inert gas occupies the interior space with the freeze-dried material to prevent deterioration of the material. The method also includes covering the gas permeable material of the second end component, to trap the oxygen-free inert gas within the interior space with the freeze-dried material. The method includes storing the freeze-dried material in the entrapped oxygen-free inert gas within the vessel for a storage period prior to introduction of the reconstituting liquid. 
     In one embodiment, the method further includes placing the covered vessel within an outer container during storage. 
     In another embodiment, there is an assembly for freeze-drying plasma, whereby the container that holds the liquid plasma is separate from the container or structure having the permeable membrane used for removal of vapor during the freeze-drying process. The two containers will be connected by a tubing that will allow vapors to pass from one of the containers to the other. The tubing will be pinched shut or clamped before being removed from the freeze-dryer to isolate the plasma containing container. After removal from the freeze-dryer, the first container can be sealed and severed from the tubing and second container. 
     These and other areas of importance and significance will become apparent from following description. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  is a front elevation view of a device for storing freeze-dried material, e.g., freeze-dried human plasma, and a reconstituting liquid for the freeze-dried material, making possible a reconstitution of the freeze-dried material within the device and an administration of the reconstituted freeze-dried material directly from the device to a recipient, the device being shown prior to the removal of an outer protective skirt. 
         FIG. 2  is side elevation view of the device shown in  FIG. 1 . 
         FIG. 3  is a front elevation view of the device shown in  FIG. 1 , showing the tearing of the outer protective skirt for its removal prior to manipulating the device to reconstitute the freeze-dried materials. 
         FIG. 4A  is a front elevation view of the device shown in  FIG. 3 , after the removal of the outer protective skirt and prior to manipulating the device to reconstitute the freeze-dried materials. 
         FIG. 4B  is side elevation view of the device shown in  FIG. 4A . 
         FIG. 5A  is a side elevation section view of the interior sealing wall and associated valve assembly formed within the device taken generally along line  5 A- 5 A in  FIG. 1 , prior to the removal of the outer protective skirt. 
         FIG. 5B  is a side elevation section view like that shown in  FIG. 5A , showing an alternative arrangement of the interior sealing wall and multiple valve assemblies. 
         FIG. 6  is a side elevation section view of the interior sealing wall and associated valve assembly formed within the device taken generally along line  6 - 6  in  FIG. 4A , after the removal of the outer protective skirt and prior to manipulating the device to reconstitute the freeze-dried materials. 
         FIG. 7  is a side elevation section view of the interior sealing wall and associated valve assembly like that shown in  FIG. 6 , after opening at least one region of interior sealing wall and prior to manipulating the device to reconstitute the freeze-dried materials. 
         FIG. 8  is a front elevation view of the device shown in  FIG. 1 , showing the removal of the outer protective skirt prior to manipulating the device to reconstitute the freeze-dried materials. 
         FIG. 9  is a front elevation view of the device shown in  FIG. 8 , showing the manipulation of the valve assembly to open at least one region of the interior sealing wall, in the manner also shown in  FIG. 7 . 
         FIGS. 10 to 15  are front elevation view of the device shown in  FIG. 9 , showing the manipulating the device to reconstitute the freeze-dried materials. 
         FIG. 16  is a front elevation view of the device shown in  FIG. 15 , showing the administration of reconstituted material directly from the device to a recipient. 
         FIGS. 17A to 17E  are diagrammatic perspective views to an illustrative process for the preparation of a freeze-dried plasma cake from liquid human plasma, prior to insertion and storage within the device shown in  FIG. 1 . 
         FIGS. 18 and 19  are front elevation views of placing a freeze-dried material (like the plasma cake formed using the process  FIGS. 17A to 17E ) in the first chamber of the device shown in  FIG. 1 . 
         FIG. 20  is a front elevation view of placing a reconstituting liquid for the freeze-dried material in the second chamber of the device shown in  FIG. 1 . 
         FIG. 21  is a front elevation view of placing the outer protective sleeve about the device, to create the device shown in  FIG. 1 . 
         FIG. 22  is a front elevation view of an alternative device for storing freeze-dried material, e.g., freeze-dried human plasma, and a reconstituting liquid for the freeze-dried material, making possible a reconstitution of the freeze-dried material within the device and an administration of the reconstituted freeze-dried material directly from the device to a recipient, the device being shown prior to the removal of an outer protective skirt. 
         FIG. 23  is a front elevation interior section view of the valve assembly formed in the device taken generally along line  23 - 23  in  FIG. 22 , prior to the removal of the outer protective skirt. 
         FIG. 24  is a front elevation view of the device shown in  FIG. 22 , after the removal of the outer protective skirt and prior to manipulating the device to reconstitute the freeze-dried materials. 
         FIG. 25  is a front elevation interior section view of valve assembly like that shown in  FIG. 23 , taken generally along line  25 - 25  in  FIG. 23  after removal of the outer protective skirt. 
         FIGS. 26 and 27  are front elevation interior section views showing the passage of materials through the valve assembly shown in  FIG. 25  by manipulating the device to reconstitute the freeze-dried materials. 
         FIGS. 28A and 28B  are largely schematic views of an alternative way of packaging the reconstituting liquid for the freeze-dried material in the second chamber of the device of the type shown in  FIG. 1  or  22 . 
         FIGS. 29A and 29B  are largely schematic views of another alternative way of packaging the reconstituting liquid for the freeze-dried material in the second chamber of the device of the type shown in  FIG. 1  or  22 . 
         FIG. 30  is a front elevation view of a system for storing freeze-dried material, e.g., freeze-dried human plasma, and a reconstituting liquid for the freeze-dried material, comprising individual first and second containers and a transfer set that makes possible a reconstitution of the freeze-dried material within the system for administration to a recipient. 
         FIG. 31  is a front elevation view of the system shown in  FIG. 30 , with the first and second containers joined in fluid communication by the transfer set to reconstitute the freeze-dried material. 
         FIG. 32  is a front elevation view of one of the containers of the system shown in  FIGS. 30 and 31 , after the freeze-dried material has been reconstituted, showing the administration of reconstituted material directly from the container to a recipient. 
         FIG. 33  is a front elevation view of a device for storing freeze-dried material, e.g., freeze-dried human plasma, and a reconstituting liquid for the freeze-dried material, the device being sized and configured for freeze-drying material in situ within the device. 
         FIG. 34  is a front elevation view of the device shown in  FIG. 33 , showing the conveyance of liquid plasma into the device for freeze-drying in situ within the device. 
         FIG. 35  is a perspective view of several devices shown in  FIG. 34  after placement in a freeze-dryer for the purpose of freeze-drying liquid plasma in situ within each of the devices. 
         FIG. 36  is a front elevation view of a device shown in  FIG. 35  after removal from the freeze-dryer, showing the freeze-dried plasma cake that has been formed in situ within the device, and prior to the conveyance of a reconstituting material into the device. 
         FIG. 37  is a front elevation view of a device shown in  FIG. 36  after the conveyance of a reconstituting material into the device. 
         FIG. 38  is a front elevation view of placing an outer protective sleeve about the device shown in  FIG. 37 , after conveyance of the reconstituting material into the device, to create the device of a type shown in  FIG. 1 . 
         FIG. 39A  is an exploded perspective view of a multifunctional device for freeze-drying, storing, reconstituting, and administering a material, such as plasma, comprises a vessel made of several components having different physical properties to thereby serve different functions. 
         FIG. 39B  is an assembled perspective view of the device shown in  FIG. 39A , showing the flexible side wall component and transparent, gas impermeable end component. 
         FIG. 39C  is an assembled perspective view of the device shown in  FIG. 39A , showing the flexible side wall component and the gas permeable end component. 
         FIG. 39D  is an assembled side elevation view of the device shown in  FIG. 39A , taken along line  39 D- 39 D of  FIG. 39C . 
         FIGS. 40 and 41A  are perspective views of a freeze-dried material storage assembly comprising the vessel shown in  FIGS. 39A to 39D  sealed within a gas-impermeable overwrap, and also showing in perspective view a rigid outer container with a lid for enclosing the freeze-dried material storage assembly during transport and storage until the instance of use, as  FIG. 41  shows. 
         FIG. 41B  shows a perspective view of a freeze-dried material storage assembly sealed within a gas-impermeable overwrap, and placed in a rigid outer container with a lid, as  FIG. 41  shows, with the outer container also including storage space for a vessel of reconstitution liquid and associated reconstitution and administration sets. 
         FIGS. 42 and 43  are perspective views of a unitary freeze-died material storage assembly comprising a vessel as shown in  FIGS. 39A to 39D  and an integral closure cover,  FIG. 42  showing the closure cover in an opened condition, and  FIG. 43  showing the closure cover in a closed condition. 
         FIGS. 44 and 45  are perspective views of the unitary freeze-dried material storage assembly shown in  FIG. 43  (with the closure cover in the closed condition) placed within a rigid outer container with a lid for enclosing the unitary freeze-dried material storage assembly during transport and storage until the instance of use. 
         FIGS. 46 and 47  are perspective views of another representative embodiment of a unitary freeze-died material storage assembly comprising a vessel as shown in  FIGS. 39A to 39D  and an integral closure cover,  FIG. 46  showing the closure cover in an opened condition, and  FIG. 47  showing the closure cover in a closed condition. 
         FIGS. 48 and 49  are perspective views of the unitary freeze-dried material storage assembly shown in  FIG. 47  (with the closure cover in the closed condition) placed within a rigid outer container with a lid for enclosing the unitary freeze-dried material storage assembly during transport and storage until the instance of use. 
         FIGS. 50 and 51  are perspective views showing the transfer of a unit of liquid plasma into a unitary freeze-dried material storage assembly of the type shown in  FIG. 42 , with the closure cover in the opened condition, which begins the process using the unitary freeze dried material storage assembly. 
         FIG. 52  is a perspective view of the placement of several unitary freeze-dried material storage assemblies shown in  FIGS. 50 and 51  into a freeze dryer, the closure covers being in the opened condition, the freeze dryer exposing the unitary freeze-dried material storage assemblies to a range of temperature and vacuum conditions to lyophilize the liquid plasma into freeze-dried plasma, the open closure cover accommodating sublimation of water vapor during drying. 
         FIG. 53  is a perspective view of the several unitary freeze-dried material storage assemblies within the freeze dryer shown in  FIG. 52 , with the closure covers still in the opened condition, the freeze dryer exposing the unitary freeze-dried material storage assemblies to a blanket of oxygen-free inert gas, the open closure covers accommodating infiltration of the oxygen-free inert gas into the freeze-dried plasma material contained within the assemblies. 
         FIG. 54  is a perspective view of the several unitary freeze-dried material storage assemblies within the freeze dryer shown in  FIG. 53 , with the closure covers being placed into the closed condition to trap the oxygen-free inert gas within the unitary freeze-dried material storage assemblies, to protect the freeze-dried plasma material from degradation during subsequent transport and storage. 
         FIGS. 55 and 56  are perspective views of the unitary freeze-dried material storage assembly shown in  FIG. 54  (with the closure cover in the closed condition) placed within a rigid outer container with a lid for enclosing the unitary freeze-dried material storage assembly during transport and storage until the instance of use. 
         FIG. 57  shows the reconstitution of the freeze-dried plasma material within a unitary freeze-dried material storage assembly after under the freeze-drying  15 - and packaging process shown in  FIGS. 50 to 56 , by transferring a reconstituting liquid from a source container into the unitary freeze-dried material storage assembly for mixing with the freeze-dried plasma material. 
         FIG. 58  shows the administration of reconstituted freeze-dried plasma material from a unitary freeze-dried material storage assembly into an individual. 
         FIG. 59  shows the mixing of freeze-dried plasma material with a reconstituting liquid, after transferring a reconstituting liquid into the unitary freeze-dried material storage assembly as shown in  FIG. 57 , by transferring the mixture of reconstituting liquid and freeze-dried plasma material back to the source container, the mixture being transferred back and forth in the manner shown in  FIGS. 57 and 59  until ready for administration. 
         FIG. 60  shows the administration to an individual of freeze-died plasma material reconstituted using a unitary freeze-dried material storage assembly as shown in  FIG. 57 , the reconstituted material being ultimately transferred after mixing as shown in  FIGS. 57 and 59  out of the unitary freeze-dried material storage assembly into the original reconstituting liquid container for administration. 
         FIG. 61  is a front elevation view depicting an alternate system and device for freeze-drying material, e.g. plasma, with the system comprising a first collapsible container that acts as a primary storage portion and a secondary lyophilizing portion, with the two portions forming a single device or assembly, connected by a tubing. 
         FIG. 62  is a front elevation view depicting the system and device depicted in  FIG. 61 , with a pH adjustment solution being aseptically added to the first container. 
         FIG. 63  is a front elevation view further depicting the system and device of  FIG. 61 , with liquid plasma being introduced into the first container. 
         FIG. 64  is a front elevation view of the system and device of  FIG. 61 , with the device being filled with plasma. 
         FIG. 65  is a perspective view of several devices shown in  FIG. 64  after placement in a freeze-dryer for the purpose of freeze-drying liquid plasma in situ within each of the devices, with the primary portion (the first collapsible container) being in contact with the heat transfer surface of the freeze-dryer. 
         FIGS. 66A-68B  provide various depictions of instruments, such as closure devices, used for closing or pinching shut a tubing that connects the secondary lyophilizing portion to the primary storage portion to form the final bag shown in  FIG. 66 , using the lyophilizers to close and pinch shut the tubing prior to being removed from the freeze-dryer. 
         FIG. 69  depicts the tubing, used to connect the first and second containers, being heat sealed to seal shut the first container. 
         FIG. 70  is a front elevation view of the device of  FIG. 64  containing plasma after it has been freeze-dried and with the secondary lyophilizing portion being removed. 
         FIG. 71  provides a front elevation view of a second arrangement of the secondary portion of the alternate system and device for freeze-drying material, discussed above with respect to  FIGS. 61-69 . 
         FIG. 72  provides a front elevation view of a further arrangement of the secondary portion of the alternate system and device for freeze-drying material, discussed above with respect to  FIGS. 61-69   
         FIG. 73  shows the mixing of freeze-dried plasma material with a reconstituting liquid so that the plasma material and the reconstituting liquid can be mixed so that they are ready for administration. 
         FIG. 74  shows the administration to an individual of freeze-died plasma material reconstituted using a unitary freeze-dried material storage assembly as shown in  FIG. 70 . 
         FIG. 75  shows the treatment of single unit plasma prior to freeze-drying. 
         FIG. 76  is a diagrammatic perspective view of an illustrative step in the process for the preparation of a freeze-dried plasma cake from liquid human plasma, similar to the step show in  FIG. 17A , with the additional step of ascorbic acid being added along with the human plasma. 
         FIG. 77  is a front elevation view of placing a reconstituting liquid for the freeze-dried material, along with ascorbic acid in the second chamber of the device shown in  FIG. 1 . 
         FIG. 78  is a graphical comparison of the pH value of lyophilized plasma prepared according to the present invention having various ascorbic acid concentrations. 
         FIG. 79  is a graphical representation comparing the thrombin time (TT) of lyophilized plasma prepared according to the present invention having various ascorbic acid concentrations to determine stability over a period of 12 weeks. 
         FIG. 80  is a graphical representation comparing the prothrombin time (PT) of lyophilized plasma prepared according to the present invention having various ascorbic acid concentrations and stored at various temperatures to determine stability over a period of 12 weeks. 
         FIG. 81  is a graphical representation comparing the activated partial thromboplastin time (aPTT) of lyophilized plasma prepared according to the present invention having various ascorbic acid concentrations and stored at various temperatures to determine stability over a period of 12 weeks. 
         FIG. 82  is a graphical representation comparing the fibrinogen concentration of lyophilized plasma prepared according to the present invention having various ascorbic acid concentrations and stored at various temperatures to determine stability over a period of 12 weeks. 
         FIG. 83  is a graphical representation comparing the Factor V stability of lyophilized plasma prepared according to the present invention having various ascorbic acid concentrations and stored at various temperatures to determine stability over a period of 12 weeks. 
         FIG. 84  is a graphical representation comparing the Factor VIII stability of lyophilized plasma prepared according to the present invention having various ascorbic acid concentrations and stored at various temperatures to determine stability over a period of 12 weeks. 
         FIG. 85  is a timeline used to demonstrate the process carried out on swine models for comparing lyophilized plasma of the present invention to other forms of plasma. 
         FIG. 86  is a graphical representation comparing clotting factor activity and coagulation assays of various qualities of lyophilized plasma. 
         FIG. 87  is a graphical representation comparing mean blood loss of various forms of plasma. 
         FIG. 88  is a graphical representation of the heart rate of swine tested according to the timeline shown in  FIG. 85  for treatment with various forms of plasma. 
         FIG. 89  is graphical representation of the mean arterial pressure (MAP) of swine tested according to the timeline shown in  FIG. 85  for treatment with various forms of plasma. 
         FIG. 90  is graphical representation of lactation levels of swine tested according to the timeline shown in  FIG. 85  for treatment with various forms of plasma. 
         FIG. 91  is a graphical representation comparing prothrombin time (PT) of swine tested according to the timeline shown in  FIG. 85  for treatment with various forms of plasma. 
         FIG. 92  is a graphical representation comparing partial thromboplastin time (PTT) of swine tested according to the timeline shown in  FIG. 85  for treatment with various forms of plasma. 
         FIG. 93  is a graphical representation of Interleukin-6 (IL-6) cytokine comparisons of swine tested according to the timeline shown in  FIG. 85  for treatment with various forms of plasma. 
     
    
    
     DESCRIPTION OF THE PREFERRED EMBODIMENT 
     Although the disclosure hereof is detailed and exact to enable those skilled in the art to practice the invention, the physical embodiments herein disclosed merely exemplify the invention which may be embodied in other specific structures. While the preferred embodiment has been described, the details may be changed without departing from the invention, which is defined by the claims. 
     I. Device for Storing and Reconstituting Freeze-Dried Plasma 
       FIGS. 1 and 2  show a device  10  for storing and administering a freeze-dried material. The device  10  comprises a flexible bag having a first collapsible chamber  12  and a second collapsible chamber  14 . 
     The first chamber  12 , also referred to as the dry chamber, contains an aliquot of a freeze-dried material  16 . The nature and type of freeze-dried material  16  can vary. It can, e.g., be single unit blood plasma, or pre-treated single unit blood plasma, or single donor unit of blood plasma. In the illustrated embodiment, the freeze-dried material comprises human plasma, and the aliquot is a single donor unit of human plasma. 
     The second chamber  14 , also referred to as the wet chamber, contains a reconstituting liquid  18  for the freeze-dried material  16 . The nature and type of the reconstituting material  18  can vary. In the illustrated embodiment, the reconstituting material  18  comprises sterile water, which may be degassed, if desired. In use, the sterile water in the wet chamber  14  is mixed with the freeze-dried plasma in the dry chamber  12  to provide plasma for transfusion. The plasma is reconstituted and administered on site using the device  10 . 
     The first chamber  12  is sized and configured to maintain the freeze-dried material  16 , prior to its reconstitution, in a vacuum packed, aseptic, moisture-free and low concentration oxygen environment, preferably accommodating long term storage, e.g., at least 2 years at room temperature. Stored in this environment, the freeze-dried material  16  retains its desired qualities for transfusion. 
     The second chamber  12  is sized and configured to maintain the reconstituting liquid  18 , prior to its mixing with the freeze-dried material  16 , in an aseptic environment and at a low gas concentration, preferably accommodating long term storage, e.g., at least 2 years at room temperature. 
     The volume of each of the chambers  12  and  14  is preferably approximately 50% larger than the volume of the freeze-dried material  16  in the first chamber  12 . This provides ample volume within the device  10  for mixing the freeze-dried material  16  and reconstituting liquid  18 , either in the first chamber  12  or the second chamber  14 , as will be described in greater detail later. 
     The device  10  may be made, e.g., of an inert medical-grade plastic material, such as polyvinyl chloride, polyethylene, polypropylene, or high density polyethylene. The device  10  can comprise a multi-laminate of polymer layers for greater durability, e.g., to resist tearing and puncturing that could be encountered in normal handling. 
     The material of the device  10  can be selected to be transparent, if desired, to allow visual inspection of the contents of the chamber  12  and  14 . The material in the first chamber  12  can be selected to provide a gas-impermeable barrier, such as a metallized, reduced gas-permeability coating, or a metal laminate. In this case, the wall of the first chamber may be opaque. 
     Furthermore, the device  10  may be enveloped prior to use by a vacuum sealed over-wrap  20  (shown in phantom lines in  FIG. 1 ), made, e.g., a metallized, gas impermeable material. The over-wrap  20  enhances shelf-stability. 
     An interior sealing wall  22  (see  FIG. 1 ) compartmentalizes the device  10  into the first and second chambers  12  and  14  (see also  FIG. 5A ). The sealing wall  22  provides a barrier between the first chamber  12  and the second chamber  14 , which normally prevents contact between the freeze-dried -material  16  and the reconstituting liquid  18  during storage, up to the instant of use. 
     As FIGS.  5 A/B and  7  show, one or more regions  24  of the sealing wall  22  may be selectively opened by a caregiver, as will be described in greater detail later. The region(s)  24 , when opened, make possible fluid communication between the two chambers  12  and  14 . The fluid communication makes it possible to mix the reconstituting liquid  18  with the freeze-dried material  16 , as will further be described in greater detail later. 
     The region(s)- 24  of the sealing wall  22  may be opened in various ways. In a representative embodiment (see  FIG. 5 ), the sealing wall  22  includes a normally closed valve assembly  26  associated with each region  24  where the sealing wall  22  is to be opened. In  FIG. 5A , a single region  24  is shown, so a single valve assembly  26  is shown. As shown in  FIG. 5B , where multiple regions  24   a  and  24   b  are provided, each region  24   a  and  24   b  would include its own dedicated valve assembly  26   a  and  26   b , respectively. 
     In the representative embodiment (see  FIGS. 5A and 5B ), each valve assembly  26  includes a primary, pressure sensitive valve  28 . The valve  28  can take the form, e.g., of a short duck bill or two way flap valve. The primary valve  28  is sized and configured to normally resist flow communication between the two chambers  12  and  14 . 
     In the representative embodiment, each valve assembly  26  also includes a normally closed septum  30  between the valve  28  and the wet chamber  14 . The septum  30  maintains closure between the two chambers  12  and  14 , independent of the valve  28 . Independent of the valve  28 , the septum  30  prevents unintended passage of material between the two chambers  12  and  14 , thereby maintaining the separate integrity of the freeze-dried material  16  and the reconstituting liquid  18  within the device  10  prior to use. 
     The septum  30  includes an integrated tear member  32  that is incorporated within the septum  30 . The integrated tear member  32  is coupled to a pull string  34  that extends through a fluid sealed pass-through or septum  36  in the wall of the second chamber  14 . As  FIG. 1  shows, the pull string terminates outside the device  10  at a pull tab  38 . 
     As  FIGS. 6 and 7  show, the tear member  32  is sized and configured to open the septum  30  when a caregiver pulls on the tab  38 . The pass-through or septum  26  seals around the pull string  34 , and also seals close after passage of the pull string  34  from the interior of the chamber  14 , maintaining in integrity of the second chamber  14 . Opening the septum  30  in this manner forms the open region  24  (see  FIG. 7 ). The open region  24  places the first and second chambers  12  and  14  into communication through the valve  28 . 
     With the region  24  opened (see  FIG. 7 ), the primary valve  28  still serves to normally resist flow communication between the two chambers  12  and  14 . However, when the region  24  is opened, the valve  28  is sized and configured to resiliently yield in response to a difference in fluid pressure between opposite sides of the valve  38  (see  FIGS. 11 and 14 ). In response to the pressure differential, the valve  28  opens in the direction of the fluid pressure differential, from the region of higher pressure toward the region of lower pressure. 
     As will be described in greater detail later (as shown, respectively, in  FIGS. 10 and 13 ), the caregiver creates the fluid pressure differential across the valve  28  by selectively squeezing one chamber and not the other chamber. Fluid is expelled in response to the fluid pressure differential through the valve  28  from the chamber that is squeezed into the chamber that is not squeezed. 
     The multi-component valve assembly  26  provides a redundant sealing capability, to assure that the chambers  12  and  14  remain separated until it is desired to reconstitute the freeze-dried material  16 . 
     In a representative embodiment (see  FIGS. 1  and  2 ), the device  10  further includes an outer tear-away skirt  40 , which provide further redundancy. As  FIGS. 1 and 2  show, the skirt  40  overlays the device  10  in the region of the sealing wall  22 . The skirt  40  serves to overlay and protect the components of the valve assembly  26  associated with the sealing wall  22 . 
     At least one region of the skirt  40  is circumferentially attached about an exterior wall of the device, e.g., by adhesive, either in the region of the first chamber, the second chamber, or both. Furthermore, as the skirt  40  is installed about the device  10 , the exterior wall of the device is desirably plicated or pleated or otherwise bunched together (as  FIGS. 1 and 2  show). Alternatively, the placations can be performed in the wall of the container. 
     The placations relieve wall stress in the region of the sealing wall  22 . The skirt  40 , once attached, maintains these placations or pleats, and thereby serves to relieve or distribute wall stresses in the region of sealing wall  22  and the components of the valve assembly  26  associated with the sealing wall  22 . Such wall stresses can arise, e.g., due to the weight of the reconstituting liquid  18  contained in the second chamber  14 , and/or by virtue of handling during transport and manipulation prior to use. The presence of the overlaying skirt  40  also serves to isolate the components of the valve assembly  26  associated with the sealing wall  22  from unintended contact during transport and prior to use. 
     As  FIG. 1  shows, the skirt  40  includes an integrated tear member  42 . The integrated tear member  42  includes a pull string  44  that terminates with a pull tab  46 , that depends outside the skirt  40 . The tear member  42  is sized and configured to tear open the skirt  40  when a caregiver pulls on the tab  46  (as  FIG. 3  shows). Upon removal of the skirt  40 , the placations of the walls of the bags  12  and  14  are relieved (as  FIGS. 4A and 4B  show), placing the components of the valve assembly  26  associated with the sealing wall  22  into condition for manipulation. 
     It should be understood that reference to the first chamber  12  and the second chamber  14  is done to distinguish one chamber from the other, and not to limit either chamber to a specific spatial relationship. For example, the chambers  12  and  14  may be arranged face to face, having vertical edges in contact. 
     The technical features of the device  10  includes separate chambers or compartments that are separated by sealing means that will allow for eventual interconnection and intercommunication, between the chambers, which can be accomplished in various ways. Furthermore, reference to a bag or chambers should not be limited to any specific structure or shape but should be understood to refer any container capable of carrying and mixing the contents  16  and  18 . 
     II. Preparing and Packaging the Freeze Dried Material and Reconstituting Liquid 
     Preparing and packaging the freeze-dried material  16  and reconstituting liquid  18  comprises two main processing steps: (i) freeze-drying the material  16 , and (ii) packaging the material  16  and the reconstituting liquid  18  within the chambers  12  and  14 . 
     A. Preparation of Freeze-Dried Plasma 
     In a representative embodiment, the freeze-dried material  16  comprises plasma. A description of an illustrative way of preparing freeze-dried plasma for packaging in the device  10  therefore follows. 
     Preparation and manufacturing of the plasma will take place in an aseptic setting. Preferably, manufacturing and preparation procedures can be done, for example, in an ISO Class 5 clean room (or better) with ISO Class 3 bio-containment hoods for aseptic handling of human plasma. Freeze drying can be done aseptically in a CIP/SIP freeze dryer. 
     Human plasma is collected from a single donor in a conventional way, e.g., by collecting a unit of whole blood from the donor in a closed system collection bag, followed by centrifugal separation of the plasma and its collection in an integrally connected transfer bag (containing one plasma unit of about 250 ml). Each unit (contained in the transfer bag) will be handled individually in the bio-containment hood. Between handling one single donor unit and another unit single donor unit from a different donor, there may be a line clearance protocol for change-over in the bio-containment hood, or a validation process for flow design and change-over can be otherwise provided. This protocol may address removal of all tools and materials associated with the previous handling. It may also address the thorough wash down of the containment work area and work area instruments (mass balances) to ensure no residues of the previous handling were left in place. The identification of single donor samples will be maintained by bar coding and other tagging of the single donor human plasma containers. 
     As shown in  FIG. 17A , prior to freeze drying, the 250 ml human plasma unit is dispensed from the transfer bag  48  into a sterile, pyrogen free, rectangular mold  50  (e.g., 4 cm×10 cm×12.5 cm—d×w×1). The mold  50  can be stainless-steel; however it can also be composed of metal with good thermal transfer properties such as aluminum, aluminum alloy, titanium or gold. The mold  50  may be coated on its inside surfaces with a tough, inert barrier film with good release properties such as PTFE or diamond. 
     As shown in  FIG. 17B , the mold  50  containing the human plasma is then placed inside a water-impermeable, vapor-permeable, sterile, heat sealable bag  52  with bar coding and tagging  54  indicative of the human plasma identification (source, blood type, date of collection, etc.). This vapor permeable bag  52  would typically be manufactured using microporous PTFE membrane material (e.g. Gore-Tex™) or microporous HDPE membranes (e.g. Tyvek™). 
     The bag  52  is heat sealed to contain the mold  50  and human plasma. The bag  52  is designed to neatly contain the mold  50  and its contents without any bunching or sagging of the bag material below the surface of the interior mold wall edge or at the base of the mold. 
     As shown in  FIG. 17C , the mold  50  inside the containment bag  52  is then placed inside a freeze dryer  56  on an aseptic freeze dryer shelf surface  58 . The freeze dryer  56  used for the lyophilization will be a validated clean in place, steam in place freeze dryer with shelf area of near 200 square feet or more. Such a freeze dryer  56  can accommodate at least 500 molds when it is fully loaded. 
     Once loaded, the freeze dryer cycle is started. This cycle generally cools the human plasma to near -45° C. and freezing for a prescribed period, e.g., 2 to 8 hours, followed by cooling of the freeze dryer condenser and application of vacuum to start the freeze drying cycle. A freeze-dried human plasma cake  60  is formed. 
     In a representative primary freeze drying cycle, the temperature of the human plasma cake  60  needs to remain below its collapse temperature (e.g., −33° C.) to maintain its integrity. When the moisture content of the cake  60  is below 5% weight per weight (w/w), a secondary drying cycle (the elevated temperature) may be used to further lower the moisture content, if desired. The combined primary and secondary freeze drying cycles may take 72 hours or more, but such times will vary with the processing conditions. At the conclusion of the freeze drying cycle, the freeze dryer vacuum may be opened to an atmosphere of an oxygen-free, high purity inert gas such as nitrogen or argon. 
     As shown in  FIG. 17D , the freeze dried cakes  60  in their molds  50  and containment bags  52  are removed to an aseptic containment cart  62  whose environment may be maintained under a nitrogen or argon blanket to exclude moisture and oxygen. The containment cart  62  may couple to the front of the freeze dryer to allow for transfer of the freeze dryer contents under a controlled inert gas blanket. 
     The containment carts  62  may be used to store human freeze dried plasma cakes (each cake within a mold  50  and enclosed within a bag  52 ) as well as allow cakes to be transferred to a device loading area, which allows loading of the freeze dried plasma cake  60  into the device  10 , as will be described in greater detail later. 
     B. Packaging Freeze-Dried Plasma and Water into the Device 
     As shown in  FIG. 1 , the device  10  comprises a first aseptic vacuum port  64 , which communicates with the first chamber  12 , and a second aseptic vacuum port  66 , which communicates with the second chamber  14 . The vacuum ports  64  and  66  are sized and configured for connection to various tubing T during final assembly (see  FIGS. 18 to 21 ) to facilitate packaging of the freeze-dried plasma material  16  and reconstituting liquid  18  (e.g., water) within the device  10 . 
     An administration port  68  is also heat sealed in communication with the second chamber  14 . The administration port  68  is used during the packaging process to convey the reconstituting liquid  18  into the second chamber  14 , as will be described in greater detail later. After the reconstituting liquid  18  is packaged within the chamber  14 , the administration port  68  is sealed with a conventional septum or frangible membrane assembly or a convention screw-lock leur fitting  70 , to accommodate its coupling to an administration set  72  to the port  28  at time of transfusion, as shown in  FIG. 16 . 
     The device  10  also comprises a heat sealable aseptic flange  74  (see  FIG. 1 ), which allows a freeze-dried plasma cake  60  to be inserted into the first chamber  12 , as shown in  FIG. 18 , and then sealed in an aseptic fashion, as shown in  FIG. 19 . 
     A slot  76  may be pre-formed on the flange  74 . The slot  76  makes it possible to hang the device  10  at a desired gravity head height for administering reconstituted plasma to an individual, as  FIG. 16  shows. 
     Individual single donor human plasma freeze dried cakes  60  are aseptically loaded into the device  10  (see  FIG. 18 ) through the flange  74 . The device loading area may be, e.g., a bio-containment hood that excludes significant oxygen and moisture contamination by inert gas blanketing. Also the device loading area may be an aseptic glove-box system with an inert gas environment. 
       FIGS. 18 and 19  depict a representative loading process. The bag  52  is opened, and the plasma cake  60  removed from the mold  50 . The plasma cake  60  is loaded through the open flange  74  into the first chamber  12 . As shown in  FIG. 17E , it is anticipated that the plasma cake  60  can be transferred into the chamber  12  directly from the mold  50  (after removal of the bag  52 ) using a single-use, aseptic, clear-plastic applicator tool  78 , similar to a large open-ended spatula. Once the chamber  12  is loaded, the flange  74  can be sealed closed using various conventional aseptic techniques, e.g., dielectric welding or heat sealing. 
     The loading of the plasma chamber  12  can be through an “oyster style” opening that comprises approximately 50% of the flange  74  of the chamber  12 , which can be readily sealed close after loading. An oyster opening would allow loading of the plasma cake  60  without concerns of damaging the first chamber  12  or the freeze-dried plasma during the process. In the case of the oyster opening, there would be sufficient excess overlay of the edge seam to allow for straightforward edge-seam alignment and contact during the sealing process. 
     Preferably, after loading and sealing of the chamber  12 , an aseptic vacuum is applied through tubing T connected to the vacuum port  64  on the first chamber  12  (see  FIG. 19 ). Upon achieving near 100 mTorr of pressure, the vacuum port  64  is heat sealed and the tubing T removed. This evacuation process provides for the eventual ability to mix and reconstitute the human freeze dried plasma without introduction of bubbles and without foaming. The vacuum would also cause the plasma cake  60  to be compacted to a fine powder, forming the freeze-dried material  16  within the chamber  12 . 
     To maintain a direct traceable link between the source plasma and the material  16  packaged into the chamber  12 , the device  10  preferably includes a bar coding and tagging  54 ′ (see  FIG. 1 ), which is indicative of the human plasma identification (source, blood type, date of collection, etc.), and which replicates or is otherwise linked to the bar coding and tagging  54  placed on the bag  52  enveloping the mold  50  at the time of freeze-drying. In this way, the device  10  maintains a traceable link back to the human donor source. 
     To assist in the reconstitution of the freeze dried plasma material  16 , an aseptic dense sphere of an inert material such as, but not limited to, glass, polyvinyl chloride or high density polyethylene may be added to the inside of the chamber  12  prior to its closure. 
     The reconstituting liquid  18  (in the representative embodiment, gas-free water) is introduced into the second chamber  14 . The vacuum port  66  and administration port  68  are connected to feed lines  80  and  82 , respectively, as  FIG. 20  shows. Gas in the chamber  14  is removed by application of aseptic vacuum. 
     The vacuum port  66  is sealed and the tubing  80  is removed. The required aliquot (e.g., approximately 250 ml) of reconstitution fluid is added to the chamber  14  through the administration port  68 . The tubing  82  is removed and the administration port  68  is then sealed with the conventional septum or frangible membrane assembly or a convention screw-lock leur fitting  70 , which accommodate coupling of the administration set  68  to the port  68  at time of transfusion. 
     To assist in the reconstitution of the freeze dried plasma, an aseptic dense sphere of an inert material such as, but not limited to, glass, polyvinyl chloride or high density polyethylene may be present inside the second chamber  14 . 
     As  FIG. 21  shows, after packaging the freeze-dried material  16  and the reconstituting liquid  18  in the manner just described, the wall of the device  10  is plicated in the region of the sealing wall  22 , as previously described, and the outer skirt  40  attached. The overwrap  418   20  can be applied, as shown in  FIG. 1 , if desired. 
     The device  10  is ready for storage, transport, and use 
     III. Reconstitution and Administration of the Freeze-Dried Material 
     The device  10  makes possible a purposeful two step manipulation in anticipation of reconstituting the freeze-dried material  16 . 
     In the first step (shown in  FIG. 8 ), the tear member  42  is pulled to open and remove the skirt  40 , which places the sealing wall  22  of the device  10  in the ready for use configuration shown in  FIG. 6 . In the second step (shown in  FIG. 9 ), the tear member  32  is pulled to open the septum  20  (which  FIG. 7  shows in greater detail). The region  24  of the sealing wall  22  is thereby opened. 
     When the region  24  is opened, the caregiver can apply pressure to the second chamber  14  to express the reconstituting liquid  18  from the second chamber  14  into the first chamber  12  (see  FIGS. 10 and 11 ), thereby beginning the reconstitution of the freeze-dried material  16 . More particularly, with the region  24  opened, the caregiver can apply pressure to the second chamber  14  (as  FIG. 10  shows) and not the first chamber  12 . As  FIGS. 10 and 11  show, the pressure differential between the second chamber  14  and the first chamber  12  expels the liquid  18  from the second chamber  14 , through the valve  28  (which yields in response to the pressure differential to open in the direction of the first chamber  12 , as  FIG. 11  shows), and into the first chamber  12 . The expelled liquid  18  mixes with the freeze-dried material  16  in the first chamber  12 , beginning the reconstitution. 
     As  FIG. 12  show, shaking the device  10  accelerates the mixing of liquid  18  and freeze-dried material  18  in the first chamber  12 . 
     When the region  24  is opened, the caregiver can subsequently apply pressure to the first chamber  12  to express the material  16 , now at least partially reconstituted in the liquid  18 , from the first chamber  12  into the second chamber  14  (see  FIGS. 13 and 14 ). Reconstitution of the freeze-dried material  16  is advanced. More particularly, as  FIG. 13  shows, the caregiver can now apply pressure to the first chamber  12  (as  FIG. 13  shows) and not the second chamber  14 . As  FIGS. 13 and 14  show, the pressure differential between the first chamber  12  and the second chamber  14  expels the mixture of the liquid  18  and the freeze-dried material  16  from the first chamber  12 , through the valve  28  (which yields in response to the pressure differential to open in the direction of the second chamber  14 , as  FIG. 14  shows), and back into the second chamber  14 . The expelled liquid  18  continues to mix with the freeze-dried plasma material  18 , furthering the reconstitution of the material  18 . 
     As  FIG. 15  shows, shaking the device  10  further accelerates the mixing of water and freeze-dried plasma in the second chamber  14 . 
     The material  16  reconstituted in the liquid  18  can be passed back and forth between the two chambers  12  and  14  by alternating pressure on the chambers  12  and  14 , with intermediate shaking, until the desired degree of mixing occurs, at which time the mixture is ready for transfusion. More particularly, the caregiver can proceed to squeeze one chamber and not the other, to expel the mixture of the liquid  18  and freeze-dried material  18  back and forth between the chambers  12  and  14 , with periodic shaking, until the desired degree of mixing and reconstitution of the plasma is accomplished. 
     At this point (as  FIG. 16  shows), the caregiver can couple the administration fitting  70  of the device  10  to the fluid administration set  72 . The reconstituted plasma is transfused by gravity flow through a phlebotomy needle  84  into the circulatory system of an individual. 
     The administration fitting  70  can further include a static mixing tube  86  (as shown in  FIG. 16 ), to assist in continued reconstitution of plasma aliquot  5  with water  7  during transfusion. 
     The device  10  as described provides: 
     i) long term stable containment of a freeze-dried material such as freeze-dried human plasma; 
     ii) eventual rapid reconstitution of the freeze-dried material with a reconstituting liquid for injection; and 
     iii) eventual delivery of the reconstituted freeze dried material to a trauma victim in a safe, aseptic manner. 
     IV. Other Representative Embodiments 
     A. Dual Containers With Intermediate Valve Passage 
       FIG. 22  shows another representative embodiment of a device  100  for storing an administering a freeze-dried material. The device  100  comprises a first collapsible container  102  and a second collapsible container  104 , joined by an intermediate normally closed valve assembly  106 . 
     The device  100  shares many of the technical features of the device shown in  FIG. 1 , albeit the particular structure differs. The first container  102  comprises the dry chamber  12  as previously described, and is sized and configured to contains an aliquot of a freeze-dried material  16 , such as a freeze-dried single donor unit of human plasma. 
     The second container  104  comprises the wet chamber  14 , as previously described, and is sized and configured to contain a reconstituting liquid  18  for the freeze-dried material  16 . As before described, the reconstituting material  18  can comprise, e.g., sterile water, which may be degassed, if desired. 
     In use, the sterile water in the wet chamber  14  is mixed with the freeze-dried plasma in the dry chamber  12  to provide plasma for transfusion. The plasma is reconstituted and administered on site using the device  10 . 
     As before described, the first container  102  is sized and configured to maintain the freeze-dried material  16 , prior to its reconstitution, in a vacuum packed, aseptic, moisture-free and low concentration oxygen environment, preferably accommodating long term storage, e.g., at least 2 years at room temperature. Stored in this environment, the freeze-dried material  16  retains its desired qualities for transfusion. 
     As also before described, the second container  104  is sized and configured to maintain the reconstituting liquid  18 , prior to its mixing with the freeze-dried material  16 , in an aseptic environment and at a low gas concentration, preferably accommodating long term storage, e.g., at least 2 years at room temperature. 
     The volume of each of the containers  102  and  104  is preferably approximately 50% larger than the volume of the freeze-dried material  16  in the first chamber  12 . This provides ample volume within the device  10  for mixing the freeze-dried material  16  and reconstituting liquid  18 , either in the first container  102 , or the second container  104 , as will be described in greater detail later. 
     The containers  102  and  104  may be made, e.g., of an inert medical grade plastic material, such as polyvinyl chloride, polyethylene, polypropylene, or high density polyethylene. One or both of the container  102  and  104  can comprise a multi-laminate of polymer layers for greater durability, e.g., to resist tearing and puncturing that could be encountered in normal handling. 
     The material of the containers  102  and  104  can be selected to be transparent, if desired, to allow visual inspection of the contents of the chamber  12  and  14 . The material in the first container  102  can be selected to provide a gas-impermeable barrier, such as a metallized, reduced gas-permeability coating, or a metal laminate. In this case, the wall of the first chamber may be opaque. 
     As before described, the device  100  may be enveloped prior to use by a vacuum sealed over-wrap  20  (shown in phantom lines in  FIG. 22 ), made, e.g., a metallized, gas impermeable material. The over-wrap  20  enhances shelf-stability. 
     In the alternative representative embodiment shown in  FIG. 22 , the valve assembly  106  includes a pressure sensitive valve  108  enclosed within a flexible tubular valve passage  110 , which extends between the two containers  102  and  104 . The valve  108  can take the form, e.g., of a short duck bill or two way flap valve. The valve  108  is sized and configured to normally resist flow communication between the two containers  102  and  104 . However, the valve  108  is sized and configured to resiliently yield in response to a difference in fluid pressure between opposite sides of the valve  108  (in the same manner as the valve  28  shown in  FIGS. 11 and 14 ). In response to the pressure differential, the valve  108 , like the valve  28 , opens in the direction of the fluid pressure differential, from the region of higher pressure toward the region of lower pressure. 
     The regions of the wall of the containers to which the valve passage  110  is joined normally close communication between the containers  102  and  104  through the valve passage  110 . 
     An outer tear-away skirt  112  is wrapped around the mid-regions of the containers  102  and  104  and the intermediate valve passage  110 . The skirt  112  serves to overlay and protect the components of the valve assembly  106  prior to use. At least one region of the skirt  112  is circumferentially attached about an exterior wall of each container  102  and  104 , e.g., by adhesive, either in the region of the first chamber, the second chamber, or both. 
     As  FIG. 23  shows, within the outer skirt  112 , the mid-regions of the containers  102  and  104 , and the valve passage  110  itself, are desirably plicated or pleated or otherwise bunched together, shortening the length of each container  102  and  104  and the valve passage  110 . Alternatively, the placations can be performed in the walls of the containers  102  and  104  and/or valve passage  110 . The presence of the overlaying skirt  112  serves to isolate the valve passage  100  from unintended contact during transport and prior to use. 
     As  FIG. 23  shows, the walls of each container  102  and  104  that overlay opposite ends of the valve passage  110  each includes an integrated tear member  112 . As  FIG. 23  shows, each integrated tear member - 112  is coupled by an internal pull string  114  to an adjacent side wall of the respective container  102  and  104 . The internal pull string  114  is normally held in slight tension when the device  100  is in the plicated condition shown in  FIG. 22  (i.e., when the mid-regions of the containers  102  and  104 , and the valve passage  110  itself, are plicated and held in this condition by the outer shirt  112 ). When the device  100  is in the plicated condition, the tension on the internal pull string  114  is not sufficient to affect the tear member  112 . The walls of each container  102  and  104  that overlay opposite ends of the valve passage  110  remain closed. When the device  100  is in the plicated condition, the chambers  12  and  14  and their contents remain isolated and separated prior to use. 
     As  FIGS. 24  shows, the skirt  112  can be torn and removed by operation of an integrated tear member  116  (in the manner shown in  FIG. 3 ), to place the device  100  in the condition shown in  FIG. 24 . As  FIG. 24  shows, upon removal of the skirt  112 , the placations of the walls of the containers  102  and  104  and valve passage  110  are relieved, and the device  100  lengthens. 
     As  FIG. 25  shows, when the device  100  lengthens, tension on the internal pull string  114  is increased. The increased tension is sufficient to activate the tear member  112 , tearing open regions  116  of the walls on opposite ends of the valve passage  110  (as  FIG. 25  shows). The open regions  116  place the first and second chambers  12  and  14  into communication through the valve passage  110 . 
     With the regions  116  opened, the caregiver can proceed to manipulate the device  100  in the same manner previously described with respect to device  10  (as shown in  FIGS. 10 to 16 ). The caregiver creates the fluid pressure differential across the valve  108  by selectively squeezing one container and not the other container. Fluid is expelled in response to the fluid pressure differential through the valve  108  from the container that is squeezed into the container that is not squeezed to mix and reconstitute the freeze-drive material for administration. Transfer of materials in opposite directions between the chambers  12  and  14  through the valve passage  110  as a result of the manipulation of the containers  102  and  104  is shown in  FIGS. 26 and 27 . 
     B. Dual Containers with Transfer Set 
       FIG. 30  shows a representative embodiment of a system  200  for storing an administering a freeze-dried material. The system  200  comprises a first collapsible container  202  and a second, separate collapsible container  204 . The system  200  further comprises a transfer set  206  for establishing fluid communication between the first and second containers  202  and  204 . 
     The system  200  shares many of the technical features of the devices shown in  FIGS. 1 and 22 , albeit the particular structure differs. 
     The first container  202  comprises the dry chamber  12  as previously described, and is sized and configured to contains an aliquot of a freeze-dried material  16 , such as a freeze-dried single donor unit of human plasma. To maintain a direct traceable link between the source plasma and the material  16  in the chamber  12 , the container  202  preferably includes a bar coding and tagging  54  (see  FIG. 30 ), which is indicative of the human plasma identification (source, blood type, date of collection, etc.). In this way, the container  202  maintains a traceable link back to the human donor source. 
     The second container  204  comprises the wet chamber  14 , as previously described, and is sized and configured to contain a reconstituting liquid  18  for the freeze-dried material  16 . As before described, the reconstituting material  18  can comprise, e.g., sterile water, which may be degassed, if desired. 
     In use (see  FIG. 31 ), using the transfer set  206 , the sterile water in the wet chamber  14  is mixed with the freeze-dried plasma in the dry chamber  12  to provide plasma for transfusion. The plasma is reconstituted and administered on site using the system  200 . 
     As before described, the first container  202  is sized and configured to maintain the freeze-dried material  16 , prior to its reconstitution, in a vacuum packed, aseptic, moisture-free and low concentration oxygen environment, preferably accommodating long term storage, e.g., at least 2 years at room temperature. Stored in this environment, the freeze-dried material  16  retains its desired qualities for transfusion. 
     As also before described, the second container  204  is sized and configured to maintain the reconstituting liquid  18 , prior to its mixing with the freeze-dried material  16 , in an aseptic environment and at a low gas concentration, preferably accommodating long term storage, e.g., at least 2 years at room temperature. 
     The volume of each of the containers  202  and  204  is preferably approximately 50% larger than the volume of the freeze-dried material  16  in the first chamber  12 . This provides ample volume within the containers  202  and  204  for mixing the freeze-dried material  16  and reconstituting liquid  18 , either in the first container  202 , or the second container  204 , or both, as will be described in greater detail later. 
     The containers  202  and  204  may be made, e.g., of an inert medical grade plastic material, such as polyvinyl chloride, polyethylene, polypropylene, or high density polyethylene. One or both of the container  202  and  204  can comprise a multi-laminate of polymer layers for greater durability, e.g., to resist tearing and puncturing that could be encountered in normal handling. 
     The material of the containers  202  and  204  can be selected to be transparent, if desired, to allow visual inspection of the contents of the chamber  12  and  14 . The material in the first container  202  can be selected to provide a gas-impermeable barrier, such as a metallized, reduced gas-permeability coating, or a metal laminate. In this case, the wall of the first chamber  12  may be opaque. 
     Each container  202  and  204  may be enveloped prior to use by a vacuum sealed over-wrap  208  (shown in phantom lines in  FIG. 30 ), made, e.g., a metallized, gas impermeable material. The over-wrap  208  enhances shelf-stability. The transfer set  206  also is desirably packaged in a sterile over-wrap  208  prior to use (as shown in phantom lines in  FIG. 31 ). 
     The transfer set  206  includes plastic needles or spikes  210  at each end. An outer tear-away skirt or cap  216  can placed or wrapped around each needle or spike  210  to preserve sterility until the instant of use. 
     In use, the needles or spikes  210  are sized and configure to puncture conventional pierceable membranes  212  located within port tubes  214  coupled in fluid communication with each container  202  and  204 . Each membrane  212  normally seals the respective container  202  and  204  until pierced by the respective needle or spike  210  of the transfer set  206 . Once pierced by the needle or spike  210 , fluid communication is opened through the port tube  214 . 
     With the port tubes opened  214  opened, the caregiver can proceed to manipulate the system  200  to transfer the reconstituting liquid  18  from the second container  204  into contact with the freeze-dried material  16 , as  FIG. 31  shows, The caregiver can create a fluid pressure differential across the transfer set  206  by selectively squeezing one container and not the other container. Fluid is expelled in response to the fluid pressure differential through the transfer set  206  from the container that is squeezed into the container that is not squeezed to mix and reconstitute the freeze-drive material for administration. Transfer of materials in opposite directions back and forth between the chambers  12  and  14  can proceed as necessary to reconstitute the freeze-dried material, at which time administration can occur. 
     At this time, the caregiver can couple the administration fitting  70  (shown coupled to the first container  202 ) to an appropriate administration set, for transfer of the reconstituted material to the circulatory system of an individual, as shown in  FIG. 31 , in the same manner as before described with reference to  FIG. 16 . The administration fitting  70  can also be coupled to the second container  204 , or both the first and second containers  202  and  204 . 
     C. Alternative Ways to Package the Reconstituting Liquid 
     FIGS.  28 A/B and  29 A/B shows alternative ways to package the reconstituting liquid  18  in a device  10  or device  100  as previously described. In these alternative ways, it is not necessary to use the administration port  68  to convey the reconstituting liquid  18 , but can be closed and sealed in a pre-packaging operation. 
     In one alternative representative embodiment (see FIG.  28 A/B), the wet chamber  14  includes two packaging ports  120  and  128 . In use (see  FIG. 28A ), the first port  120  is coupled to a source  124  of the reconstituting liquid  18  via a first inline valve  122 . The second port  128  is coupled to a vacuum source  125  via a second inline valve  126 . 
     As shown  FIG. 28A , the first valve  122  is closed and the second valve  126  is opened. A vacuum is applied to the interior of the chamber  14 . As shown in  FIG. 26B , the first valve  122  is opened and the second valve  126  is closed. The reconstituting liquid  18  is conveyed by gravity flow into the chamber  14 . Both packaging ports  120  and  128  are sealed. 
     In another alternative representative embodiment (see FIGS.  29 A/B), the wet chamber  14  includes a single packaging port  130 . In use (see  FIG. 29A ), the port  130  is coupled to a source  132  of the reconstituting liquid  18  and a vacuum source  134  through a two way valve  136 . 
     As shown  FIG. 29A , the two way valve  136  is operated to close communication with the liquid source  132  and to open communication with the vacuum source  134 . A vacuum is applied to the interior of the chamber  14 . As shown in  FIG. 29B , the two way valve  136  is operated to open communication with the liquid source  132  and to close communication with the vacuum source  134 . The reconstituting liquid  18  is conveyed by gravity flow into the chamber  14 . The packaging port  130  is sealed. 
     In both arrangements, the administration port  68  can be inserted and sealed close in a pre-packing operation. The administration port  68  is not used until it is time to administer the reconstituted freeze-dried material, as shown in  FIG. 16 . 
     D. Alternative Ways to Package the Freeze-Dried Material 
     In an alternative embodiment, the material  16  can be freeze-dried in situ within the chamber  12 . In this arrangement, as  FIG. 33  shows, a device  300  is compartmentalized by a sealing wall  22  into a chamber  12  and a chamber  14 , in the manner previously described. The sealing wall  22  includes a septum  26  with pull string  34  and tab  38 , as previously described. 
     To accommodate freeze-drying of the plasma within the chamber  12 , the device  300  is made of a material that resists cracking at the low temperatures (e.g., below −33° C.) encountered during freeze-drying. Candidate materials include polyolefin materials, polyurethane materials, polyurethane, elastomer materials, and polysilicone materials. Polyvinyl chloride materials treated to withstand low temperatures can also be used. 
     The device  300  also includes first and second aseptic ports  302  and  304 , which communicate with the first chamber  12 . The first aseptic port  302 , in use, conveys liquid plasma into the chamber  12  for freeze-drying. The first port  302  is desirably normally closed by a pierceable membrane or septum  314 . The second aseptic port  304  is normally closed by a gas permeable membrane such as a gas permeable membrane  316 . In use, the gas permeable membrane  316  accommodates the transport of vapors and gases into and out of the chamber  12  during and after the freeze-drying process, but otherwise prevents liquid from leaving the chamber  12 . The gas permeable membrane  316  can comprise, e.g., a nylon material, a polytetrafluoroethylene (PTFE) material, or a polypropylene material. 
     The device  300  also includes an aseptic port  306 , which communicates with the second chamber  14 . The port  306 , in use, conveys a reconstituting fluid into the second chamber  14 , as previously described (e.g., see  FIGS. 29A and 29B ). The first port  302  can also be normally closed by a pierceable membrane or septum  314 . 
     An administration port  310  is also heat sealed in communication with the second chamber  14 . The administration port  310 , in use, conveys reconstituted material from the second chamber  14  for administration to an individual, as previously described. 
     As  FIG. 34  shows, the first port  302  is sized and configured to be attached to tubing T coupled to a source of liquid plasma  312 . In the illustrated embodiment, the tubing T includes a spike or needle  318  that pierces the membrane  314  in the port  302 , to open fluid communication through the port  302  into the chamber  12 . 
     Through the tubing T, a desired volume of liquid plasma is conveyed from the source  312  into the first chamber  12 . Following the conveyance of liquid plasma into the first chamber  12 , the tubing T is removed, and the port  302  is sealed closed. At this stage of processing, the second chamber  14  remains empty, as  FIG. 34  shows. 
     To maintain a direct traceable link between the source plasma and the material  16  that will be freeze-dried in the chamber  12 , the device  300  preferably includes a bar coding and tagging  54 ′ (see  FIG. 31 ), which is indicative of the human plasma identification (source, blood type, date of collection, etc.), and which replicates or is otherwise linked to the bar coding and tagging  54  placed on the source plasma bag  312 . In this way, the device  300  maintains a traceable link back to the human donor source. 
     As shown in  FIG. 35 , one or more devices  300 , with each chamber  12  filled with liquid plasma, is placed inside a freeze dryer  320  on an aseptic freeze dryer shelf surfaces  322 . Once loaded, the freeze dryer cycle is started. This cycle generally cools the human plasma to near −45° C. and freezing for 2 to 8 hours, followed by cooling of the freeze dryer condenser and application of vacuum to start the freeze drying cycle. As a result, a freeze-dried human plasma cake  324  is formed in situ within the chamber  12  of each device  300  (see  FIG. 36 ). 
     The representative parameters for the freeze-drying process have been previously described and are incorporated herein by reference. 
     Throughout the freeze drying process, the gas permeable membrane  316  within the port  304  accommodates passage of gases, e.g., water vapor as it sublimates from the liquid plasma during freeze-drying, but otherwise prevents passage of liquid plasma from the chamber  12 . 
     As shown in  FIG. 36 , after freeze-drying, the devices  300  with the freeze dried cakes  324  in their chambers  12  are removed from the freeze dryer  320 . 
     Preferably, an aseptic vacuum is applied through the port  304 . Upon achieving near 100 mTorr of pressure, the port  304  is heat sealed closed. This evacuation process provides for the eventual ability to mix and reconstitute the human freeze dried plasma without introduction of bubbles and without foaming. The vacuum would also cause the plasma cake  324  to be compacted to a fine powder, forming the freeze-dried material  16  within the chamber  12 . The devices  300  can be maintained under a nitrogen or argon blanket to exclude moisture and oxygen until subsequent processing. 
     Next (see  FIG. 37 ), the reconstituting liquid  18  is introduced into the second chamber  14  through the port  306 , for example, in manner shown in  FIGS. 29A and 29B . The port  306  is then sealed. 
     As  FIG. 38  shows, after packaging the freeze-dried material  16  and the reconstituting liquid  18  in the manner just described, the wall of the device  300  is plicated in the region of the sealing wall  22 , as previously described, and an outer skirt  40  (with pull string  44  and tab  46 ) attached, as also previously described. An overwrap  20  can be applied, as shown in  FIG. 1 , if desired. 
     The device  300  is ready for storage, transport, and use. 
     It should be appreciated that liquid plasma could be freeze-dried in situ within the container  202  shown in  FIG. 30  in the same manner as just described. 
     V. Devices, Systems and Methods for Freeze-Drying and Storing Materials for Reconstitution 
     A. Multifunctional Freeze-Drying and Storage Vessel 
       FIGS. 39A to 39D  show a representative embodiment of a multifunctional device  400  for freeze-drying, storing, reconstituting, and administering a material, such as plasma. The device  400  is sized and configured to receive the material while it undergoes freeze-drying within the device  400 . The device  400  is also sized and configured to serve as a vessel for the freeze-dried material while it undergoes transport, handling, and storage prior to reconstitution at an intended site. The device  400  is also sized and configured to further serve as a vessel in which the freeze-dried material can be reconstituted. The device  400  is also sized and configured to also serve as a vessel from which the freeze-dried material, after being reconstituted, can be delivered to an individual in a safe and aseptic manner. Using the multifunctional device  400 , a given material can be freeze-dried, transported, stored, reconstituted, and administered in a single vessel. 
     As shown in the exploded view of  FIG. 39A , the device  400  comprises a vessel  402  made of several components having different physical properties to thereby serve different functions. As shown, the vessel  402  includes a side wall component  404  that peripherally encircles an open interior space  406 . The vessel  402  also includes first and second end components  408  and  410  that overlay the side wall component  404 , enclosing the interior space  406 . The vessel  402  also includes first, second, and third port components  412 ,  414 , and  416  that pass through regions of the side wall component  404  to provide fluid communication into the interior space  406  bounded by the side wall component  404  and the first and second overlaying end components  408  and  410 . 
     Assembled together (as  FIGS. 39B to 39C  show), the various components form a unitary, multifunctional vessel  402  in which a given material can be freeze-dried, then transported and stored, and then reconstituted, and then administered. 
     As shown in  FIGS. 39A to 39D , the first and second end components  408  and  410  comprise frames made of a rigid or semi-rigid material selected to form a lightweight, yet durable structural skeletons for the ends of the vessel  402 . The material for the first and second end components  408  and  410  can comprise, e.g., non-plasticized polyvinyl chloride, or polyethylene, or polypropylene, or high density polyethylene. The material is desirably inert and of a medical grade sufficient for contact with animal tissue and fluids. The frames defined by the first and second end component  408  and  410  can, e.g., be molded in the desired shape and size. 
     The frames defined by the first and second end components  408  and  410  define and maintain a shape for the vessel  402 , as well as provide overall structural support and attachment sites for other components of the vessel  402 . The frames defined by the first and second end components  408  and  410  provide for the vessel  402  uniting structural elements that withstand pressure conditions and other forces imposed upon the vessel  402  during freeze-drying and subsequent handling. 
     The frames defined by the first and second end components  408  and  410  each supports a panel of material, respectively  408 ′ and  410 ′. In the illustrated embodiment, the panels of material  408 ′ and  410 ′ span horizontally across the respective end component  408  and  410 . The panels of material  408 ′ and  410 ′ are peripherally sealed to the frames defined by the end components  408  and  410 , e.g., by adhesives or heat sealing techniques. 
     The materials  408 ′ and  410 ′ selected for the panels differ, because they serve different functions. This technical feature will be described in greater detail later. 
     The side wall component  404  is appended to the frames defined by the first and second end components  408  and  410 . The side wall component  404  spans in a vertical direction between the side edges of the end components  408  and  410 . The side wall component  404  is peripherally sealed to the side edges of the end components  408  and  410 , e.g., by adhesives or heat sealing techniques. 
     The sidewall component  404  and the first and second end components  408  and  410  provide a closed, sealed integrity to the interior space  406 . 
     The side wall component  404  comprises a flexible, gas impermeable material. The material is also desirably inert and of a medical grade sufficient for contact with animal tissue and fluids. The material for the side wall component  404  can comprise, e.g., plasticized polyvinyl chloride, or polyethylene film, or polypropylene film, or high density polyethylene film. The side wall component  404  can comprise a continuous film of flexible material, as shown in  FIG. 39A , or comprise shorter lengths of flexible film material sealed together. 
     The flexibility of the side wall component  404  accommodates expansion and contraction and flexure of the vessel  402  in response to pressure conditions encountered during freeze-drying and subsequent handling. Desirably, the material of the side wall component  404  also provides resistant to tearing or puncturing during freeze-drying and subsequent handling of the vessel  402 . The material for the side wall component  404  is desirably transparent, thereby allowing a user to visually see and inspect the contents of the vessel  402 , without allowing gas transmission between the interior space  406  and the ambient environment. 
     The material  408 ′ of the first end component  408 , like the side wall component, is also selected to be gas impermeable, to complement the side wall component  404  in this function. Desirably, the material  408 ′ is also selected to be transparent, to thereby contribute to the visible view into the interior space  406 . The material  408 ′ of the first end component  408  can be flexible or rigid, as desired. 
     It should be appreciated that not all of the material  408 ′ of the first end component  408  need be transparent. The material  408 ′ can include a region of transparency sufficient to permit viewing the interior space  406 , with the remainder of the material  408 ′ being gas impermeable and non-transparent. 
     Like the material for the side wall component  404 , the material  408 ′ for the first end component  408  is desirably inert and of a medical grade sufficient for contact with animal tissue and fluids. The material  408 ′ for the first end component  408  can comprise, e.g., plasticized polyvinyl chloride film, or polyethylene film, or polypropylene film, or high density polyethylene film. 
     The material  410 ′ of the second end component  410  desirably possesses, at least in part, physical characteristics that are different than the physical characteristics of the side wall component  404  and the first end component  408 , because the component  410  serves a different function. More particularly, the material  410 ′ of the second end component  410  is selected to be gas permeable; for example, hydrophobic. The gas permeable material  410 ′ accommodates the transport of vapors and gases into and out of the interior space  406  during and after the freeze-drying process, but, if hydrophobic, otherwise prevents liquid from entering or leaving the interior space  406 . The gas permeable material  410 ′ can comprise, e.g., a nylon film material, a polytetrafluoroethylene (PTFE) film material or other fluoropolymer film materials, a polypropylene film material, or a polyurethane film material. 
     The presence of the gas permeable material  410 ′ of the second end component  410  allows water vapor to sublimate from material within the interior space  406  during the freeze-drying process. The presence of the gas permeable material  410 ′ of the second end component  410  also allows inert gases to be introduced into the interior space  406  after the freeze-drying process, if desired, to provide a protective atmosphere within the vessel  402  conducive to long term storage of the material. This technical feature will be described in greater detail later. 
     The surface area of the gas permeable material  410 ′ of the second end component  410  may affect the rate of sublimation during the freeze-drying process, i.e., the greater the surface area the greater the rate of sublimation. In  FIGS. 39A to 39D , the gas permeable material  410 ′ of the second end component  410  overlays the entirely of the end component  410 . Alternatively, and as will be described later with respect to the embodiment shown in  FIGS. 46 and 47 , the gas permeable material  410 ′ of the second end component  410  can comprise a smaller region of the end component  410 , with the remainder of the second end component  410  being gas impermeable and, desirably, transparent. 
     The first, second, and third port components  412 ,  414 , and  416  are sealed within regions of the side wall component  404 . The ports  412 ,  414 , and  416  comprise, e.g., extruded or molded medical grade plastic tubes that are sealed, e.g., by heat or adhesive, to the adjacent material of the side wall component  404 . The ports  412 ,  414 , and  416  provide fluid communication into the interior space  406  formed by the side wall component  404  and the first and second overlaying end components, as described. 
     Each port component  412 ,  414 , and  416  is desirably initially sealed with a conventional septum or frangible membrane assembly or by a convention screw-lock luer fitting. Each port component  412 ,  414 , and  416  is sized and configured to be coupled to transfer tubing to enable transfer of materials into and out of the interior space  406 , as will be described in greater detail later. 
     For example, in a representative arrangement, the first port component  412  can be sized and configured, in use, to accommodate introduction of a material in liquid form into the interior space  406  for freeze-drying in situ within the vessel  402 . The second port component  414  can be sized and configured, in use, to accommodate introduction of a reconstituting liquid into the interior space  406  for mixing with and reconstituting the freeze-dried material. The third port component  416  can be sized and configured, in use, to accommodate transfer of reconstituted material from the interior space  406 . The use of the port components for these purposes will be described in greater detail later. 
     In the illustrated embodiment, the first port component  412  occupies a different side wall region than the second and third port components  414  and  416 . This separation segregates the port component  412  dedicated to the freeze-drying function from the port components  414  and  416  dedicated to the reconstitution and administration functions. 
     As best shown in  FIG. 39D , at least the first port component  412  is desirably oriented at a non-perpendicular angle relative to the side wall component  404 . More particularly, the port component  412  angles away from the first end component  408 , presenting a high-gravity position above the plane of the gas permeable material  410 ′ of the second end component  410 . This orientation minimizes wetting of the gas permeable material  410 ′ of the second end component  410  during introduction of the liquid material into the interior space  406  through the port component  412 . Although the freeze-drying process will ultimately dry a wetted material  410 ′ of the second end component  410 , prevention of wetting in the first instance may nevertheless be desirable, to maximize the rate of sublimation throughout the freeze-drying process. 
     During the freeze-drying process, the vessel  402  sits on a shelf within a freeze-dryer in the orientation shown in  FIG. 39D  (as also shown in  FIG. 52 . In this orientation, the gas-impermeable material  408 ′ of the first end component  408  rests on the shelf. The frame defined by the first end component  408  provides a stable platform of support for the liquid material as it undergoes freeze-drying, keeping the vessel  402  upright in this desired orientation. 
     In this desired upright orientation, the gas permeable gas permeable material  410 ′ of the second end component  410  faces upward into the freeze-drying environment. In this orientation, during drying, sublimating water vapor will escape upward from the material through the gas permeable material  410 ′ of the second end component  410 . 
     Specific details of the use of the vessel  402  before, during, and after the freeze-drying process will be described in greater detail later. 
     B. Freeze-Dried Material Storage Assembly 
     As will also be described in greater detail later, after completion of the freeze-drying process, the vacuum condition existing during the drying process may, if desired, be opened to an atmosphere of an oxygen-free, high purity inert gas, such as nitrogen or argon. The oxygen-free inert gas enters the interior space  406  through the gas permeable material  410 ′ of the second end component  410 , to exclude moisture and oxygen. 
     In this arrangement, while the vessel  402  (now containing the freeze-dried material) is maintained under the blanket of the oxygen-free inert gas, the vessel  402  is placed in a vacuum sealed, transparent vapor barrier or overwrap  418 , as shown in  FIG. 40 . The overwrap  418  is made from a gas-impermeable material and is desirable flexible, e.g., plasticized polyvinyl chloride film, or polyethylene film, or polypropylene film, or high density polyethylene film, as previously described in connection with the first end component  408 . Such materials may be used in combination with metallized, reduced gas-permeability coatings, or metal laminates. The vapor barrier or overwrap  418  traps the oxygen-free gas environment within the vessel  402  during transportation and storage. 
     The vessel  402  and overwrap  418  comprise a freeze-dried material storage assembly  420 . The exclusion of moisture and oxygen in the presence of the oxygen-free inert gas trapped by the overwrap  418  prevents degradation of the freeze-dried material carried within the vessel  402  during subsequent transport and storage. 
     The freeze-dried material storage assembly  420  can be further protected during transportation and storage by placement within a rigid outer container or can  422  as shown in  FIGS. 40 and 41A . The outer container  422  may comprise, e.g., of metal or high impact plastic material. The outer container  422  provides further protection against tearing, puncturing, or collapse of the overwrap  418  and vessel  402  during subsequent handling and storage. As  FIG. 41B  shows, the outer container  422  can, if desired, include additional compartments to hold, along with the freeze-dried material vessel  402 , a vessel filled with a reconstitution liquid, as well as associated reconstitution and administration sets. 
     In the illustrated embodiment (as shown in  FIG. 41A ), the outer container  422  includes a lid  424  that closes and, desirably, seals the container  422 . The lid  424  can be removed to provide access to the vessel  402  and overwrap  418  at the instance of use. 
     If desired (as shown in  FIGS. 42 and 43 ), one or more integrity marker elements  426  can be placed within or on the interior of the overwrap  418 . The integrity marker elements  426  carry a material sensitive to the presence of oxygen and/or moisture, or combinations thereof, and/or other pre-selected conditions adverse to or possibly adverse to the integrity or efficacy of the freeze-dried material. For example, the sensitive material can change color to visibly indicate through the overwrap  418  when a predetermined threshold level of oxygen and/or moisture, or combination thereof, exists within the overwrap  418 . The markers  426  provide further visual indications of the integrity and efficacy of the freeze-dried material within the freeze-dried material storage assembly  420  prior to reconstitution. 
     C. Unitary Freeze-Dried Material Storage Assemblies 
       FIGS. 42 and 43  show a representative embodiment of a unitary freeze-dried material storage assembly  428 . In this representative embodiment, the vessel  402  as above described and shown in  FIGS. 39A to 39D  further includes a pivotally mounted closure cover  430 . The closure cover  430  is made from a gas-impermeable material, e.g. polyvinyl chloride, or polyethylene, or polypropylene, or high density polyethylene. Such materials may be used in combination with metallized, reduced gas-permeability coatings, or metal laminates. 
     In the illustrated embodiment, the closure cover  430  is made from a generally rigid material. In this arrangement, a hinge assembly  432  on the frame defined by the second end component  410  couples the closure cover  430  on the vessel  402  for movement between an opened condition, as shown in  FIG. 42 , and a closed condition, as shown in  FIG. 43 . 
     In the opened condition (shown in  FIG. 42 ), the closure cover  430  is spaced away from the gas permeable material  410 ′ of the second end component  410 , permitting unrestricted gas transmission through the gas permeable material  410 ′ of the second end component  410  for the purposes previously described. 
     In the closed condition (shown in  FIG. 43 ), the closure cover  430  covers the entirety of the gas permeable material  410 ′ of the second end component  410 , substantially blocking gas transmission through it. 
     Desirably, the edges of the closure cover  430  and frame defined by side end component  410  are sized and configured, e.g., by interference fit and/or by use of gasket assembly, to form a gas-impermeable seal assembly about the entirety of the gas permeable material  410 ′ of the second end component  410  when the closure cover  430  is in the closed condition. If a vapor barrier overwrap is to be used, the seal assembly need not be “air tight” or aseptic, but instead provide sufficient gas holding capacity to accommodate handling in the time period between removal from the freeze dryer and the application of the vapor barrier overwrap. 
     Desirably, a latch assembly  434  on the closure cover  430  and the second end component  410  forms a lock when the closure cover  430  is in the closed condition, resisting inadvertent opening the closure cover  430 . 
     Alternatively, the closure cover  430  can comprise a more flexible material attached to the frame defined by the second end component  410 , which is normally rolled or folded away from the gas permeable material  410 ′ of the second end component  410  (i.e., the opened condition). In this arrangement, the more flexible closure cover  430  is unrolled or unfolded and drawn over the gas permeable material  410 ′ of the second end component  410  (i.e., the closed condition). The more flexible closure cover  430  is then peripherally sealed about the gas permeable material  410 ′ of the second end component  410 , e.g., by heat sealing. 
     In the arrangement shown in  FIGS. 43 and 43 , the unitary freeze-dried material storage assembly  428  undergoes the freeze-drying process in the orientation shown in  FIG. 42 , with the gas permeable material  410 ′ of the second end component  410  facing upward, and the closure cover  430  being in the opened condition (this is also shown in  FIG. 52 ). In this orientation, during drying, water vapor will sublimate and escape upward from the material within the vessel  402  through the gas permeable gas permeable material  410 ′ of the second end component  410 . 
     As previously described, after drying, a blanket of oxygen-free inert gas may, if desired, be introduced over the unitary freeze-dried material storage assembly  428  in the orientation shown in  FIG. 42 . The oxygen-free inert gas enters the interior space  406  through the gas permeable gas permeable material  410 ′ of the second end component  410 , to infiltrate and exclude moisture and oxygen in the interior space  406 , as previously described. 
     In this arrangement, while the unitary freeze-dried material storage assembly  428  (now containing the freeze-dried material) is maintained under the blanket of the oxygen-free inert gas, the closure cover  430  is placed into its closed condition (see  FIG. 54 ), and the latch assembly  434  is engaged, as shown in  FIG. 43 . The closure cover  430  traps the oxygen-free gas environment within the unitary freeze-dried material storage assembly  428  during subsequent transportation and storage. As before described, the exclusion of moisture and oxygen in the presence of the oxygen-free inert gas trapped within the unitary freeze-dried material storage assembly  428  prevents degradation of the freeze-dried material carried within the vessel  402  during subsequent transport and storage. 
     As shown in  FIGS. 44 and 45 , the unitary freeze-dried material storage assembly  428  can be placed within a rigid outer container or can  422  with a lid  424 , as previously described, made e.g., of metal or high impact plastic material. The outer container  422  provides further protection against tearing, puncturing, or collapse of the unitary freeze-dried material storage assembly  428  during subsequent handling and storage. As earlier described, if desired, the outer container  422  can include one or more separate compartments to hold a vessel containing a reconstitution liquid, as well as associated reconstitution and administration sets. 
     If desired, the unitary freeze-dried material storage assembly  428  shown in  FIG. 43  can also be placed a vacuum sealed, transparent gas-impermeable vapor barrier or overwrap  418 , of the type shown in  FIG. 40 , prior to placement in the rigid outer container. The optional overwrap  418  is shown in phantom lines in  FIG. 43 . 
       FIGS. 46 and 47  show an alternative representative embodiment of a unitary freeze-dried material storage assembly  428 . In this representative embodiment, the vessel  402  as above described and shown in  FIGS. 39A to 39D  includes a region of gas permeable material  436  that does not extend over the entire area of the second end component  410 . In this arrangement, the remaining region  438  of the second end component  410  comprises a gas-impermeable material, examples of which have already been described. 
     As shown in  FIG. 46 , the region of gas permeable material  436  is supported by and sealed to a frame  440 , which is itself joined to the second end component  410 . The sealing can be accomplished, e.g., by adhesives or heat. 
     As  FIG. 46  shows, the frame  440  rises slightly above the plane of the remainder  438  of the second end component  410 . Stand-offs  442  extend from the frame  440  into the vessel  402 , to moderate inward flexure of the frame  440  relative on the second end component  410 , e.g., when closing the closure cover  430 , as will be described. Upon an initial amount of inward flexure of the frame  440  under such conditions, the stand-offs  442  will move into contact with the first end component  408  and will thereby resist further inward flexure. 
     In this arrangement, the frame  440  carries a pivotally mounted closure cover  430 . The closure cover  430  is made from a gas-impermeable material, e.g. polyvinyl chloride, or polyethylene, or polypropylene, or high density polyethylene. Such materials may be used in combination with metallized, reduced gas-permeability coatings, or metal laminates. 
     In the illustrated embodiment, the closure cover  430  is made from a generally rigid material. In this arrangement, A hinge assembly  432  on the frame  440  couples the closure cover  430  for movement between an opened condition, as shown in  FIG. 46 , and a closed condition, as shown in  FIG. 47 . 
     In the opened condition (shown in  FIG. 46 ), the closure cover  430  is spaced away from the region of gas permeable material  436  carried by the frame  440 , permitting unrestricted gas transmission through the region of gas permeable material  436  during and after the freeze-drying process for the purposes previously described. 
     In the closed condition (shown in  FIG. 47 ), the closure cover  430  covers the entirety of the region of gas permeable material  436  carried by the frame  440 , substantially blocking gas transmission through it. 
     Desirably, the edges of the closure cover  430  and the frame  440  are sized and configured, e.g., by interference fit and/or by use of gasket assembly, to form a gas-impermeable seal about the entirety of the frame  440  when the closure cover  430  is in the closed condition. If a vapor barrier overwrap is to be used, the seal need not be “air tight” or aseptic, but instead provide sufficient gas holding capacity to accommodate handling in the time period between removal from the freeze dryer and the application of the vapor barrier overwrap. 
     Desirably, a latch assembly  434  on the closure cover  430  and the frame  440  forms a lock when the closure cover  430  is in the closed condition, resisting inadvertent opening the closure cover  430 . 
     Alternatively, the closure cover  430  can comprise a more flexible material attached to the frame  440 , which is normally rolled or folded away from the gas permeable second end component  410  on the frame  440  (i.e., the opened condition). In this arrangement, the more flexible closure cover  430  is unrolled or unfolded and drawn over the gas permeable second end component  410  on the frame  440  (i.e., the closed condition). The more flexible closure cover  430  is then peripherally sealed about the frame, to cover the gas permeable second end component  410 , e.g., by heat sealing. 
     In the arrangement shown in  FIGS. 46 and 47 , the unitary freeze-dried material storage assembly  428  undergoes the freeze-drying process in the orientation shown in  FIG. 46 , with the region of gas permeable material  436  carried by the frame  440  facing upward and the closure cover  430  in the opened condition. In this orientation, during drying, water vapor will sublimate and escape upward from the material through the region of gas permeable material  436  carried by the frame  440 . As previously described, after drying, a blanket of oxygen-free inert gas is introduced over the unitary freeze-dried material storage assembly  428  while maintained the orientation shown in  FIG. 46 . The oxygen-free inert gas enters the interior space  406  through the region of gas permeable material carried by the frame  440 , to infiltrate and exclude moisture and oxygen in the interior space  406 , as previously described. 
     In this arrangement, while the unitary freeze-dried material storage assembly  428  (now containing the freeze-dried material) is maintained under the blanket of the oxygen-free inert gas, the closure cover  430  is placed into its closed condition and the latch assembly  434  engaged, as shown in  FIG. 47 . The closure cover  430  traps the oxygen-free gas environment within the unitary freeze-dried material storage assembly  428  during subsequent transportation and storage. As before described, the exclusion of moisture and oxygen in the presence of the oxygen-free inert gas trapped within the unitary freeze-dried material storage assembly  428  prevents degradation of the freeze-dried material carried within the vessel  402  during subsequent transport and storage. 
     As shown in  FIGS. 48 and 49 , the unitary freeze-dried material storage assembly  428  can be placed within a rigid outer container or can  422  with a lid  424 , as previously described, made e.g., of metal or high impact plastic material. The outer container  422  provides further protection against tearing, puncturing, or collapse of the unitary freeze-dried material storage assembly  428  during subsequent handling and storage. 
     If desired, the unitary freeze-dried material storage assembly  428  shown in  FIG. 43  can also be placed a vacuum sealed, transparent gas-impermeable overwrap  418 , of the type shown in  FIG. 40 , prior to placement in the rigid outer container. The optional overwrap  418  is shown in phantom lines in  FIG. 47 . 
     D. Using the Unitary Freeze-Dried Material Storage Assemblies 
     1. Freeze-Drying a Material within a Freeze-Dried Material Storage Assembly 
     In a representative embodiment, the freeze-dried material comprises plasma. A description of an illustrative way of preparing freeze-dried plasma for packaging in a representative freeze-dried material storage assembly as disclosed in  FIGS. 42 and 43  therefore follows. 
     Preparation and manufacturing of the plasma will take place in an aseptic, clean room setting. The manufacturing and preparation procedures can be done, for example, in an ISO Class 5 clean room (or better) with ISO Class 3 bio-containment hoods for aseptic handling of human plasma. Freeze drying can be done aseptically in a CIP/SIP freeze dryer. 
     Human plasma is collected from a single donor in a conventional way, e.g., by collecting a unit of whole blood from the donor in a closed system collection bag, followed by centrifugal separation of the plasma and its collection in an integrally connected transfer bag  444  (containing one plasma unit of about 250 ml). Each unit (contained in the transfer bag  444 ) will be handled individually in the bio-containment hood. Between handling one single donor unit and another unit single donor unit from a different donor, there will be a line clearance protocol for change-over in the bio-containment hood, or a validation process for flow design and change-over can be otherwise provided. This protocol may address removal of all tools and materials associated with the previous handling. It may also address the thorough wash down of the containment work area and work area instruments (mass balances) to ensure no residues of the previous handling were left in place. The identification of single donor samples will be maintained by bar coding and other tagging of the single donor human plasma containers. 
     The freeze-dried material storage assembly  428  is subjected to a pre-processing protocol to provide a sterile, pyrogen free assembly. A representative size for the assembly  420  for freeze-drying about 250 ml of plasma is about 10 cm×12 cm×2 cm (l×w×d). 
     As shown in  FIG. 50 , the 250 ml human plasma unit is dispensed from the transfer bag  444  into the freeze-dried material storage assembly  428 . Flexible medical grade tubing  446  coupled integrally to the transfer bag  444  is coupled to the first port component  412  in an aseptic manner, e.g., using known aseptic coupling techniques well know in blood component processing or a spike or a leur fitting coupling under aseptic conditions. The plasma can be transferred from the transfer bag  444  into the freeze-dried material storage assembly  428  through the tubing  446  and the first port component  412  by gravity flow. 
     As shown in  FIG. 51 , the transfer tubing  446  is then disconnected in an aseptic fashion either under the conditions described above or using, e.g., a Hematron® Dielectric Sealer to provide snap-apart aseptic seals well known in blood component processing. 
     Bar coding and tagging  448  is applied to freeze-dried material storage assembly  428 . The bar coding and tagging  448  reflects the human plasma identification  450  carried by the transfer bag  444  (source, blood type, date of collection, etc.). 
     As shown in  FIG. 52 , the freeze-dried material storage assembly  428  (now containing the liquid plasma) is then placed inside a freeze dryer  452  on an aseptic freeze dryer shelf surface  454 . The freeze dryer  452  used for the lyophilization is desirably a validated clean in place, steam in place freeze dryer. 
     As shown in  FIG. 52 , the freeze-dried material storage assembly  428  is oriented with the gas permeable material  410 ′ of the second end component  410  facing upward, and the closure cover  430  placed in the opened condition. 
     Once loaded, the freeze dryer cycle (controlled by a processor  456 ) is started. This cycle generally cools the human plasma to near −45° C. and freezing for 2 to 8 hours, followed by cooling of the freeze dryer condenser and application of vacuum to start the freeze drying cycle. A freeze-dried human plasma cake is formed within the freeze-dried material storage assembly  428 . 
     In the primary freeze drying cycle, the temperature of the human plasma cake needs to remain below −33° C. (the collapse temperature) to maintain its integrity. When the moisture content of the cake is below 5% weight per weight (w/w), a secondary drying cycle (the elevated temperature) is used to further lower the moisture content. Generally the combined primary and secondary freeze drying cycles will take at least 72 hours. As before described, in the orientation shown in  FIG. 52 , during the drying cycle, sublimating water vapor will escape upward from the frozen plasma material through the gas permeable material  410 ′ of the second end component  410 , unrestricted by the opened closure cover  430 . 
     The flexible side wall component  404  accommodates flexure of the vessel due to pressure conditions encountered during the freeze drying cycle. 
     At the conclusion of the freeze drying cycle (see  FIG. 53 ), the freeze dryer vacuum is opened (by operation of the controller  456 ) to an atmosphere  458  of an oxygen-free, high purity inert gas such as nitrogen or argon. As before described, the blanket of oxygen-free inert gas enters the interior space of the freeze-dried material storage assembly  428  through the gas permeable material  410 ′ of the second end component  410 , unrestricted by the opened closure cover  430 , to infiltrate and exclude moisture and oxygen in the interior space, as previously described. 
     As shown in  FIG. 54 , while the unitary freeze-dried material storage assembly  428  (now containing the freeze-dried material) is maintained under the blanket of the oxygen-free inert gas, the closure cover  430  is placed into its closed condition and the latch assembly  434  engaged. 
     In a representative embodiment shown in  FIGS. 52 and 53 , the freeze-dryer  452  includes means  460  for providing aseptic access into the freeze-dryer  452 , so that the closure cover  430  can be manually closed, as  FIG. 54  shows. Alternatively, the means  460  can comprise remotely actuated mechanical or robotic means within the freeze dryer, to close the closure covers  430  of the unitary freeze-dried material storage assemblies  428 . 
     Still alternatively, the freeze-dried material storage assemblies  428  can removed to an aseptic containment area or cart (e.g., as generally shown  FIG. 17D ) having a contained environment maintained under an oxygen-free inert gas blanket to exclude moisture and oxygen. The containment area or cart may couple to the front of the freeze dryer to allow for transfer of the freeze dryer contents under a controlled inert gas blanket. The closure covers  430  of the freeze-dried material storage assemblies  428  can be closed within the environment provided by the aseptic container area or cart. 
     It should be appreciated that, instead of providing a unitary closure cover  430 , or in combination with a unitary closure cover  430 , a vacuum sealed, transparent overwrap  418 , as shown in  FIG. 40 , made from a gas-impermeable material can be placed over the vessel  402  in the presence of an oxygen-free inert gas environment. 
     Regardless, the closure cover  430  and/or overwrap  418  traps the oxygen-free inert gas environment within the unitary freeze-dried material storage assembly  428  (or a vessel  402  with an overwrap  418 ) during subsequent transportation and storage. As before described, the exclusion of moisture and oxygen in the presence of the oxygen-free inert gas trapped within the unitary freeze-dried material storage assembly  428  prevents degradation of the freeze-dried material carried within the vessel  402  during subsequent transport and storage. 
     As shown in  FIGS. 55 and 56 , the closed unitary freeze-dried material storage assembly  428  (with or without an overwrap  418 ) can be placed within a rigid outer container or can  422  with a lid  424 , as previously described, made e.g., of metal or high impact plastic material. The outer container  422  provides further protection against tearing, puncturing, or collapse of the unitary freeze-dried material storage assembly  428  during subsequent handling and storage. 
     2. Reconstituting and Administering Freeze-Dried Plasma from a Unitary Freeze-Dried Material Storage Assembly 
     In use at a remote site (see  FIG. 57 ), the freeze dried material storage assembly  428  (or vessel  402  with overwrap  418 , as shown in  FIGS. 40 and 41 ) is removed from the outer container  422 . After removal of the overwrap  418  (if provided), a transfer set  462  is coupled to a container  464  of sterile reconstituting liquid (e.g., water) and the second port component  414  of the respective unitary freeze-dried material storage assembly  428  (or vessel  402 ). The transfer set  462  can include plastic needles or spikes at each end to make the coupling, e.g., as shown in  FIG. 30 . The transfer set  462  may be long and flexible (as shown in  FIG. 57 ). Alternatively, the transfer set  462  can be short and rigid, to reduce storage space and simplify handling. 
     The caregiver can now proceed to manipulate the freeze dried material storage assembly  428  (or vessel  402 ), together with the container  464  of reconstituting liquid to transfer the reconstituting liquid from the container  464  into contact with the freeze-dried material within the freeze dried material storage assembly  428 , as  FIGS. 57 and 59  shows. The caregiver can create a fluid pressure differential across the transfer set  462  by selectively establishing head height differentials. Fluid can be expelled in response to the fluid pressure differential through the transfer set  462  back and forth the between the freeze dried material storage assembly  428  (or vessel  402 ) and the container  464  of reconstituting liquid  206 , as necessary to reconstitute the freeze-dried material, in preparation for administration to an individual. 
     In the embodiment shown in  FIG. 58 , the reconstituted material is administered from the freeze dried material storage assembly  428  (or vessel  402 ). In this arrangement, the administration set  462  used for mixing is uncoupled from the second port component  414 , and the second port component  414  is closed (as before described, the second port component  414  can include a septum that automatically closes upon the removal of the transfer spike or needle). At this time, as shown in  FIG. 58 , the caregiver couples the third port component  416  to an administration set  466 , for transfer of the reconstituted material into the circulatory system of an individual, as shown in  FIG. 58 . The administration set  466  includes a phlebotomy needle  468  for insertion into a vein, in the same manner as before described with reference to  FIG. 16  or  32 . The flexible side wall component  404  accommodates the collapse of the vessel  402  as the reconstituted material is administered into the circulatory system of an individual. 
     In the embodiment shown in  FIG. 60 , the reconstituted material is administered from the container  464  that initial contained the reconstituting liquid. In this arrangement, after mixing, the reconstituted material is finally transferred from freeze dried material storage assembly  428  (or vessel  402 ) to the reconstituting liquid container  464 . In this arrangement, the administration set  462  used for mixing is uncoupled from the reconstituting liquid container  464 , and the associated port  470  closed. At this time, as shown in  FIG. 60 , the caregiver couples another port  472  on the reconstituting liquid container  464  to an administration set  466 , for transfer of the reconstituted material to the circulatory system of an individual, as shown in  FIG. 60 . The administration set  466  includes a phlebotomy needle  468  for insertion into a vein, in the same manner as before described with reference to  FIG. 16  or  32 . 
     VII. Further Embodiment of a Device and System for Freeze-drying, Storing, and Administering Plasma 
       FIGS. 61-71  depict an alternate embodiment of a freeze-drying container and process, that allows freeze-drying of liquid plasma directly in the container that will be used to transfer the reconstituted plasma to a patient, without the container being placed within a vapor permeable bag or membrane during the freeze-drying process. That is, the container that contains the plasma is in communication with a vapor permeable membrane during the freeze-drying process, but is not required to be placed within a vapor permeable bag during the freeze-drying process. 
     Many freeze drying plasma processes require placing plasma within a lyophilization unit, normally with the plasma in a first container, and then placing that first container within a vapor permeable bag, made of a microporous PTFE or HDPE membrane. After freeze-drying, the vapor permeable membrane bag is discarded and the freeze-dried plasma is transferred to a container that can be used by an end user, such as a medic, with such a container preferably being of a likeness to blood bags normally used by medics and the like. An issue with such a process is that the expansion of the vapor permeable bag may cause the permeable bag to pull away from the heat transfer surface during lyophilization, which could result in less than optimal freeze-drying. The process described below can minimize such an issue. 
       FIG. 61  depicts a system  500  for freeze-drying, storing and delivering plasma to a patient, without the need to transfer the freeze-dried plasma to another container or system. The system  500  generally comprises a first collapsible container  502  and a second container  504  that contains a membrane material  506  that would typically be manufactured using microporous PTFE membrane material (e.g. Gore-Tex™) or microporous HDPE membranes (e.g. Tyvek™). The second container  502  may or may not be collapsible. The first container  502  and the second container  504  are connected by tubing  508  having an open first end  510 , with the tubing  508  allowing vapors to be transferred from the first container  502  to the second container  504  during the freeze-drying process. The tubing  508  can be of any diameter, but preferably has a diameter of approximately 1-2 cm. The tubing  508  can also be any length, but should be sufficiently long so that the tubing  508  can be pinched, closed, sealed, and severed, as will be discussed below with respect to  FIGS. 66A-69 . 
     Still referring to  FIG. 61 , the second collapsible container  504  also has an opening  512 , which receives the open first end  510 . Normally, the system  500  is provided in an assembled device, and it is preferred that the system  500  is provided as an assembled device, most specifically for sterility issues. However, if assembly is necessary, it is preferable that a heat seal will be applied to the second collapsible container  504  so that the opening  512  is sealed around the open first end  510 , providing the assembled system  500  shown in  FIG. 63 . It should be noted that the tubing  508  will then provide an open vapor or gas pathway  514  from the first collapsible container  502  to the second collapsible container  504 . The tubing  508  may be integrally formed with the first container  502 , or be a separate element that would be connected to the first container  502 , similarly to how the second container  504  is connected to the tubing  508 . 
       FIG. 62  depicts an empty assembly  500 . Due to carbonate removal during the freeze-drying process, generally in the form of carbon dioxide (CO 2 ), the pH of the eventual reconstituted plasma is elevated, i.e. higher than desired. Thus, the pH within the assembly  500  may need to be adjusted, either by adding a pH adjustment solution prior to adding plasma to the first container  502 , or backfilling CO 2  during lyophilization.  FIG. 62  demonstrates a pH adjustment solution being introduced to the assembly  500  prior to the addition of plasma. An aseptic adjustment solution port  515 , preferably a spike connection, located on the first container  502 , allows the aseptic addition of a pH adjustment solution. Preferable adjustment solutions include acids (ascorbic acid, etc.) or a buffer solution. Once the desired amount of buffer solution is added to the assembly, the solution port  515  will be sealed shut, preferably with the solution port  515  being heat sealed. 
       FIG. 63  depicts the assembly  500  after the adjustment solution has been added to the first container  502 . As stated, the solution port  515  is sealed shut. Liquid plasma is then added to the first container  502  by way of an aseptic plasma addition port  517 . Once a predetermined volume or weight of plasma has been added to the first container  502 , the port  517  will be closed and sealed, preferably heat sealed. 
       FIG. 64  displays the assembly  500 , filled with liquid plasma, with the ports  515  and  517  sealed Once the first collapsible container  502  is filled with a predetermined amount of liquid plasma  16 , the system  500  will be subjected to the freeze-drying process. As shown in  FIG. 65 , one or more devices  500 , with each first collapsible container  502  filled with liquid plasma, is placed inside a freeze dryer  320  on an aseptic freeze dryer shelf surfaces  322 . The assembly  500  is placed within a reusable stoppering mechanism, discussed further with respect to  FIGS. 66A-69B , that allows and does not restrict the interface between the first container  502  and the heat transfer membrane  506  and the shelf surface  322 . The reusable mechanism is designed to pinch close the connection tubing  508  when the lyophilizer mechanism activates at the end of the drying cycle. The freeze dryer  452  used for the lyophilization may be a validated clean in place (CIP), steam in place (SIP) freeze dryer, but the described closed system of the assembly allows for operation in a non-CIP/non-SIP lyophilization environment. Once loaded, the freeze dryer cycle is started. This cycle generally cools the human plasma to near −45° C. and freezing for 2 to 8 hours, followed by cooling of the freeze dryer condenser and application of vacuum to start the freeze drying cycle. To insure that the first collapsible container  502  stays in sufficient contact with the heat transfer surface  322  during the freeze-drying process, the first collapsible container  502  may contain an internal support structure (not shown), and/or the container  502  may be produced from a thicker, more resilient material than previously used. As a result, freeze-dried human plasma is formed in situ within the first collapsible container  502  (see FIG.  66 ). 
     The representative parameters for the freeze-drying process have been previously described and are incorporated herein by reference. 
     Throughout the freeze drying process, the gas permeable membrane  506  located on the second collapsible container  502  accommodates passage of gases, e.g., water vapor as it sublimates from the liquid plasma during freeze-drying, but otherwise prevents passage of liquid plasma from the first collapsible container  502 . The second collapsible container  504  can expand or collapse without effecting or altering the contact of the first collapsible container  502  with the freeze dryer shelf  322 . The container  502  does not have the semiporous membrane  506  located directly on the container  502 , thereby providing a permanent, air-tight seal for the container  502  through administration of reconstituted plasma, without having to transfer freeze-dried plasma from the first container  502  to an alternate container for reconstitution or administration. This can lead to increased sterility insurance. 
     The container  502  may also be vacuum packed and a sealing mechanism, as shown by example in  FIGS. 67A-69B , could be used while the first collapsible container  502  is still under vacuum ( FIG. 66 ), which would result in a very tightly packaged container. At the end of the freeze-drying process, the stoppering mechanism closes off the tubing  508  while there still is a vacuum within the freeze-dryer or the stoppering mechanism closes off the tubing  508  after the vacuum within the freeze-dryer has been broken by an inert gas, CO 2 , or a combination of the two. 
     Once the freeze-drying process has been completed, the system  500  will be removed from the freezer and the second collapsible container  504  will be removed from the first collapsible container  502  in a manner that will insure the tubing  508  and the fluid pathway  514  are sealed.  FIGS. 66A-69  depict various methods and closure devices for sealing and closing the tubing  508  after the system  500  has been through the freeze-drying process, with each of the processes providing a permanent and air-tight seal for the first collapsible container  502 . It should be noted that the assembly  500  will be removed from the freeze-dryer with the first container  502 , the second container  504 , and the tubing  508  still connected, with the desired closure mechanism in place on the tubing and maintaining positive closure between the containers  502  and  504 . 
       FIG. 66A  shows a cross-sectional view of the tubing  508  being arranged to receive a pinch-point mechanism  530 . The pinch-point mechanism  530  generally comprises a pinching component  532  and an inset  534  having a surface  536  that receives the pinching component  532 . The surface  536  can be a softer material than the rest of the mechanism  530  to ease in the pinching process. 
       FIG. 66B  shows the mechanism  530  pinching down on the tubing  508  to seal shut the tubing  508  and the fluid pathway  530 . Once the mechanism  530  is secured in place, the tube  508  can be heat sealed and severed, as shown in  FIG. 70 , to further insure an air-tight seal for the container  502 . 
       FIG. 67A  shows an alternate mechanism  540  for pinching and sealing the tubing  508 . The mechanism  540  generally comprises a pinching member  542  and a hinged member  544 . The hinged member  544  has two arms  546 , pivotally connected by a hinge  548 . Rolling members  550  are positioned along the arms  546  to assist in the mechanism  540  providing a tight seal on the tubing  508 . 
     As shown in  FIG. 67B , the pinching member  542  is forced downwardly into the tubing  508  into the hinged member  544 . The arms  546  pivot upwardly, with the rolling members  550  assisting in the arms moving inwardly toward one another, thereby providing the necessary seal. The tubing  508  can then be subjected to further heat sealing, as discussed with respect to  FIG. 69 . 
       FIG. 68A  demonstrates another pinching mechanism  560  used for sealing the first collapsible container  502  and the tubing  508 . The tubing  508  is folded over itself while it is being pinched, which may provide an easier pinching process. The pinching mechanism  560  generally forms a ratcheting mechanism that comprises a fixed portion  562  and a movable portion  564 , and a guide member  566 . 
     As shown in  FIG. 68B , the movable portion  564  is moved downwardly towards the fixed portion  562 , with the guide member  566  providing support for the movable portion  564  and closing off the tubing  508 . As with the other noted pinching and sealing mechanisms, the tubing  508  can then be subjected to further heat sealing, as discussed with respect to  FIG. 69 . It should be noted that a ratcheting mechanism as discussed with respect to  FIGS. 68A-68B  could also be incorporated into the pinching mechanisms  530  and  540 , discussed in  FIGS. 66A-B  and  FIGS. 67A-B , respectively, to provide the necessary air-tight seal of the tubing  508 . 
     It should be noted that the first collapsible container  502  is independently shaped and formed from the second collapsible container  504  and the permeable membrane  506  that is supported by the second container  504 . Conversely, the design of the second container  504  and the permeable membrane  506  can be altered as well.  FIGS. 71 and 72  show alternate embodiments for the system  500 , where the second collapsible container is not used, but tubing  508  is directly connected to a permeable membrane  570  ( FIG. 71 ) or the tubing  508  is mated with a filter  572  ( FIG. 72 ), with the filter possibly being a commercially available filter known in the art. For example, the filter media may comprise a hydrophobic polymer, such as a polypropylene, polyester, polyethylene, polyurethane, polyvinylidene fluoride or polytetrafluoroethylene material. The permeable membrane  570  will be sealed around the tubing  508  or the filter  572 , preferably by heat sealing so that The permeable membrane  570  will function similarly to the membrane  506  and the previous membranes discussed in the freeze-drying process, and can be severed from the first collapsible container  502  as discussed with respect to  FIGS. 67A-69B . 
       FIG. 70  provides the first collapsible container  502  filled with plasma after the freeze-drying and sealing process. The collapsible container  502  can be stored within a pouch  580  designed with a minimum or low moisture vapor transmission rate (MVTR). The pouch may also have oxygen absorbers  582  and water absorbers  584  to aid in protecting the dried plasma from exposure to water and oxygen during storage. An administration port  518  is located on the first collapsible container  502 , for delivery of reconstituted plasma to a patient, generally as depicted and discussed previously (see e.g.,  FIGS. 16 ,  32 , and  60 ). The administration port  518  preferably has a standard blood bag aseptic connection arrangement, with a typical blood bag spike  520 . The port  518  is sealed until the container  502  will be administered to a patient. The first collapsible container  502  further supports a reconstitution port  522 , also preferably with a standard blood bag aseptic connection arrangement for attaching the container  502  to a liquid container, as similarly described with respect to  FIGS. 31 and 60 . 
     In use at a remote site (see  FIG. 73 ), the transfer set  462  is coupled to the container  464  of sterile reconstituting liquid (e.g., water) and the first collapsible container  502 . The transfer set  462  can include plastic needles or spikes at each end to make the coupling, e.g., as shown in  FIG. 30 . The transfer set  462  may be long and flexible (as previously shown in  FIG. 57 ). Alternatively, the transfer set  462  can be short and rigid, to reduce storage space and simplify handling. 
     The caregiver can now proceed to manipulate the container  464 , to transfer the reconstituting liquid from the container  464  into contact with the freeze-dried material within the first collapsible container  502  to mix the reconstituting liquid  206  and the freeze-dried material within the first collapsible container  502 , in preparation for administration to an individual. 
       FIGS. 73 and 74  demonstrate that the first collapsible container  502  is similar in appearance to a typical blood bag known and used in the industry, thereby providing a familiar container for a caregiver to administer the reconstituted plasma. In  FIG. 73 , the reconstituted material is administered from the first collapsible container  502  that initially contained the liquid plasma prior to freeze-drying, which then subsequently contained the freeze-dried plasma after freeze drying. A pinch valve  574  located on the administration set  462  used for mixing is closed, thereby preventing further transfer of fluid from the associated port  470 . The administration set  462  may then be severed and sealed. At this time, as shown in  FIG. 74 , the caregiver couples the administration port  522  on the first collapsible container  502  to the administration set  466 , for transfer of the reconstituted material to the circulatory system of an individual, as shown in  FIG. 74 . The administration set  466  includes the phlebotomy needle  468  for insertion into a vein, in the same manner as before described with reference to  FIG. 16 ,  32  or  60 . 
     VIII. Treatment of Single Unit Plasma Before Freeze-Drying 
     In the previously described embodiments, single unit human plasma is dispensed from a source transfer bag into a receptacle in which the plasma under goes freeze-drying, which (depending upon the particular embodiment) comprises a mold  50  (see  FIG. 17A ); or a compartmentalized container device  300  (see  FIG. 34 ); or a multifunctional device  400  (see  FIG. 50  or  FIG. 64 ). Whatever the particular embodiment, during the dispensing of the plasma from the source transfer container  600  into the freeze-drying receptacle  602  (see  FIG. 75 ), an inline treatment device  604  can be provided for treating the plasma prior to freeze-drying. 
     The form of treatment can vary. For example, the plasma can carry residual cellular blood components such as platelets, red blood cells, and/or leukocytes that were not removed during processing of the plasma from whole blood. The plasma can also carry, for example, viral or bacterial agents that reside in the plasma or are carried on or within cellular blood components in the plasma. The plasma can also include blood group specific antibodies, e.g., anti A IgM/IgG or anti B IgM/IgG. 
     The inline treatment device  600  can likewise vary according to the form of treatment desired. For example, for the removal of cellular blood components, the inline device  604  can include an appropriate conventional filtration media having an affinity for the blood component or components. For the removal of pathogens or viral or bacterial agents, the inline device  604  can include a convention pathogen inactivation mechanism, e.g., actinic radiation, photo-deactivation, pasteurization, solvent detergent treatment, and the like. For the removal or neutralization of blood group specific antibodies, the inline device  600  can include an antigen having an affinity for the particular antibody. 
     For example, in a representative embodiment, the source transfer container  600  contains single unit blood plasma, e.g., derived from a single donor plasma of a specified blood group, either type A plasma or type B plasma. If the specified blood group is type A plasma, the inline treatment device  604  includes an antigen for removing or neutralizing B antibodies from the type A plasma. Likewise, if the specified blood group is type B plasma, the inline treatment device  604  includes an antigen for removing or neutralizing A antibodies from the type B plasma. The antigen can be bound to a resin material housed within the inline treatment device  604 , through which the plasma is passed. This has the effect of making possible the processing freeze-dried plasma obtained from a single unit source of either Type A or Type B (which comprise more than two-thirds of all blood donors), and providing a freeze-dried plasma product that is universally applicable to patients of all different blood groups. 
     It should be appreciated that plasma can be treated in any of the manners described during blood processing before being transfer into the source container, and thereby comprise pre-treated single unit blood plasma. 
     IX. Addition of Ascorbic Acid to Plasma for Stability and Efficacy Purposes 
     Regardless of the processing and storage of the type of plasma being used or administered, it must be as utile as requirements for plasma currently being used. That is, freeze-dried or lyophilized plasma prepared and stored according to the present invention, must be comparably stable and efficacious as plasma currently being used, such as fresh, frozen plasma. It has been determined that plasma prepared and stored according to the present invention compares to currently used plasma and, in fact, has additional, unexpected benefits compared to prior forms of plasma. Particularly, the stability and efficacy of plasma of the present invention has been improved by the addition of ascorbic acid, i.e. vitamin c, to the plasma, prior to reconstitution of the plasma. 
     A. Addition of Ascorbic Acid to Plasma 
       FIG. 76  demonstrates one step for the addition of ascorbic acid (AA) to plasma, prior to freeze drying. The 250 ml human plasma unit is dispensed from the transfer bag  48  into a sterile, pyrogen free, rectangular mold  50  (e.g., 4 cm×10 cm×12.5 cm—d×w×1). The mold  50  can be stainless-steel; however it can also be composed of metal with good thermal transfer properties such as aluminum, aluminum alloy, titanium or gold. The mold  50  may be coated on its inside surfaces with a tough, inert barrier film with good release properties such as PTFE or diamond. An ascorbic acid solution  700  is added to the human plasma, either directly into the transfer bag  48  or into the mold  50  by way of delivery line  702 . Preferably, about 600 mg of AA is added to the plasma, thereby having a concentration of AA of about 16 mmol. The in-line treatment device  604  can also be incorporated for further filtering of the plasma prior to transferring the material to the mold  50 . Once added, the mold  50  can be further processed to lyophilize and freeze-dry the plasma, as described above with respect to  FIGS. 17B-17E . 
     Alternatively, the ascorbic acid  700  may be added to the plasma after the plasma has been freeze-dried and/or lyophilized.  FIG. 77  depicts the addition of ascorbic acid, along with a reconstitution fluid to the device  10 , similarly to the addition of reconstitution fluid as discussed with respect to  FIGS. 18-21 , above. The reconstituting liquid  18  (in the representative embodiment, gas-free water) is introduced into the second chamber  14 . Ascorbic acid  700  is also introduced into the second chamber  14 , through administration port  768  and feed line  782 . The vacuum port  66  and administration ports  68 ,  768  are connected to feed lines  80  and  82 ,  782  respectively, as  FIG. 77  shows. Gas in the chamber  14  is removed by application of aseptic vacuum. 
     The vacuum port  66  is sealed and the tubing  80  is removed. The required aliquot (e.g., approximately 250 ml) of reconstitution fluid is added to the chamber  14  through the administration port  68 , as well as the required amount of ascorbic acid. The tubing  82  and the tubing  782  are removed and the administration ports  68  and  768  are then sealed. Alternatively, the ascorbic acid  700  may be added to the reconstitution fluid  18  prior to introducing the reconstitution fluid into the chamber  14  so that the fluid  18  and the ascorbic acid  700  both are introduced into the port  68 , as shown in  FIG. 20 . The chamber  14  would then be sealed as described with respect to  FIG. 20 , above. Once sealed, the device  10  can then be transported, stored, and reconstituted as discussed above. It should also be understood that the structures of  FIGS. 76 and 77  are exemplary, and ascorbic acid could be added to other structures shown and discussed, above. 
     B. Stability of Plasma Containing Ascorbic Acid 
     As noted above, the plasma of the present invention must perform sufficiently as well as plasma that is currently being used and administered. As such, plasma of the present invention was tested to determine the stability and other factors of the plasma. In carrying out the tests, fresh frozen plasma (FFP) were pooled, than aliquotted into molds  50 , as described above, with each mold containing 40 ml of plasma. Various amounts of 200 mM ascorbic acid solution were added to each of the molds  50 , and the molds  50  were lyophilized, as described above. Table 1, below, depicts the various amounts of ascorbic acid introduced into the plasma. 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Summary of the various amounts of aqueous ascorbic 
               
               
                 acid solution added to pooled plasma 
               
            
           
           
               
               
               
            
               
                   
                 Final Ascorbic 
                 Volume of 200 mM of 
               
               
                   
                 Acid Concentration 
                 aqueous Ascorbic Acid 
               
               
                   
                 in FFP 
                 added (mL) 
               
               
                   
               
            
           
           
               
               
               
            
               
                 0 
                   
                 0.00 
               
               
                 7 
                 mM 
                 1.45 
               
               
                 10 
                 mM 
                 2.11 
               
               
                 15 
                 mM 
                 3.24 
               
               
                   
               
            
           
         
       
     
     When the lyophilization cycle was complete, the individual lyophilized plasma cakes were reconstituted with Sterile Water for Injection (SWFI) (i.e. reconstitution fluid). pH readings were measured for each condition at room temperature (25° C.). The results are depicted in  FIG. 78 . The addition of ascorbic acid to the plasma has a positive effect on the plasma, as it down regulates the pH to a more physiological acceptable level. As shown in  FIG. 78 , addition of 15 mM of ascorbic acid reduces the pH from above 8.9 to a more neutral pH of ˜7.5. 
     Because the addition of 15 mM of ascorbic acid produced a more physiological acceptable pH level compared to the other tested amounts, the stability of lyophilized plasma containing no ascorbic and 15 mM ascorbic acid was evaluated at three different storage conditions (i.e., Room Temperature (RT), 4° C. and 42° C.). Ascorbic acid is a well known antioxidant that has a wide range of applications. It was proposed that its functions in lyophilized plasma are not only to aid in adjustment of pH to physiological pH upon reconstitution, but also to act as an antioxidant in the lyophilized plasma cake. 
     Reconstituted lyophilized plasma at various conditions was evaluated to ensure that the addition of ascorbic acid does not affect certain coagulation factors:
         coagulation assays (thrombin time (TT), prothrombin time (PT), activated partial thromboplastin time (aPTT))   fibrinogen   Factor V   Factor VIII       

     Testing was done at various time intervals (pre-lyophilization, at lyophilizations, and 2, 4, 8, and 12 weeks after lyophilization) to determine whether the addition of ascorbic acid had any effect on the stability of the plasma. Testing was also done at various temperatures (room temperature (RT), 4° C., and 42° C.) The results are shown in  FIGS. 79-84 . 
       FIG. 79  shows that the thrombin time (TT) is not deleteriously affected by the addition of ascorbic acid (AA) and, in fact, at elevated temperatures (42° C.), stability is greatly improved. 
     Similarly, in  FIG. 80 , prethrombin time (PT) is also not deleteriously affected by the addition of ascorbic acid (AA) and, at elevated temperatures (42° C.), stability is greatly improved. 
       FIG. 81  demonstrates that the activated partial thromboplastin time (aPTT) is not deleteriously affected by the addition of ascorbic acid (AA). At elevated temperatures (42° C.), the stability of the plasma is also shown to perform better than the plasma without any ascorbic acid added. 
     In  FIG. 82 , the fibrinogen data for each sample with and without the addition of ascorbic acid performs similarly. That is, at room temperature, 4° C., and 42° C., the samples containing ascorbic acid performed similarly to those without any ascorbic acid. 
       FIG. 83  compares the loss of Factor V from the plasma at the various time intervals. The results show that the addition of ascorbic acid to the plasma at each time interval results in substantially less than the plasma without any additional ascorbic acid. After 12 weeks, at room temperature and at 4° C., the plasma with ascorbic acid added retained over 50% of Factor V, while retention of the plasma without ascorbic acid was around 20%. At elevated temperatures (42° C.), plasma with ascorbic acid still retained about 25% of Factor V, while the plasma without ascorbic acid had lost 100% of the Factor V. 
     Similarly, in  FIG. 84 , the retention of Factor VIII was higher at each temperature for each sample containing ascorbic acid compared to the respective sample without ascorbic acid. 
     The results show that, not only does ascorbic acid not negatively affect the stability of the plasma, but has positive effects on the plasma, as well. 
     C. Comparison of Plasma to Other Types of Plasma 
     The lyophilized plasma containing ascorbic acid (vitamin C) (LP) prepared according to the present invention, and discussed above, was tested to determine the efficacy of the plasma compared to the efficacy of fresh frozen plasma (FFP), as well as both types of plasma in combination with packed red blood cells (PBRC), i.e. LP:PBRC and FFP:PBRC. The results indicated that LP had similar or better results than the FFP for various factors, such as clotting, coagulation, and post-injury blood loss. The testing is presented, below. 
     Materials and Methods 
     Preparation of Lyophilized Plasma 
     All experimental procedures were done in accordance with the guidelines of the Institutional Animal Care and Use Committee at Oregon Health &amp; Science University and the United States Army Institute of Surgical Research. Blood products used in the study were obtained from juvenile Yorkshire crossbred swine. Using sterile precautions, a cervical cut-down was performed and the external jugular vein was cannulated with an 8Fr Introducer (Argon Medical Devices, Athens, Tex.). Animals were exsanguinated and blood was collected in citrated Terumo Teruflex (Terumo Medical Corporation, Tokyo, Japan) triple blood donation bags. Whole blood was centrifuged at 5000 g for 9 minutes at 4° C. and plasma was removed using a Baxter Plasma Extractor (Baxter Healthcare, Deerfield, Ill.). Plasma was stored at −20° C. for transport to HemCon Medical Technologies, Inc. (Portland, Oreg.) for lyophilization. Sterile LP was then stored at room temperature for up to one month. Immediately prior to use, LP was reconstituted to its original volume using sterile water containing ascorbic acid for pH adjustment. 
     Serum Clotting Factor Level Measurements 
     Samples of plasma were analyzed for levels of Factors II, V, VII, VIII, IX, X, XI, XII, fibrinogen, protein C, antithrombin III using a BCS Coagulation System machine (Dade Behring Inc, Marburg, Germany) at the time of initial plasma preparation and after LP was fully reconstituted. Functional clotting assays partial thromboplastin time (PTT) and prothrombin time (PT) were performed at the same two time points. 
     Animal Model 
     The swine model used is a well-validated model of severe injury and hemorrhagic shock described by Cho, et al., and depicted in  FIG. 85 . Briefly, 32 juvenile Yorkshire crossbred swine (8 per group) were anesthetized mechanically ventilated, and had invasive lines placed. All subjects underwent femur fracture using a Schermer captive bolt gun (Karl Schermer Co, Ettlinger, Germany) to produce a comminuted long bone fracture with severe overlying soft tissue injury. After laparotomy, subjects were cooled to 33° C. with intraperitoneal saline and underwent controlled hemorrhage by removing 60% of their estimated blood volume via a central line followed by 30 minutes of shock. Subjects were infused with 0.9% normal saline at volumes three times the controlled hemorrhage volume in order to induce acidosis and coagulopathy. To mimic operative rebleeding and to produce an injury allowing measurement of blood loss after randomization to treatment, subjects received a grade V liver injury followed by 30 seconds of uncontrolled hemorrhage. Following the uncontrolled hemorrhage period, the liver was packed tightly with laparotomy sponges. Swine were randomized to receive either FFP, LP, 1:1 ratio of FFP:PRBC, or 1:1 ratio of LP:PRBC at 50 mL/min, infusing volumes equal to the blood removed during controlled hemorrhage. Resuscitation was initiated at the time of the liver packing. Subjects were then monitored for 4 hours after resuscitation and subsequently chemically euthanized. Hemodynamic data (heart rate and blood pressure) were recorded continuously throughout. Blood loss after liver injury was carefully recorded using pre-weighed laparotomy sponges and a pre-weighed suction canister. Serum samples were tested for prothrombin time (PT), partial thromboplastin time (PTT) and lactate at baseline, after femur fracture, before controlled hemorrhage, before liver injury, and hourly for 4 hours after resuscitation with study fluid. In order to quantify levels of (Interleukin) IL-6, IL-8, and TNF-α, serum samples were collected prior to liver injury and 2 and 4 hours after administration of study fluids and quantified by commercially available enzyme linked immunosorbent assay (ELISA, R&amp;D Systems, Minneapolis, Minn.). 
     Statistical Analysis 
     Data were analyzed using SPSS software version 16.0 (SPSS, Chicago, Ill.). Variables were assessed for normal distribution Comparisons between groups at the various time points were analyzed using independent t-tests. Paired samples t-tests were used to compare same-group samples across various time points. Significance was denoted at a p&lt;0.05. 
     Results 
     Effect of Lyophilization on Factor Function 
     On average, for the lyophilized plasma (with ascorbic acid) clotting factor levels were decreased to 84% of their pre-lyophilization values, as shown in  FIG. 86 . Compared to pre-lyophilization values, factor V retained 84% activity, factor VIII retained 84% activity, factor IX retained 100% activity, and antithrombin III retained 93% activity. PTT was prolonged by 9% and INR was prolonged by 13% compared to pre-lyophilization. 
     Effect of Different Resuscitation Regimens on Outcomes 
     No pigs died prior to the end of the study in any group. There were no differences between the four resuscitation groups with respect to blood loss after liver injury ( FIG. 87 ) or heart rate (HR) after resuscitation ( FIG. 88 ). The mean arterial pressure (MAP) was lower at various time points after resuscitation in the FFP group compared to the other  3  groups ( FIG. 89 ). The MAP in the LP:PRBC group was higher than the LP group at 3 hours post resuscitation. Lactate levels and PT were similar in all groups at all four post-resuscitation time points ( FIG. 90  and  FIG. 91 ). PTT in the FFP group was lower than the LP:PRBC group and the FFP:PRBC group at all four post-resuscitation time points ( FIG. 92 ). PTT in the LP group was lower than LP:PRBC and the FFP:PRBC group at variable time points post-resuscitation. 
     There were no differences between LP and FFP or between the 1:1 groups with respect to IL-8 and TNF-α at any time point in the study. IL-6 levels in the LP group were less than the FFP:PRBC group at 2 hours post injury and less than the FFP group at 4 hours post injury ( FIG. 93 ). IL-6 levels in the LP:PRBC group were less than the FFP:PRBC group at 2 hours. 
     Analysis 
     The lyophilization process of plasma according to the present invention, LP (lyophilized plasma with ascorbic acid), results in a modest reduction in clotting factor activity in vitro, and that LP is as safe and effective as FFP (fresh frozen plasma) for resuscitation after severe multi-system injury. On average, clotting factors were decreased by 14% for LP, which compares favorably with the standard 25-40% reduction caused by freezing and thawing FFP. Industry standards for assessing the quality of FFP require the thawed FFP to maintain factor V, factor VIII and AT III levels of at least 70%. The LP retained these factors at levels much greater than the required 70%. Additionally, the overall 14% reduction in clotting factor levels seemed to have only a minor effect on the functional clotting assays PTT and INR. Even more importantly, LP was at least as effective as FFP in reversing the coagulopathy induced in the animal model. 
     The combined data on mortality, blood loss after liver injury, coagulation parameters and lactate demonstrate that LP is as effective as FFP for resuscitation after severe injury. LP performed as well or better than FFP in all of the above areas. Further, the data suggests that LP has great promise as a resuscitation fluid in the combat and pre-hospital settings as well as in the hospital. 
     X. Conclusion 
     The foregoing is considered as illustrative only of the principles of the invention. Furthermore, since numerous modifications and changes will readily occur to those skilled in the art, it is not desired to limit the invention to the exact construction and operation shown and described. While the preferred embodiment has been described, the details may be changed without departing from the invention, which is defined by the claims.