A spray drying process that reduces pathogens

Abstract

The spray dried plasma by the nature of its characteristics and the process by which it is made allows for a dried plasma with reduced pathogens. It has at least three aspects of pathogen reduction that results in a plasma product with reduced or no pathogens. The first aspect relates to a spray drying disposable and machine that eliminates or minimizes the introduction of pathogens into the spray drying process and is referred to as the anti-contamination process. The second aspect of pathogen reduction relates to the spray drying process and parameters that reduces pathogens that may be present in the donor plasma, and / or inhibit pathogen growth and / or pathogen proliferation introduced by the donor plasma. The third aspect results in reduced pathogens occurring during storage of the dried plasma wherein the manufacturing and storage conditions are such that pathogens are reduced, and / or pathogen growth and / or pathogen proliferation are inhibited.

Description

E:\Office Documents\corporate_Docs\Antoinette's corporate\client\0118 Velico Medical Inc\0118.0141-002 PCT N1 Pathogen Reduction AntiContamination\N1PCTFINALPathogenReduction.docxAGGPATENT APPLICATION September 14, 2023 Docket No.: 0118.0141002 N1 -1- FILED ELECTRONICALLYA Spray Drying Process That Reduces Pathogens RELATED APPLICATION(S)

[0001] This application claims the benefit of U.S. Provisional Application No 63406747, entitled, “A Spray Drying Process That Reduces Pathogens” by Kyle D. Erickson et al., filed September 15, 2022.

[0002] The entire teachings of the above application(s) are incorporated herein by reference.

[0003] GOVERNMENT SUPPORT This invention was made with Government support under contract Nos. and 75A50121C00059 awarded by the Biomedical Advanced Research and Development Authority (BARDA). The Government has certain rights in the invention.

[0005] BACKGROUND OF THE INVENTION

[0006] Dried plasma has great medical importance, and its development has been the subject of of development and expense. Known methods for drying blood plasma include freeze drying, also known as lyophilization, and spray drying. Dried plasma has applications in the military, mass casualty events, and in the treatment of certain disorders.

[0007] With respect to plasma, transfused human blood plasma is often crucial to bleeding and wound treatment in trauma victims, and in surgery. Unfortunately, plasma is not readily available in many circumstances world-wide including battlefield, first responder, and rural settings, remote from large hospitals, and in the second and third world.

[0008] In general, blood plasma can be stored frozen for long periods which reduces the load but does not eliminate any pathogens that may be present. Liquid plasma can only be stored for very short periods in part because pathogens can grow or proliferate in theDocket No. 0118.014100 -2- plasma. Freeze dried plasma can also preserve any pathogens that may be present in it and when later rehydrated, the pathogens may be viable along with the plasma.

[0009] Hence, a need exists for dried plasma that has reduced pathogens, as compared to its liquid, donor plasma. A further need exists for a spray dried plasma process reduces or minimizes the introduction of pathogens such as a method which is aseptic. Finally, a need exists for dried plasma, that when stored, has reduced pathogens or does not allow them to proliferate or grow. SUMMARY OF THE INVENTION

[0010] The spray dried plasma by the nature of its characteristics and the process by which it is allows for a dried plasma with no or reduced pathogens. It has at least three aspects of pathogen reduction that results in a plasma product with reduced or no pathogens. The first aspect relates to a spray drying disposable and machine that reduces or minimizes the introduction of pathogens into the spray drying process and is referred to as the aseptic or anti-contamination process. The second aspect of pathogen reduction relates to the spray drying process and parameters that reduces the concentration of pathogens that may be present in the donor plasma, and / or inhibit pathogen growth and / or pathogen proliferation introduced by the donor plasma. The third aspect results in reduced pathogens occurring during storage of the dried plasma wherein the manufacturing and storage conditions are such that pathogens are reduced, and / or pathogen growth and / or pathogen proliferation are inhibited.

[0011] The present invention includes a spray dried plasma that is spray dried from a donor wherein the spray dried plasma has a reduced amount of pathogens, as compared to donor plasma. The present invention also includes a reconstituted previously spray dried plasma that is spray dried from a donor plasma, wherein the reconstituted spray dried plasma has a reduced amount of pathogens, as compared to donor plasma and has one or more plasma proteins that induce clot formation. In an embodiment, the reconstituted previously spray dried plasma has the one or more plasma proteins having a level within a corresponding clinical reference range. In another embodiment, the one or more plasma proteins that induces clot formation is von Willebrand Factor (vWf). In one aspect, vWF canDocket No. 0118.014100 -3- be measured by von Willebrand Factor assay or von Willebrand Factor Ristocetin Cofactor assay or Willebrand Factor antigen assay. In an aspect, the reconstituted previously spray dried plasma has von Willebrand Factor Antigen or von Willebrand Factor Ristocetin Cofactor in an amount ranging about 50 IU / dL and about 200 IU / dL.

[0012] The present invention involves a spray dried plasma having a reduced amount of pathogens. The spray dried plasma is made using a spray drying disposable device, having a spray drying head and a plasma drying chamber, in a spray drying system having a drying gas source, a plasma source and a pressurized aerosol gas source, as described herein. The spray drying disposable device has a spray drying head having a spray dry nozzle assembly in fluid communication with the plasma source and the pressurized aerosol gas source, wherein the pressurized aerosol gas flows in a vortex pattern, wherein, when in use, the pressurized aerosol gas atomizes the plasma in the drying chamber to obtain atomized plasma droplets; wherein, when in use, the spray drying head source supplies drying gas to the drying chamber at a temperature of between about 90° C and about 130° C. The spray drying disposable device further includes a drying chamber, attached to a baffle plate, wherein atomized plasma droplets evaporate in the presence of the drying gas emitted from the one or more drying gas jets to thereby obtain dried plasma particles and humid air; a capture filter, residing in the drying chamber, wherein the capture filter captures the dried plasma particles and allows the humid air to pass; and a gas outlet, wherein said gas outlet is attached to the exhaust port of the spray drying apparatus, wherein the humid air flows through the gas outlet; wherein an amount of pathogens is reduced in the spray dried plasma, when reconstituted, as compared to spay dried plasma not spray dried using the spray dried disposable or to that present in the donor plasma. In an embodiment, the atomized plasma droplets have a temperature between about 20oC and about 65oC.

[0013] In another embodiment, the spray dried plasma having a reduced amount of pathogens is sprayed using a spray drying disposable device a spray drying head with a spray dry nozzle assembly in fluid communication with the plasma source and the pressurized aerosol gas source, wherein the pressurized aerosol gas flows in a vortex pattern, wherein, when in use, the pressurized aerosol gas atomizes the plasma in the drying chamber to obtain atomized plasma droplets; a plenum having a drying gas inlet in communication with the drying gas source, wherein, when in use, the drying gas resides in the plenum with uniform air pressure,Docket No. 0118.014100 -4- wherein the plenum supports the nozzle assembly; and a baffle plate forming the floor of the plenum having one or more drying gas jet, wherein drying gas jet provides drying gas to the drying chamber. The spray drying disposable device also has drying chamber, attached to the baffle plate, wherein atomized plasma droplets evaporate in the presence of the drying gas emitted from the one or more drying gas jet to thereby obtain dried plasma particles and humid air; a capture filter, residing in the drying chamber, wherein the capture filter captures the dried plasma particles and allows the humid air to pass; and a gas outlet, wherein said gas outlet is attached to the exhaust port of the spray drying apparatus, wherein the humid air flows through the gas outlet; wherein an amount of pathogens is reduced in the spray dried plasma, when reconstituted, as compared to spay dried plasma not spray dried using the spray dried disposable or to that present in the donor plasma. In an embodiment, the drying gas has a temperature of between about 90° C and about 130° C. In yet another embodiment, the atomized plasma droplets have a temperature between about 20oC and about 65oC. The spray drying disposable device further comprises a baffle plate filter in communication with the baffle plate, wherein the baffle plate filter comprises 0.2 micron filter.

[0014] The present invention further includes methods of spray drying plasma wherein the spray plasma, when reconstituted, has a reduced amount of pathogens and has one or more plasma proteins that include clot formation. The method includes the step spray drying plasma from a donor plasma; the spray dried plasma is made with a spray drying disposable device, as described herein. When in use, the spray drying head source supplies drying gas to the drying chamber at a temperature of between about 90° C and about 130° C. The reconstituted spray dried plasma has a reduced amount of pathogens, as compared to donor plasma and has one or more plasma proteins that induce clot formation.

[0015] In an embodiment, the pathogen reduction method includes using spray drying head with dry nozzle assembly in fluid communication with the plasma source and the pressurized aerosol gas source, wherein the pressurized aerosol gas flows in a vortex pattern, wherein, when in use, the pressurized aerosol gas atomizes the plasma in the drying chamber to obtain atomized plasma droplets; a plenum having a drying gas inlet in communication with the drying gas source, wherein, when in use, the drying gas resides in the plenum with uniform air pressure, wherein the plenum supports the nozzle assembly; a baffle plateDocket No. 0118.014100 -5- forming the floor of the plenum having one or more drying gas jets, wherein the one or more gas jets provide drying gas to the drying chamber. The disposable further includes the drying chamber, attached to the baffle plate, wherein atomized plasma droplets evaporate in the presence of the drying gas emitted from the one or more drying gas jet to thereby obtain dried plasma particles and humid air; a capture filter, residing in the drying chamber, wherein the capture filter captures the dried plasma particles and allows the humid air to pass; and a gas outlet, wherein said gas outlet is attached to the exhaust port of the spray drying apparatus, wherein the humid air flows through the gas outlet. The amount of pathogens is reduced in the spray dried plasma, when reconstituted, as compared to spay dried plasma not spray dried using the spray dried disposable or to that present in the donor plasma. The methods described herein involve using donor plasma that is pretreated with an amount of an spray dry stable acidic substance (SDSAS) and one or more amino acids, as described herein..

[0016] The present invention further includes methods for storing spray dried plasma with reduced pathogens, the steps of the method include subjecting or storing the dried plasma to a temperature between about -80ºC and about 45oC for a period of time ranging from about 1 day and about 48 months; wherein the amount of pathogens are not increased, as compared to that before storage. The amount of pathogens is decreased after storage, and the level of one or more plasma proteins induces clot formation.

[0017] Advantageously, the rehydrated previously dried plasma of the present invention has reduced pathogens, as compared to its corresponding donor plasma. The spray drying system of the present invention reduces and minimizes the introduction of pathogens during the spray drying process and therefore the resulting dried plasma has less pathogens as compared to plasma processed in other ways. Finally, the low residual moisture of the dried plasma allows it to be stored and stable with a reduction in pathogens or without pathogen growth or proliferation.

[0018] BRIEF DESCRIPTION OF THE DRAWINGS and other objects, features and advantages of the invention will be from the following more particular description of preferred embodiments of the invention, as illustrated in the accompanying drawings in which like reference charactersDocket No. 0118.014100 -6- refer to the same parts throughout the different views. The drawings are not necessarily to scale, emphasis instead being placed upon illustrating the principles of the invention.

[0020] Fig. 1 is a black and white photograph showing phase contrast microscopy at 100X and 400X magnification of cholesterol crystals in freeze-dried plasma known as LYOPLAS TM plasma.

[0021] Fig. 2 is a black and white photograph showing phase contrast microscopy at 100X and 400X magnification of cholesterol crystals in freeze-dried plasma known as FLYP TM plasma.

[0022] Fig. 3 is a panel of black and white photographs showing phase contrast microscopy at 100X magnification of cholesterol crystals in freeze-dried plasma with LYOPLAS TM plasma (left) and FLYP TM plasma (right).

[0023] Fig. 4 is a panel of black and white photographs showing of phase contrast microscopy of spray dried plasma with no visible cholesterol crystals observed as follows: 100x magnification of donor, pre- spray dried, plasma (upper left panel), 100x magnification of spray dried, plasma (upper right panel), 400x magnification of donor, pre- spray dried, plasma (lower left panel), and 400x magnification of spray dried, plasma (lower right panel),

[0024] Fig. 5 is a composite bar / line graph showing particulate concentration (particulates / mL) and particulate size (logarithmic scale (µm)) of single unit dried plasma after 7.5 months of room temperature storage at initial rehydration and 4 hours post rehydration compared to its paired thawed frozen control plasma as follows: control plasma (CP) at time zero (T=0), spray dried plasma of the present invention (ODP) at time zero (T=0), control plasma (CP) at time of 4 hours (T=4), spray dried plasma of the present invention (ODP) at time of 4 hours (T=4).

[0025] Fig. 6 is a composite bar / line graph showing particulate concentration (particulates / mL) and particulate size (logarithmic scale (µm)) of single unit dried plasma after 12 months of refrigerated storage at initial rehydration and 4 hours post rehydration compared to its paired thawed frozen control plasma as follows: control plasma (CP) at time zero (T=0), spray dried plasma of the present invention (ODP) at time zero (T=0), control plasma (CP) at time of 4 hours (T=4), spray dried plasma of the present invention (ODP) at time of 4 hours (T=4).

[0026] Fig. 7 is a panel of black and white photos of Scanning Electron Microscopy (SEM) of spray dried plasma particles of Run #3, upper left panel at 2000X, upper right panel atDocket No. 0118.014100 -7- 5000X, middle left panel at 5000X, middle right panel at 1000X, lower left panel at 5000X with measurements overlaid thereon showing sizes between .99 µm and 7.87 µm and lower right panel at 2000X. These photos illustrate the small size and amorphous character of the present invention.

[0027] Fig. 8A is a panel of black and white photos of Scanning Electron Microscopy (SEM) of spray dried plasma particles of Run #7, upper left panel at 2000X, upper right panel at 1000X, middle left panel at 5000X, middle right panel at 5000X, lower left panel at 1000X and lower right panel at 2000X.

[0028] Fig. 8B is a panel of black and white photos of Scanning Electron Microscopy (SEM) of spray dried plasma particles of Run #7 at 5000X with measurements overlaid thereon showing sizes between 1.46 µm and 6.53 µm.

[0029] Fig. 9A is a bar graph showing results of Clot Time (R) in minutes from a Thromboelastography TEG study with Rebuilt WB and Simulated Resuscitation (abbreviations: WB:FFP = whole blood (WB) rebuilt with Fresh Frozen Plasma (FFP); WB:ODP _ whole blood rebuilt with spray dried plasma of the present invention (ODP); WB+10% FFP = whole blood resuscitated with 2 units of FFP; WB+10% ODP = whole blood resuscitated with 2 units of ODP). Reference ranges are shown by dotted lines.

[0030] Fig. 9B is a bar graph showing results of Clot Rate (angle) in degrees from a Thromboelastography TEG study with Rebuilt WB and Simulated Resuscitation (abbreviations: WB:FFP = whole blood (WB) rebuilt with Fresh Frozen Plasma (FFP); WB:ODP _ whole blood rebuilt with spray dried plasma of the present invention (ODP); WB+10% FFP = whole blood resuscitated with 2 units of FFP; WB+10% ODP = whole blood resuscitated with 2 units of ODP). Reference ranges are shown by dotted lines.

[0031] Fig. 9C is a bar graph showing results of Clot Strength (MA) in Maximum Amplitude (mm) from a Thromboelastography TEG study with Rebuilt WB and Simulated Resuscitation (abbreviations: WB:FFP = whole blood (WB) rebuilt with Fresh Frozen Plasma (FFP); WB:ODP _ whole blood rebuilt with spray dried plasma of the present invention (ODP); WB+10% FFP = whole blood resuscitated with 2 units of FFP; WB+10% ODP = whole blood resuscitated with 2 units of ODP). Reference ranges are shown by dotted lines.Docket No. 0118.014100 -8-

[0032] Fig. 9D is a bar graph showing results of Lysis index (30 minutes) in LY30 in percentage(%) from a Thromboelastography TEG study with Rebuilt WB and Simulated (abbreviations: WB:FFP = whole blood (WB) rebuilt with Fresh Frozen Plasma (FFP); WB:ODP _ whole blood rebuilt with spray dried plasma of the present invention (ODP); WB+10% FFP = whole blood resuscitated with 2 units of FFP; WB+10% ODP = whole blood resuscitated with 2 units of ODP). Reference ranges are shown by dotted lines.

[0033] Fig. 9E is a bar graph showing results of Lysis index (60 minutes) in LY60 in (%) from a Thromboelastography TEG study with Rebuilt WB and Simulated Resuscitation (abbreviations: WB:FFP = whole blood (WB) rebuilt with Fresh Frozen Plasma (FFP); WB:ODP _ whole blood rebuilt with spray dried plasma of the present invention (ODP); WB+10% FFP = whole blood resuscitated with 2 units of FFP; WB+10% ODP = whole blood resuscitated with 2 units of ODP). Reference ranges are shown by dotted lines.

[0034] Fig. 10A is a bar graph showing results of Clot Time (R) in minutes from a TEG study with plasma only (abbreviations: FFP = Fresh Frozen Plasma (FFP); ODP - spray dried plasma of the present invention (ODP); WB+10% FFP = whole blood resuscitated with 2 units of FFP; WB+10% ODP = whole blood resuscitated with 2 units of ODP). Reference ranges are shown by dotted lines.

[0035] Fig. 10B is a bar graph showing results of Clot Rate (angle) in degrees from a TEG study with plasma only (abbreviations: FFP = Fresh Frozen Plasma (FFP); ODP - spray dried plasma of the present invention (ODP); WB+10% FFP = whole blood resuscitated with 2 units of FFP; WB+10% ODP = whole blood resuscitated with 2 units of ODP). Reference ranges are shown by dotted lines.

[0036] Fig. 10C is a bar graph showing results of Clot Strength in Maximum Amplitude (MA) (mm) from a Thromboelastography TEG study with plasma only (abbreviations: FFP = Fresh Frozen Plasma (FFP); ODP - spray dried plasma of the present invention (ODP); WB+10% FFP = whole blood resuscitated with 2 units of FFP; WB+10% ODP = whole blood resuscitated with 2 units of ODP). Reference ranges are shown by dotted lines.

[0037] Fig. 11 is a bar graph showing results of vWF (von Willebrand Factor): ristocetin (Rist) Activity in percent (5) from a ristocetin cofactor assay. (abbreviations: WB:FFP =Docket No. 0118.014100 -9- whole blood (WB) rebuilt with Fresh Frozen Plasma (FFP); WB:ODP _ whole blood rebuilt with spray dried plasma of the present invention (ODP); WB+10% FFP = whole blood resuscitated with 2 units of FFP; WB+10% ODP = whole blood resuscitated with 2 units of ODP). Reference ranges are shown by dotted lines.

[0038] Fig. 12 is a bar graph showing results of ADAMTS-13 (von Willebrand factor-cleaving protease) activity from ADAMTS-13 assay. (abbreviations: FFP = Fresh Frozen Plasma (FFP); ODP - Spray dried plasma of the present invention (ODP)..

[0039] Fig. 13A is two bar graphs showing results of from a platelet adhesion Bioflux study showing arterial shear at 900s-1of intensity NS (10 minutes) in Fluorescence Intensity Units (FIU) and Area NS (10 minutes) coverage in percentage (%). (abbreviations: NS=Normal Shear conditions; WB:FFP = whole blood (WB) rebuilt with Fresh Frozen Plasma (FFP); WB:ODP _ whole blood rebuilt with spray dried plasma of the present invention (ODP); WB+10% FFP = whole blood resuscitated with 2 units of FFP; WB+10% ODP = whole blood resuscitated with 2 units of ODP). Reference ranges are shown by dotted lines.

[0040] Fig. 13B includes two bar graphs showing results of from a platelet adhesion Bioflux study showing pathological shear at 4000s-1of intensity HS (10 minutes) in Fluorescence Intensity Units (FIU) and Area HS (10 minutes) coverage in percentage (%). (abbreviations: HS= High Shear conditions; WB:FFP = whole blood (WB) rebuilt with Fresh Frozen Plasma (FFP); WB:ODP - whole blood rebuilt with spray dried plasma of the present invention (ODP); WB+10% FFP = whole blood resuscitated with 2 units of FFP; WB+10% ODP = whole blood resuscitated with 2 units of ODP). Reference ranges are shown by dotted lines.

[0041] Fig. 14A includes two bar graphs showing results of from a CAT (Calibrated Automated Thrombogram) Thrombin Generation Assay showing lag time in minutes and Endogenous Thrombin Potential (ETP) (nM. Min). (abbreviations: WB:FFP = whole blood (WB) rebuilt with Fresh Frozen Plasma (FFP); WB:ODP - whole blood rebuilt with spray dried plasma of the present invention (ODP); WB+10% FFP = whole blood resuscitated with 2 units of FFP; WB+10% ODP = whole blood resuscitated with 2 units of ODP). Reference ranges are shown by dotted lines. ETP is a measurement that reflects coagulation status and if someone is prone to bleeding (reduced ETP) or clotting (elevated ETP).

[0042] Fig. 14B includes two bar graphs showing results of from a CAT (Calibrated Automated Thrombogram) Thrombin Generation Assay showing peak thrombin (nM) and time to peakDocket No. 0118.014100 -10- (minutes). (abbreviations: WB:FFP = whole blood (WB) rebuilt with Fresh Frozen Plasma (FFP); WB:ODP - whole blood rebuilt with spray dried plasma of the present invention (ODP); WB+10% FFP = whole blood resuscitated with 2 units of FFP; WB+10% ODP = whole blood resuscitated with 2 units of ODP). Reference ranges are shown by dotted lines.

[0043] Fig. 15A includes three bar graphs showing results of Flow Cytometry of Residual Cell Matter in total events, Total labeled events and CD41A (platelet) events. (abbreviations: FFP = Fresh Frozen Plasma (FFP); ODP - Spray dried plasma of the present invention (ODP)..

[0044] Fig. 15B includes three bar graphs showing results of Flow Cytometry of Residual Cell Matter in CD45 (WBCs) and CD235 (RBCs). (abbreviations: FFP = Fresh Frozen Plasma (FFP); ODP - Spray dried plasma of the present invention (ODP); WBCs – White Blood Cells; RBCs – Red Blood Cells)

[0045] Fig. 16 panels A - C are schematic illustrations depicting unfolding / refolding model of the vWF A2 domain and protelolysis by ADAMTS13. (A) Cartoon of the vWF A2 domain in its native folded state. (B) The first step of unfolding occurs from the C- terminal end of the vWF A2 domain, influenced by the presence of the vicinal disulfide bond (cysteines depicted by C). Initial unfolding occurs up to, or including, the central b4 sheet in which the scissile bond (YM) is contained. This unfolding intermediate step exposes the high-affinity ADAMTS13 spacer-binding site. (C) Once the stabilizing effect of the calcium-binding site (CBS) is overcome this results in the complete unfolding of the vWF A2 domain and the positioning of the ADAMTS13 active site for nucleophilic attack of the Y1605-M1606 scissile bond.

[0046] Fig. 17 is a bar graph showing that formulation of plasma with citric acid stabilizes during spray drying ~50% von Willebrand Factor: Ristocetin Cofactor (vWF:RCo) activity without any impact of other coagulation factors (Prothrombin (PT), Activated Partial Thromboplastin Time (aPTT), Fibrinogen (FGN), Factor V (FV), Factor VII (FVII), Factor VIII (FVIII), Factor IX (FIX), vWF antigen (vWF-Ag), % von Willebrand Factor: Ristocetin Cofactor (vWF:RCo)). This is done at time zero, time upon completion of spray drying, normalized to Control Plasma (Fresh Frozen Plasma (FFP)). CP indicates Control Plasma; SpDP indicates Spray-Dried Plasma; PreT indicates plasma formulation with SDSAS.

[0047] Fig. 18 is a bar graph showing that formulation of plasma with citric acid confersDocket No. 0118.014100 -11- stability to vWF and all other coagulation factors (Prothrombin (PT), Activated Partial Thromboplastin Time (aPTT), Fibrinogen (FGN), Factor V (FV), Factor VII (FVII), Factor VIII (FVIII), Factor IX (FIX), vWF antigen (vWF-Ag), % von Willebrand Factor: Ristocetin Cofactor (vWF:RCo)). This is done at during storage at 6 weeks at 4°C . SpDP indicates Spray-Dried Plasma; PreT indicates plasma formulation with SDSAS. CP indicates Control Plasma and FFP is Fresh Frozen Plasma.

[0048] Fig.19 is a bar graph showing that pre-treatment of plasma with citric acid confers to vWF and all other coagulation factors (Prothrombin (PT), Activated Partial Thromboplastin Time (aPTT), Fibrinogen (FGN), Factor V (FV), Factor VII (FVII), Factor VIII (FVIII), Factor IX (FIX), vWF antigen (vWF-Ag), % von Willebrand Factor: Ristocetin Cofactor (vWF:RCo)). This is done during storage after 2 weeks at 25°C . SpDP indicates Spray-Dried Plasma; PreT indicates plasma formulation with SDSAS. CP indicates Control Plasma and FFP is Fresh Frozen Plasma.

[0049] Fig. 20 is a bar graph showing that formulation of plasma with citric acid confers to coagulation factors (Prothrombin (PT), Activated Partial Thromboplastin Time (aPTT), Fibrinogen (FGN), Factor V (FV), Factor VII (FVII), Factor VIII (FVIII), Factor IX (FIX), vWF antige (vWF-Ag)). This is done during storage at 2 weeks at 37°C. SpDP indicates Spray-Dried Plasma; PreT indicates plasma formulation with SDSAS. CP indicates Control Plasma and FFP is Fresh Frozen Plasma.

[0050] Fig. 21 is a photographic image showing that formulation of plasma with citric acid vWF during SpD (spray drying). CP indicates Control Plasma; SpDP indicates Spray-Dried Plasma; PreT indicates plasma formulation with SDSAS; FFP indicates Fresh Frozen Plasma.

[0051] Fig.22A is a line graph showing pH for CP / FFP and the fed plasma under constant feeding rate of 10 mL / min, but variable aerosol gas flow rates (0, 5, 10, 15, and 20 L / min). CP indicates Control Plasma; FFP indicates Fresh Frozen Plasma; vWF indicates von Willebrand Factor.

[0052] Fig.22B is a line graph showing the results activity (%, IU / dL) of vWF:RCo activity CP / FFP and Fed Plasma under constant plasma feeding rate of 10 mL / min, but variable aerosol gas flow rates (0, 5, 10, 15, and 20 L / min). CP indicates Control Plasma; FFP indicates Fresh Frozen Plasma.Docket No. 0118.014100 -12-

[0053] Fig.23A is a line graph showing pH for CP / FFP and Fed Plasma at Aerosol gas flow rates of 10 L / min; fluid =2 mL / min, 10 L / min; fluid=4 mL / min, 10 L / min; fluid=6 mL / min, L / min; fluid= 8 mL / min, and 10 L / min; fluid= 10 mL / min. CP indicates Control Plasma; FFP indicates Fresh Frozen Plasma; vWF indicates von Willebrand Factor; vWF:RCo (vWF activity measured by vWF ristocitein assay).

[0054] Fig.23B is a line graph showing the results activity (%, IU / dl) of vWF:RCo for and Fed Plasma at Aerosol gas flow rates of 10 L / min; fluid = 2 mL / min, 10 L / min; 4 mL / min, 10 L / min; fluid= 6 mL / min, 10 L / min; fluid= 8 mL / min, and 10 L / min; fluid =10 mL / min. CP indicates Control Plasma; FFP indicates Fresh Frozen Plasma.

[0055] Fig. 24 is a bar graph showing the effect of different SDSAS-formulations on the recovery and pH during spray. The pH levels prior to and post spray were shown on the top of the bar graph. CP indicates Control Plasma; FFP indicates Fresh Frozen Plasma; vWF indicates von Willebrand Factor.

[0056] Fig.25A- C are bar graphs showing the effect of different SDSAS- formulationson therecovery and pH during spray drying. {A) citric acid, (B) lactic acid, and (C) pH. The pH levels prior to and post spray were shown on the top of the bar graph. vWF indicates von Willebrand Factor; vWF:RCo (vWF activity measured by vWF ristocitein assay).

[0057] Fig. 26 is a line graph, in color, showing the amount of C3a ng / mL and pH for batches 1542, 1541 and the respective control plasma (CP) for experiments performed in Example 16.

[0058] Fig. 27 is a line graph showing the amount of C5a ng / mL and pH for batches 1543, 1541 and the respective control plasma (CP) for experiments performed in Example 16.

[0059] Fig. 28 is a bar graph showing C5a analysis of control plasma (CP) / Never Frozen Plasma (NFP) rapidly and slowly pretreated with 400 mM glycine HCl, 400 mM glycine HCl + 1 M glycine, 148 mM citric acid, 148 mM citric acid + 1 M glycine in tube (n=7) and Control Plasma / Never Frozen Plasma (CP / NFP).

[0060] Fig. 29 is a bar graph showing C3a measurement in ng / mL of CP / FFP (control plasma / fresh frozen plasma), CP / FFP / PreT (Control plasma / Fresh Frozen Plasma / Pre- treated), and ODP / NFP / PreT (On demand plasma (Applicant’s inventive spray dried plasma)Docket No. 0118.014100 -13- never frozen, pretreated) average of n=20 ± 1 SD.

[0061] Fig. 30 is a bar graph showing C5a measurement in ng / mL of CP / FFP, CP / FFP / PreT, and ODP / NFP / PreT average of n=20 ± 1 SD. Fig. 31 is a bar graph showing ODP / NFP / PreT activity and antigen measurements normalized to CP / FFP. Fig. 32 is a bar graph showing aPTT (activated partial thromboplastin time), PT (prothrombin time), and TT (thrombin time) of ODP / NFP / PreT normalized to CP / FFP. Fig. 33 is a bar graph showing vWF Antigen (von Willebrand Factor), vWF:RCo (vWF activity measured by vWF ristocitein assay), and vWF Activity of ODP / NFP / PreT (On demand plasma (Applicant’s inventive spray dried plasma) never frozen, pretreated) normalized to CP / FFP (Control plasma / Fresh Frozen Plasma).

[0065] Fig. 34 is a bar graph showing activation marks D-Dimer, TAT, and F1+2 of ODP / NFP / PreT (On demand plasma (Applicant’s inventive spray dried plasma) never frozen, pretreated) normalized to CP / FFP (Control plasma / Fresh Frozen Plasma).

[0066] Fig. 35 is a bar graph showing chemistry analyzer results of ODP / NFP / PreT (On demand plasma (Applicant’s inventive spray dried plasma) never frozen, pretreated) for IgG, IgM, IgA, total protein, albumin, Triglycerides, Cholesterol, LDL Cholesterol, HDL Cholesterol and Calcium, normalized to CP / FFP(Control plasma / Fresh Frozen Plasma).

[0067] Fig. 36 is a bar graph showing Thrombelastography Hemostasis System (TEG) results for R-Reaction Time (min) K (min), α (angle) and MA (Maximum Amplitude (mm)) of ODP / NFP normalized to CP / FFP(Control plasma / Fresh Frozen Plasma).

[0068] Fig. 37A is a bar graph showing shows coagulation profile results and ELISA assay results for various clotting factors and compliment activation for never frozen plasma pretreated with 400 mM glycine HCl and spray dried at 49, 50, 51, and 52°C exhaust gas temperatures.

[0069] Fig. 37B is a bar graph showing shows coagulation activation and complement activation marks (D-Dimer, TAT, PF1.2, C3a and C5a) for fresh frozen plasma normalized to control plasma pretreated with 400 mM glycine HCl and spray dried at 49, 50, 51, and 52°C exhaust gas temperatures.

[0070] Fig. 38A is a bar graph showing C5a levels in ng / mL at 10 min, 1 hour, 2 hours, and 21 hours, of rehydrated plasma pretreated with Glycine HCl (GlyHCl) at 400 mM, 280 mM,Docket No. 0118.014100 -14- 140 mM and Citric Acid (CA) at 148 mM, 100 mM and 50 mM.

[0071] Fig. 38B is a bar graph showing the pH of plasma pretreated with Glycine HCl (GlyHCl) at 400 mM, 280 mM, 140 mM and Citric Acid (CA) at 148 mM, 100 mM and 50 mM. Fig. 39 is a line graph showing the pH levels of the pretreatment solutions and pre- treated plasma samples when plotted against the glycine concentration (mM). Fig. 40 is a bar graph showing C5a levels (ng / mL) for 400 mM glycine HCl, supplemented with increasing concentrations of glycine (0, 400, 600, 800, 1000, 1200, 1400 and 1600 mM) at 10 minutes and 60 minutes using rapid mixing (e.g., when larger volume of plasma is rapidly added to a relatively smaller volume of the pretreatment solution).

[0074] Fig. 41A is a line graph showing the pH of pretreatment solution and plasma (batches 1949 & 1950) pretreated with 400 mM lactic acid, and supplemented with increasing concentrations of glycine (0, 400, 600, 800, 1000, 1200, 1400 and 1600 mM).

[0075] Fig. 41B is a bar graph showing the C5a levels in ng / mL of plasma (batches 1949 & 1950) pretreated with 400 mM lactic acid and supplemented with increasing concentrations of glycine wherein the pretreated plasma had concentrations of 0, 20, 30, 40, 50, 60, 70, and 80 mM glycine.

[0076] Fig. 42A is a schematic showing a perspective view of the spray drying disposable device, which includes the liquid plasma bag, spray drying head and spray drying chamber, wherein the disposable has alignment elements which allow it to align with a spray drying apparatus and a finishing apparatus.

[0077] Fig. 42B is a model representation of the three-dimensional flow geometry of the flow model of the disposable during operation. This model is used to create the computer flow models described herein.

[0078] Fig. 43A is a schematic showing a perspective view of the spray drying head of the spray drying disposable device shown in Fig. 42A.

[0079] Fig. 43B is a schematic showing an exploding view of the spray dry nozzle assembly and the spray drying head of spray drying disposable device shown in Fig. 42A.

[0080] Fig. 43C is a schematic showing a perspective view of the spray dry nozzle assembly from the spray drying head of spray drying disposable device.

[0081] Fig. 43D is a schematic showing a perspective view of the spray dry nozzle assembly of Fig. 43C but with the aerosol reservoir housing being transparent to show the inner structuresDocket No. 0118.014100 -15- of the assembly.

[0082] Fig. 43E is a schematic showing a perspective view of the spray dry nozzle assembly of Fig. 43C but with the aerosol reservoir housing, the nozzle cap and nozzle cap insert being removed and showing the manifold and the cannula.

[0083] Fig. 43F is a schematic showing a front view of an embodiment of the angled edge cannula that is part of the spray dry nozzle assembly. Fig. 43G is a schematic showing a perspective, top view of nozzle cap insert that guides the cannula and aerosolized air.

[0085] Fig. 43H is a schematic showing a perspective, bottom view of nozzle cap insert having the cannula inserted therein.

[0086] Fig. 43I is a schematic showing a top view of the nozzle cap. Fig. 43Ia is schematic showing a bottom view nozzle cap of Fig. 43I with the cannular residing within the cap opening.

[0088] Fig. 43Ib schematic showing three possible vortex generator flow patterns that could be used with the nozzle cap insert of Fig. 43I.

[0089] Fig. 43Ic is a schematic showing a cross-sectional view of nozzle cap insert of Fig. 43H residing within nozzle cap of Fig. 43I.

[0090] Fig. 43J is a schematic showing a perspective, bottom view of the plenum of the spray drying head.

[0091] Fig. 43K is a schematic showing a partial front view of the spray dryer showing, in part, the drying gas deflector.

[0092] Fig. 43Ka is a model representation showing the modeled drying gas flow within the plenum chamber using a constant velocity magnitude surface of 15 m / s.

[0093] Fig. 43L is a schematic showing a perspective, top view of the baffle plate of the spray drying head.

[0094] Fig. 43La is a schematic showing a cut out section of the rib design of the baffle plate shown in Fig. 43L and a cut out section of another variation of the baffle plate rib design. Fig. 43La also shows a cross section of one of the ribs.

[0095] Fig. 43M is a schematic showing a perspective, bottom view of the baffle plate of the spray drying head.

[0096] Fig. 43Ma is a model representation showing uniform jet penetration and drying gasDocket No. 0118.014100 -16- distribution at constant velocity of 25 m / s and hence results in a circumferentially uniform introduction of the drying gas around the spray plume.

[0097] Fig. 43N is a schematic showing a schematic showing the droplet plume formation, aerosol gas flow and drying gas flow that promotes rapid mixing in the disposable of the present invention.

[0098] Fig. 43Na is a model representation displaying the gas velocity magnitude contours within the disposable’s center plane indicating the drying jet penetration into the drying chamber and the effect of baffle plate flow channels and interaction with the high velocity spray plume which act to pull the drying gas jets radially inward to assist in the desired rapid mixing of the droplets and gas flows.

[0099] Fig. 43O is a model representation showing the gas pressure flow (psig) (top) and tangential velocity flow (m / s) (bottom) of the vortex generated in the nozzle insert and cap assembly.

[0100] Fig. 43P is a model representation showing gas velocity magnitude flow (m / s) in parts of the vortex generator.

[0101] Fig. 43Q is a schematic showing the transformation of a liquid droplet to a dried particle using the disposable of the present invention.

[0102] Fig. 43R is a line graph showing the droplet wet bulb temperature inoC and drying gas temperature inoC of water droplets dried to particles having 0% Relative Humidity (RH), 10% RH and 20% RH. This particular graph illustrates the concept but is not specific to plasma.

[0103] Fig. 43S is a line graph showing the evaporation mass transfer of droplet temperatures over time for all the averaged droplet trajectories average with three simulated drying gas inlet temperatures of 80oC, 100oC, and 114oC in the model. Note the evaporation process cools the droplet to keep the delicate liquid protein below 30oC.

[0104] Fig 43Sa is a line graph temperature inoC and time (seconds) of a plasma droplet as it becomes a particle in the model. Once the evaporation is complete, the protein encased in a solid particle is more tolerant of elevated temperature as it equilibrates with the dryer outlet temperature. In this case, evaporation occurs in less than fractions of a second (e.g., 0.01 to 0.05 seconds).

[0105] Fig. 43T is a model representation showing the path of droplets having a size of 5Docket No. 0118.014100 -17- microns, 15 microns and 25 microns, during evaporation, and the figure shows that smaller droplet size allows for more rapid evaporation mass transfer in a shorter path, enabling a physically smaller drying chamber.

[0106] Fig. 44A is a schematic showing an exploding view of the drying chamber of the spray drying disposable shown in Fig. 42A. Fig. 44B is a schematic showing a front view of a separator or spacer that can be used in the drying chamber of the spray drying disposable. Fig. 45A is a schematic showing a front view of the spray drying apparatus with the door closed.

[0109] Fig. 45B is a drawing of a partial front view of the spray drying apparatus without the door to reveal the drying chamber housing having alignment elements that allow for alignment with the spray drying disposable device.

[0110] Fig. 45C is a schematic showing a front view of the spray drying apparatus with handle of the door being engaged and the door being opened.

[0111] Fig. 46A shows the alignment elements aligning the spray drying disposable device and the spray drying apparatus.

[0112] Fig. 46B is a schematic showing a partial front view of the spray drying apparatus without the door and with the spray drying disposable device installed and deflector engaged.

[0113] Fig. 46C is a schematic showing the architecture of dryer 200. Abbreviations included are: AM – Air Manifold, B - Transfer Bag, CN – Connector, DPT - Differential Pressure Transducer, F – Filter, FS - Flow Sensor, H – Heater, MFC - Mass Flow Controller, OS – Sensor, P - Peristalic Pump, PP - Pneumatic Piston, PR - Pressure Regulator, PRV - Pressure Relief Valve / Rupture Disk, PT - Pressure Transducer, PV - Valve (Arrow Down Is Fail Closed, Arrow Up Is, Fail Open), S – Scale, SS – Solenoid, TC – Thermocouple, TS - Thermocouple Sensor, and TT - Temperature Transducer.

[0114] Fig. 46D is a flow chart showing the steps of the leak detection method employing the spray dryer and spray drying disposable.

[0115] Fig. 46E is a flow chart showing the steps of the pressure detection method employing the spray dryer and spray drying disposable.

[0116] Fig. 46F is a flow chart showing the steps of the method for detecting integrity of filters and disposable interface employing the spray dryer and spray drying disposable.Docket No. 0118.014100 -18-

[0117] Fig. 46G is a line graph showing the slope employing the method for detecting integrity of filters and showing that the filters are intact by measuring Pressure (psig), elapsed time (min) and pressure rate of change (psi / min)..

[0118] Fig. 46H is a line graph showing the slope employing the method for detecting integrity of filters and showing that the capture filter has failed intact by measuring Pressure (psig), elapsed time (min) and pressure rate of change (psi / min).

[0119] Fig. 46I is a line graph showing the slope employing the method for detecting integrity of filters and showing that the baffle filter has failed intact by measuring Pressure (psig), elapsed time (min) and pressure rate of change (psi / min).

[0120] Fig. 47A is a schematic showing a front view of the finishing apparatus in the loading position and without the spray drying disposable attached.

[0121] Fig. 47B is a schematic showing a front view of the finishing apparatus of Fig. 47A but without the front cover with the shuttle in the lowered position.

[0122] Fig. 47C is a schematic showing a front view of the finishing apparatus of Fig. 47B with the shuttle in the raised position and without the disposable attached.

[0123] Fig. 47D is a schematic showing a front view of another embodiment of the finishing apparatus in the raised position and without the spray drying disposable attached.

[0124] Fig. 47E is a schematic showing a front view of the finishing apparatus of Fig. 47D but without the front cover with the shuttle in the raised position.

[0125] Fig. 47F is a schematic showing a front view of the finishing apparatus of Fig. 47D but without the front cover with the shuttle in the lowered position.

[0126] Fig. 47G is a schematic showing a front view of the finishing apparatus of Fig. 47D but without the front cover with the shuttle in the raised and inverted position.

[0127] Fig. 47H is a schematic showing a front view of the finishing apparatus of Fig. 47D but without the front cover with the shuttle in the lowered and inverted position.

[0128] Fig. 47I is a schematic showing a close-up perspective view of a portion of the rail system of the finishing apparatus of Fig. 47D

[0129] Fig. 48A is a schematic showing a front view of the finishing apparatus in the loading position with the spray drying head of the disposable aligned thereto.

[0130] Fig. 48B is a schematic showing a front view of the finishing apparatus in the raised position with the disposable aligned thereto.Docket No. 0118.014100 -19-

[0131] Fig. 48C is a schematic showing a front view of the finishing apparatus in the raised position with a portion of the disposable attached thereto, after the first seal and separate step is completed and the frame is rotated into position.

[0132] Fig. 48D is a schematic showing a front view of another embodiment of the finishing apparatus in the raised position and with the spray drying disposable attached. Fig. 48E is a schematic showing a front view of the finishing apparatus shown in Fig. 48D in the lowered position and with the spray drying disposable attached. Fig. 48F is a schematic showing a front view of the finishing apparatus shown in Fig. 48D in the raised and inverted position and with the spray drying disposable attached.

[0135] Fig. 48G is a schematic showing a close up, top view of the receiver of the finishing apparatus shown in Fig. 48D.

[0136] Fig. 48H is a schematic showing a close up, perspective view of the receiver of the finishing apparatus shown in Fig. 48D.

[0137] Fig. 48I is a schematic showing a close up, perspective view of the tensioner of the finishing apparatus shown in Fig. 48D.

[0138] Fig. 48J is a schematic showing a close up, perspective view of the separator having a roller that engages the tensioner of the finishing apparatus shown in Fig. 48D.

[0139] Fig. 48K is a schematic showing a close up, perspective view of the impactor, separator and sealer of the finishing apparatus shown in Fig. 48D.

[0140] Fig. 48L is a schematic showing a side view of the impactor, separator and sealer of the finishing apparatus shown in Fig. 48D.

[0141] Fig. 49A is a schematic showing a front view of the spray dry plasma unit obtained from the spray drying disposable device after processed by the finishing apparatus.

[0142] Fig. 49B is a schematic showing the architecture of finisher 400’. Abbreviations included are: AM -Air Manifold, CN – Connector, CV - Check Valve, EM - Electric Motor, FS - Flow Sensor, FR - Flow Restrictor, PP - Pneumatic Piston, PR - Pressure Regulator, PT - Pressure Transducer, PV - Valve Arrow (Down Is Fail Closed, Arrow Up Is Fail Open), SC - Speed Controller, SR – Sensor, and VG - Vacuum Generator

[0143] Fig. 50 is a bar graph comparing vWF% ratio of reconstituted plasma (to a never dried control aliquot) that was dried using disposable devices with composite nozzle assembly without a chamfer, with a chamfer and a benchmark stainless-steel nozzle.Docket No. 0118.014100 -20-

[0144] Fig. 51A is a flow chart showing the steps of the pretreatment methodology.

[0145] Fig. 51B is a flow chart showing the steps of the spray drying methodology employing the spray dryer and spray drying disposable. Fig. 51C is a flow chart showing the steps of the finishing methodology employing the finisher and spray drying disposable to create a dried plasma unit. Fig. 51D is a flow chart showing the steps of the storage methodology once the spray dried unit is made. Fig. 52 is a schematic showing the geometry of the cannula to show sheer on vWF protein when exiting the cannula. The curved arrow shows the aerosol gas vortex direction within the annulus. In order to show the amount sheer impact on the liquid plasma at the exit area of the cannula, the figure shows the cannula edges at 15, 90, 45 (with chamfer and sharp edges) and 60 degrees and shows how the sheering contact of the cannula is reduced by the angled edge. DETAILED DESCRIPTION OF THE INVENTION

[0149] A description of preferred embodiments of the invention follows. The present invention relates to spray dried plasma with reduced or no pathogens. Spray dried plasma is obtained using the spray drying devices and disposables, as further described herein. The spray dried plasma of the present invention by the nature of its characteristics and the process, such as an aseptic process, by which it is made allows for a dried plasma with reduced or no pathogens.

[0151] The present invention relates to three aspects of pathogen reduction that results in a plasma product with reduced or no pathogens.

[0152] The first aspect relates to a spray drying disposable and spray drying machine that eliminates or minimizes the introduction of pathogens into the spray drying process and is referred to as the aseptic or anti-contamination process. Relatedly, the spray drying process also aids in reducing or eliminating any pathogens that become attached to the baffle filter located in the plenum, the surface of the plenum interior or capture filter by subjecting that baffle filter, the plenum interior, and / or capture filter to high temperature drying gas at the beginning and during the drying cycle.Docket No. 0118.014100 -21-

[0153] The second aspect of pathogen reduction relates to the spray drying process and parameters that reduces pathogens that may be present in the donor plasma, and / or inhibit pathogen growth and / or pathogen proliferation introduced by the donor plasma.

[0154] The third aspect results in reduced pathogens occurring during storage of the dried plasma wherein the manufacturing and storage conditions are such that pathogens are reduced, and / or pathogen growth and / or pathogen proliferation are inhibited. The “reduction” in pathogens refers to the reduction in the number of pathogens and / or the reduction in the proliferation (e.g., its ability to split into two daughter cells) of a pathogen.

[0155] The spray drying plasma of the present invention is for use in transfusion medicine and otherwise, is an alternative to, liquid plasma, frozen plasma or freeze-dried plasma. In an embodiment, the dried plasma product of the present invention is a standardized dose of dried plasma, referred to herein as a “plasma unit,” “dried plasma unit,” “plasma product,” or “unit”, intended for rapid rehydration at the point of care. In an embodiment, the standardized single unit of dried plasma originates from a single unit of donor plasma. The dried plasma with reduced pathogens of the present invention allows for transfusion of plasma into recipients with reduced risks.

[0156] The dried plasma unit of the present invention is, in part, a source of plasma proteins for patients who are deficient or have defective plasma proteins for which there are no specific replacement factors available. The dried plasma unit of the present invention can also be used for those who are in need of blood volume and / or coagulation factor treatment such as external or internal bleeding or disease state. The plasma of the present invention reduces risk in these vulnerable patients and does so where conventional plasma (such as fresh frozen plasma (FFP) or plasma frozen within 24 hours (PF24)) is unavailable or impractical for use such as pre-hospital transfusions, rural / austere hospitals and military applications.

[0157] Additional indications for use of the dried plasma unit of the present invention include the management of preoperative or bleeding patients who require replacement of multiple coagulation factors (e.g., liver disease, Disseminated Intravascular Coagulation (DIC)) as well as patients undergoing massive transfusion who have clinically significant coagulation deficiencies.Docket No. 0118.014100 -22-

[0158] The dried plasma unit of the present invention is suitable for use for massive transfusion as it does not expose the recipient to risks of pathogen contaminated plasma which introduces the risk of sepsis, as compared to other blood products now available or expected to be available for transfusion.

[0159] To better understand the reduced pathogen characteristics of the dried plasma and how the dried plasma is made, a description of the spray drying system, disposable and methods are later described.

[0160] The first aspect: a spray drying disposable, machine and system that provides an aseptic anti-contamination process.

[0161] The spray drying system is described in detail later herein. The following points out the specific elements of the system that contribute to this anticontamination process that may be aseptic.

[0162] The spray drying system including the disposable device, drying machine and the finisher, and the spray drying process of the present invention provides a robust system that reduces, minimizes or eliminates the introduction of pathogen contamination from the environment. The overall concept of having a sterile, single use disposable having a drying chamber within which the drying of the plasma occurs and which becomes the actual plasma unit product reduces, minimizes or eliminates pathogen contamination. The design of the disposable itself is one that creates a barrier between the plasma and the dryer / outside air. The design of the disposable also prevents the plasma itself from coming in contact with any dryer parts. The disposable is sterilized, and the spray drying system ensures aseptic processing, meaning it does not introduce contaminants into the plasma. The aseptic contamination resistant overall design ensures reduced or no pathogen introduction to the spray dried product of the present invention when producing dried plasma units at a blood center or other such environments.

[0163] Several attributes and features of the spray drying system and disposable contribute to aseptic processing and prevents pathogen introduction into the system during spray drying. One or more, or any combination, of the following features contribute to the reduced or no contamination of pathogens from outside into the spray drying system of the present invention:Docket No. 0118.014100 -23- a. One or more filters (e.g., redundant HEPA / ULPA filters) in communication with the drying gas source (e.g., Millipore Series 30000.2 micron filter CTGB71TP3 from Millipore Sigma of Danvers MA USA); b. One or more filters (e.g., redundant HEPA / ULPA filters) (e.g., 0.2 micron filters) in communication with the pressurized / aerosolized gas source (e.g., Millipore Series 30000.2 micron filter CTGB71TP3 from Millipore Sigma of Danvers MA USA); c. Use of Clean Dry Air (CDA) for a source of drying gas and aerosolized / pressurized gas provided into the disposable; d. Covers at the connection ports for the disposable (e.g. drying gas inlet port 22 and gas outlet port 30) and the dryer (e.g., drying gas source inlet deflector 242 and gas exhaust port 208); e. Baffle filter 94 in spray drying head 2; f. One or more filters in communication with humid air after exiting through exhaust port 208 (e.g., consumable exhaust line to outside air - Saint Gobain 1000 SLPM 0.1 micron filter LI2F010NTCTC from Saint Gobain Performance Plastics of Worcester MA USA and Enclosure exhaust line to outside air - Saint Gobain 50 SLPM 0.1 micron filter LI2F010NCTC from Saint Gobain Performance Plastics of Worcester MA USA); g. Lower filter / capture filter 36 (e.g., 0.2 micron filter) which is in plasma drying chamber 28 of disposable 100 and captures dried plasma while filtering humid air; h. Sterilization of single use components: Disposable 2, Plasma Pretreatment Container (PPC), Sterile Water for Injection (SWFI); i. Sterilized Connections Devices (SCD) are used with liquid plasma before drying and with dried plasma during rehydration (e.g., SCD between disposable and liquid plasma unit); j. Constant positive air flow is maintained for cleanliness and to reduce or minimize ingestion of pathogens from the environment from the CDA system to the air inlet to the disposable; and k. Heated drying gas at an inlet temperature in the range of about 100-130° C (e.g., 114Docket No. 0118.014100 -24- oC) aids in reducing or eliminating pathogen growth or proliferation of any pathogens that may have entered the plenum, become attached to the plenum filter or capture filter.

[0164] These features reduce, minimize or eliminate the introduction of pathogens into the spray drying system. These features are described in detail herein as parts of the spray drying system, dryer, disposable or finisher.

[0165] Testing demonstrates the spray drying system of the present invention maintains product integrity and reduces, minimizes or eliminates pathogen contamination into the spray dried plasma unit. Test results were reviewed and accepted by regulatory authorities for use in human clinical safety trials. In particular, process simulation studies demonstrated the bioburden in rehydrated spray dried plasma unit at time zero (t=0) and at 4 hours (t=4) after rehydration is comparable or about the same to the starting plasma material that was not spray dried. The starting plasma from a donor is not a sterile composition. The bioburden, the presence and quantity of bacteria, in the reconstituted previously dried plasma sample and paired control was evaluated. See Example 35. The data show that the spray drying process does not introduce pathogens into the system or into spray dried plasma unit.

[0166] The heated gas from the drying cycle (an inlet temperature in the range of about 90-130° C (e.g., 114oC)) aids in reducing or eliminating pathogen growth or proliferation of any pathogens that may have entered the plenum, baffle filter, or capture filter during mounting of the disposable 100 to the dryer 200 by subjecting the plenum’s interior surfaces and the baffle filter to the heated gas before and throughout the drying cycle.

[0167] Similarly, Bacterial Filtration Efficiency (BFE) testing was performed on baffle filter 94 in spray drying head 2, and the lower capture filter 36 in plasma drying chamber 28, which are the filters that interface between the disposable and the dryer at the inlet and outlet. The testing demonstrates that these filters maintain their integrity during and after exposure to the cumulative stresses of temperature and pressure of the spray drying process. All 20 baffle filters 94 and lower capture filter 36 passed the ASTM F2101 test for Bacterial Filtration Efficiency. See Example 8.

[0168] In particular, these test results from increased challenge BFE demonstrate that baffle filters 94 and lower capture filter 36 maintain their bacterial retentive or captureDocket No. 0118.014100 -25- properties throughout the process of spray drying a unit and reduce or eliminate the introduction of contamination from outside environment. Data from this process simulation study showed no bacterial growth in the spray dried plasma unit. See Example 35.

[0169] The dried plasma of the present invention ensures dried plasma product protection when producing spray dried plasma. The system includes a number of system level features, individually or in combination, that eliminate, impede, or prevent the introduction of contaminants.

[0170] Studies of the spray dried plasma anti-contamination system of the present invention have demonstrated that the spray drying system of the present invention does not introduce contamination, and is aseptic even when plasma is spray dried in an unclassified environment (e.g., Bio Safety Level 2 (BSL2) or less).

[0171] The present invention is an integrated, self-contained, anti-contamination system. This permits it to be used in unclassified environments.

[0172] The Centers for Disease Control and Prevention (CDC) and the National Institutes of Health (NIH) established criteria for levels of containment. The term “containment” is used in the field to describe safe methods for managing biological materials in the laboratory environment where they are being handled or maintained. The purpose of containment is to reduce or eliminate exposure of laboratory workers, other persons, and the outside environment to potentially hazardous or detrimental materials. These criteria consist of combinations of practices and techniques, safety equipment, and laboratory facilities. Each combination is specifically appropriate for the operations performed, biological materials to be used, and the function or activity to be performed.

[0173] Primary concerns in many blood environments are largely related to the safety of the site personnel. These include accidental needlesticks or cuts or mucous membrane exposures, or ingestion of infectious materials. The spray dried plasma process and system of the present invention does not use needles, or open bladed cutting tools. Even though organisms routinely manipulated in blood plasma processing are not known to be transmissible by the aerosol route, procedures with aerosol or high splash potential can increase the risk of personnel exposure. The spray drying plasma system of the present invention contains the aerosolized plasma in filter-protected disposable 2 and, during spray drying, is protected by the housing of the spray dryer, which is a robust container withDocket No. 0118.014100 -26- redundant leak prevention controls. High splash potential is the same or less than routine blood plasma manufacture by present methods.

[0174] The second aspect: pathogen reduction that may be present in the donor plasma, as it relates to the spray drying process and parameters

[0175] The second aspect of pathogen reduction relates to the spray drying process and parameters that reduce pathogens that may be present in the donor plasma, and / or inhibit pathogen growth and / or pathogen proliferation introduced by the donor plasma.

[0176] In the instance when the donor plasma has a pathogen within it, the spray drying system of the present invention reduces pathogens and / or inhibits pathogen growth and proliferation. Pathogens may be introduced while obtaining the donor plasma. Obtaining the donor plasma involves a needle stick through skin and / or hair on the skin that can have pathogens on it. As the needle pushes through the skin, it can collect pathogens during insertion and introduce those pathogens into the donor plasma. Additionally, the donor may have pathogens in its native plasma as well.

[0177] The spray drying system of the present invention subjects plasma droplets and / or dried plasma articles to heat energy and / or air-liquid interfacial stress such that pathogens are reduced and / or their growth and proliferation are inhibited. In an embodiment, the spray dried plasma of the present invention has been exposed to time and temperature during drying that is within the range which diminishes pathogens while preserving plasma protein function. In another embodiment, the spray dried plasma of the present invention has been exposed to spray drying conditions (e.g., aerosolization, rapid mixing, evaporation forces) that impose air-liquid interfacial stress which diminishes pathogens while maintaining plasma protein function (e.g., maintaining the functionality of the plasma proteins without resulting in protein aggregates or formation of neoantigens).

[0178] In the spray dry system of the present invention, as a droplet becomes a particle, it is subjected to heat and air-liquid interfacial stress. There are generally three stages as the droplet becomes a dried plasma particle. Fig. 43Q is a schematic that shows the liquid plasma droplet undergoing the drying process. In the middle portion of Fig. 43Q, a schematic of the droplet becoming a particle is shown during rapid mixing. The three stages are: the liquid droplet (labeled “constant rate” drying), the partially dried droplet (the middle schematic labelled “falling rate” (diffusion limited)), and the dried particle. During rapidDocket No. 0118.014100 -27- mixing while the droplet becomes a particle, each stage is subjected to heat and / or interfacial stress for a period of time sufficient to reduce pathogens and / or their proliferation.

[0179] The plasma droplet and the partially dried droplet are subjected the temperature of the drying gas which is between about 90oC to about 130oC (e.g., between about 100oC to about 114oC). During the initial phase of drying the plasma droplet absorbs the energy of the hot drying gas thereby evaporating the bulk of the moisture from the droplet. The resulting temperature of the plasma droplet is between about 20oC and about 65oC. See Fig 43Sa. Without being held to any particular theory, the temperature of the droplet goes from about room temperature to about 65oC during evaporation. The drying rate is constant and as the liquid particle evaporates and loses moisture, the moisture transfers from the liquid plasma droplet to the drying gas, and the heat from the drying gas transfers to the plasma droplet making it into a dried particle. The plasma droplet enters the drying chamber essentially at room temperature and the temperature stays constant the majority of the evaporation period. See Fig. 43S. Once most all of the moisture leaves the particle, the temperature of the particle increases to equilibrate with the dryer chamber exit temperature of 65oC. In an embodiment, the evaporation process also applies interfacial stress on the droplet as it turns into a dried particle. Therefore, the dried particle, during the spray drying, is subjected to or maintained at a temperature between about 55°C and about 85°C for a time period between about 5 minutes to about 40 minutes. The particles dried first are subjected to this temperature for longer than those dried at the end of the spray drying process. The average temperature to which the plasma particle is exposed is about 65°C for a minimum of about 5 minutes.

[0180] During evaporation, the droplet is maintained at a lower temperature thereby protecting heat sensitive proteins such as vWF. See Fig. 43S. But even these lower temperatures are sufficient for reducing pathogens and / or their proliferation.

[0181] According to the following tables from Food Safety and Inspection Service (FSIS) bacteria is reduced by about 6.5-Log10 Lethality or by about 7-Log10 Lethality. (FSIS Cooking Guideline for Meat and Poultry Products (Revised Appendix A) GUIDELINE ID FSIS-GD-2021-0014 ISSUE DATE December 2021 https: / / www.fsis.usda.gov / guidelines / 2021-0014) For example, the charts below are theGenerally, salmonella is one of the moreDocket No. 0118.014100 -28- difficult bacteria to reduce to safe levels and is used as benchmark for determining the safety. Often, if one is able to reduce salmonella by at least about 6.5 (e.g., or about 7) on a log10 scale, then other harmful bacteria are also considered to be reduced (excluding harmful spores which are reduced by sterilization). Id.

[0182] Both time and temperature parameters are met to achieve the Log reduction target. Table 1 . Table 1 FSIS, Appendix A for Beef Minimum Internal TemperatureMinimum processing time in minutes or seconds afterDocket No. 0118.014100 -29-

[0183] As can be seen in Table 1, the pasteurization temperature ranges from about 65oC for 85-95 seconds will achieve 6.5 log10 (106.5) reduction and about 7 log10107.0) reduction, respectively, of salmonella bacteria. The plasma of the present invention is subjected to this temperature for a minimum of 5 minutes and much of the plasma particles are subjected to this temperature for up to 40 minutes, more than enough time to reduce bacteria by about or less 7 log10 (107) (e.g., 6 log10 (106), 5 log10 (105), 4 log10 (104), 3log10 (103), or 2 log10(102)).

[0184] As can also be seen from the table above temperatures above 71oC instantly reduce salmonella to 6.5 and 7 log lethality. Portions of the interior of the disposable of the present invention including the plenum, the baffle filter and the capture filter are subjected to temperatures for time points that are lethal to salmonella, for example, temperatures in the range of about 100-130° C (e.g., 114oC) for 1 -10 minutes before plasma flow begins.

[0185] The third aspect: pathogen reduction occurs during storage, as it relates to dried plasma storage conditions

[0186] The present invention embraces a method for increasing the storage time of spray dried plasma. During storage of spray dried plasma of the present invention, spray dried plasma can be stored with reduced pathogen proliferation, and without substantial loss of plasma protein function. Since there is such a low residual moisture in the dried plasma of the present invention, pathogens do not proliferate and in fact, in an embodiment, are inhibited.

[0187] As described herein, the spray drying system of the present invention is designed to produce a low residual moisture dried plasma powder. The low residual moisture reduces bacteria and / or inhibits pathogen proliferation and / or pathogen growth and does so during storage at various temperatures and environments.

[0188] The low residual moisture plasma unit is obtained by using the disposable and system as described herein and dried with the parameters as described herein. In particular, the low residual moisture of the dried plasma is attributable to a number of aspects of the spray drying system that promote rapid mixing of the drying gas and the plasma droplets during spray drying. Rapid mixing allows for rapid mass transfer and evaporation which results in a low residual moisture dried plasma product.Docket No. 0118.014100 -30-

[0189] For example, as described herein, the design of liquid nozzle cap insert 80 and nozzle cap 76 allow the pressurized aerosol gas to flow though annulus 81 in a vortex pattern to maximize aerosolization and promote rapid mixing of the aerosolized plasma droplets with the drying gas, as further described herein.

[0190] Also as described herein, small droplet size which is enhanced by the angled edge of the cannula, promotes rapid mixing, faster evaporation, reduced drying time and, in part, provides a low residual moisture plasma product. See Fig. 43T which shows that the larger the droplet size the longer it takes for the droplet to evaporate with higher drying gas temperatures. Droplet size is also impacted by the ratio of the pressurized gas flow rate to the liquid feed rate and nozzle design.

[0191] Additional factors involved in the rapid mass transfer and evaporative drying stage of the plasma droplet also include the temperature of the plasma and the drying gas, the surface area of the droplet, the humidity in the drying gas and the air circulation within the plasma drying chamber. As described herein, when initially exiting the nozzle assembly, the temperature of the drying gas is between about 90oC to about 130oC (e.g., between about 100oC to about 114oC).and the temperature of the plasma droplet is between about 20oC and about 65oC in the plume as shown in Fig 2Sa. The heat flows from a point of higher temperature to that of a lower temperature, and in this case the drying gas heat flows to the plasma droplet. With respect to the surface area, the droplet is spherical thereby maximizing its surface area and the droplet size is very small so the mass and heat transfer can happen quickly. The relative humidity in the drying gas is very low and therefore the low humidity of the surrounding drying gas promotes evaporation of the plasma particle. Finally, as described in more detail herein, the drying gas is emitted using several drying gas jets in an angled and downward direction into the plasma drying chamber and into the plume of atomized droplets to initiate rapid mixing of the drying air and the atomized droplets, which increases the rate of evaporation of the liquid droplets.

[0192] Yet another factor, as described herein, that contributes to a low residual moisture product is the starting liquid droplet size produced by the nozzle assembly. This impacts the residence time in the drying chamber needed to complete evaporation and a small droplet size allows evaporation to happen more quickly. The smaller the liquid droplet, the larger the ratio of evaporation surface area to droplet mass and the faster the mass transfer rate from theDocket No. 0118.014100 -31- droplet. As seen from Fig. 43T, the drying chamber can be shortened to that where the majority of the evaporation occurs while still allowing a dried particle to achieve less than 2.5% residual moisture before being deposited on lower filter 36.

[0193] The percent residual moisture in the plasma dried with the disposable and dryer of invention is very low, e.g., below 2.5%, 2%, 1%, e.g., about 1.46% residual as measured by Karl Fischer moisture sensor, Model No. C30S Compact KF Coulometer (Mettler Toledo Billerica Massachusetts USA). This is a very low moisture level which is due to effective and efficient evaporation of the plasma droplet occurring in the upper portions of drying chamber 28 and the process conditions. In this aspect, powder moisture level is in equilibrium with chamber outlet air stream relative humidity.

[0194] Dried plasma with low moisture reduces pathogens or their growth / proliferation, as promotes protein stability during storage.

[0195] The reduction of pathogens or their growth / proliferation is achieved in part by its low residual moisture. The low residual moisture enables storage and stability of spray dried plasma without pathogen growth / proliferation at various temperatures and for longer periods of time.

[0196] “Stability” or “Shelf-life Stability” or “Unit Stability” refers to stored spray dried when reconstituted, behaves comparably to plasma that has been dried but not stored. Stability includes comparing amounts of various plasma proteins and their function (e.g., vWF, Factor V, Factor VIII, etc.) and / or plasma characteristics (e.g., pH, particle size, etc.) present in reconstituted spray dried plasma before (time =0 seconds) and after storage (e.g., up to 48 months at refrigeration, up to 12 months at room temperature). Stability involves comparing the respective value of one or more of these proteins / characteristics before and after storage to determine similarity to one another or to determine if the value is within the clinical range. In an embodiment, the values a plasma protein / characteristic after storage are within about 25% or less (25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, and 1%), as compared that before storage. In an embodiment, the values of plasma proteins / plasma characteristics after storage are within the clinical range for that plasma protein. Such plasma characteristics include pH, Osmolality (mOsm / kg), particulate size, particulate quantity and particulate distribution. Plasma proteins and their function include e.g., Total Protein (mg / ml), Activated Partial Thromboplastin Time (aPTT)(s), ProthrombinDocket No. 0118.014100 -32- Time (s), International Normalized Ratio (INR (s)), Thrombin Time (s), Factor V (%), Factor VII (%), Factor VIII (%), Factor IX (%), Factor X (%), Factor XI (%), Factor XIII Activity (%), Factor XIII Antigen (%), Protein C Activity (%), Protein S Activity (%), Fibrinogen (mg / dL), Plasminogen (%),Plasmin InhibitorPI (%), Antithrombin III (%), von Willebrand Factor Antigen (% or IU / dL), von Willebrand Factor Ristocetin Cofactor (% or IU / dL), C5a (ng / mL), Prothrombin Fragment F 1+2 (pmol / L), Thrombin-Antithrombin Complex (TAT) (µg / L), Protein (mg / mL) and the like. Measuring such plasma characteristics and plasma proteins are known in the art. In another embodiment, the acceptable or clinical range for von Willebrand Factor Ristocetin Cofactor (VWF:RCo) is between about 50 and about 200 IU / dL, or about 50 and about 200 IU / dL, and von Willebrand Factor Antigen (VWF:Ag) value is between about 50 and 200 IU / dL.

[0197] In an embodiment, the clinical reference range for determining shelf life or unit stability is as follows:

[0198] Table 2Docket No. 0118.014100 -33- Factor XI Activity FXI Act (%) (50 - 200)

[0199] The data shows that the dried plasma is stable after being stored as a dried plasma for a time period up to about 24 months at refrigerated temperatures (2-8°C), 6 months at a Room Temperature (20-25°C) and 3 months elevated Room Temperature (30°C). Example 5 describes experiments performed that verified that the spray dried plasma unit remained stable during storage.

[0200] The spray dried plasma of the present invention, after release testing is complete and the product is determined safe for use, as described herein, can be stored at elevated temperatures, room temperature, refrigerated temperatures, freezing temperatures or a combination thereof. In certain aspects, spray dried plasma is refrigerated to enable storage for longer periods of time.Docket No. 0118.014100 -34-

[0201] The dried plasma of the present invention can be stored between a temperature of -80ºC and 45ºC. Room temperature is considered between about 20ºC and 30ºC. In an aspect, refrigerated temperatures are between about 1°C and about 6°C. The terms “cooling,” “cold temperature,” “temperature below room temperature,” and “temperature below ambient temperature,” interchangeably refer to any temperature between 1ºC and 20ºC. In any of the embodiments of the invention described herein, the temperature is selected from the group of temperatures consisting of 45ºC, 44ºC, 43ºC, 42ºC, 41ºC, 40ºC, 39ºC, 38ºC, 37ºC, 36ºC, 35ºC, 34ºC, 33ºC, 31ºC, 30ºC, 29ºC, 28ºC, 27ºC, 26ºC, 25ºC, 24ºC, 23ºC, 22ºC, 21ºC, 20ºC, 19ºC, 18ºC, 17ºC, 16ºC, 15ºC, 14ºC, 13ºC, 12ºC, 11ºC, 10ºC, 9ºC, 8ºC, 7ºC, 6ºC, 5ºC, 4ºC, 3ºC, 2ºC, 1ºC, 0ºC , -1ºC, -5ºC, -10ºC, -20ºC, -30ºC, -40ºC, -50ºC, - 60ºC, -70ºC, and -80ºC. In some embodiments, the spray dried plasma of the present invention is stored at a temperature of less than about 15°C, preferably less than 10°C, and more preferably less than 5°C. In some other embodiments, the spray dried plasma of the present invention is stored at room temperature. In other embodiments, the spray dried plasma of the present invention is stored at warm temperatures, e.g., above 23ºC. In further embodiments, the spray dried plasma of the present invention can be stored in freezing temperatures (e.g., 0ºC to - 80ºC). It is noted that although the dried plasma product can be stored in freezing temperatures, it does not need to be maintained in freezing temperatures to be stable for transfusion. On one hand, storage conditions can be carefully monitored, and on the other, storage conditions can be an array of temperatures depending on circumstances on the ground. In yet another embodiment, the dried plasma can be stored at a combination of temperatures.

[0202] As used in all of the aspects and embodiments of the invention herein, the term “period of time” or “time period” refers to a duration of time during which spray dried plasma of the present invention is stored at any given temperature. The term “period of time” can range from seconds to minutes to hours to days to weeks to months to years. In preferred embodiments, the term “period of time” refers a number of hours including about 3 to about 120 hours, e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97,Docket No. 0118.014100 -35- 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, and 120 hours. In certain embodiments the period of time for which spray dried plasma of the present invention can be stored include about 1 and about 30 days (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, and 30). In certain embodiments the period of time for which spray dried plasma of the present invention can be stored include about 1 and about 48 months (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48 months).

[0203] In an embodiment, spray dried plasma of the present invention can be stored at room temperature for at least about 1 hour to about 7.5 months. In an embodiment, spray dried plasma of the present invention can be stored at refrigerated temperature for about 1 hour to about 48 months. In an aspect, the spray dried plasma of the present invention can be refrigerated on any day or days during storage.

[0204] In various other embodiments, the spray dried plasma of the present invention is stable and preserves the functionality of plasma proteins and / or plasma characteristics during the time period, when compared to corresponding clinical reference ranges or that prior to storage, or compared to thawed previously frozen plasma over the same period of time following spray drying. In an embodiment, spray dried plasma samples stored between a temperature of –80ºC and 45ºC are suitable for transfusion after extended periods of storage time, in an embodiment, for at least about 2 days, at least about 7 days, at least about 2 weeks, at least about 3 weeks, at least about 1 month, at least about 2 months, at least about 3 months, at least about 6 months, at least about 9 months, at least about 1 year, or at least about 1 ½ years, about 2 years, about 3 years, about 4 years, or longer.

[0205] The invention, in certain aspects, provides a novel method of storing a spray dried plasma without pathogen growth or proliferation in which the steps include obtaining plasma from a donor and drying the plasma using the methods and systems described herein, and storing the dried plasma for a period of time without a substantial loss or subclinical loss of plasma protein amounts and / or plasma characteristics and without pathogen growth or proliferation.

[0206] Definitions of pathogens / inhibition / proliferationDocket No. 0118.014100 -36-

[0207] The term “pathogen” as used herein, refers to one or more microorganisms or the like that cause infection as described in (Dodd, R. Y. New Engl. J. Med. 327:419-421 (1992); Soland, E. M., et al. J. Am. Med. Assoc. 274:1368-1373 (1995) and Schreiber, G. B., et al. New Engl. J. Med. 334:1685-1690 (1996)). Exemplary pathogens include, but are not limited to a virus, bacteria, parasite, protozoa, or fungus. Examples of viruses include, but are not limited to Herpes simplex virus, HIV, hepatitis, hepatitis A, hepatitis B, hepatitis C, Respiratory syncytial virus, blue tongue virus, and bovine diarrhea virus. Virus also includes Cytomegalovirus, Epstein-Barr virus, Herpes Simplex type I and II viruses, and other viruses that circulate freely in the blood, as well as cell-associated viruses. Fungus includes, but is not limited to e.g., Aspergillus. And typical parasites include, but are not limited to, for example: Ameoba, Plasmodiunm, Leishmania, Mycosus profundus, Trypanosoma, Spirochete, and Arbovius.

[0208] Bacteria commonly associated with blood products including plasma and whose growth or proliferation is inhibited by the spray drying system of the present invention include, but are not limited to Aspergillus, Bacillus sp, Bacteroides eggerthii, Candida albicans, Citrobacter sp, Clostridium perfringens, Corynebacterium sp, Diphtheroid, Enterobacter aerogenes, Enterobacter amnigenus, Enterobacter cloacae, Enterococcus avium, Enterococcus faecalis, Escherichia coli, Fusobacterium spp., Granulicatella adiacens, Heliobacter pylori, Klebsiella sp, (K. pneumonia, K. oxytoca), Lactobacillus sp, Listeria sp, Micrococcus sp, Peptostreptococcus, Proteus vulgaris, Pseudomonas sp, Pseudomys oralis, Propionibacterium sp, Salmonella sp, Serratia sp, Staplhylococcus sp (Coagulase-negative Staphylococcus, Staphylococcus epidermidis, Staphylococcus aureus), Streptococcus sp, (S. gallolyticus, S. bovis, S. pyogenes, S. viridans), Serratia marcescens, and Yersinia enterocolitica.

[0209] The term “detrimental effect” as used herein, can refer to a detrimental effect the viability or usability of plasma) that is caused by one or more pathogens. The term “detrimental effect” as used herein, can also refer to the detrimental effect upon the patient (e.g., the consequences of the infection itself) that is caused by one or more pathogens such as sepsis.

[0210] The term “bacterial contamination” as used herein, refers to contamination by any pathogen or by non-pathogenic bacteria in the dried plasma or the reconstituted previouslyDocket No. 0118.014100 -37- dried plasma. “Inhibiting bacterial proliferation” refers to reducing and / or inhibiting the growth of bacteria in the plasma.

[0211] The invention, in other aspects, provides a novel method of inhibition of bacterial proliferation in plasma by obtaining a plasma from a donor and spray drying the plasma using the spray drying systems described herein.

[0212] Bacterial contamination of plasma is a concern because it may cause sepsis in recipients receiving it. Bacterial contamination can result from the donor having pathogens in their plasma or from non-sterile techniques in obtaining blood from the donor, or in poor handling of the plasma after donation. Despite good sterile techniques in obtaining donated blood or plasma, bacteria can still persist in the donor plasma. For example, even though a technician uses an antibacterial agent to clean the skin at the site of donation, bacteria can be embedded within the layers of the skin, i.e., intradermally. So, upon penetration of the skin with a needle, bacterial contamination of the plasma donation can occur. As a result, bacterial testing at the blood center before the plasma is released is performed to reduce the risk of sepsis.

[0213] Encompassed in the method of the present invention is testing for bacterial proliferation in reconstituted previously spray dried plasma at one or more time points to determine that bacterial proliferation is in fact inhibited before being transfused into the recipient. Bacterial testing can occur at a single time point (e.g., at the point of care) and the results can be compared to a standard to determine if bacterial proliferation has occurred in the reconstituted previously spray dried plasma to be transfused. Additionally, bacterial testing of the reconstituted previously spray dried plasma can occur at more than one time point to assess if the particular sample has exhibited inhibition of bacterial proliferation. An increase in bacterial proliferation or the presence of bacterial proliferation indicates that the reconstituted previously spray dried plasma are contaminated and cannot be used for transfusion. The absence of bacterial proliferation indicates that the reconstituted previously spray dried plasma can be used for transfusion. Using reconstituted previously spray dried plasma of the present invention results in plasma that is suitable for transfusion.

[0214] A number of tests exist to determine the presence of bacterial contamination in reconstituted previously spray dried plasma. Bacteria can be tested by the presence of a polypeptide or protein that is common to bacteria and not found in plasma, by cultureDocket No. 0118.014100 -38- techniques, Gram staining, scanning techniques, the presence of nucleic acid that is conserved in bacteria, scans, and the like.

[0215] A commonly used test in determining bacterial contamination of reconstituted previously spray dried plasma is the Pan Genera Detection (PDG® test) (Verax Biomedical, Incorporated, Worcester Massachusetts). The PGD® test can detect an array of bacteria in blood components. This broad detection is based on the existence of shared, or conserved, antigens that are common to the cell walls of the two broad classes of bacteria: lipoteichoic Acids on Gram-positive bacteria and lipopolysaccharides on Gram-negative bacteria. The test targets these conserved Gram-positive and Gram-negative antigens to test biological samples for a broad range of bacterial contaminants by using binding agents to directly bind to these targets. Although the level or presence of the specific bacteria is not determined by this test, the test does determine the presence of a number of bacteria in the reconstituted previously spray dried plasma.

[0216] Culture methods can be employed to determine the presence or absence of bacterial contamination and / or bacterial proliferation. One commercially available test is referred to as the BacT / ALERT® test (bioMérieux, Inc., Durham, NC). Bacterial detection is based on the evolution of carbon dioxide by proliferating bacteria. A carbon-dioxide- sensitive liquid emulsion sensor at the bottom of the culture bottle changes color and is detected through alteration of light reflected on the sensor. BacT / ALERT® test detects the presence of a number of bacteria, fungi, and yeasts.

[0217] Another method for bacterial detection involves measuring the oxygen content in a reconstituted previously spray dried plasma sample. An example is the Pall eBDS test (Pall Corporation, Port Washington, NY). The approach to detection measures the oxygen content of air within the sample pouch as a surrogate marker for bacteria. An oxygen analyzer is used to measure the percent of oxygen in the headspace gas of the pouch or bag having the reconstituted previously spray dried plasma. If bacteria are present in the reconstituted previously spray dried plasma sample collected, an increasing amount of oxygen is consumed through the metabolic activity and proliferation of the bacteria in the sample during incubation, resulting in a measurable decrease in oxygen content of the plasma as well as the air within the sample pouch.Docket No. 0118.014100 -39-

[0218] A more conventional method for determining the presence of bacterial proliferation is a reconstituted previously spray dried plasma is Gram staining. Gram staining allows one to differentiate bacterial species into classes (Gram-negative or Gram- positive) in an effort to begin to identify the microorganism. The test detects peptidoglycan, a glycan in the cell wall of the bacteria.

[0219] A sample from the reconstituted previously spray dried plasma can be obtained and cultured to determine if any bacteria are present. The growth media is inoculated or plated with the sample and under controlled conditions suitable for bacterial growth. Bacteria can be grown and identified.

[0220] Other methods known in the art or developed in the future can be used to determine bacterial proliferation in the reconstituted previously spray dried plasma of the present invention.

[0221] The methods of the present invention involve reducing bacterial proliferation in spray -dried plasma using the system described herein. The bacterial proliferation is reduced, as compared to a standard or to another assessment taken at a different time point. The methods described herein reduce bacterial proliferation by at least about 5% (e.g., by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%). In an embodiment, the methods of the present invention completely inhibit bacterial proliferation, as compared to that at in the donor plasma just before spray drying.

[0222] Subjecting A Low pH Pretreatment Solution To Plasma To Heat Conditions Of Spray Drying Further Reduces Pathogens and / or Their Proliferation

[0223] In an embodiment, the pretreatment of plasma with the pretreatment solution (e.g., spray dry stable acidic substance (SDSAS), and one or more amino acids) described herein allows for the plasma proteins to be protected during the drying process. Additionally, the pretreatment of plasma allows for the pH of the reconstituted plasma to be near normal pH, also as further described herein. However, in another aspect, the pretreatment of the plasma with a solution having a low pH allows for more effective reduction of pathogens while being subjected to the heat of the spray drying process.

[0224] In particular, the pH of the pretreatment solution is in a range between about 2.0 and about 4.0 (e.g., 2.0, 2.2, 2.4, 2.6, 2.8, 3.0, 3.2, 3.4, 3.6, 3.8, 4.0). The low pHDocket No. 0118.014100 -40- pretreatment solution assists in reducing pathogens and / other their proliferation. While not bound by a particular theory, it is believed that pH affects the structure of all macromolecules that compose bacteria and pathogens. Lipids are hydrolyzed by low pH levels and they undergo chemical reactions and fall apart into constituent molecules, which may be toxic to the pathogen. The membrane is made of lipids; if it is disrupted, the cell ceases to exist.

[0225] When subjected to the low pH along with the heat of the spray drying process, a reduction in pathogens and / or their proliferation occurs. When the pretreatment solution combined with plasma results in a pretreated plasma having a pH of between about 6.0 and about 6.6 (e.g., 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6). In an embodiment, it is believed that subjecting a lower pH pretreated or formulated plasma to the heat of spray drying, the heat is more effective in reducing pathogens and / or their proliferation.

[0226] A description of the plasma, the spray drying device and the spray drying disposable follows.

[0227] Detailed Description of the Plasma A description of preferred embodiments of the invention follows. The present invention relates to spray dried plasma. Spray dried plasma is obtained using the spray drying devices and disposables, as further described herein. The spray dried plasma of the present invention has one or more or any combination of the following characteristics: 1) is largely amorphous, as defined as lacking definite form, e.g., with small particles (<20 microns) of proteins and lipids with high specific surface area and small diffusion length, the largely amorphous dried particles differ from lyophilized plasma which typically forms a consolidated, porous “cake”; 2) when reconstituted has very few or no crystals (e.g., cholesterol) made by the drying process; 3) when reconstituted, because the system of the present invention does not create particulates, the number of larger particulates is reduced, and in particular, mean size of particulates is reduced when measuring with a Coulter Multisizer 4 by theDocket No. 0118.014100 -41- electrical sensing zone method for particulates having a size between about 2 µm and 60 µm, as compared to particulates in the donor plasma; 4) has low residual moisture; 5) reconstitutes rapidly in under four minutes, as measured from first touch by a user to completed reconstitution with no visible clumps; 6) is stable when stored under refrigeration, at room temperature, at elevated temperatures or a combination of those and allows for storage for longer periods of time, as compared to liquid plasma; 7) is stable for 4 hours or greater (e.g., up to 26 hours) after reconstitution and product stays aseptic; 8) when reconstituted, exhibits protein functionality of the most plasma proteins including fragile proteins, such as von Willebrand’s factor, and other active proteins in the donor plasma; 9) when reconstituted with Sterile Water for Injection (SWFI), reconstituted plasma is at a near normal plasma pH (not abnormally high (alkalotic)) which is preferable for use in transfusion, and is ready for transfusion without treatment with or storage in CO2 or other post-drying pH adjustment; and 10) when reconstituted, does not activate complements (C5a, C3a) as compared to apheresed plasma (Complement activation is involved in inflammation).

[0230] The spray drying plasma of the present invention is for use in transfusion and otherwise, and is an alternative to liquid plasma, frozen plasma or freeze-dried plasma. In an embodiment, the dried plasma product of the present invention is a standardized dose of dried plasma, referred to herein as a “plasma unit,” “dried plasma unit,” “plasma product,” or “unit”, intended for rapid rehydration at the point of care. The dried plasma of the present invention allows regional blood centers and others to prepare a single donor unit of plasma or pooled plasma in a dried form that can be stored, shipped, and transfused more easily than frozen plasma.

[0231] The dried plasma unit of the present invention is, in part, a source of plasma proteins for patients who are deficient or have defective plasma proteins for which there are no specific replacement factors available. The dried plasma unit of the present invention can also be used for those who are in need of blood volume and / or coagulation factor treatmentDocket No. 0118.014100 -42- such as external or internal bleeding or disease state. The dried plasma unit of the present invention provides a plasma product where conventional plasma (such as fresh frozen plasma (FFP) or plasma frozen within 24 hours (PF24)) is unavailable or impractical for use such as pre-hospital transfusions, rural / austere hospitals and military applications.

[0232] Additional indications for use of the dried plasma unit of the present invention include the management of preoperative or bleeding patients who require replacement of multiple coagulation factors (e.g., liver disease, Disseminated Intravascular Coagulation (DIC)) as well as patients undergoing transfusion who have clinically significant coagulation deficiencies. The plasma of the present invention, when reconstituted, allows for clot formation.

[0233] The dried plasma unit of the present invention is suitable for use for massive transfusion as it can induce clot formation and does not expose the recipient to risks of high pH, cholesterol or other crystals, particulates, or complement activation, as compared to other blood products now available or expected to be available for transfusion.

[0234] To better understand the characteristics of the plasma, a description of the spray drying system, disposable and methods are described herein.

[0235] Plasma characteristics: Dried plasma is amorphous and has few or no crystals

[0237] The present invention relates to a reconstituted previously spray dried plasma which is made by a process which creates very few or no crystals (e.g., cholesterol crystals). “Reconstituted plasma” as used herein refers to reconstituted plasma that was spray dried using the methods, disposable and spray drying system described herein. In a particular embodiment, the reconstituted plasma, in an embodiment, has no or few crystals visible with a phase contrast microscope set at 100X, 400X or both. Cholesterol crystals are cholesterols that form a type of lattice structure, often an elongated rod-like crystal morphology or thin quadrilateral plate morphology. In another embodiment, the reconstituted plasma has less crystals (e.g., cholesterol crystals), as compared to reconstituted plasma that has previously been lyophilized. Crystals in reconstituted lyophilized plasma have been observed in a range between about 1 and about 3 µm. The formation of dried plasma with little or no crystals isDocket No. 0118.014100 -43- the result, in part, of the spray drying process, the design of the nozzle of the disposable, the rapid mixture of the liquid plasma droplet and the drying gas during spray drying, the droplet size, and the parameters at which the plasma is dried. These aspects are further described herein.

[0238] While not adhering to any particular theory, it is believed that the spray drying plasma using disposable 100 and dryer 200, as described herein, allows for rapid mass transfer (e.g. less than 1 second) to convert the liquid plasma to an amorphous dried material. As a result, crystalline solid production for many molecules including cholesterol is suppressed as the minimum required time and mobility needed for the molecules to orient themselves in a low energy crystalline state is not present during the manufacture of the spray dried plasma of the present invention. In contrast, lyophilization has a long-time window (e.g., hours or days) allowing time for crystal formation including cholesterol crystals, to form and stabilize. See Examples 1 and 2.

[0239] Examples 1 and 2 describe the observance of no crystal formation in the rehydrated spray dried plasma of the present invention but present in lyophilized plasma. Crystals can be identified with microscopic examination, and namely through compound microscopy, phase contrast microscopy, or electron microscopy at various magnifications (e.g. 40X, 80X, 100X, 200X, 400X, 800X, 1000X, and 2000X). In an embodiment and in the Examples, crystals are seen in all of the lyophilized plasma samples at 100X and 400X.

[0240] In an aspect, cholesterol crystals are undesirable in blood plasma. The process of freeze drying / lyophilization creates such crystals in the rehydrated freeze-dried plasma product. Lyophilized human blood plasma has crystal formation when reconstituted and so the recipient receives cholesterol crystals when transfused with reconstituted plasma that was previously freeze-dried. Cholesterol crystals are undesirable in the human body because they induce inflammation in atherosclerosis which could lead to heart attacks and strokes. Furthermore, cholesterol cannot be easily degraded by mammalian cells and so they have a lasting effect. In particular, cholesterol crystals are known to have the following undesirable characteristics: 1) Excess cholesterol is excreted in the bile and transported in specialized transporter molecules (HDL, LDL) and modified by cells, but not degraded. In certain instances, cholesterol crystals are formed in the bile and are involved in the formation of stones;Docket No. 0118.014100 -44- 2) Cholesterol crystals are formed during the formation of the atherosclerotic plaque. Excessive crystal growth is implicated in plaque rupture and severity of symptoms is related to cholesterol crystal content; 3) Cholesterol crystals are spread through the body as a result of plaque rupture. Cholesterol crystals released from the atherosclerotic plaque can spread to almost any organ. Effects can vary between relatively benign skin manifestations to multi organ failure and such symptoms are exhibited within a few hours to 5 months after exposure; 4) Cholesterol crystals derived from human atheroma, a degeneration of the walls of the arteries caused by accumulated fatty deposits and scar tissue, can still be found well after vessel repair. Furthermore, it has been shown that large crystals damage the endothelial cells, whereas small crystals seem to have fewer damaging effects; and 5) Cholesterol crystals activate the immune system and induce inflammation processes.

[0241] The effects of cholesterol crystals can be immediate or late, and can vary from to patient. If cholesterol crystal related effects (mild or serious) are seen in the patient they could be interpreted as part of the patient’s disease or due to dietary behavior, and not as caused by the treatment with reconstituted lyophilized plasma.

[0242] Based on the undesirable effects of cholesterol crystals, the dried plasma of the present invention is preferable for transfusion, as compared to lyophilized plasma. Reconstituted plasma of the present invention does not contain crystals (e.g., cholesterol crystals) visible at 100X and 400X and is largely amorphous. “Amorphous” refers to a noncrystalline solid in which the atoms and molecules are not organized in a lattice pattern and lacking long-range order. While freeze drying promotes cholesterol crystal formation by super cooling, the spray drying process of the present invention minimizes or avoids crystal formation by promoting rapid mixing and rapid mass transfer in which a plasma droplet is dried into an amorphous plasma particle in less than a second. Note that crystals originating from a donor may be present in the finished product. The spray drying process does not create additional crystals in the plasma. If certain crystals are present in the donor plasma, then they may also be present in the reconstituted previously spray dried plasma. For example, it is possible that uric acid crystals or calcium phosphate crystals present in the donor plasma may also be found in the reconstituted plasma unit. As such, the reconstitutedDocket No. 0118.014100 -45- previously spray dried plasma of the present invention is largely amorphous or substantially amorphous (e.g., at least about 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90% amorphous). As a result, crystalline solid production for many molecules including cholesterol is suppressed as the conditions needed for crystalline formation is not present during the spray drying of the present invention. In contrast lyophilization process promotes crystal formation including cholesterol crystal formation by providing time and conditions for crystalline formation. See Figs. 1-4. Fig. 7 provides photos that illustrate the small size and amorphous character of the present invention.

[0243] As a consequence, the dried plasma of the present invention is largely, substantially, or wholly amorphous in structure with few or no crystals (e.g., cholesterol crystals). Since the spray drying process described herein does not produce crystals, the reconstituted plasma of the present invention has few or no crystals, except for those crystals present in the corresponding donor plasma.

[0244] The Mean Size of Particulates In Reconstituted Plasma Of The Present Invention Is Reduced

[0245] Particulates of various kinds such as protein aggregates, platelets and microparticles naturally occur in blood plasma including that of humans. When plasma containing excessive or large particulates is transfused to a recipient, these protein aggregates or particulates may react with the recipient’s body to cause inflammation or other immunological reactions, even though those particulates may have been benign to the donor.

[0246] As described herein, the term “particulates” differs from the term “particles.” "Particulates” refer to protein aggregates, platelets and microparticles in reconstituted plasma. “Particles” refer to the individual discreet components that make up the dried plasma.

[0247] Particulate studies have been done for the dried plasma of the present invention and it has been determined that there is no increase in particulate size or distribution in the rehydrated units of the dried plasma of the present invention compared to its paired control. The particulate size analysis data supports that the rehydrated plasma of the present invention does not exhibit protein aggregates or other particulates after storage and rehydration.

[0248] In other words, when reconstituted, the rehydrated plasma exhibits a reduced mean size of particulates when measuring with a Coulter Multisizer 4 by the electricalDocket No. 0118.014100 -46- sensing zone method for particulates having a size between about 2 µm and 60 µm, as compared to particulates in the donor plasma. Unlike the spray dried plasma of the present invention, lyophilized plasma has not been reported to reduce particulate size or quantity in the resulting, reconstituted plasma for transfusion.

[0249] The reconstituted plasma of the present invention has a reduced mean size of particulates, as compared to that found in the originating donor plasma. Without being committed to a particular theory, it is believed that the spray drying process of the present invention reduces the larger particulates in donor plasma into smaller sizes but the proteins, even delicate proteins, remain functional and integral. See Example 3.

[0250] A particulate analysis characterization study was conducted to evaluate spray drying manufacturing effects, in-use stability and product shelf-life effects on protein aggregation and other particulates. See Example 3. The particulate size analysis data supports that rehydrated dried plasma units of the present invention do not exhibit protein aggregates or other particulates due to manufacture and storage up to 7.5 months at room temperature, up to 24 months refrigeration or after a combined storage temperature of 1 year refrigerated + 6 months room temperature when compared to (1) its paired control and (2) after t=4 hours post rehydration .

[0251] Particulates, as measured by the electrical sensing zone method in the range of 2 to 60 microns, are not increased in mean size when compared to a paired control that has not been spray dried. As measured, particulates in reconstituted plasma ranged in size between about 2 µm and about 3.5 µm, as compared to donor plasma which range in size between about 2 µm and about 7 µm. See Figs. 5 (7.5 months room temp) and Fig. 6 (12 months refrigeration). In an embodiment, the largest particulate size of reconstituted previously spray dried plasma is about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10% 5%, 4%, 3%, 2%, 1%, as compared to largest particulate size found in donor plasma. See Tables 11 and 12 of Example 3. As described further herein, plasma proteins participate in and allow for clot formation including fragile proteins such as von Willebrand factor (vWF). In all instances, the mean size of particles in the 2 µm to 60 µm range was reduced in the reconstituted spray dried plasma of the present invention as compared to the paired, not sprayed dried plasma control. Additionally, the mean size of particulates is reduced, as compared to donor plasma when measured in the range of in the about 2 µm to about 60 µm range. The percentage ofDocket No. 0118.014100 -47- particulates having a mean size ranging between about 2 µm to about 60 µm range in reconstituted plasma of the present invention is reduced as compared to donor plasma. In an embodiment, the particulates having a mean size ranging about 2 µm to about 60 µm range found in reconstituted previously spray dried plasma is about 60%, 50%, 40%, 30%, 25%, 20%, 15%, 10% 5%, 4%, 3%, 2%, 1% less as compared to the number found donor plasma.

[0252] Particulate analysis for determining particulate size, particulate quantity and distribution can be obtained using a Beckman Coulter Multisizer 4 (electrical sensing zone method) e.g., performed in the range of 2 to 60 µm.

[0253] The spray dried plasma of the present invention has low residual moisture, e.g., less than 2.5% residual moisture

[0254] As described herein, the spray drying system of the present invention is designed to produce a low residual moisture dried plasma powder. The low residual moisture 1) reduces any potential clogging of the lower filter, 2) facilitates faster reconstitution to create a plasma unit suitable for transfusion, and 3) contributes to stability during storage, including storage time, storage temperature and for storage in different environments.

[0255] The low residual moisture plasma unit is obtained by using the disposable described herein and dried with the parameters described herein. In particular, the low residual moisture of the dried plasma is attributable to a number of aspects of the spray drying system that promote rapid mixing of the drying gas and the plasma droplets during spray drying. Rapid mixing allows for rapid mass transfer / evaporation which results in a low residual moisture plasma product and contributes to the very compact dimensions of the spray drying disposable of the present invention, especially the plasma drying chamber.

[0256] For example, as described herein, the design of liquid nozzle cap insert 80 and nozzle cap 76 allow the pressurized aerosol gas to flow though annulus 81 in a vortex pattern to maximize aerosolization and promote rapid mixing of the aerosolized plasma droplets with the drying gas, as further described herein.

[0257] Also as described herein, small droplet size which is enhanced by the angled edge of the cannula, promotes rapid mixing, faster evaporation, reduced drying time and, in part, provides a low residual moisture plasma product. See Fig. 43T which shows that the larger the droplet size the longer it takes for the droplet to evaporate with higher drying gasDocket No. 0118.014100 -48- temperatures. Droplet size is also impacted by the ratio of the pressurized gas flow rate to the liquid feed rate and nozzle design.

[0258] Additional factors involved in the rapid mass transfer and evaporative drying stage of the plasma droplet also include the temperature of the plasma and the drying gas, the surface area of the droplet, the humidity in the drying gas and the air circulation within the plasma drying chamber. As described herein, when initially exiting the nozzle assembly, the temperature of the drying gas is between about 90oC to about 130oC (e.g., between about 100oC to about 114oC).and the temperature of the plasma droplet is between about 20oC and about 65oC in the plume as shown in Fig 43S and 43Sa. The simulated droplet temperatures in Fig. 43S were obtained by averaging over all droplet trajectories during the period of constant rate evaporation and indicate that over the range of drying gas inlet temperatures during the early near nozzle evaporation the proteins do not experience those high drying gas inlet temps. In fact, plasma proteins are protected by the energy loss due to evaporation when the proteins are most vulnerable, which is in the liquid state. Fig.43Sa focusses on a single droplet to aerosol particle pathway. Fig. 43Sa shows a longer time-line and indicates the highest temperature the protein will experience is in the solid state, after evaporation and while in the capture filter. The heat flows from a point of higher temperature to that of a lower temperature, and in this case the drying gas heat flows to the plasma droplet. With respect to the surface area, the droplet is spherical thereby maximizing its surface area and the droplet size is very small so the mass and heat transfer can happen quickly. The relative humidity in the drying gas is very low and therefore the low humidity of the surrounding drying gas promotes evaporation of the plasma particle. Finally, as described in more detail herein, the drying gas is emitted using several drying gas jets in an angled and downward direction into the plasma drying chamber and into the plume of atomized droplets to initiate rapid mixing of the drying air and the atomized droplets, which increases the rate of evaporation of the liquid droplets.

[0259] Yet another factor, as described herein, that contributes to a low residual moisture product is the starting liquid droplet size produced by the nozzle assembly. This impacts the residence time in the drying chamber needed to complete evaporation and a small droplet size allows evaporation to happen more quickly. The smaller the liquid droplet, the larger the ratio of evaporation surface area to droplet mass and the faster the mass transfer rate from theDocket No. 0118.014100 -49- droplet. As seen from Fig. 43T, the drying chamber can be shortened to that where the majority of the evaporation occurs while still allowing a dried particle to achieve less than 2.5% residual moisture before being deposited on lower filter 36.

[0260] The percent residual moisture in the plasma dried with the disposable and dryer of present invention is very low, e.g., below about 2.5%, 2%, 1%, preferably about 1.46% residual moisture, as measured by Karl Fischer moisture sensor, Model No. C30S Compact KF Coulometer (Mettler Toledo Billerica Massachusetts USA). This is a very low moisture level which is due to effective and efficient evaporation of the plasma droplet occurring in the upper portions of drying chamber 28 and the process conditions. In this aspect, powder moisture level is in equilibrium with chamber outlet air stream relative humidity.

[0261] Dried plasma with low moisture improves reconstitution time and protein stability during storage.

[0262] The dried plasma of the present invention reconstitutes rapidly e.g., under four minutes

[0263] The principal reason human plasma is not available as widely as needed is that, before the present invention, blood plasma, in general, could only be stored frozen for long periods or as a liquid for very short periods. Accordingly, if a large amount of plasma is needed (e.g., such as in a mass casualty event), it may not be available in such quantities, or if plasma is needed in an emergency, it may not be available in time since it has to be thawed which can take 30-45 minutes or more.

[0264] The spray dried plasma of the present invention can be stored for longer periods of time, as described herein, and can be rapidly reconstituted in under four minutes from first contact by the user to completed reconstitution. The phrase “reconstituted” or “rehydrated” are used interchangeably herein and refers to mixing the dried plasma of the present invention with a reconstitution solution (e.g., Sterile Water for Injection (SWFI)) to obtain a liquid plasma suitable for transfusion.

[0265] Rapid reconstitution is attributable, in part, to the amorphous nature, low residual moisture, small particle size, increased surface area of the particles of the dried plasma and the amount of air present in the plasma unit bag (e.g., between about 3 and about 15 mL volume of air). Low residual moisture levels of about 2.5% or less of the dried plasma assist in faster reconstitution because the low residual moisture does not contribute to clumping ofDocket No. 0118.014100 -50- the plasma particles into larger agglomerates with less surface area. In the case of a dried plasma (e.g., solute), the interactions between the dried plasma particles and the solvent (e.g., SWFI) are strong such that the individual solute particles separate from each other and, surrounded by solvent molecules, enter the solution to form liquid plasma. The drier the particles, the faster they will enter the solution. Similarly, the smaller the dried plasma particle and more exposed surface area of the particles exist, the easier it is for the solution (e.g., SWFI) to surround the particle and for the particle to enter into solution. In an embodiment, particle size of the dried plasma is between about 1 and about 7 microns. Particle analysis for determining particle size of the dried plasma, particle quantity and distribution can be obtained using a Scanning Electron Microscopy (SEM). See Example 7.

[0266] The methods of the present invention further include reconstituting the dried using a physiologically compatible reconstitution solution. The reconstitution solution can be mixed with the dried plasma using one of spike ports 42A or 42 B of the dried plasma unit 60. The reconstitution solution is connected to the dried plasma bag with one of the spike ports and the water is manually pushed into the dried plasma bag and mixed. Further, the spray dried formulated plasma of the present invention may be reconstituted with sterile water (e.g., sterile water for injection (SWFI) or similar) or clean, non-sterile water and, if desired, filtered after reconstitution. In normal circumstances the clinician / healthcare provider / end user rehydrates a unit with a unit of the supplied system SWFI. In a preferred embodiment, sterile water for injection is used for the reconstitution solution. In the case in which SWFI is not available, in an aspect, distilled water may be used. In other embodiments, for example when a pretreatment step is not performed, the reconstitution solution further includes amino acid (e.g., glycine), or a buffered solution (e.g., acid such as hydrochloric acid or citric acid). The plasma unit having the dried plasma and the bag weighs between about 45 grams and about 55 grams (e.g., about 50 grams). The dried plasma weighs between about 15 grams and about 25 grams (e.g., about 17 grams). The plasma unit bag weighs about 28 grams to about 38 grams (e.g., 33 grams). The amount of reconstitution solution used to rehydrate the dried plasma is in a range between about 175 mL and about 230 mL (e.g., 200 mL and about 225 mL). When the dried plasma is rehydrated with the reconstitution solution to thereby create a plasma unit ready for transfusion, the rehydrated plasma unit has a final volume of between about 180 mL andDocket No. 0118.014100 -51- about 236 mL (e.g., about 205 mL and about 231 mL) or about 197 grams and about 247 (e.g., about 217 grams and about 242 grams). In a further embodiment the reconstitution solution is pre-measured to 208 mL or other volume to allow for potential evaporative loss in storage.

[0267] It is contemplated that the reconstituted spray dried formulated plasma of the present invention has a pH of about 6.5 to about 7.8, or about 6.9 to about 7.5 (e.g., 6.5, 6.6, 6.7, 6.8.6.9.7.0.7.1.7.2.7.3.7.4.7.5, 7.6, 7.8). The amounts of the pretreatment components, further described herein, can be adjusted to achieve the designed pH of the reconstituted spray dried formulated plasma.

[0268] Additionally, once the reconstitution solution is added, a step of the method includes shaking, rocking and / or agitating the reconstitution plasma unit to ensure the mixing and uniformity of the reconstitution solution and dried plasma. The reconstituted plasma is ready for transfusion into a recipient. Examples of recipients include human, primate, mammal, animal and the like. In an embodiment, this method of reconstituting a dried plasma unit can be done in under 7 minutes (e.g., 7, 6, 5, 4, 3, and 2 minutes) and preferably under about 3.5 minutes, measured at the initial timepoint, time 0, (the time of the first contact of the reconstitution solution with the dried plasma unit) until the final time point which is the time that the plasma is reconstituted with the reconstitution solution having no visible clumps that can be seen with the naked eye (total reconstitution time).

[0269] This method was carried out by various individuals, some trained as a medic and others untrained and timed. Rehydration was achieved on average in less than about 4 minutes.

[0270] In a preferred embodiment the SWFI is provided in a pre-measured container supplied as part of a kit with the other components of the system. However, if needed, SWFI from any source can be substituted for the pre-packed SWFI of the kit so long as the amount of SWFI used in the rehydration is the same as that specified. In an embodiment, the single use container is 200 mL of sterile water for injection (SWFI) is packaged in a 250 mL bag within an overwrap pouch. The kit can further include a rehydration Tubing Set e.g., a commercially approved standard sterile fluid transfer set (e.g. FENWAL TM Plasma Transfer Sets with Two Spikes 4C2243 or equivalent) to transfer the SWFI into the unit. Additionally, a transfusion tubing set can also be included to transfuse rehydrated plasmaDocket No. 0118.014100 -52- into a patient. An example of a commercially approved standard sterile transfusion set / administration set is FENWAL TM Blood Component Recipient Set with Standard Blood Filter and Luer Adapter 4C2160 or equivalent.

[0271] The spray dried plasma of the present invention is stable when stored under refrigeration, at room temperature or at elevated temperatures and allows for storage for longer periods of time

[0272] The spray dried plasma stability is achieved in part by its low residual moisture. low residual moisture in the spray dried plasma enables for storage at various temperatures and for longer periods of time.

[0273] “Stability” or “Shelf-life Stability” or “Unit Stability” refers to stored spray dried when reconstituted, behaves comparably to plasma that has been dried but not stored. Stability includes comparing amounts of various plasma proteins and their function (e.g., vWF, Factor V, Factor VIII, etc.) and / or plasma characteristics (e.g., pH, particle size, etc.) present in reconstituted spray dried plasma before (time =0 seconds) and after storage (e.g., up to 48 months at refrigeration, up to 12 months at room temperature). Stability involves comparing the respective value of one or more of these proteins / characteristics before and after storage to determine similarity to one another or to determine if the value is within the clinical range. In an embodiment, the values a plasma protein / characteristic after storage are within about 25% or less (25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, and 1%), as compared that before storage. In an embodiment, the values of plasma proteins / plasma characteristics after storage are within the clinical range for that plasma protein. Such plasma characteristics include pH, Osmolality (mOsm / kg), particulate size, particulate quantity and particulate distribution. Plasma proteins and their function include e.g., Total Protein (mg / ml), Activated Partial Thromboplastin Time (aPTT)(s), Prothrombin Time (s), International Normalized Ratio (INR (s)), Thrombin Time (s), Factor V (%), Factor VII (%), Factor VIII (%), Factor IX (%), Factor X (%), Factor XI (%), Factor XIII Activity (%), Factor XIII Antigen (%), Protein C Activity (%), Protein S Activity (%), Fibrinogen (mg / dL), Plasminogen (%),Plasmin InhibitorPI (%), Antithrombin III (%), von Willebrand Factor Antigen (% or IU / dL), von Willebrand Factor Ristocetin Cofactor (% or IU / dL), C5aDocket No. 0118.014100 -53- (ng / mL), Prothrombin Fragment F 1+2 (pmol / L), Thrombin-Antithrombin Complex (TAT) (µg / L), Protein (mg / mL) and the like. Measuring such plasma characteristics and plasma proteins are known in the art. In another embodiment, the acceptable or clinical range for von Willebrand Factor Ristocetin Cofactor (VWF:RCo) is between about 50 and about 200 IU / dL, or about 50 and about 200 IU / dL, and von Willebrand Factor Antigen (VWF:Ag) value is between about 50 and 200 IU / dL.

[0274] In an embodiment, the clinical reference range for determining shelf life or unit stability is as follows:

[0275] Table 2Docket No. 0118.014100 -54- Protein S PS Act (%) (60 - 150)

[0276] The data shows that the dried plasma is stable after being stored as a dried plasma for a time period up to about 24 months at refrigerated temperatures (2-8°C), 6 months at a Room Temperature (20-25°C) and 3 months elevated Room Temperature (30°C). Example 5 describes experiments performed that verified that the spray dried plasma unit remained stable during storage.

[0277] The spray dried plasma of the present invention, after release testing is complete and the product is determined safe for use, as described herein, can be stored at elevated temperatures, room temperature, refrigerated temperatures, freezing temperatures or a combination thereof. In certain aspects, spray dried plasma is refrigerated to enable storage for longer periods of time.

[0278] The dried plasma of the present invention can be stored between a temperature of -80ºC and 45ºC. Room temperature is considered between about 20ºC and 30ºC. In an aspect, refrigerated temperatures are between about 1°C and about 6°C. The terms “cooling,” “cold temperature,” “temperature below room temperature,” and “temperature below ambient temperature,” interchangeably refer to any temperature between 1ºC andDocket No. 0118.014100 -55- 20ºC. In any of the embodiments of the invention described herein, the temperature is selected from the group of temperatures consisting of 45ºC, 44ºC, 43ºC, 42ºC, 41ºC, 40ºC, 39ºC, 38ºC, 37ºC, 36ºC, 35ºC, 34ºC, 33ºC, 31ºC, 30ºC, 29ºC, 28ºC, 27ºC, 26ºC, 25ºC, 24ºC, 23ºC, 22ºC, 21ºC, 20ºC, 19ºC, 18ºC, 17ºC, 16ºC, 15ºC, 14ºC, 13ºC, 12ºC, 11ºC, 10ºC, 9ºC, 8ºC, 7ºC, 6ºC, 5ºC, 4ºC, 3ºC, 2ºC, 1ºC, 0ºC , -1ºC, -5ºC, -10ºC, -20ºC, -30ºC, -40ºC, -50ºC, - 60ºC, -70ºC, and -80ºC. In some embodiments, the spray dried plasma of the present invention is stored at a temperature of less than about 15°C, preferably less than 10°C, and more preferably less than 5°C. In some other embodiments, the spray dried plasma of the present invention is stored at room temperature. In other embodiments, the spray dried plasma of the present invention is stored at warm temperatures, e.g., above 23ºC. In further embodiments, the spray dried plasma of the present invention can be stored in freezing temperatures (e.g., 0ºC to - 80ºC). It is noted that although the dried plasma product can be stored in freezing temperatures, it does not need to be maintained in freezing temperatures to be stable for transfusion. On one hand, storage conditions can be carefully monitored, and on the other, storage conditions can be an array of temperatures depending on circumstances on the ground. In yet another embodiment, the dried plasma can be stored at a combination of temperatures.

[0279] As used in all of the aspects and embodiments of the invention herein, the term “period of time” or “time period” refers to a duration of time during which spray dried plasma of the present invention is stored at any given temperature. The term “period of time” can range from seconds to minutes to hours to days to weeks to months to years. In preferred embodiments, the term “period of time” refers a number of hours including about 3 to about 120 hours, e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, and 120 hours. In certain embodiments the period of time for which spray dried plasma of the present invention can be stored include about 1 and about 30 days (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, and 30). In certain embodiments the period of time for which spray dried plasmaDocket No. 0118.014100 -56- of the present invention can be stored include about 1 and about 48 months (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48 months).

[0280] In an embodiment, spray dried plasma of the present invention can be stored at room temperature for at least about 1 hour to about 7.5 months. In an embodiment, spray dried plasma of the present invention can be stored at refrigerated temperature for about 1 hour to about 48 months. In an aspect, the spray dried plasma of the present invention can be refrigerated on any day or days during storage.

[0281] In various other embodiments, the spray dried plasma of the present invention is stable and preserves the functionality of plasma proteins and / or plasma characteristics during the time period, when compared to corresponding clinical reference ranges or that prior to storage, or compared to thawed previously frozen plasma over the same period of time following spray drying. In an embodiment, spray dried plasma samples stored between a temperature of –80ºC and 45ºC are suitable for transfusion after extended periods of storage time, in an embodiment, for at least about 2 days, at least about 7 days, at least about 2 weeks, at least about 3 weeks, at least about 1 month, at least about 2 months, at least about 3 months, at least about 6 months, at least about 9 months, at least about 1 year, or at least about 1 ½ years, about 2 years, about 3 years, about 4 years, or longer.

[0282] The invention, in certain aspects, provides a novel method of storing a spray dried plasma in which the steps include obtaining plasma from a donor and drying the plasma using the methods and systems described herein, and storing the plasma for a period of time without a substantial loss or subclinical loss of one or more plasma protein amounts and / or plasma characteristics.

[0283] The spray dried plasma of the present invention is stable after reconstitution for up to 26 hours prior to use

[0284] The spray dried plasma stability also refers to the characteristics of spray dried plasma that has been reconstituted for a period of time and which is suitable for transfusion. The reconstituted previously dried plasma of the present invention remains stable and ready for transfusion for up to 26 hours, when reconstituted plasma is stored at room temperature, at refrigerated temperature, or at elevated temperature.Docket No. 0118.014100 -57-

[0285] “Stability” or “In Use Stability” also refers to previously spray dried plasma that has been reconstituted for a period of time (e.g., at 2 hours, 4, hours, 6 hours, 8 hours, 10 hours, 12 hours, 14 hours, 16 hours, 18, hours, 20, hours, 22 hours, 24, hours or 26 hours) and behaves comparably to spray dried plasma immediately or contemporaneously after reconstitution (e.g., from 0-5 hours, 1-15 minutes). Stability includes comparing amounts of various plasma proteins, their function (e.g., vWF, Factor V, Factor VIII, etc.) and / or plasma characteristics (e.g., pH, particle size, etc.) present in reconstituted previously spray dried plasma before (time =0 seconds) and after storage (e.g., time= 26 hours) of reconstituted plasma. In another embodiment, stability includes determining the amounts of various plasma proteins, their function (e.g., vWF, Factor V, Factor VIII, etc.) and / or plasma characteristics (e.g., pH, particle size, etc.) present reconstituted previously spray dried plasma after storage to determine that one or more fall within the corresponding clinical reference range. Stability involves comparing the respective value of one or more of these proteins / characteristics before and after reconstituted storage to determine similarity to one another, or the respective value to its clinical reference range. In an embodiment, the values of a plasma protein / characteristic after reconstituted storage is within about 25% or less (25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, and 1%), as compared that before reconstituted storage. When one or more values of plasma proteins after storage are within 25% of that prior to storage or within the clinical reference range, the reconstituted previously dried plasma is suitable for transfusion. Such plasma characteristics include pH, Osmolality (mOsm / kg), particulate size, particulate quantity and particulate distribution. Plasma proteins and their function include e.g., Total Protein (mg / ml), Activated Partial Thromboplastin Time (aPTT (s)), Prothrombin Time (s), International Normalized Ratio (INR (s)), Thrombin Time (s), Factor V (%), Factor VII (%), Factor VIII (%), Factor IX (%), Factor X (%), Factor XI (%), Factor XIII Activity (%), Factor XIII Antigen (%), Protein C Activity (%), Protein S Activity (%), Fibrinogen (mg / dL), Plasminogen (%), Plasmin Inhibitor (%),Antithrombin III (%), von Willebrand Factor Antigen (% or IU / dL), von Willebrand Factor Ristocetin Cofactor (% or IU / dL), C5a (ng / mL), Prothrombin Fragment F 1+2 (pmol / L), Thrombin-Antithrombin Complex (TAT) (µg / L), and the like. Measuring such plasma characteristics and plasma proteins are known in the art. In another embodiment, the acceptable or clinical range for von Willebrand Factor Ristocetin CofactorDocket No. 0118.014100 -58- (VWF:RCo) is between about 50 and about 200 IU / dL, or about 50 and about 200 IU / dL, and von Willebrand Factor Antigen (VWF:Ag) value is between about 50 and 200 IU / dL.

[0286] In an embodiment, the clinical reference range for In-Use stability is as follows:

[0287] Table 3Docket No. 0118.014100 -59- AT III Act (%) Plasminogen *Clinical refer

[0288] Thedata shows that the dried plasma when reconstituted is stable for transfusion after being stored as a reconstituted plasma for a time period ranging between about 1 hours to about 26 hours. Example 4 describes experiments performed that verified that the reconstituted previously spray dried plasma unit remained stable during storage and prior to transfusion.

[0289] The reconstituted spray dried plasma of the present invention can be stored for transfusion at room temperature or be refrigerated. In certain aspects, reconstituted previously spray dried plasma is refrigerated to enable storage prior to transfusion for longer periods of time.

[0290] The reconstituted previously dried plasma of the present invention can be stored for transfusion between a temperature of 1ºC and 45ºC. Room temperature is considered between about 20ºC and 30ºC. In an aspect, refrigerated temperatures are between about 1°C and about 6°C. terms “cooling,” “cold temperature,” “temperature below room temperature,” and “temperature below ambient temperature,” interchangeably refer to any temperature between 1ºC and 20ºC. In any of the embodiments of the invention described herein, the temperature is selected from the group of temperatures consisting of 45ºC, 44ºC, 43ºC, 42ºC, 41ºC, 40ºC, 39ºC, 38ºC, 37ºC, 36ºC, 35ºC, 34ºC, 33ºC, 31ºC, 30ºC, 29ºC, 28ºC, 27ºC, 26ºC, 25ºC, 24ºC, 23ºC, 22ºC, 21ºC, 20ºC, 19ºC, 18ºC, 17ºC, 16ºC, 15ºC, 14ºC, 13ºC, 12ºC, 11ºC,Docket No. 0118.014100 -60- 10ºC, 9ºC, 8ºC, 7ºC, 6ºC, 5ºC, 4ºC, 3ºC, 2ºC, and 1ºC. In some embodiments, the reconstituted previously spray dried plasma of the present invention is stored at a temperature of less than about 15°C, preferably less than 10°C, and more preferably less than 5°C. In some other embodiments, the reconstituted previously spray dried plasma of the present invention is stored at room temperature. In other embodiments, the reconstituted previously spray dried plasma of the present invention is stored at warm temperatures, e.g., above 23ºC. In yet other embodiments, the dried plasma is stored at a combination of these temperatures.

[0291] As used in all of the aspects and embodiments of the invention herein, the term of time” or “time period” refers to a duration of time during which reconstituted previously spray dried plasma of the present invention are stored prior to transfusion at any given temperature. The term “period of time” can range from seconds to minutes to hours to days to weeks to months. In an embodiment, the term “period of time” refers a number of hours including about 3 to about 30 hours, e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 hours.

[0292] In various other embodiments, the reconstituted previously spray dried plasma of present invention is stored prior to transfusion at room temperature. The reconstituted previously spray dried plasma of the present invention is stable and preserves the functionality of plasma proteins and / or plasma characteristics during the time period, when compared to corresponding clinical reference ranges or that prior to storage, or compared to thawed previously frozen plasma over the same period of time. Reconstituted previously spray dried plasma samples stored at or below room temperature are thus suitable for transfusion after extended periods of storage time, in an embodiment, for at least about 1 hour, at least about 2 hours, at least about 3 hours, at least about 4 hours, at least about 5 hours, at least about 6 hours, at least about 7 hours, or at least about 8 hours and up to about 26 hours.

[0293] The invention, in certain aspects, provides a novel method of storing a reconstituted spray dried plasma in which the steps include obtaining plasma from a donor and drying the plasma using the methods and systems described herein, and reconstituting the plasma and storing the reconstituted plasma for a period of time without a substantial loss (e.g., about 25% or less) or without a subclinical loss of one or more plasma protein amountsDocket No. 0118.014100 -61- and / or plasma characteristics. The method further includes transfusing the stored reconstituted plasma into a recipient.

[0294] The spray dried plasma of the present invention exhibits functionality of the most fragile of proteins, including von Willebrand’s factor, and other active proteins, absence of complement activation (C5a, C3a) and a pH which is near normal without buffering or treatment with CO2

[0295] The spray dried plasma of the present inventions is able to preserve the most fragile proteins and functions of the donor plasma, reduce complement activation and having a pH upon reconstitution that is near normal without additional buffering or treatment with CO2. This is accomplished with pretreating the liquid plasma with pretreatment solution (e.g., spray dry stable acidic substance (SDSAS) and one or more amino acids).

[0296] The present invention provides efficacy preservation and includes the preservation of the function of clotting factors in the plasma in a manner that does not otherwise harm the plasma or the transfused patient. During spray drying, some blood plasma proteins degrade to some extent, due to shear stress, surface stress (e.g., air-liquid interfacial stress), exposure to extreme pH, thermal stress, dehydration stress, and other environmental stresses.

[0297] The methods and compositions of the present invention recognize that pH and associated stresses can be reduced or the effects of which can be ameliorated by the use of formulations of the liquid plasma prior to or contemporaneously with spray drying. Formulation of the liquid plasma by glycine HCl or a similar spray dry stable acidic substance (SDSAS), and one or more amino acids (e.g., glycine), at novel concentrations, maintains the pH of the plasma at a non-alkaline level during the spray drying process. Adding an amino acid to the pretreatment solution results in higher recovery and better subsequent storage stability of plasma protein functions when compared to unformulated plasma

[0298] In a particular embodiment, the pretreatment solution is added to the donor plasma, wherein the pretreatment solution when added to the plasma has glycine in an amount ranging between about 10 µmole / mL of plasma and about 110 µmole / mL of plasmaDocket No. 0118.014100 -62- (e.g., about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110 µmole / mL of plasma), and hydrochloric acid (HCl) in an amount ranging between about 10 µmole / mL of plasma and about 30 µmole / mL of plasma (e.g., about 10, 15, 20, 25, and 30 µmole / mL of plasma), to thereby obtain formulated plasma. The formulated plasma has a pH in a range between about 6.0 and about 6.6 which offsets spray drying impacts on pH to yield a final rehydrated product that is at normal physiologic pH, a pH range between about 6.5 to about 7.8. In an embodiment, pretreatment of the plasma is optional.

[0299] In an embodiment, the amounts of an SDSAS and amino acid for the pretreatment step ranges as follows:

[0300] Table 4 Glycine (µmol / mL) HCl (µmol / mL)

[0301] mM mM 9 5 0

[0302] Proteins and their functionality are also protected in part because mass transfer occurs at lower temperatures, as described herein. As described, the drying rate is constant and as the liquid particle evaporates and loses moisture, the moisture transfers from the liquid plasma droplet to the drying gas, and the heat from the drying gas transfers to the plasma droplet making it into a dried particle. The plasma droplet enters the drying chamber essentially at room temperature and the temperature stays constant the majority of the evaporation period. See Fig. 43S. Once most all of the moisture leaves the particle, the temperature of the particle increases to equilibrate with the dryer chamber exit temperature of 65oC. During evaporation, the droplet is maintained at a lower temperature thereby protecting heat sensitive proteins such as vWF. See Fig. 43S. During the evaporation process, the temperature of the liquid droplet and the proteins therein experience a lower temperature, the thermodynamic wet bulb temperature, compared to the inlet drying gas temperature, thereby protecting the proteins. See Fig. 43R. Evaporation reduces protein temperature to near the thermodynamic wet bulb value and when the evaporation slows the particle temperature rises. See Fig. 43Sa.Docket No. 0118.014100 -63-

[0303] The term "recovery" is defined herein as referring to the percentage of an analyte preserved after spray drying compared with the analyte in a sample of the same native plasma that may have been frozen (the same sample before spray drying); the analyte is analyzed on native plasma and / or rehydrated plasma at the same protein concentrations. The analyte can be any known plasma substance such as a plasma protein (e.g., vWF antigen or fibrinogen), as described herein, and can be measured by concentration or activity of the analyte (e.g., vWF:RCo activity), also as described herein. The amount of the analyte can be compared to its corresponding clinical reference range.

[0304] A spray dry stable acidic substance (SDSAS) as used herein is any substance such as an acid or acidic salt or other substance that effectuates pH and is physiologically suitable for addition to the plasma being spray dried and physiologically suitable to the subjects (human or otherwise) to which the reconstituted plasma is to be administered (transfused). The SDSAS remains sufficiently stable (e.g., does not materially evaporate or chemically breakdown) during the spray drying process. The SDSAS effectuates the pH adjustment described herein which results, for example, a maintained or an improved von Willebrand's factor recovery or functionality in the reconstituted plasma described herein, as compared to non-pretreated spray dried plasma. Specific examples of spray dry stable acidic substances include glycine HCl, HCl, citric acid, lactic acid, monosodium citrate and other SDSAS's described herein. Other SDSAS's may be known in the art or may be determinable by straightforward experimentation.

[0305] In an embodiment, pretreatment solution of the present invention used to obtain the spray dry formulation includes one or more SDSAS and one or more amino acids. Addition of the amino acid allows for protection of the plasma proteins during spray drying without lowering the pH of the pre-treatment solution. The addition of an amino acid increases the pH of the pretreatment solution, but surprisingly does not affect the pH of pretreated spray dried plasma or the rehydrated spray dried plasma. Further, in an embodiment, using an amino acid along with a SDSAS provides a spray dried plasma, once rehydrated, with reduced levels of C5a, an anaphylatoxin, or levels of C5a that are similar to never frozen plasma (NFP) or FDA approved apheresed plasma products. In an embodiment, complement activation is associated with inflammation and should be kept low within a clinically acceptable range. In particular, the pH of the pretreatment solutionDocket No. 0118.014100 -64- is in a range between about 2.0 and about 4.0 (e.g., 2.0, 2.2, 2.4, 2.6, 2.8, 3.0, 3.2, 3.4, 3.6, 3.8, 4.0) and when combined with plasma results in a pretreated plasma having a pH of between about 6.0 and about 6.6 (e.g., 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6) , and once the previously spray dried plasma is reconstituted, results in a reconstituted plasma having a pH of between about 6.5 to about 7.8 (e.g., 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.8). The total concentration of the one or more amino acid(s) in the reconstituted plasma is present in an amount between about 1 mM and about 150 mM. Examples of amino acids that can be combined with the SDSAS for the pretreatment solution include glycine, alanine, asparagine, glutamine, isoleucine, leucine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine. In a particular embodiment, glycine is an amino acid added to the SDSAS composition.

[0306] Accordingly, spray drying formulation, i.e., treatment of feed plasma prior to or contemporaneously with spray drying, preserves and allows recovery or functionality of active clotting factors of rehydrated plasma that has undergone the spray drying process as well as long term stability during storage after drying. As further discussed below, these improvements to certain embodiments of spray drying of blood plasma involving formulation with a SDSAS and / or an amino acid, also improve the ease and lower the cost of rehydration of the plasma product by allowing the spray dried plasma to be rehydrated with sterile water (e.g., water for injection: WFI or sterile water for injection: SWFI). The spray dried plasma of the present invention may be reconstituted with sterile water for injection without the need for a buffered rehydration solution or treatment or storage with CO2 or other treatment to control the pH of the reconstituted plasma.

[0307] The spray dried plasma of the present invention, in an embodiment, has improved functionality or recovery of active plasma proteins, long term stability of plasma proteins and a reduction in anaphylatoxins. In an embodiment, the method to obtain the dried plasma of the present invention includes combining donor plasma with a pretreatment solution having a SDSAS and an amino acid, and a spray drying system. The invention further contemplates adjusting the pH of the donor plasma with the SDSAS by bringing the concentration of the SDSAS to about 1 mM to about 50 mM, which lowers the pH of the plasma to about 5.0 to about 6.5 to create formulated plasma. In another embodiment, the invention further contemplates adjusting the pH of the plasma to be sprayDocket No. 0118.014100 -65- dried with a pretreatment solution having a SDSAS and an amino acid by bringing the concentration of the SDSAS in the formulated plasma to about 1 mM to about 50 mM and the amino acid compound to about 1 mM and about 150 mM , which lowers the pH of the plasma to about 6.0 to about 6.6 to create formulated plasma.

[0308] In an embodiment, to obtain the dried plasma of the present invention, the methods include methods for producing spray dried plasma by combining plasma with a pretreatment solution, wherein the pretreatment solution comprises an amino acid (e.g., glycine) in an amount ranging between about 10 µmole / mL of plasma and about 110 µmole / mL of rehydrated plasma (e.g., about 10, 20, 30, 40, 50, 60, 70, 80. 90. 100110 µmole / mL of plasma), and SDSAS (e.g., hydrochloric acid (HCl)) in an amount ranging between about 10 µmole / mL of plasma and about 30 µmole / mL of rehydrated plasma (e.g., about 10, 15, 20, 25, and 30 µmole / mL of plasma), to thereby obtain formulated plasma. The method also includes drying the formulated plasma with a spray drying system to create spray dried formulated plasma, as described herein. In an embodiment, the pretreatment solution has glycine in an amount of about 84 µmole / mL of plasma and HCl in an amount of about 20 µmole / mL of plasma.

[0309] The pretreatment solution, in an embodiment, has glycine in an amount ranging about 15 mmol and about 30 mmol (e.g., about 15, 20, 25, and 30 mmol) or between about 43 µmol / mL and about 473 µmol / mL, and HCl in an amount ranging between about 3 mmol and about 7 mmol (e.g., about 3, 4, 5, 6, and 7 mmol) or between about 43 µmol / mL and 129 µmol / mL, to thereby obtain formulated plasma; and drying the formulated plasma with a spray drying system to create spray dried formulated plasma. In a certain embodiment, the pretreatment solution has glycine in an amount of about 22 mmol and HCl in an amount of about 5.3 mmol.

[0310] After drying the formulated plasma with the spray drying system to create spray dried formulated plasma, the spray dried formulated plasma had a recovery of functional von Willebrand factor (vWF) of at least 10 to at least 100 percentage points greater than the recovery of functional von Willebrand factor obtained from an otherwise identical spray dried plasma that has not undergone acid formulation with the pretreatment solution of the present invention. In another embodiment, the spray dried formulated plasma has a recovery of functional von Willebrand factor (vWF) that are similar to or within about 20% (e.g.,Docket No. 0118.014100 -66- about 15%, 10%, 5%) of never frozen plasma or FDA approved plasma products to fresh frozen plasma. The SDSAS may be selected from any known in the art, however, glycine HCl, HCl, citric acid and lactic acid are preferred substances for use in the present invention. When adding an amino acid to the SDSAS to form the pretreatment solution, in an embodiment, glycine is a preferred substance of the present invention (e.g., HCl / glycine or citric acid / glycine combinations). The physiologically compatible pretreatment solution is added to the plasma before spray drying and preferably shortly before spray drying or contemporaneously with spray drying. Additionally, the pH of the plasma may be determined before the addition of a SDSAS and an amino acid to the plasma to determine an appropriate amount of acid to add. In an embodiment, about 7.4 mM of citric acid is added to the CPD plasma or WB plasma. In an embodiment, the pH of the formulated plasma is about 5.5 to about 6.5. The present invention further contemplates that the recovery of functional vWF may be from about 10 to about 100 percentage points to about 40 percentage points (e.g., about 10, 15, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 percentage points) greater than the recovery of functional von Willebrand factor obtained from an otherwise identical spray dried plasma that has not undergone pretreatment with a SDSAS and an amino acid . In an embodiment, the data show that levels of Ristocetin Cofactor (vWF:RCo) were non-existent or undetectable when plasma is spray dried without pretreatment. See Figs. 17-19. In this embodiment, Ristocetin Cofactor (vWF:RCo) of pretreated spray dried plasma was 2X, 3X, 4X, 5X or greater, as compared to non-pretreated spray dried plasma.

[0311] It has been discovered that desirable C5a levels result from a pretreatment having an SDSAS and an amino acid addition, and optionally rapid mixture / agitation of the pretreatment components. In an embodiment, desirable C5a levels are those that are similar to C5a levels in NFP or an FDA approved apheresis plasma product. In an embodiment C5a levels are reduced, as compared to C5a levels from rehydrated plasma that underwent pretreatment only with a SDSAS, and optional rapid mixture / agitation of the pretreatment components. In particular, levels of C5a for reconstituted plasma resulting from the pretreatment solution of the present invention can be between about 0.1 ng / mL to about 74 ng / mL and in particular between 20 ng / mL and 70 ng / mL (e.g., about 30 ng / mL). C5a levels are reduced when treated with a pretreatmentDocket No. 0118.014100 -67- solution having at least one SDSAS and at least one amino acid, as compared to plasma subjected to a pretreatment solution having only an SDSAS. In an embodiment, the C5a levels are reduced by about 20% or less (e.g., 20%, 15%, 10%, 5%, 1%), as compared to plasma subjected to the pretreatment solution of the present invention with acid alone. In another embodiment, C5a levels are about the same as that of never frozen plasma, or within about 20% or less (e.g., 20%, 15%, 10%, 5%, 1%) of C5a in never frozen plasma or approved FDA apheresis plasma products.

[0312] The present invention contemplates a reconstituted spray dried plasma product for human transfusion (administration), the reconstituted spray dried plasma product having been reconstituted with, for example, sterile water for injection and the reconstituted spray dried plasma product having a pH of about 6.5 to about 7.8. The reconstituted plasma of the present invention has functional von Willebrand factor of greater than 5 percentage points as compared to the recovery of functional von Willebrand factor obtained from an otherwise identical spray dried plasma that has not undergone formulation with a SDSAS and / or an amino acid; or about 5 percentage points to about 40 percentage points (e.g., about 25 percentage points to about 35 percentage points) greater than the recovery of functional von Willebrand factor obtained from an otherwise identical spray dried plasma that has not undergone pretreatment with a SDSAS and an amino acid. Additionally, the present invention relates to reconstituted spray dried plasma having levels of C5a between about 0.1 ng / mL to about 74 ng / mL and in particular between 20 ng / mL and 40 ng / mL (e.g., about 30 ng / ml). In an embodiment, the present invention pertains to reconstituted spray dried plasma pretreated with an SDSAS and an amino acid have levels the C5a levels that are reduced e.g., by about 20% or less (e.g., 20%, 15%, 10%, 5%, 1%), as compared to plasma subjected to the pretreatment solution having an SDSAS . In another embodiment, C5a levels of reconstituted plasma of the present invention are about that approved FDA plasma products.

[0313] Autologous uses of the dried plasma unit of the present invention The dried plasma of the present invention can be from a third-party donor or from the intended recipient. The latter is known as autologous plasma. Autologous refers to plasma taken from and provided to the same individual. Autologous spray dried plasma is desirable because it is compatible with recipient’s plasma, e.g., there is no need to beDocket No. 0118.014100 -68- ABO / Rh type matched before use, and it is unlikely to carry pathogens or immunogens allergenic to the recipient.

[0315] The autologous use of spray dried plasma of the present invention is characterized by at least two major components – the dried plasma unit and a reconstitution fluid. In an embodiment, the reconstitution solution is sterile water (SWFI) and the resultant reconstituted plasma has a near physiologic pH. In an embodiment, the sterile water comes as a premeasured, standard amount in a pouch.

[0316] In an embodiment, an intended recipient who may be at increased risk for a plasma transfusion (e.g., soldier, policeman, fire fighter, adventurer) can provide / donate their own plasma and one can dry the plasma using the methods described herein to obtain a dried plasma unit, having the one or more of the characteristics described herein. One or more dried plasma units dried from plasma provided by the intended recipient can be carried by or on behalf of the intended recipient so that it is readily available for their use. In this embodiment, autologous uses are encompassed by the present invention because the dried plasma units are light weight, whose packaging is rugged, and storable in various temperatures (including room temperature and / or warmer temperatures). When the intended recipient is in need of a plasma transfusion, a health care provider (e.g., doctor, nurse, first responder) carries the reconstitution solution (e.g., SWFI) and can reconstitute the intended recipient’s own dried plasma for emergency transfusion. In an emergency, instant plasma with its trove of clotting factors would be available to the bleeding recipient. Early delivery of much needed plasma in a trauma situation increases the likelihood of a favorable result.

[0317] In an embodiment, the reconstitution solution (e.g. SWFI) can be carried by either the health care provider or the intended recipient. However, in situations like a battlefield, it is preferable for the health care provider to carry the reconstitution solution because it tends to be relatively heavy and SWFI is readily available to health care providers. The advantage of an autologous dried plasma unit is the elimination of compatibility issues and reduction in infectious transmission. Autologous dried plasma does not need to undergo ABO Rh matching and reduces infection transmission by a non-autologous or potentially allogeneic donor.

[0318] The manufacturing system for the dried plasma of the present invention is relatively compact and can be deployed in a very wide variety of environments. TheDocket No. 0118.014100 -69- manufacturing process for the spray dried plasma of the present invention is automated and easy to use by a person with minimal education and straight forward training.

[0319] Accordingly, the steps of providing an autologous dried plasma unit include the step of providing liquid plasma by the intended recipient (which can be obtained using methods known in the art and / or those described herein) and drying the intended recipient’s liquid plasma using the spray drying system described herein. Finally, the method includes the step of reconstituting the recipient-provided dried plasma unit and, when in need, transfusing the recipient-provided reconstituted dried plasma in the recipient. The recipient provided dried plasma unit can be reconstituted by a health care provider or other qualified person.

[0320] This autologous dried plasma system can readily be adapted to other situations or available for use by any person that wants their own plasma nearby and readily available for future use. Other situations in which autologous dried plasma can be used include those who are residents of rural areas and are far from sources of plasma, or those that live anywhere but prefer to have their own plasma transfused into them in the case of an emergency. Intended recipients can store their own dried plasma units in the home, in safe rooms, vehicles, or with the health care providers, and the local health care providers can carry the reconstitution solution. In accordance with the storage data herein, such dried plasma units for autologous use can be replenished every 1-4 years. Wide deployment of the dried plasma unit of the present invention can be done as a prophylactic measure against death or permanent injury from bleeding trauma or disease.

[0321] A recipient can be human or mammal. Pretreatment Formulation and Process - Detailed Description

[0322] Embodiments of the present disclosure are directed to methods and compositions relating to a spray dried liquid sample. In certain embodiments, the liquid sample is plasma obtained from a blood donor. In a preferred embodiment, the blood donor is human. However, it may be understood that the disclosed embodiments may be employed to spray dry any biological mixture of solid particles and / or molecules in a continuous liquid medium, including, but not limited to, colloids, suspensions and sols (a colloidal suspension of very small particles).

[0323] The control of reconstituted dried plasma pH by the invention described hereinDocket No. 0118.014100 -70- which utilizes spray drying is an improvement over the reported pH control of reconstituted plasma made by freeze drying or lyophilization. Excessively high or low pH of blood plasma is associated with increased morbidity or mortality including pH above 7.8 (“alkalosis”). In this respect the present invention is superior to the freeze-dried products and processes of the prior art. Drying processes result in loss of CO2 which causes the pH of the dried product to increase unless controlled in some manner. The present invention does that with no extra processing steps, reconstitution with sterile water only and has been approved for clinical trials by the United States Food and Drug Administration. Reconstitution with sterile water only is highly desirable in dried plasma products. In the event of loss or damage to the pre- measured, pre-packaged sterile water for reconstitution provided as part of a kit for emergency, ER, OR or other urgent use of any dried plasma, a measured amount of readily available sterile water can be used for reconstitution.

[0324] “Human lyophilized plasma is …alkalotic with a pH near 8, ….” Zaza M, Kalkwarf KJ, Holcomb JB. Dried Plasma. Damage Control Resuscitation. 2019;145-162. Published 2019 May 6. doi:10.1007 / 978-3-030-20820-2_8, page 8, second full paragraph. Zaza, et al excuse this by saying “however, [lyophilized plasma] is well tolerated clinically in humans”, citing solely the 2013 article of Saillol, et al,. “The evolving role of lyophilized plasma in remote damage control resuscitation in the French Armed Forces Health Service.” Transfusion. 2013;53:65S–71S. The Saillol article concerns the French Army’s lyophilized dried plasma known as French LYophilized Plasma (FLYP). Saillol, et al admit that “the pH upon reconstitution [of FLYP”] is close to 8.” Id. at 67S. The Saillol, et al. report is limited to combat situations of severe bleeding in which the protocol included tranexamic acid, FLYP with red blood cells in a 1:1 ratio and other actions to control the patient’s blood pH. Id. at 66S. Salliol, et al, admit that “further research is needed to determine specific indications for FLYP in the therapeutic management of civilian patients with severe hemorrhage.” Id. at Abstract last sentence, see p. 65S. In contrast, the lower, well-controlled pH of the present invention is expected to be suitable for use in all situations where plasma transfusion is indicated in any amount under any circumstance.

[0325] The pH of reconstituted dried plasma made by the freeze-drying process of BCT has been reported to be high at 7.66-7.94. 7.94 is very close to 8.0. The so- called French Lyophilized Plasma (FLYP) plasma product made by the French Army isDocket No. 0118.014100 -71- reported to also have a reconstituted pH “close to 8.0.” See Flaumenhaft, et al, Retention of Coagulation Factors and Storage of Freeze-Dried Plasma, Military Med., Vol. 6, January / February Supplement, pp. 400-407, 403 (2021). “TFDP [Terumo Freeze Dried Plasma] units exhibited a significant elevation in pH after freeze drying, as expected based on other lyophilized plasma products….” Sheffield WP, et al., “Retention of hemostatic and immunological properties of frozen plasma and COVID-19 convalescent apheresis fresh- frozen plasma produced and freeze-dried in Canada” Transfusion. 2021 Dec 14. doi: 10.1111 / trf.16772. Epub ahead of print. PMID: 34907536.). There is no report that the Terumo freeze-dried plasma product has been approved for clinical trials in the United States or elsewhere. The FLYP plasma has not been the subject of clinical trials in the United States. The authors of Flaumenhaft dismiss the high pH of the Terumo-BCT reconstituted freeze-dried plasma material as “within the Terumo-BCT required range of 7.0-8.0” and “aligned” with the FLYP plasma pH of close to 8.0. In contrast, the pH of the spray dried plasma product of the present invention does not exceed 7.8 and in general has a pH range closer to physiological pH.

[0326] The reported pH of the reconstituted dried plasma made by the freeze-drying process of Teleflex has not been reported. However, it is evidently higher (more alkaline) than physiological pH at the end of the freeze-drying process such that, in a clinical trial investigator’s contract with United States Food and Drug Administration, Teleflex described its REPLASTMfreeze dried plasma as requiring extra processing and equipment to reduce the pH of the Teleflex product when reconstituted. In particular, an acidic reconstitution fluid must be used to restore pH of the reconstituted [lyophilized plasma] to a physiological pH before infusion. Van, et al, J Trauma Injury, Infection and Critical Car Vol 71 No 1, p22 (July 2011). In fact, according to Van, preliminary studies in our laboratory revealed that LP reconstituted without an acid has a pH of ~9 and its infusion resulted in rapid death. ID. at 20. The REPLASTMfreeze dried process is described as including the following steps: “vacuum chamber is broken with medical grade carbon dioxide (C02) gas to correct for loss of dissolved CO2 from the starting plasma material during the freeze-drying process” and that “in addition, REPLASTMis packaged in an outer foil pouch that is flushed with a fixed amount of CO2 gas, which results in a near neutral pH in the reconstituted … product.” Jose A. Cancelas, Investigator’s Agreement A Phase 1, Single-Center, Partial Doubleblind,Docket No. 0118.014100 -72- Randomized, Controlled (Versus Fresh Frozen Plasma [Ffp] In Cohort 3 Only) Clinical Study Of The Safety Of Ascending Doses Of Autologous Freeze Dried Plasma (Fdp) In Healthy Volunteers, Apr19, 2018; pp-24.25. Date of download Dec 62021 https: / / clinicaltrials.gov / ProvidedDocs / 26 / NCT02930226 / Prot_000.pdf The system of spray drying plasma of the present invention does not require elaborate, expensive use of CO2 gas treatment of the dried plasma or CO2 storage of the dried plasma to control pH in the reconstituted plasma product or the equipment needed to effect these extra, pH correction processes.

[0327] Plasma Plasma is the fluid that remains after blood has been centrifuged (for example) to remove cellular materials such as red blood cells, white blood cells and platelets. Plasma is generally yellow-colored and clear to opaque. It contains the dissolved constituents of the blood such as proteins (6-8%; e.g., serum albumins, globulins, fibrinogen, etc.), glucose, clotting factors (clotting proteins), electrolytes (Na+, Ca2+, Mg2+, HCO3-, Cl, etc.), hormones, etc. Whole blood (WB) plasma is plasma isolated fromwhole blood with noadded agents except anticoagulant(s). Citrate phosphate dextrose (CPD) plasma, as the name indicates, contains citrate, sodium phosphate and a sugar, usually dextrose, which are added as anticoagulants. The level of citrate in CPD plasma, derived from whole blood, is about 20 - 30 mM. Thus, the final citrate concentration in the whole blood derived CPD plasma formulated with 7.4 mM citric acid will be about 27.4 - 37.4 mM.

[0329] The plasma of the present invention may be dried after pooling or unit-by-unit. Pooling of multiple plasma units has some benefits. For example, any shortfall in factor recovery on an equal-volume basis can be made up by adding volume from the pool to the finished product. There are negative features as well. Making up volume from the pool to improve factor recovery is expensive. Importantly, pooled plasma must be constantly tested for pathogens as any pathogens entering the pool from, for example, a single donor, runs the risk of harming hundreds or thousands of patients if not detected. Even if detected, pathogen contamination of pooled plasma would render the whole pool valueless. Testing can be obviated by pathogen inactivation of the plasma by irradiation or chemically such as solvent detergent treatment; however, each such treatment adds cost and complexity to pooled plasma processing. In any event, pooled plasma processing is generally unsuitableDocket No. 0118.014100 -73- to the blood centers and generally only really suitable to an industrial, mass production environment.

[0330] Conversely, unit-by-unit (unit) collection and processing is well-suited to the blood center environment and eliminates the risk of pooled plasma pathogen contamination by allowing for pre-processing testing for pathogens and tracking of the unit to ensure that each unit leaves the blood center site pathogen free. The inventors have discovered that efficient and effective preservation and recovery of functional clotting factors is the standard by which successful unit blood plasma processing should be measured. Such efficiency is also very helpful in the pooled plasma environment as well.

[0331] Clotting Factors There are many blood plasma factors associated with clotting. The methods and compositions of the present invention include recovering amounts of functional fibrinogen, Factor V, Factor VII, Factor IX and vWF from rehydrated plasma that has undergone the spray drying process. Such blood plasma factors are important in-patient treatment especially after trauma injuries to promote clotting of wounds. Thus, rapid administration of plasma is an important factor contributing to positive clinical outcomes. The spray dried plasma of the present invention can be readily reconstituted in a few minutes at the location of the trauma event without moving the patient and without time delay. Further, the spray dried plasma of the present invention has high levels of functional proteins that are stable for extended periods of time without freezing.

[0333] Functional vWF has generally been difficult to recover and has become one indicator for preservation of all factors. The present invention includes recovering amounts of functional vWF, in an amount in rehydrated spray dried plasma that is at least about 5 percentage points or greater (e.g., about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60 or greater percentage points) as compared to amounts of functional vWF of rehydrated spray dried plasma that do not undergo the pre-treatment steps of the present invention. The present invention includes recovering amounts of functional vWF, in an amount in rehydrated spray dried plasma that is at about 5 percentage points to about 40 percentage points or about 10 percentage points to about 35 percentage points higher as compared to amounts of functional vWF of rehydrated spray dried plasma that do not undergo the pretreatment step of the present invention. vWF activity is typically assayed with an assay called the vonDocket No. 0118.014100 -74- Willebrand factor: Ristocetin cofactor (vWF:RCo) assay, as is known to those of skill in the art. The vWF:RCo assay measures the ability of a patient's plasma to agglutinate platelets in the presence of the antibiotic Ristocetin. The rate of Ristocetin induced agglutination is related to the concentration and functional activity of the plasma von Willebrand factor. Another assay, the vWF antigen assay, measures the amount of vWF protein present in a sample. Yet another assay to determine if functional clotting plasma proteins exist in reconstituted previously spray dried plasma is to perform a Bioflux study. See Example 9 and 10.

[0334] In an embodiment, one or both von Willebrand Factor Antigen (% or IU / dL), Willebrand Factor Ristocetin Cofactor (% or IU / dL) are measured before and after spray drying to determine the recovery of vWF. In an embodiment, the acceptable or clinical range for von Willebrand Factor Ristocetin Cofactor (VWF:RCo) is between about about 50 and about 200 IU / dL, and von Willebrand Factor Antigen (VWF:Ag) value is between about 50 and 200 IU / dL. In an embodiment, the present invention includes determining amounts of vWF using a VWF:RCo assay or a VWF:Ag assay in rehydrated spray dried plasma that is at least about 5 percentage points or greater (e.g., about 5, 10, 15, 20, 25 or greater percentage points) as compared to those amounts in rehydrated spray dried plasma that do not undergo the pre-treatment steps of the present invention.

[0335] Pretreatment solution having one or more Spray Dry Stable Acidic Substances (SDSAS) and one or more amino acids

[0336] The present invention contemplates the use of a pretreatment solution that includes one or more physiologically compatible spray dry stable acidic substances (SDSAS) combined with one or more amino acids, as a formulation agent for plasma prior to being spray dried. As used herein, the phrase “SDSAS and amino acid” and the like refers to a pretreatment solution that has at least one SDSAS and / or at least one amino acid. Similarly, the use of “the SDSAS” or “an amino acid” refers to one or more SDSAS or one or more amino acids, respectively. The phrase “formulated plasma” or “pretreated plasma” or “PreT” refers to the mixture of the pretreatment solution (e.g., at least one SDSAS and / or at least one amino acid) and plasma prior to spray drying. Dried formulatedDocket No. 0118.014100 -75- plasma refers to spray dried plasma that was pretreated with the pretreatment solution.

[0337] While the present invention is not limited by theory, it is presumed by the inventors that the SDSAS of the present invention (e.g., citric acid, lactic acid, hydrochloric acid, etc.) exerts its effects because it prevents or alleviates the rising of the pH of the plasma during the spray drying process. Addition of an amino acid to the SDSAS still allows the pre-treatment solution to have an acidic pH, but not be so low as to harm the plasma proteins. Non-limiting examples of suitable SDSAS are hydrochloride (HCl), citric acid and lactic acid. When an SDSAS is combined with an amino acid, an example includes glycine HCl. The SDSAS (e.g., (HCl, citric acid or lactic acid) and amino acid (e.g., glycine) can be added to the plasma in a combined form (e.g., glycine HCl) or as separate compounds (e.g., glycine and HCl). Other non-limiting examples of suitable acids are ascorbic acid and gluconic acid. Because CO2 is lost from plasma during spray drying, the reaction generating bicarbonate and H+from CO2and H2O is shifted away from H+, thereby increasing the pH (i.e., Chatelier's principle). Human blood / plasma contains a buffer system comprised of carbonic acid (H2CO3) and bicarbonate anion (HCO3-), which is important for maintaining blood pH between 7.35 and 7.45, as a value higher than 7.8 can lead to death. In this buffer, hydronium (H3O+) and bicarbonate anion are in equilibrium with carbonic acid (Equation 1). Furthermore, the carbonic acid in the first equilibrium can decompose into CO2 gas and water, resulting in a second equilibrium system between carbonic acid and water (Equation 2).

[0338] In summation, the blood buffer is: H2CO3 + H2O ⇌ H3O++ HCO3- (Equation 1)

[0339] With the following simultaneous equilibrium: H2CO3 ⇌ H2O + CO2 (Equation 2)

[0340] Spray drying drives off CO2leading to the reduction of the levels of H2CO3and and thereby drives up the pH level of the drying plasma. Consequently, the pretreatment solution of the present invention helps to safely lowers the pH of the formulated plasma prior to spray drying to result in a spray dried plasma, that when reconstituted with Sterile Water for Injection, has a resulting physiologically compatible pH.Docket No. 0118.014100 -76-

[0341] Glycine addition helps offset this change. Amino acid addition prevents the pH from going too low. Therefore, in an embodiment, the plasma is formulated with a pretreatment having the SDSAS and an amino acid. Because of the formulation / pretreatment step, vWF activity loss is reduced and / or the amount of undenatured vWF is increased, as compared to spray dried plasma not subjected to the formulation steps of the present invention. The SDSAS is present in the pretreatment solution in an amount between about 1 mM to about 50 mM, which lowers the pH of the formulated plasma to about 5.5 to about 6.5 or to about 7.2 to create formulated plasma. When the amino acid such as glycine is also present along with the SDSAS in the pretreatment solution in an amount between about 1 mM to about 150 mM, the pH of the formulated plasma is about 6.0 to about 6.6.

[0342] Because the physiologically compatible SDSAS and amino acid of the present invention is included in this manner, the inventors further determined that the rehydration step can be performed by water alone (e.g., SWFI). Alternatively, sodium phosphate or other agents can optionally be added to the rehydration solution. Further, any other suitable rehydration fluid as can be determined by one of ordinary skill in the art may be used.

[0343] From experiments conducted by the inventors with spray drying, it has been discovered that the von Willebrand factor activity level in plasma dried by spray drying is affected, in part, by the shear forces generated during the aerosolization process (see, Examples, below) and an increase in the pH of the plasma. The present invention shows that the utilization of a step wherein the plasma is formulated with at least one SDSAS and at least one amino acid greatly improves the recovery and stability of active vWF over conditions where the SDSAS and the amino acid is not used as formulation agents.

[0344] A SDSAS is a substance which does not evaporate easily at room temperature at atmospheric pressure. Typically, the boiling point of the SDSAS will be greater than about 150OC at atmospheric pressure. In addition to glycine HCl, non-volatile acids that are suitable of use as the SDSAS of the present invention include phosphorus-containing acids such as, for example, ortho-phosphoric acid, pyrophosphoric acid, meta-phosphoric acid, poly phosphoric acid, alkyl- and aryl-substituted phosphonic and phosphinic acids, phosphorous acid, and the like, and mixtures thereof. Other non-volatile acids suitable for use as the SDSAS of the present invention include, but are not limited to ascorbic acid,Docket No. 0118.014100 -77- citric acid, lactic acid, gluconic acid, oxalic acid, halogenated acetic acids, arene sulfonic acids, molybdic acid, phosphotungstic acid, tungstic acid, chromic acid, sulfamic acid, and the like.

[0345] In an embodiment, the pre-treatment solution of the present invention can include one or more SDSAS and one or more amino acids. Addition of the amino acid to the SDSAS allows for protection of the plasma proteins without allowing the pH to go too low and cause protein damage and other deleterious effects such as complement activation. The addition of an amino acid increases the pH of the pretreatment solution, but surprisingly does not affect the pH of rehydrated spray dried plasma (ODP). In particular, the pH of the pretreatment solution is in a range between about 2.0 and about 4.0 and results in a formulated plasma (e.g., prior to spray drying) having a pH of between about 6.0 and about 6.6, results in a rehydrated plasma having a pH of between about 6.5 to about 7.8. In an embodiment, the pre-treatment solution of the present invention includes the SDSAS and at least one (e.g., one or more) amino acids. The total concentration of the amino acid(s) present in the pretreatment solution is an amount between about 1 mM and about 150 mM. Examples of amino acids that can be added to the SDSAS of the pretreatment solution include alanine, asparagine, glutamine, glycine, isoleucine, leucine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine. In a particular embodiment, glycine is added to the SDSAS composition as shown in the examples. The addition of an amino acid increases the pH of the pretreatment solution, but surprisingly does not affect the pH of rehydrated spray dried plasma (ODP). See Example 17. Even more surprisingly, addition of an amino acid mitigates C5a elevation. See Example 16. These amino acids have at least two pKa values, as follows:

[0346] Table 5 Amino acid pKa1 pKa2 pKa3 pI Glycine 2.34 9.60 --- 5.97 Alanine 2.34 9.69 --- 6.00 Valine 2.32 9.62 --- 5.96 Leucine 2.36 9.60 --- 5.98 Isoleucine 2.36 9.60 --- 6.02Docket No. 0118.014100 -78- Amino acid pKa1 pKa2 pKa3 pI Methionine 2.28 9.21 --- 5.74 Proline 1.99 10.60 --- 6.30 Phenylalanine 1.83 9.13 --- 5.48 Tryptophan 2.83 9.39 --- 5.89 Asparagine 2.02 8.80 --- 5.41 Glutamine 2.17 9.13 --- 5.65 Serine 2.21 9.15 --- 5.68 Threonine 2.09 9.10 --- 5.60 Tyrosine 2.20 9.11 --- 5.66

[0347] SDSAS useful in the process of the invention are capable of replacing (or compensating for) the volatile acid, i.e. CO2 that escapes from the plasma during spray drying. As indicated above, examples or suitable acids include, but are not limited to, glycine HCl, ascorbic acid, citric acid, gluconic acid, and lactic acid.

[0348] A volatile acid as defined herein has a pKa less than about 3 and a boiling point less than about 150°C at atmospheric pressure. Typically, the pKa of the volatile acid is within the range of about 1 to about 15. Non-limiting examples of volatile acids are hydrogen chloride, hydrogen bromide, hydrogen iodide, hydrogen fluoride, acetic acid, formic acid, hydrogen sulfide, hydrogen selenide, sulfur dioxide, fluorosulfonic acid, methane sulfonic acid, trifluoroacetic acid, trifluoromethanesulfonic acid, and the like.

[0349] A volatile strong acid can be fixed with an amino acid or like to render it non- volatile, making it easier to use. For example, volatile hydrogen chloride can be converted to glycine hydrogen chloride (glycine HCl, glycine hydrochloride). To distinguish this from adding an amino acid to the pretreatment solution, this describes how an SDSAS is made. The pretreatment solution includes the addition of an amino acid that is not bound to an acid with an already conjugated and stable SDSAS. Alternatively, glycine and HCL can be added to the pretreatment solution in such amounts to form the SDAS (e.g., glycine HCL) and free amino acid (e.g., glycine) in solution. Since there is a 1:1 relationship between glycine and HCl, one can add more glycine than HCl to obtain a solution with free glycine and glycine HCl. In an embodiment of the present invention, HCl and glycine are added to a solvent, such as SWFI, to create the pretreatment solution such that the final concentration in the formulated plasma is about 16.8 mM HCl and about 69.6 mM glycine. In other words, in anDocket No. 0118.014100 -79- embodiment of the present invention, 5.2 mM HCl and 21.5 mM glycine is added to 50 mL of solvent, such as SWFI, to create the pretreatment solution. The present invention includes adding the following to a solvent, such as SWFI, to create a pretreatment solution: between about 3.0 to about 7.0 mmol (e.g., 3.0, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, and 7.0) mmole HCl and about 15 mmole and about 30 mmole glycine (e.g., about 15, 17, 20, 22, 25, 27, 30 mmoles glycine) in 50 mL of solvent to obtain 260 of formulated plasma. In yet other words, glycine in an amount of about 440 mM, and HCl in an amount of about 106 mM is present in the pretreatment solution. In an embodiment between about 290 mM to about 570 mM (e.g., about 290, 300, 350, 400, 450, 500, 550, and 570) glycine and about 70 mM to about 140 mM (e.g., 70, 80, 90, 100, 110, 120, 130, 140 mM) HCl is present in the pretreatment solution.

[0350] In this case, 16.8 mM glycine HCl and 86.4 (69.6-16.8) glycine are present in the formulated plasma of the present invention, which is within the range of a SDSAS of about 1 mM to about 50 mM and amino acid of about 1 mM and about 150 mM (e.g., 50mM to about 100 mM). The following table shows how the calculations above were obtained.

[0351] Table 6 Experiment #1 formulation - M= Pre-T # of mmoles Final con. Plasma Glycine, 1.0 M and glycine HCl, mmole / m Sol in In (mL) e(add the volume of pretreatment solution and plasma)

[0352] In an embodiment, the pretreatment solution of the present invention can have a formulation ratio of 405 mM glycine to 98 mM HCl. This embodiment may be advantageously used to treat approximately 266 mL of plasma to be dried with 53 mL ofDocket No. 0118.014100 -80- pretreatment solution made from 1.61 g of glycine and 0.52 g of HCl 36.5 or the equivalent amounts. This formulation results in approximately 67 mM of glycine and approximately 16 mM HCL in the approximately 319 mL of formulated plasma before that plasma is spray dried.

[0353] A study of pooled, ABO matched, never frozen plasma (NFP) derived from whole blood treated with the citrate phosphate dextrose (CPD) anticoagulation regimen and pretreated with pretreatment solution of the present invention demonstrated that dilution of the pretreatment solution of the present invention by plasma to be dried by +20%, + 10%, +5%, -5%, -10%, and -20% had no meaningful impact on the characteristics of the dried plasma when reconstituted and then assayed by a 25 assay panel. In particular, the impact of the pretreatment dilution range on the sensitive vWFRCo assay showed normalized percent recovery of vWF by the assay to be within the error range displayed by the control plasma. pH was also well controlled, ranging between 7.32 and 6.84 with the pH of the control plasma being 7.11.

[0354] These results demonstrate that the pretreatment solution of the present invention reliably permits the production of spray dried plasma which displays, after reconstitution, assay panel comparable results to that of NFP, FFP and PF24 and with assay characteristics essentially the same as NFP, FFP and PF24.

[0355] The pretreatment solution of the present invention can have a ratio by weight of to HCl of between approximately 5 and 3. The formulated plasma which has been treated with the pretreatment solution of the present invention can have a ratio by mmol / ml of glycine to HCl of between approximately 5 and 3.

[0356] In an embodiment, the present invention involves adding a volatile acid and an amino acid as separate compounds (e.g., not as a salt) to create the solution. The volatile acid and amino acid should be added in amounts that results in a SDSAS of about 1 mM to about 50 mM and amino acid of about 1 mM and about 150 mM. In the case where there is a 1:1 relationship between the volatile acid and the amino acid, such as is the case with HCl and glycine, each are added in equal amounts ranging between about 1 mM to about 50 mM and additional amounts of amino acid is added to achieve the free amino acid concentration of about 1 mM and about 150 mM in solution. In other embodiments, a corrosive strong acid can be converted to an acidic salt for use in pretreating plasma priorDocket No. 0118.014100 -81- to spray-drying. Examples include NaHSO4and NaH2PO4: namely the acidic salts of sulfuric acid.

[0357] In an embodiment, the pretreatment solution has glycine in an amount ranging between about 10 µmole / mL of plasma and about 110 µmole / mL of plasma (e.g., about 10, 20, 30, 40, 50, 60, 70, 80. 90. 100110 µmole / mL of plasma), and hydrochloric acid (HCl) in an amount ranging between about 10 µmole / mL of plasma and about 30 µmole / mL of plasma (e.g., about 10, 15, 20, 25, and 30 µmole / mL of plasma), to thereby obtain formulated plasma. In an embodiment, the pretreatment solution has glycine in an amount of about 84 µmole / mL of plasma and HCl in an amount of about 20 µmole / mL of plasma.

[0358] In another embodiment, the pretreatment solution has an amount of glycine and an amount of HCl that forms a ratio that allows for free glycine to be present in the pretreatment solution, In one aspect, the ratio of glycine to HCl is between about 1.5 and about 8.0 (e.g., 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.5, 7.0, 7.5, 8.0). In a certain embodiment the ratio of glycine to HCl is 4.15. In yet another embodiment, the ratio of glycine to HCl effects a pH of the pretreatment solution to be between about 2.0 and about 4.0, or results in a formulated plasma of step a) has a pH of about 6.0 to about 6.6. Once reconstituted with sterile water, the formulated plasma having the above-referenced ratio of glycine to HCl results in a pH of about 6.7 to about 7.8.

[0359] The present invention further includes a method of producing spray dried plasma by combining plasma with a pretreatment solution, wherein the pretreatment solution has glycine in an amount ranging between about 15 mmol and about 30 mmol (e.g., about 15, 20, 25, and 30 mmol), and HCl in an amount ranging between about 3 mmol and about 7 mmol (e.g., about 3, 4, 5, 6, and 7 mmol). In a certain embodiment, the pretreatment solution has glycine in an amount of about 22 mmol and HCl in an amount of about 5.3 mmol.

[0360] Non-volatile acids and acidic salts are collectively defined as and included as spray dry stable acidic substance (SDSAS's) in this invention. The pretreatment solution of the present invention includes, in an embodiment, the SDSAS and one or more amino acids.

[0361] In an embodiment, the SDSAS and / or one or more amino acids of the present invention is added to the plasma within about 30 minutes, about 20 minutes, about 15 minutes, about 10 minutes, about 5 minutes, about 1 minute or time zero (0 minutes) of spray drying the plasma. In an embodiment, the SDSAS of the present invention is addedDocket No. 0118.014100 -82- contemporaneously to the plasma as the plasma is being pumped into the spray drying apparatus. The term "contemporaneously" shall be defined herein as meaning within about 60 seconds, about 50 seconds, about 40 seconds, about 30 seconds, about 20 seconds, about 10 seconds, about 5 seconds, about 1 second and about 0 seconds. With the addition of the amino acid to the SDSAS which increases the pH of the pretreatment solution, in an embodiment, the plasma formulation (e.g., pretreatment solution and plasma to be spray dried) may be able to be stored or allowed to sit for up to about 24 hours (e.g., 1, 5, 10, 15, 20, or 24 hours) before spray drying.

[0362] In an embodiment, the present invention includes mixing the pretreatment and the plasma to be spray dried using a technique called rapid mixing. The rapid mixing step is optional. One of the inventive discoveries includes that rapid or instant mixing of the pretreatment composition and the plasma. It was discovered that slowly mixing the pretreatment solution with the plasma allows localized contact or pockets of unmixed acid to contact the plasma proteins, which can harm these proteins and specifically increase C5a. In contrast, when rapidly mixing and / or agitating the pretreatment solution with the plasma, in an embodiment, amounts of C5a are similar that of fresh frozen plasma or other similar FDA approved products on the market. Rapid mixture and / or agitation allows for instant, thorough and rapid mixing of the pretreatment solution (e.g., having a SDSAS and one or more amino acids) and the plasma. See Examples 17 and 18. Rapid mixture is defined as adding a large volume of plasma to a relatively small volume of a pretreatment solution, prior to spray drying the plasma. In general, when adding a large volume to a much smaller volume (e.g., a volume that is between about 10 and about 30% (about 10, 15, 20, 25, 30%) of the large volume), the mixing of the two volumes results in a rapid and thorough mixture of the two volumes. In a preferred embodiment, 260 mL of plasma is added to 50 mL of the pretreatment solution. In an embodiment, once rapid / instant mixing occurs, the operator can gently invert the bag having both the pretreatment solution and the plasma a few times (e.g., 1-5 times) to further mix the two together. By contrast, with respect to mixing the pretreatment solution with plasma, when pouring a small volume of pretreatment solution into a large volume of plasma to be spray dried, it takes longer for the small volume to be well mixed into the larger volume and pockets of the small volume can form within the larger volume. During this time it wasDocket No. 0118.014100 -83- discovered that the localized contact or pockets of unmixed acid formed within the mixture caused an increase in the amount of C5a in the resulting reconstituted plasma. Agitation is defined as a constant shaking or movement of components (e.g., SDSAS, amino acid, and plasma) of a pretreatment solution. Rapid mixture or agitation results in a uniformly mixed plasma formulation with little or no localized contact or pockets of unmixed acid.

[0363] The present inventions relate to adding SDSAS and at least one amino acid to blood plasma to be spray dried in a time period prior to spray drying short enough to obtain a formulation with the desired pH ("plasma formulation") and to prevent denaturing or damage of certain plasma protein(s) such as von Willebrand's factor due to prolonged exposure to the low pH condition or prevent the increase of C5a. In an embodiment, keeping the time delay to 30 minutes or less between formulation of the plasma with SDSAS and spray drying, as described below, results in improved recovery of plasma proteins, including von Willebrand factor, without undesirable protein damage due to prolonged exposure to the low pH condition prior to spray drying.

[0364] The time period between pretreatment formulation and spray drying will depend on the pH / acidity of the plasma formulation created by the mixing of the SDSAS, an amino acid, and the plasma. In an embodiment, the time period between contacting the SDSAS and an amino acid, with the blood plasma and spray drying the plasma is in a range between about 0 seconds (e.g., at the time aerosolization occurs: time 0) and about 30 minutes. In an embodiment, to minimize protein denaturing, the time between adding of the pretreatment solution to the plasma and spray drying should be kept to minimum. The actual maximum time between formulation and spray drying is determined empirically. This close-in-time formulation at time 0 is referred to herein as "contemporaneous formulation."

[0365] There are a number of methods by which contemporaneous formulation may be carried out. In one embodiment a formulation station is provided in association with the spray dryer. In conjunction with the formulation station, the weight or volume of the pre- spray dried plasma is determined and an SDSAS and amino acid dose measured to obtain the desired pH of the plasma formulation. The dose may be introduced into the plasma by any convenient method including by injection through a port on the plasma bag. In an embodiment, the bag containing the plasma and the bag containing the pretreatment solution are sterilely connected by tube using a tube sealer that can sterilely heat seal twoDocket No. 0118.014100 -84- ends of a tube together. In such a case, the transfer of the plasma to the pretreatment bag can be done manually or with the use of a collection monitor or scale. Gravity can be used to assist the transfer by hanging the plasma bag higher than the pretreatment bag. A formulation station may be manually, semi-automatically or automatically operated. Naturally, the timing of the dosing should be controlled as described above. Timing control may be manual, semi-automatic or automatic.

[0366] In another embodiment, an appropriate dose of SDSAS and one or more amino acids is introduced into the plasma flow channel of the spray dryer prior to the spray drying head. The pretreatment solution introduction is controlled manually, semi- automatically or automatically to result in the desired plasma formulation.

[0367] In a further embodiment, an appropriate dose of the pretreatment solution (e.g., SDSAS and one or more amino acids) is introduced into the spray drying chamber sufficiently close to the spray drying nozzle so that the pretreatment solution and plasma are mixed together to form a plasma formulation before spray drying occurs in the spray drying chamber connected to the spray drying head. Pretreatment solution (e.g., SDSAS one or more amino acids) introduction is controlled manually, semi-automatically or automatically to result in the desired plasma formulation.

[0368] In yet another embodiment, the pretreatment solution is combined with the donor plasma using a sterile connection device and a scale, as further described herein.

[0369] C5a G protein-coupled receptors are prevalent throughout the human body, comprising approximately 60% of known cellular receptor types, and mediate signal transduction across the cell membrane for a very wide range of endogenous ligands. They participate in a diverse array of physiological and pathophysiological processes, including, but not limited to those associated with cardiovascular, central and peripheral nervous system, reproductive, metabolic, digestive, immunological, inflammatory, and growth disorders, as well as other cell-regulatory and proliferative disorders. One of the most intensively studied G protein-coupled receptors are the complement (C) system of humans and other mammals that involves more than 20 components that participate in an orderly sequence of reactions resulting in complement activation. The blood complement system has a wide array of functions associated with a broad spectrum of host defense mechanismsDocket No. 0118.014100 -85- including anti-microbial and anti-viral actions. Products derived from the activation of C components include non-self-recognition molecules C3b, C4b and C5b, as well as the anaphylatoxins C3a, C4a and C5a that influence a variety of cellular immune responses. These anaphylatoxins molecules are involved in pro-inflammatory actions, both acute and chronic inflammation, and its accompanying pain and tissue damage.

[0371] The data reveals the surprising result that the addition of an amount of glycine to an acid pretreatment solution increases the pH of the pretreatment solution, providing additional protection to plasma proteins and mitigating C5a elevation. The addition of appropriate level of glycine does not affect the ultimate pH of the rehydrated spray dried plasma (ODP).

[0372] It is desirable for the pretreatment solution of present invention to result in rehydrated plasma that has reduced levels of C5a or levels in FDA approved apheresed plasma products. In particular, the present invention involves a pretreatment solution that results in C5a levels similar to fresh frozen or never frozen plasma or available FDA approved apheresed plasma products. As shown in the results of Fig. 28, Example 17, if localized contact with a low pH acid can be avoided, C5a levels are similar to that of Never Frozen plasma. Localized contact with a low pH acid can be avoided by increasing the pH using an amino acid, such as glycine, or utilizing a rapid mixing technique described above, or a combination thereof. As can be seen from Fig. 28, when using Glycine HCl having a pH of 1.32 without the addition of a more basic amino acid such as glycine, the rapid mixture technique reduces the C5a levels from about 64 ng / mL to about 31 ng / ml. When lowering the pH by adding glycine to the SDSAS (e.g., the Glycine HCl / Glycine pretreatment solution), the rapid mixture technique resulted in a C5a level to about 12 ng / mL, close to the level of Never Frozen Plasma (NFP) which is about 10 ng / ml. Using citric acid, having a pH of 2.28 by itself without an amino acid addition and using the rapid mixture technique also results in C5a level similar (12.66 ng / mL) to NFP. The pH of citric acid is higher than that of glycine HCl. When pretreatment solution of citric acid is combined with an amino acid (glycine), the pH of the solution is 3.4 and the rapid mixture does not really affect the C5a level, as both are close to NFP, e.g., about 10 ng / mL. It has been discovered that desirable C5a levels result from a pretreatment solution having an SDSAS and an amino acid addition, rapid mixture / agitation of the pretreatment components, or the combination of both. InDocket No. 0118.014100 -86- particular, levels of C5a for reconstituted plasma resulting from the pretreatment solution of the present invention can be between about 4.7 ng / mL to about 74 ng / mL and in particular between 8 ng / mL and 12 ng / mL (e.g., about 10 ng / ml). C5a levels are reduced, as compared to plasma not subjected to a pretreatment solution having at least one SDSAS and at least one amino acid. In an embodiment, the C5a levels are reduced by about 20% or less (e.g., 20%, 15%, 10%, 5%, 1%), as compared to plasma not subjected to the pretreatment solution of the present invention. In another embodiment, referring to Fig. 28, C5a levels, with rapid mixing and with certain pretreatment formulations, result in about levels the same as that of never frozen plasma, or within about 20% or less (e.g., 20%, 15%, 10%, 5%, 1%) of C5a in never frozen plasma or already approved FDA apheresed plasma products.

[0373] Similarly, in an embodiment, rapid mixture and / or agitation are not necessary for pretreatment solutions having a pH of about 3 to about 6 (e.g., 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0). An important discovery is that although the pH of the pretreatment solution changes e.g., ranges from about 1 to about 4, the pH of the plasma stayed about the same e.g., about 6.2.

[0374] Protein Stability of Pre-Treated Plasma Proteins potentially undergo physical degradation (e.g., unfolding, aggregation, insoluble particulate formation) by a number of mechanisms. Many proteins are structurally unstable in solution and are susceptible to conformational changes due to various stresses encountered during purification, processing and storage. These stresses include temperature shift, exposure to pH changes and extreme pH, shear stress, surface adsorption / interface stress, and so on. Proteins in solutions can be converted to solid formats (i.e., converted to a powder or other dry format by having the water and other volatile components of the protein solution greatly reduced or removed) for improved storage using a number of methods.

[0376] Freeze drying (also known as lyophilization) is the most common processing method for removing moisture from biopharmaceuticals, and can increase the stability, temperature tolerance, and shelf life of these products. It is a process wherein a suspension, colloid or solid is frozen and then "dried" under a vacuum by sublimation (phase transition). In this process, proteins can suffer from cold denaturation, interface stress ( adsorption at the water / ice-interface), exposure to increasing alkaline pH (CO2loss), and dehydration stress. Freeze drying is well established within the industry.Docket No. 0118.014100 -87- However, it requires expensive equipment that takes up a great deal of space within a production facility. Freeze drying also can take days to complete, and manufacturers that need a powdered product must incorporate a granulation step to the process. In an environment where budgets are tightening, and where time and facility space are at a premium, freeze drying might be a difficult option for some companies. Because of the space needed, drying plasma by freeze-drying technology is limited to plasma manufacturers, and cannot be implemented in blood centers.

[0377] Because of the difficulties inherent with freeze drying of plasma with regard to time, space and cost, the present invention is directed towards an improved spray drying process for plasma that overcomes the known difficulties related to the spray drying of plasma.

[0378] In the spray-drying process, the viscous liquid is pumped through the feeding line to the nozzle, where the exiting fluid stream is atomized into numerous droplets under aerosol gas. The liquid droplets are met with dry gas and turned into dry particles. It is a much shorter and less expensive process than the freeze drying process, allowing it to be implemented in research labs and blood centers. However, before the present invention, in this process, plasma proteins can suffer from extensive shear stress, interface stress, thermal stress, dehydration stress and exposure to extreme pH.

[0379] Aerosolization exposes the liquid sample to shear stress and produces an extremely rapid and very large expansion of the air-liquid interface. The synergistic effects of shear stress and air-liquid interfacial stress can cause severe detrimental effects on labile compounds such as proteins. Complex biological molecules are difficult to spray dry because they are very sensitive to high shear stress. Although some control relating to the amount of shear stress encountered can be obtained by, for example, choice of the type of atomizer used and the aerosolization pressure used, it is very challenging to apply spray drying technology to human plasma because it contains so many diverse proteins. The diverse proteins may be susceptible to different stresses and this can make it difficult determine processing conditions suitable for all of the types of proteins found in plasma. In particular, vWF, which is designed by nature to be shear sensitive for its biological functions, is the most shear-force sensitive human plasma protein. Most of the other plasma proteins remain largely intact after spray dryingDocket No. 0118.014100 -88- except vWF. As shown in the Examples section, spray-drying diminished vWF activity to below the level of detection (see, Example 27, Fig.17).

[0380] Ionizable amino acid residues have been shown to play important roles in the binding of proteins to other molecules and in enzyme mechanisms. They also have a large influence on protein structure, stability and solubility. The types of interactions these side chains will have with their environment depend on their protonation state. Because of this, their pKa values and the factors that influence them are a subject of intense biochemical interest. Strongly altered pKa values are often seen in the active sites of enzymes, to enhance the ability of ionizable residues to act as nucleophiles, electrophiles or general bases and acids. As a consequence of the change in protonation of these residues, the stability of proteins is pH-dependent. Therefore, it is believed that inhibition of the alkalization of plasma during spray drying can potentially improve the processing and storage stabilities of many plasma proteins.

[0381] As mentioned above, the spray drying process subjects plasma proteins to different forces than those are found in the lyophilization process. First, spray drying exposes plasma proteins to high stress forces during the aerosolization process as the plasma is forced through the narrow orifice exposed to high rate of air flow that is necessary to create suitably sized droplets for drying. Second, the spray drying process exposes plasma proteins to high temperatures that are necessary to force the water from the aerosolized droplets. Third, the spray drying process subjects the plasma proteins to dramatic and rapid increases in pH as a result of the rapid release of CO2during drying. Since lyophilization does not subject plasma proteins to these forces, and especially to this unique combination of forces, one of ordinary skill in the art would not look to nor find suggestion or motivation in the lyophilization art with regard to improving the spray drying process for plasma.

[0382] In spite of the difficulties associated with the spray drying of plasma, a spray drying process of the present invention results in high recovery and high stability of functional plasma proteins, especially, but not limited to vWF, wherein the recovery of vWF is in an amount in rehydrated spray dried plasma that is at least about 5 percentage points or greater (e.g., about 5, 10, 20, 30, 40, 50, 60, 70, 80 percentage points or greater) as compared to amounts of active / undenatured vWF of rehydrated spray dried plasma that does not undergo the pretreatment steps of the present invention.Docket No. 0118.014100 -89-

[0383] The compositions and steps of the present invention relate to the impact of the formulation of liquid plasma with a SDSAS, for example, glycine HCL alone or in conjunction with an amino acid on the recovery from the spray drying process and stability (during storage of dried and rehydrated plasma after spray drying) of functional vWF and other coagulation factors. This can be done by adding a SDSAS such as, for example, glycine HCl, citric acid or lactic acid alone or in conjunction with an amino acid such as glycine to the liquid plasma before spray drying begins or contemporaneously with the spray drying process. During the spray drying process, CO2 loss occurs which causes the pH of the plasma composition to become more alkaline (e.g., to increase) and adding SDSAS and an amino acid thereby maintain the plasma pH in a range to prevent significant denaturing of the clotting factors, esp. vWF. Thus, the pretreatment of plasma with citric acid, glycine HCl or other SDSAS, or in conjunction with an amino acid serves at least three main purposes: 1) increases in-process recovery of plasma proteins; 2) increases stability of plasma proteins during storage; and 3) allows spray dried plasma to be rehydrated with water (e.g., sterile water, WFI), eliminating the need for a specific rehydration solution.

[0384] When liquid plasma is formulated with SDSAS and an amino acid before it is dried, the acid resides in the dried plasma product at a level consistent to improved storage lifetime and reduced degradation of clotting factors during storage. A "level consistent to improve storage lifetime" also means, herein, at a level that results in a physiological pH upon reconstitution of the spray dried plasma. The use of the SDSAS and the amino acid also permits simple rehydration by low cost, readily available sterile water for injection or, in an emergency, plain water at a physiological pH. The convenience, lowered cost and improved safety associated with direct rehydration by water is evident. Advantages include savings in being able to ship dried plasma product without the weight and bulk of rehydration fluid and savings in the cost from not having to specially formulate rehydration fluid and reduction or elimination of refrigeration or freezing during storage.

[0385] Thus, the inventors have discovered that plasma formulation by a SDSAS and an amino acid results in spray dried plasma that has high recovery or functionality of plasma proteins, especially vWF, highly improved storage properties of the dried plasma and approximately neutral pH when rehydrated with water without a buffering rehydration fluid. Thus, the present invention permits spray dried plasma to be manufactured withoutDocket No. 0118.014100 -90- the additional expense and complexity of pretreatment with additional stabilizers such as polyols and others known in the art. However, the use of stabilizers is not contraindicated and may be beneficial in some instances.

[0386] In a further embodiment, a new composition of matter for blood plasma spray drying is created by dosing by any means the blood plasma prior to spray drying with added citrate (i.e., citric acid) or other suitable SDSAS and amino acids at appropriate concentrations, as disclosed herein.

[0387] In a further embodiment the newly dosed citrate formulated blood plasma before spray drying has a concentration of citrate of about 27.5 mM and about 40.4 mM, or of about 31.6 mM and 34.2 mM.

[0388] In a further embodiment a new spray dried blood plasma product is created by spray drying blood plasma formulated with appropriate levels of a suitable SDSAS (e.g., citric acid) and a suitable amino acid prior to or contemporaneously with drying and then drying the blood plasma to the desired level of moisture. The desired level of moisture is generally less than 2%.

[0389] In various embodiments, citric acid or other SDSAS and amino acids are added to the plasma as a formulation. Experiments relating to the effect of SDSAS and amino acids on protection of the activities of proteins found in plasma are explained further in the exemplification section of this specification. The concentrations at which citric acid, for example, is used are between about 1 to about 15 mM. or between about 5 mM to about 10 mM (e.g., 7.4 mM). Accordingly, plasma proteins can be preserved better when citric acid, at the indicated concentrations, is added to it prior to or contemporaneously with spray drying. The activity of vWF is provided in the exemplification because this factor is especially sensitive to denaturing and damage by spray drying (See, Fig. 17 and Fig.18) and, thus, is a good indicator protein to show the beneficial effects of SDSAS and amino acids with regard to recovery and stability of the spray dried plasma proteins.

[0390] Examples of other physiologically compatible SDSAS and amino acids are known to those of ordinary skill in the art and described herein.

[0391] In an embodiment, single donor plasma expressed from collected whole blood or by apheresis which has never been frozen and is less than 24 hours old from collection is desirably utilized for this process. The plasma is collected from blood by standardDocket No. 0118.014100 -91- techniques known to those of ordinary skill in the art, as described herein. Plasma is collected through a process call plasmapheresis. Plasmapheresis refers to a procedure in which the plasma is separated from the blood either by centrifugation or membrane filtration. The system process is also usable with pooled plasma if such is desired and with starting blood plasma material made with any currently available anti-coagulation system such as those known as CPD, CP2D, ACD-A and ACD-B. A sterile, non-pyrogenic, single-use container with SDSAS e.g., a 50 ml solution glycine and hydrochloric acid packaged in a 500 ml container within an overwrap pouch. In an embodiment, the process of the present invention includes converting a single donor unit of plasma which is collected by standard procedures into a single unit of spray dried plasma.

[0392] The dried plasma of the present invention can be from a third-party donor or from the potential recipient himself / herself. The latter is known as autologous plasma. Autologous plasma is highly desirable because it resolves compatibility issues, e.g., it does not need to be ABO / Rh typed before use and is unlikely to carry any foreign substance such as pathogens or immunogens to the recipient whose plasma it is.

[0393] The in vitro characterization data demonstrate that the manufacturing effects of the system are comparable between units manufactured with different starting materials. Units manufactured from apheresed plasma (ACD-A anti-coagulation treatment) showed similar percent change due to manufacturing effects on the starting material as compared to units manufactured from whole blood derived plasma (CPD anti-coagulation treatment). A statistical analysis (ANOVA) was performed on the percent change pre and post manufacturing between the two starting materials across 20 assays including clotting times, coagulation function, and activation markers. Of the 20 assays, total protein concentration, PT, TT, and Factor VIII and XIII activities were determined to be statistically significantly different, however, the mean percent change is similar, and the mean values are all within the clinical reference range. In summary, the in vitro test results support the conclusion that the manufacturing impact on both apheresed and whole blood plasma is comparable, and the coagulation profile is within ±20% of their paired control or within the normal reference range.

[0394] Spray Dryer and the Spray Drying Process In general, a spray dryer system (spray dryer device) is provided for sprayDocket No. 0118.014100 -92- dryinga liquid sample such as blood plasma.

[0396] The pretreated plasma is dried with the components and system for using a spray drying disposable device. The spray drying system include a spray drying apparatus (hereinafter referred to as “drying apparatus,” “machine,” “spray dryer” or “dryer”), a spray drying finishing apparatus (hereinafter referred to as “finishing apparatus” “seal and separator,” or “finisher”) and a spray drying disposable device (hereinafter referred to as “disposable device” or “disposable”). The present invention includes a system that allows the spray drying disposable device having a liquid atomization nozzle and drying chamber that efficiently dries liquids including liquid human or animal blood plasma while protecting the active components such as plasma proteins. The spray drying disposable device is installed in the spray dryer that controls plasma flow, pressurized aerosol gas flow, drying gas flow, temperatures, pressures, etc. within the disposable. Once the spray drying process is complete, the disposable having dried plasma powder is aligned and processed by a spray drying finishing apparatus in which a portion of the disposable is sealed and separated to become the dried plasma unit. Moreover, the invention advantageously provides apparatuses for carrying out functions of spray drying and finishing products including dried human blood plasma.

[0397] The spray drying disposable of the present invention has compact drying chamber producing dried powder (<2% residual moisture) with a high powder production rate. The disposable is small, readily handled, and easy to use drying chamber with high performance. The drying systems of the present invention are a significant improvement providing a removable, disposable drying chamber for spray drying suitable for small batch size processing, such as individual blood units.

[0398] Certain older disposable drying chambers of the Applicant were quite long, being between 58” and 66” or more in length, to allow enough time (flight path) for the plasma to be dried to an acceptable residual moisture level. See Applicant’s Patent Nos. 8533971, 8595950, 8434242, 8601712, 8533972, and 10843100. However, their length made those prior art disposables unacceptable in practice for use because they were difficult and inefficient to handle during installation in the spray dryer instrument. The shorter disposable of the present invention, as further described herein, is more easily handled than these prior art disposables which required reaching and stooping distances for users of overDocket No. 0118.014100 -93- 6’ and under 5’ respectively. The shorter disposable makes the spray drying of human blood plasma practical in real world applications by real world people. Also, the disposable drying chamber of the present invention is a removable, disposable drying chamber that preserves quality and integrity of the plasma while improving processing time and product quality at reduced cost.

[0399] Several challenges were overcome to shorten the drying chamber of the present invention. For example, drying any product to a given degree of dryness involves exposing the material to be dried with enough heat energy to obtain the desired drying level while maintaining the functionality of the substance being dried. However, shortening the drying chamber also reduces the drying pathway.

[0400] The disposable drying chamber of the present invention is improved by:

[0401] Plasma being more efficiently manufactured;

[0402] Being considerably shorter;

[0403] Being readily usable by persons of a wide range of statures;

[0404] Drying material in less time;

[0405] Reducing the inlet air temperature;

[0406] Achieving a nozzle assembly and drying environment to obtain rapid mixing of the atomized droplets with the drying gas and rapid evaporation;

[0407] Achieving a lower level of residual dryness e.g., less than 2.5% residual moisture; and

[0408] Utilizing a specially designed, cost-efficient composite spray drying nozzle, as described herein.Docket No. 0118.014100 -94-

[0409] Overview Of Spray Dry Disposable

[0410] In particular, disposable 100 has two general areas, the spray drying head 2 and the plasma drying chamber 28. Spray drying head overview Spray drying head 2 of disposable 100 that has guide 4 that is offset as positioned on plenum 6, and baffle plate 8 having ridge 9 (Figs. 42A and 43A). Plenum 6 has guide 4 on top of spray drying head 2. Within guide 4 is spray dry nozzle assembly 20 which has plasma flow inlet 18 connected to the liquid plasma via plasma tube 16 and pressurized aerosol gas inlet 14 connect to the pressurized gas via aerosol tube 10 and aerosol filter 12. Additionally, drying gas inlet port 22 is shown and is in communication with the drying gas source (not shown) which may be a source of air, nitrogen or other drying gas. Optionally, drying gas inlet port 22 may be covered by a removable cover such as a self- adhesive paper label or similar. This cover should be removed just prior to installation of disposable 100 into the spray dryer 200. The drying case source can optionally be in communication with a moisture reducing drying system. In one embodiment, the drying gas source is an Atlas-Copco SF 22+ compressor (Atlas Copco Nacka Municipality, Sweden) in conjunction with an Atlas-Copco CD45 desiccant drying system supplying clean dry air (CDA) to the spray dryer and heats air to the appropriate temperature for spray drying. In an embodiment, the drying gas flows through a filter from the CDA and, for example, is a Millipore Series 30000.2 micron filter CTGB71TP3 from Millipore Sigma of Danvers MA USA. The CDA supply is used, in an embodiment, for the supply for the drying gas and for the pressurized gas. In certain embodiments spray drying nozzle assembly 20 includes a “manifold” that coordinates the plasma and aerosol lines. When the plasma source, pressurized gas source, and drying gas source combine, the liquid plasma droplets are formed and dried into dried plasma (e.g., a fine, amorphous plasma powder). Plenum 6 has a notch, which is a locator referred to herein as locator 26 or a second locator, as further described herein.

[0413] Briefly, guide 4 fits into receiver 204 of spray drying apparatus 200 which also properly aligns disposable 100 with drying apparatus 200 (Figs. 45B and 45C). Guide 4 also aligns spray drying head 2 with respect to spray dryer 200 in a specific orientation such thatDocket No. 0118.014100 -95- drying gas inlet 22 receives the drying gas source (not shown). Ridge 9 fits into ridge receiver 207 of spray dryer 200 and provides support. Guide 4 along with ridge 9 allows alignment of disposable 100 with spray dryer 200 in a latitudinal orientation (e.g., in a plane defined by the top surface and bottom surface of the spray drying apparatus) which keeps the disposable secured so it does not move up and down within the spray drying chamber housing of the dryer. Additionally, ridge 9 of disposable 100 fits into receiver 404 of finisher 400 to secure disposable 100 to finisher 400 while finisher 400 is moving the plasma and sealing and separating the disposable to turn it into dried plasma unit 60. See Figs. 46A-C. This alignment arrangement also provides for easy, universal attachment of the disposable to both the dryer and the finisher.

[0414] First locator, locator 206 (Figs. 45B, 45C and 46A), is positioned on spray drying apparatus 200 and the second locator, locator 26 (Figs. 42A and 43A) is positioned on spray drying disposable 100 such that the first and second locator engage during installation of disposable 100 into spray drying apparatus 200 to allow for alignment of the disposable with the spray drying apparatus. The same locator, locator 26 (the second locator), on the disposable also is used to align the disposable with a third locator, locator 452 (see Figs., 47A-C), on spray drying finishing apparatus 400, the apparatus that directs the dried plasma into specific compartments of the disposable, seals and separates the dried plasma into a plasma unit having the dried plasma. This locating arrangement aligns the disposable to the spray drying apparatus axially, e.g., about an axis defined by the center of a receiver of guide 4 (see Axis A of Fig. 43A). This locating arrangement allows for easy universal attachment of the disposable to both the drying apparatus and the finishing apparatus.

[0415] As part of the disposable, spray drying head 2 includes nozzle assembly 20. This nozzle assembly allows the spray drying of the plasma to occur within the disposable. Overall, the design of the system has a spray dryer and disposable modified to have a nozzle as part of the disposable instead of the spray dryer so that spray drying occurs entirely within the disposable. This design helps keep the plasma in the disposable throughout the drying and finishing process, and out of the parts of the dryer or finisher which would require decontamination between each use. The design also minimizes external pathogen contamination by keeping the plasma within the disposable during the entire process. The nozzle assembly coordinates the plasma flow and the pressurized / aerosolized gas flow suchDocket No. 0118.014100 -96- that both are emitted at the proper rates and air flow to atomize the liquid plasma at tip of the nozzle where it is ready for rapid mixing with the drying gas. Spray drying head 2 of disposable 100 further includes plenum 6 and baffle plate 8 that guides the drying air for rapid mixing with aerosolized plasma and creates an air curtain to minimize buildup of dried plasma on the drying chamber wall.

[0416] Plasma drying chamber overview Drying chamber 28 is the area of the disposable where the plasma dries. The drying chamber is designed to capture the dried plasma while allowing the humid air to exit. The design of the drying chamber also allows the drying chamber to be sealed and separated in such a way as to form the commercial dried plasma unit.

[0418] Drying chamber 28 has three general areas, the upper portion defined by Dimension X (See Figs. 44 and 46A), the mid-section defined by Dimension U, the area between locations 44A and 44B, and the bottom portion defined by Dimension V, the portion below location 44B, that includes filter 36 and a separator 38. The upper portion is a space in which the atomized liquid plasma hits the drying gas and evaporates the liquid within the droplet and dries. In particular, the atomized plasma rapidly mixes with the drying gas and dries, as further described herein. As the plasma rapidly mixes and dries, it circulates and moves in a downward direction toward the filter. Most of the evaporation occurs in the upper portion of drying chamber 28 (Dimension X) but it does continue to dry as the plasma falls into the midsection portion (Dimension U) and the lower portion (Dimensions V) of drying chamber 28.

[0419] Drying chamber 28 also includes midsection 46, defined by Dimension U, that has “seal and separate” locations 44A and 44B, label 40, spike ports 42A and 42B and hanging slot 34. Midsection 46 also includes locator pin openings 32C. The mid-section is later processed by the spray drying finishing apparatus which involves moving dried plasma into certain locations of the plasma drying chamber and sealing and separating at or near cut locations 44A and 44B. The section between locations 44A and 44B becomes dried plasma unit 60 that will eventually be rehydrated and transfused into patients.

[0420] Disposable 100 further includes a positioning arrangement to reversibly attach the outer wall of disposable 100 to finishing apparatus 400. Positioning openings 32A, 32B,Docket No. 0118.014100 -97- and 32C are present on the outer edge of the wall of spray drying disposable device 100. (Fig. 42A, 48A). Positioning pins 432A, 432B and 432C are located on finishing apparatus 400 such that when positioning openings 32 A, 32B, and 32C are placed around positioning pins 432A, 432B and 432C of finishing apparatus 400, drying chamber 28 of disposable 100 is aligned with on the finisher apparatus. See Fig. 47C, 48B, 48C.

[0421] The lower section of drying chamber 28 includes lower filter 36 (also referred to herein as a “capture filter”), lower filter separator 38, drying gas outlet port 30, and locator pin openings 32A and 32B. Optionally, gas outlet 30 may be covered by a removable cover such as a self-adhesive paper label or similar. In an embodiment, this cover should be removed just prior to installation of the drying chamber into the spray dryer 200. Briefly, the lower filter allows for separation of the dried plasma from the humid air and the separator acts as a spacer between the drying chamber wall and the filter to allow air to more easily pass and prevent pressure buildup. Humid air refers to the air traveling through the drying chamber and includes the combination of the drying gas, the aerosolized gas and the moisture that has been removed from the plasma droplets. During the drying of the plasma, the humid air passes through lower filter 36 and lower filter separator 38, through air flow channels, and out of gas outlet 30 leaving dried plasma in lower filter 36.

[0422] Disposable 100 further includes another alignment arrangement that relates to gas outlet 30 of disposable 100 and gas exhaust port 208 of dryer 200. The spray drying apparatus has gas exhaust port 208 to allow the drying gas to exit and the bottom portion of disposable 100 has gas outlet 30 that fits into the exhaust port 208 of dryer 200. (Fig. 45B, and 46A). Additionally, spray drying finishing apparatus 400 has receiver 414 for the drying gas outlet 30 to secure the bottom of disposable 100 to finishing apparatus 400. (Fig. 48B, 48C). Again, this drying gas arrangement allows for universal attachment of the disposable to both the drying apparatus and the finishing apparatus.

[0423] Additionally, the entire length of the disposable (as measured from the top of the spray drying head to the very bottom of the drying chamber) is limited to about 40 inches or less (e.g., about 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, or 24 inches or less) and preferably about 34.8 inches. A disposable having a length of about 40 inches or less was difficult to achieve because the drying of the plasma occurs in a smaller space and smaller volume but does so gently without degrading plasma proteins. The disposableDocket No. 0118.014100 -98- length, as measured from the bottom of spray drying head 2 or bottom of baffle plate 8 to bottom of filter 36, shown as dimension Y in Fig. 46A, is about 31 inches or less (e.g., about 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19 inches or less) and in an embodiment preferably about 25.9 inches. In another aspect, the area of disposable 100 encompassed by Dimension Z, the length from the bottom of spray drying head 2 and the top of filter 36, is about 22 inches or less (e.g., about 22, 21, 20, 19, 18, 17, 16, 15, 14 inches) and preferably about 19.11 inches. In yet another, the length of Dimension X, the length between the bottom of spray drying head 2 and the top section 46, is less than about 16 inches (e.g., about 16, 15, 14, 13, 12, 11, 10, 9, 8 inches) and preferably about 12.14 inches. In an embodiment, the length of disposable can be modified or shortened. For example, the length of the disposable of the present invention can be further shortened along dimension X by about 1 inch to about 8 inches (e.g., by 1, 2, 3, 4, 5, 6, 7, or 8 inches) thereby reducing the overall length by the same amount. In other embodiments, the disposable can also be shortened anywhere along Dimension Y and Z by the same amount.

[0424] Computational Model For some of the figures, a computational model was used to show flow paths, particle evaporation and the like. Fig. 42B shows the three-dimensional flow geometry of the disposable during operation that was used for the model.

[0426] The three-dimensional model showing in Fig. 42B was based on the disposable shown in Fig. 42A and the dryer shown in Figs. 45A-C. These computer simulations show flows and mixing processes were created by first building the three-dimensional flow domain geometry. See Fig 42B. This geometry was extracted from a computer aid design (CAD) model of the system hardware to create a high-fidelity representation of the flow region inside the ODP system. The flow domain of 0.0195m3volume was discretized into 3.3M spatial cells to generate a computational mesh using the commercially available Ansys- Gambit mesh meshing software. The flow model was calculated using a commercially available computer code; Ansys-Fluent version 2019-R1 running on an HPZ840 multi- processor workstation.

[0427] The simulation utilized a steady-state segregated solver assuming ideal gas properties, K-E turbulence model and the following:

[0428] Drying gas inlet temperature = 114CDocket No. 0118.014100 -99-

[0429] Drying chamber exhaust temperature = 65C

[0430] System heat loss = 0.18kW

[0431] Drying gas flow = 750 slpm

[0432] Atomizer aerosol gas flow = 40 slpm

[0433] Feed rate = 13.5 mL / min, and varies with exhaust gas temperature

[0434] Liquid water droplets with 8.5% non-volatile mass, 5 micron diameter (monodispersed size)

[0435] Exhaust port pressure = 2.76 kPa (0.4 psig)

[0436] The inlet and product capture filters are modeled using a ‘porous zone’ function with flow resistance values set to match the measured pressure during operation in the drying gas manifold of 71.7 kPa (10.4 psig) and 27.6 kPa (4 psig) in the drying chamber at the start of a batch.

[0437] To calculate the average droplet diameter and temperature during the constant- rate evaporation period for a given set of process conditions, two customized c programs, “prsc_udf_multi_2017.c” and “processdata_multi_2017.c”, are developed at PARSEC to obtain an averaged droplet drying pathway from a converged Fluent coupled dpm solution. The program “prsc_udf_multi_2017.c” is used to export droplet tracking data step by step for information interested. The program “prsc_udf_multi_2017.c” reads exported data file generated from the first program, and then get averaged pathway from all tracked particles. Its output file can be read into Excel file.

[0438] The data shown in Figs. 42B, 43Ka, 43Ma, 43Na, 43O, 43P, 43S, 43Sa, and 43T were generated using this model.

[0439] Overview Of The Spray Dryer

[0440] Spray dryer 200 of the present invention provides a donor plasma liquid flow (e.g., pretreated), a drying air flow, a pressurized aerosol gas flow, disposable deflation air line, a disposable exhaust line, a housing exhaust line, and a leak detection line.

[0441] Along with these flow lines, spray dryer 200 of the present invention further includes a leak detection methodology 1000 that utilizes a series of pressure transducers, flow sensors and valves to assess if disposable 100 has a leak during spray drying.

[0442] Spray dryer 200 further includes a pressure detection methodology 1200 thatDocket No. 0118.014100 -100- allows pressure transducers residing outside the wall of disposable 100 to measure pressure inside of disposable 100. This process involves allowing the disposable to heat up which allows the disposable wall to soften. During use, the disposable wall exerts force against force sensors PT08224A and PT09224B which measures such force. The amount of force is used to calculate the amount of pressure within the disposable. If the pressure within disposable 100 is above a set amount (e.g., greater than 7.02 psi), then the computer system causes dryer 200 to enter fail safe mode. If the pressure is within a set acceptable amount (e.g., between about 6.7 and about less than 7.02 psi), then the computer system determines if enough plasma has been dried, by determining the amount of donor plasma left in the donor plasma bag. If a sufficient amount of plasma has been dried, then the computer system communicates that the drying run is a success and the finishing process can begin. If an insufficient amount has been dried, then the drying run fails and disposable 100 is discarded.

[0443] Additionally, dryer 200 includes methodology 1400 that determines the integrity of lower filter 36, baffle filter 94 and / or the plenum interface. This methodology utilizes pressure transducers at the inlet and in the housing, and determines the slope of the pressures during spray drying. The slope is compared against a model and when the slope deviates from the model, then the computer system determines that disposable 100 fails.

[0444] Overview of Finisher Once spray drying is completed on dryer 200, finisher 400 or 400’ of the present invention moves the plasma to the desired compartment, and then seals the walls of the disposable and cuts the walls of the disposable to form a dried plasma unit. Finisher 400 or 400’ provides an impactor, a sealer, a separator, and air extraction.

[0446] Overview of Workflow An overview of the process to use the disposable, spray dryer and finisher described herein, is as follows. The spray drying plasma methodology of the present invention includes pretreating a donated liquid plasma unit or defrosted previously frozen liquid plasma unit, drying the liquid plasma using a spray drying apparatus with the spray drying disposable device that results in a disposable having the dried plasma, finishing the disposable using the finishing apparatus that is designed to seal and separate the disposable, and transform the disposable into a dried plasma unit. The unit can be used or stored. When ready for use, the plasma unit is rehydrated and ready for transfusion into a recipient.Docket No. 0118.014100 -101-

[0448] With respect to pretreatment, the pretreatment process involves adding biocompatible components (e.g., a spray dry stable acidic substance) to the liquid plasma (or defrosted fresh frozen plasma) that protect the plasma proteins during the spray drying process which involves high temperatures and pressures.

[0449] In an embodiment, making the pretreatment solution includes adding the following to a solvent, such as SWFI: between about 3.0 to about 7.0 (e.g., 3.0, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, and 7.0) mmol HCl and about 15 mmol and about 30 mmol glycine (e.g., about 15, 17, 20, 22, 25, 27, 30 mmol glycine) in 50 mL of solvent to obtain 260 mL of formulated plasma. In yet other words, glycine in an amount of about 440 mM, and HCl in an amount of about 106 mM is present in the pretreatment solution. In an embodiment between about 290 mM to about 570 mM (e.g., about 290, 300, 350, 400, 450, 500, 550, and 570) glycine and about 70 mM to about 140 mM (e.g., 70, 80, 90, 100, 110, 120, 130, 140 mM) HCl is present in the pretreatment solution. The pretreatment container is commercially available and can be formulated, filled and finished by e.g., Berkshire Sterile Manufacturing (Lee Massachusetts USA). In an embodiment the pretreatment solution has about 440 mM / 50 ml of glycine and 106 mM / 50 ml of hydrochloric acid. (The United States Pharmacopeial Convention ("USP") monograph (12601 Twinbrook Parkway Rockville, MD 20852-1790, USA)). The pretreatment solution, when combined with liquid plasma to form a formulated plasma, protects the plasma proteins during the drying process. The formulated plasma has a pH in a range between about 5.5 and about 7.2 which offsets spray drying impacts on pH to yield a final rehydrated product that is at normal physiologic pH, a pH range between about 6.5 and 7.8 (e.g., about 6.5, 6.66.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8). pH lower than 6.5 or higher than 7.8, in certain instances, can be detrimental to the recipient. The resulting dried plasma product from the present invention is a plasma that retains its von Willebrand Factor and other blood proteins, and has fewer cholesterol crystals, less particles, less pathogens and a well-controlled pH with the aforementioned pretreatment step. Moreover, the resulting dried plasma has certain properties which are different from and superior to that of freeze-dried plasma.

[0450] A sterile connecting device (SCD), as is known in the art, is used to connect the plasma unit to the pretreatment container and the liquid plasma and in an embodiment, a fixed volume of plasma is transferred utilizing, for example, a blood collectionDocket No. 0118.014100 -102- monitor / mixer. After the liquid plasma is transferred to the pretreatment container, in an embodiment, it is gently mixed in the pretreatment container by inversion. Other mixing methods such as rocking, shaking and agitating, can be used. Additionally, the mixing can be done by the operator or a device known in the art. The bag that contained the liquid plasma is tube sealed, separated, and discarded. Pretreatment container 64 having the pretreatment solution and the liquid plasma (i.e., formulated plasma 66) is then connected to the disposable device at plasma tube 16 utilizing an SCD, resulting in a modified spray drying disposable device, shown in Fig. 42A.

[0451] Spray drying disposable device 100 is a sterile, non-pyrogenic, single user container (e.g., about 35 inches long) which utilizes a pathogen retentive filter to filter air before it enters the drying chamber and as air exits the drying chamber. See Fig. 42A. The spray drying disposable is aseptically connected to the liquid plasma at the plasma tube, tube 16.

[0452] Briefly, the drying process is as follows. See Figs. 45A-4C, 46A-5B. In an embodiment, pretreated plasma is aseptically spray dried in spray drying disposable device 100. See Fig. 45A-4C. During the process, in an embodiment, a positive airflow is maintained. Pretreated plasma is atomized using a nozzle contained within the single use spray drying disposable device creating fine plasma droplets. These droplets are then exposed to heated air. The resulting dried plasma particles are captured in filter 36 of drying chamber 28. The spray drying disposable device is then undocked from the spray drying apparatus and taken to the finishing apparatus.

[0453] An overview of the finishing process is as follows. See Figs. 47A-47C, 48A-C, 34. Once undocked from spray drying apparatus 200, disposable device 100 having dried plasma is transferred to finishing apparatus 400. Finishing apparatus 400 mechanically, acoustically or otherwise impacts or agitates the spray drying disposable device containing dried plasma to eventually consolidate dried plasma powder in the portion of the spray drying disposable device that becomes the spray dry plasma unit. The finishing apparatus utilizes an impactor to first assist the dried plasma in moving to the bottom of the disposable, and then in a second instance to the compartment that becomes spray dried plasma unit 60. Spray dry plasma unit 60 is sealed and separated from the rest of the disposable device utilizing impulse sealing. This step is the final closure step to create plasma unit 60. In anDocket No. 0118.014100 -103- embodiment, the seals are visually inspected and excess portions of the disposable device are discarded and dried plasma unit 60 is produced (see Fig. 49).

[0454] With respect to an overview of the dried plasma storage process, dried plasma unit 60 is removed from the finishing apparatus and stored in a re-sealable moisture barrier foil pouch containing a desiccant. See Patent No. 9561184. In an embodiment, the dried plasma unit is quarantined until completion of all required blood screening tests and stored at refrigeration. Upon meeting final release criteria, the pouch is opened and the dried plasma unit is relabeled for release. The dried plasma unit then placed in a resealable or other pouch, sealed, and stored following storage protocols.

[0455] In an embodiment, the dried plasma unit is compatible with commercially available fluid or other transfer sets for rehydration with sterile water for injection (SWFI). Dried plasma unit 60, once rehydrated, is also compatible with blood administration sets for transfusion. In a particular embodiment, spray dried plasma unit 60 is rehydrated within its existing container using an appropriate amount of sterile water (e.g., 200 mL, 208 mL) for injection prior to transfusion.

[0456] Detailed Description Of Spray Dry Disposable Detailed description of the spray drying head Referring to Fig. 42A, a perspective view of spray drying disposable device 100 is shown. As described above, the disposable has generally two portions, spray drying head 2 and drying chamber 28. The spray drying head includes plenum 6, guide 4, baffle plate 8, baffle filter 94 , nozzle 20, and locator notch 26 (also referred to as a “second locator” herein).

[0459] In an aspect, the purpose of spray drying head 2 is, in part, to A) assist in securing disposable 100 to dryer 200, B) coordinate the flow of the drying air, the aerosolized pressurized gas and the plasma flow, C) house the nozzle assembly, and D) house the baffle filter.

[0460] With respect to securing disposable 100 to dryer 200, the system of the present invention includes an integrated and universal alignment system. In an embodiment, locator notch 26 on plenum 6 is shown in Fig. 43A. Fig. 43A also better shows plenum 6, guide 4, baffle plate 8 and ridge 9. Locator notch 26, also referred to as a second locator, aligns with locator projection 206 (shown in Fig. 43K, 45B, 45C, and 46A), also referred to as a firstDocket No. 0118.014100 -104- locator, on spray drying apparatus 200. This locating arrangement allows for spray drying head 2 of disposable 100 to be aligned axially with spray drying apparatus 200. The locating arrangement can include any arrangement that attaches, fits, complements or otherwise communicates the locator on the disposable with the locator on the drying apparatus. Examples of locating arrangements can include a recess / projection arrangement, complementing shape arrangement, hook / receiver arrangement, channel and groove arrangement, a latch and catch arrangement, a magnetic arrangement, and the like. In Fig. 46A, the male locator is on the spray drying apparatus and a complementing female locator is on the disposable, but the arrangement can be reversed. The complementing nature of the arrangement allows for easy matchup and alignment by the operator and can prevent the door from closing unless the disposable is aligned in the spray drying apparatus. In an embodiment, the locating arrangement can include any arrangement that allows for alignment between the locator on the disposable and the locator on the drying apparatus and also allows for alignment between the disposable and the finishing apparatus. In another embodiment, the drying apparatus and the finishing apparatus have the same locator that fits the locator on the disposable to create a universal alignment. Having a universal arrangement reduces the training needed and increases muscle memory because the operator inserts the disposable into the spray dryer and the finisher in a similar way.

[0461] When the first locator of the spray drying apparatus and the second locator of the disposable are aligned, in an embodiment, the system of the present invention provides positive feedback to the operator. In an embodiment, spray drying apparatus 200 has spring clip 232 mounted to the top of the drying chamber housing and engages guide 4 when the disposable is aligned and secured in the spray drying apparatus. See Fig. 46A. Spring clip 232 is optional. In this case, the positive feedback to the operator is an audible “click”. Such feedback can include an audible indicator (e.g., an audible click) or a visual indicator (e.g., a sensor providing a communication to the display indicating alignment). Retention clip 232 is an alignment element as well since it aligns with ridge 9, further described below.

[0462] Fig. 43A also shows guide 4 which is off set from the center of baffle plate 8. The off-set design of the guide on plenum 6 allows disposable 100 to be attached to the receiver 204 (shown in Figs. 45B and 46A) of spray drying machine 200 in a specific orientation. Prior to inserting the disposable device into the dryer, the operator removes andDocket No. 0118.014100 -105- discards the adhesive covers, if present, from the top, exposing drying gas inlet port 22, and the bottom, exposing gas outlet 30. Use of such covers is optional. In a preferred embodiment, the operator removes and discards the adhesive cover from drying gas inlet port 22 only and inserts spray drying head 2 into spray drying head receiver 404. The cover the drying gas outlet 30 at the bottom of the disposable can be removed later, just before it is ready to be attached to the gas exhaust port 208. The operator generally aligns and inserts ridge 9 formed by baffle plate 8 on spray drying head 2 of disposable 100 into groove 207 of spray dryer 200. See Fig. 46A. Once engaged, the operator can use his / her hands to further push spray drying head 2 inward and it will self- align with groove receiver 207 so long as notch locator 26 on spray drying head 2 is within about 30 degrees (e.g., within about 30, 25, 20, 15, 10, 5 degrees) with respect to alignment with projection locator 206 on dryer 200. The insertion and alignment of the spray drying head can be done rapidly e.g., within 10 seconds (2-5 seconds). Receiver 204 of guide 4 also serves as drying gas inlet on the spray dryer and provides the drying gas source (not shown). Ridge 9 of spray drying head 2 also provides support and fits complementarily into groove 207 of receiver 210. It also allows spray drying head 2 of the disposable 100 to be aligned latitudinally with respect to dryer 200.

[0463] In an embodiment, receiver 210 has groove 207, as shown in Fig. 45B. The ridge and groove arrangement between the spray drying head and the dryer can be any arrangement that allows the spray drying head to fit within the drying chamber housing 202 such that the arrangement provides support and latitudinal alignment. In addition to groove 207, the receiver can be a shelf, ledge, arm, stopper, base or other structure that engages the baffle plate and allows the spray drying head to remain stable throughout the spray drying process.

[0464] The operator then inserts the disposable device by placing guide 4 into receiver 204 of the spray drying apparatus 200. Once inserted and aligned, the spray drying disposable can no longer move up and down. When using this guide and the locating arrangement, described above, they align the disposable so that it cannot move up and down and cannot move axially about the axis defined by the center of guide 4. As shown in Figs. 45B and 46A, the guide fits into receiver 204 and does so such that the fit is snug or tight. In this embodiment, once the spray drying end is aligned and in an engaged position, then theDocket No. 0118.014100 -106- operator can remove the bottom adhesive cover at drying chamber gas outlet 30 and attach it to the gas exhaust port 208, as further described herein.

[0465] When the locating arrangement (locators 26 and 206) is aligned, guide 4 is inserted into receiver 204, ridge 9 is inserted into groove receiver 207, and retention clip 232 is engaged, in an embodiment, spray drying head is inserted, secured and aligned. Specifically, in an embodiment, retention clip 232 engages ridge 9 to hold the spray drying head 2 in place. Retention clip 232 provides an audible indicator that the spray drying head is properly aligned and inserted. In the embodiment shown in Fig. 45B, 45C and 31A, the retention clip is a spring clip. The retention clip can be any type of retainer that engages ridge 9 and include, for example, a fastener, pin, clasp, slide and the like. The retainer can be made from metal, plastic, rubber and the like. The retainer that engages the spray drying head is optional.

[0466] Although in the embodiment shown in Figs. 45B, 45C, and 46A, a spring clip is used as an audible indicator to allow the operator to know that spray drying head 2 of disposable 100 is properly inserted and aligned with dryer 200, any type of indicator can be provided. The indicator can be audible, vision or tactile. In an embodiment, a sound indicator provides audible feedback mechanically or otherwise of correctly completed loading of spray drying head 2 of disposable 100. An audible indicator can be mechanical, like the sound of a spring clip locking into place, or can be generated by sensor (mechanical or pressure / contact sensor) that receives a signal of correct positioning of spray dry head 2 in dryer 200 and communicates with an actuator or processor that provides a visual indicator to the operator, for example, on display 212 or on indicator light 234. In an alternative embodiment, the sensor can send feedback to a processor that causes a sound indicator to a speaker to inform the operator of proper placement. In yet another embodiment, the feedback can be in the form of a tactile response, e.g., a vibration to inform the operator of incorrect or correct placement. Feedback can include an audible indicator (e.g., an audible click) or a visual indicator (e.g., a sensor can provide a communication to the display indicating alignment).

[0467] Fig.43B is an exploding view of spray drying head 2 and shows the parts of spray dry nozzle assembly 20 along with plenum 6, outer filter sealing ring 90, inner filter sealing ring 92, plenum filter 94 and baffle plate 8 having ridge 9. Nozzle assembly 20Docket No. 0118.014100 -107- includes, going from top to bottom in Fig. 43B, strain relief valve 75, plasma and pressurized aerosol gas manifold 72, aerosol reservoir 74, cannula 78 having opening 79, liquid nozzle cap insert 80, and nozzle cap 76. The nozzle cap 76 has opening 110 whose inner wall has a diameter, defined by Diameter Do(See Fig. 43Ia). Fig. 43Ia also shows cannula 78 that has an outer wall (outer diameter) defined by Diameter Dc. Diameter Dois slightly larger than Diameter Dc and the difference is defined by Distance Dd. The resultant difference in diameter, Distance Dd, creates annulus 81 through which pressurized air received from aerosol reservoir housing 74 forms a vortex and flows to the drying chamber 28 to facilitate the formation of small droplets of fluid to be dried. See Example 27.

[0468] Accordingly, the length of the cannula ranges between about 2 and about 5 inches, and in an embodiment, is 3.500 inches + / - .005 inches.

[0469] More specifically, referring to Fig. 43A and Fig. 43B, spray dry nozzle assembly 20 has plasma flow inlet 18 connected to pretreated liquid plasma 66 (shown in Fig. 42A) via plasma tube 16 and pressurized aerosol gas inlet 14 connect to the pressurized gas source (not shown) via aerosol tube 10 and aerosol filter 12. Additionally, drying gas inlet port 22 is shown in Figs. 42A and 43A and communicates with the drying gas source (not shown). When the plasma source, pressurized gas source, and drying gas source combine, the liquid plasma particles are formed under pressurized (aerosolized) gas and dried into a fine dried plasma (e.g., a plasma powder).

[0470] Figs. 43C and 43D show a detailed perspective view of spray dry nozzle assembly 20. In particular, Figs. 43B and 43J show where and how the spray dry nozzle assembly fits within assembly opening 96 of plenum 6 of spray dry head 2. Plasma and pressurized aerosol gas manifold 72 coordinates and directs the plasma source via inlet 18 and the pressurized aerosol gas source via inlet 14. Strain relief 75 fits and communicates with manifold 72 to provide support to tubes 10 and 16 and prevent them from collapsing under pressuring during packaging, transport and spray drying. Strain relief 75 also prevents the tubes from collapsing in the packaging and in transit. Spray dry nozzle assembly 20 includes aerosol gas reservoir housing 74 through which the pressurized aerosol gas is held and builds before being released through liquid nozzle cap insert 80 and nozzle cap opening 110 (shown in Figs. 43G, 43H, 43I, 43Ia, and 43Ic). Nozzle assembly 20 is housed by aerosol gas reservoir housing 74 and secured by nozzle cap 76. Liquid nozzle cap insert 80Docket No. 0118.014100 -108- guides cannula 78 and holds the cannula in place during use. Annulus 81 is disposed between the outer surface of cannula 78 and inner surface of opening 110. The design of liquid nozzle cap insert 80 and nozzle cap 76 allow the pressurized aerosol gas to flow though annulus 81 in a vortex pattern to maximize aerosolization and promote rapid mixing of the aerosolized plasma droplets with the drying gas, as further described herein. The entire nozzle assembly 20 is secured to opening 96 of plenum 6 which includes baffle plate 8 having ridge 9, with filter 94 therebetween and sealed by inner filter sealing ring 92 and outer filter sealing ring 90. See Fig. 43B.

[0471] Fig.43D shows the aerosol gas reservoir housing 74 as transparent so that cannula 78 and attachment to liquid nozzle cap insert 80 and nozzle cap 76 can be seen and Fig. 43E shows manifold 72 and cannula 78 with the aerosol gas reservoir housing 74, liquid nozzle cap insert 80 and nozzle cap 76 removed. Fig. 43F shows the bottom tip, the end opposite the manifold, of cannula 78 having outer wall surface 84, inner wall surface 86, flat edge 88, and beveled or angled edge 82 (e.g., a chamfer) at the bottom surface of the cannula.

[0472] It has been discovered that a cannula with an angled edge (e.g., chamfer) on the inside diameter, when used in spray drying to create the atomized plasma particles, assists or allows many of the proteins in the plasma to remain intact, functional, or both. Hence, the angled edge cannula of the present invention reduces the amount a protein degrades during spray drying because the angled edge cannula reduces shear on the passing liquid plasma film.

[0473] In a particular embodiment, a blood protein, vWF, was measured. vWF is considered a more fragile, easily degradable protein, as further described herein. In an embodiment, using spray dry nozzle with the angled cannula of the present invention, vFW recovery is maintained, as compared to a nozzle with a non-angled cannula. In fact, based on the data described in Example 28, using a composite nozzle with a chamfered cannula resulted in an increase in vFW recovery, as compared to both a composite nozzle having non- angled cannula and to a benchmark stainless steel nozzle (as Buchi Model no. 4244 Buchi Corporation of New Castle, Delaware United States). In an embodiment, using a nozzle with an angled cannula resulted in an increase in an amount at least ranging between about 1% and 25% (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, and 25%) in functional vFW recovery, as compared to a nozzle having non-angled cannula.Docket No. 0118.014100 -109- In particular, as described in Example 28, the data show that spray drying with a chamfered cannula having an angle of 45 degrees and length of 0.005” increased the vWF RCO assay result by about 9%-22%, as compared to the same system operated with a composite nozzle having a cannula without the angled edge and, surprisingly, 3.7% better as compared to the benchmark control Buchi nozzle.

[0474] Among the plasma proteins maintained throughout the spray drying process using an angled-edge cannula, includes von Willebrand Factor (vWF). vWF is involved in clotting, repairing vascular injury and platelet adhesion. In particular, vWF is a large adhesive glycoprotein with established functions in hemostasis. It serves as a carrier for factor VIII and acts as a vascular damage sensor by attracting platelets to sites of vessel injury. The regulation of vWF multimeric size and platelet-tethering function is carried out by ADAMTS13, a plasma metalloprotease that is constitutively active. It is secreted into blood and degrades large vWF multimers, decreasing their activity. Unusually, protease activity of ADAMTS13 is controlled not by natural inhibitors but by conformational changes in its substrate, which are induced when vWF is subject to elevated rheological shear forces. This transforms vWF from a globular to an elongated protein. This conformational transformation unfolds the vWF A2 domain and reveals cryptic exosites as well as the scissile bond. To enable vWF proteolysis, ADAMTS13 makes multiple interactions that bring the protease to the substrate and position it to engage with the cleavage site as this becomes exposed by shear forces. ADAMTS 13 (a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13), also known as von Willebrand factor-cleaving protease (vWFCP), is a zinc- containing metalloprotease enzyme.

[0475] Without being restricted to a theory of operation, it is believed that during spray drying, the plasma proteins are subject to considerable shear forces due to the spraying mechanism as the solutions are fluidized out of the end of a fine nozzle to form the droplets in contact with drying air. The process of unfurling multimeric vWF is expected to be triggered by the hydrodynamic forces of elevated shear stress during spray drying in combination with air-liquid interface stress. The shear-induced structural change of vWF, when combined with other physical factors associated with spray drying, such as high temperature and / or unfavorable pH as well as the air- liquid interface stress, may lead to protein denaturation (if unfolded vWF fails to refold properly post- spray drying) andDocket No. 0118.014100 -110- proteolytic degradation (unfolded vWF exposes proteolytic sites for ADMATS13), impairing the vWF activity in the spray dried plasma, as well as other proteins.

[0476] Spray drying system of the present invention can be optimized to reduce the protein damage caused by shear force and temperature and the specially designed cannula of the present invention helps to minimize shear and damage to the proteins include vWF.

[0477] The cannula of present invention, in an embodiment, has a bottom edge wherein at least a portion of the bottom edge is angled, referred to herein as an angled edge cannula. In an instance, the entire bottom edge can be angled or a portion of the bottom edge can be a flat edge (e.g., about a 90oangle from the outer wall surface or the inner wall surface). In another embodiment, a portion of the bottom edge of the cannula is a flat edge, like flat edge 88, (e.g., about 90ofrom the outer wall surface or inner wall surface) and a portion of the bottom edge of the cannula is angled, like angled edge 82, (e.g., 45oangle from the outer wall surface, or 135oangle from the inner side wall surface), as shown in Fig. 43F. This embodiment shown in Fig. 43F can edge having a flat edge (90ofrom the outer wall) from which a 45oangle is formed is referred to as a “chamfer” or as having a “chamfered edge.”

[0478] In the case in which the cannula has a bottom edge and the entire bottom edge is angled from the outer wall to the inner wall, the angle as measured from the outer wall surface ranges from about a 30oangle to about a 60oangle (e.g., about a 30o, 35o, 40o, 45o, 50o, 55o, 60oangle) and as measured from the inner wall surface ranges from about a 120oangle to about a 150oangle (e.g., about 120o, 125o, 130o, 135o, 140o, 145o, 150oangle). The length of the angled bottom edge ranges between .001 inches and about.010 inches (e.g., about .001,.002, .003, .004, .005, .006, .007, .007, .008, .009, .010 inches).

[0479] In the case in which the cannula has a bottom edge having a portion that is a flat edge and a portion that is angled, the flat edge is about 90oangle (e.g., between about 85% to about 95%) from the outer wall surface. The angled edge has an angle, as measured from the outer wall surface (imagining that the angled edge intersects the outer wall surface) ranges from about a 30oangle to about a 60oangle (e.g., about a 30o, 35o, 40o, 45o, 50o, 55o, 60oangle), and in an embodiment, is 45° + / - 5° and as measured from the inner wall surface ranges from about a 120oangle to about a 150oangle (e.g., about 120o, 125o, 130o, 135o, 140o, 145o, 150oangle), and in an embodiment, is 135° + / - 5°. See Fig. 52 for an example of a 45oand a 30oangled edge. The length of the flat edge portion ranges between .001 inchesDocket No. 0118.014100 -111- and about .009 inches (e.g., about .001,.002, .003, .004, .005, .006, .007, .007, .008, .009 inches) and the length of the angled edge portion ranges between about .001 inches and about .009 inches (e.g., about .001,.002, .003, .004, .005, .006, .007, .007, .008, .009 inches), and in an embodiment, is .005 + / - .003. The ratio between the length of the flat edge and the length of the angle edge has a range between about 5 and about 500 percent. In an embodiment, the flat edge adjoins the outer wall surface and the angled edge adjoins the inner wall surface.

[0480] The angled edge cannula, accompanied with or without a flat edge, results in less stress / shear on the plasma droplet exiting the cannula, as compared to a non-angled cannula, having a 90oangle. While not being bound to any particular theory, it is believed that when a plasma droplet exits a 90onon-angled cannula edge, a portion of the plasma droplet or plasma film undergoes a shearing effect and, in the process, degrades a high percentage of the plasma proteins therein. In this case, the 90onon-angled cannula exerts a shearing force on the droplet, thereby degrading the proteins in the plasma. As the plasma droplet exits cannula having an angled edge, as in the present invention, less sheer on the plasma droplet is exerted. As the plasma is drawn out by the air flow of an angled edge cannula, it accelerates based upon the plasma feed rate and the plasma gets pulled around the cannula edge. Unlike a cannula having a 90onon-angled edge, the plasma is not forced to make a 90 degree turn. By softening the turn that the plasma makes as it exits the cannula by angling the edge of the cannula, less shear is exerted upon the liquid film as it is drawn out. The liquid plasma film that exits out of an angled cannula is thicker and accelerates more slowly thereby reducing shear exerted on the liquid.

[0481] The inner diameter of the cannula ranges between about 0.010 inches and about 0.040 inches, and in an embodiment, is .030 inches + / - .002 inches and the outer diameter ranges between about 0.030 inches and about 0.060 inches, and in an embodiment, is .050 inches + / - .0005 inches. The angled edge of the cannula impacts the size of the atomized droplet. When exiting an angled cannula, the droplet sizes in this range is between about 5 microns and about 35 microns and in an embodiment the droplet size is about 10 microns. Small droplet size which is defined in part by the angled edge of the cannula, promotes rapid mixing, faster evaporation and reduced drying time. See Fig. 43T which shows that the larger the droplet size the longer it takes for the droplet to evaporate with higher drying gas temperatures. The shape of the droplet is created by its surface tension dominates and createsDocket No. 0118.014100 -112- a sphere after exiting the cannula. Droplet size is also primarily impacted by the pressurized gas rate ratioed to the liquid feed rate (ALR) and nozzle design.

[0482] The cannula of the present invention can be made from a stainless-steel material suitable for medical devices. Examples of the grade of stainless steel that can be used is grade 304 and 316 stainless steels. The stainless steel used for the cannula of the present invention is commercially available e.g., from Bergsen Metals (Santa Fe Springs, California, USA) or Fort Wayne Metals (Fort Wayne, Indiana, USA). The nozzle assembly (except for the cannula), nozzle insert, nozzle cap, plenum and baffle plate, outer filter ring, inner filter ring and the like can be made from plastic used in medical devices, such as a polycarbonate, polypropylene, polysulfone or combination thereof. Each aforementioned part can be made from the same material, from different materials or a combination thereof. Such plastic is commercially available and can be purchased from e.g., Covestro AG (Kaiser-Wilhelm-Allee 6051373 Leverkusen, Germany), Teknor Apex (Pawtucket, Rhode Island USA), Colorite Plastics of NJ Inc (101 Railroad Ave, Ridgefield, New Jersey USA), American RENOLIT Corporation (301 Berkeley Drive, Suite B, Swedesboro, New Jersey USA), and Exxon Mobile (Technology Centers, Baytown, TX USA 77520, United States), ), or molded from e.g., Egli Machine (Sidney, NY USA) Co, and Southwest Mold, Inc. (Tempe, AZ USA). Other materials now know or later developed can be used for the cannula and / or nozzle so long as when combined result in a maintenance or increase in vWF recovery in plasma after spray drying.

[0483] A stainless-steel nozzle, such as Buchi Model no. 4244 (Buchi Corporation of New Castle, Delaware United States), is often used in spray drying but it is expensive to manufacture or buy, especially for a disposable device that is discarded after each spray drying run. For example, a common Buchi stainless steel nozzle body, part No. 4244, costs between $1000 and $2000. The nozzle assembly of the present invention is a composite nozzle for use in spray drying and especially spray drying of delicate materials such as human blood plasma at a cost of less than $30.00, orders of magnitude less than stainless steel nozzles, such as the Buchi Model no. 4244. The described Buchi nozzle serves as a useful benchmark for a composite nozzle as it had been used by the applicant to make dried human blood plasma that preserved the proteins in blood plasma to a regulatorily acceptable level.Docket No. 0118.014100 -113-

[0484] As indicated, most of the nozzle assembly, except for the cannula which is made from a stainless-steel material, is made from a less expensive plastic material, as described above. As such, the nozzle assembly is also referred to as a "composite nozzle” or “composite nozzle assembly” to refer to the two or more different types of materials used to make the nozzle assembly (e.g., a stainless-steel cannula and a polycarbonate nozzle insert and nozzle cap). Example 28 shows that a chamfered cannula of a composite nozzle assembly provides for improved vWF recovery as compared to one that has a unchamfered cannula, and vWF recovery about as good as an expensive stainless-steel nozzle.

[0485] When the plasma exits the tip of the cannula, it is exposed to the pressurized aerosol gas at nozzle cap 76. More particularly, the pressurized aerosol gas exits in a vortex pattern through the annulus 81 and hits the liquid plasma droplets flowing from the chamfer edge / angled edge 82 of cannula 78 and the plasma atomizes to form a plume. When the atomized plasma exits the spray dry nozzle assembly, it is exposed to the drying gas and dries into plasma powder in the drying chamber. In an embodiment, the tip of cannula 78 is flush with the distal end of opening 110.

[0486] Liquid nozzle cap insert 80 secures the bottom portion of cannula 78 and guides the pressurized aerosolized gas flow. Fig. 43G shows a perspective top view liquid nozzle cap insert 80. As can be seen, cap insert 80 has insert wall 116 and cannula anchor 120 which has an opening (not shown) through which cannula 78 extends. The top of insert wall 116 forms a ridge and the side of insert wall 116 defines a series of recesses 118 and projections 119. Cannula anchor 120 supports the tip of cannula 78 during plasma flow. Angled edge 82 of cannula 78 through which the plasma exits and annulus 81 through which the pressurized aerosol gas is emitted create the actual nozzle. Cannula anchor 120 is a hollow, cylindrical base but can be of any shape so long as the cannula is supported, and its position maintained during spray drying. The recesses of the wall, recesses 118, allow for the pressurized air to pass from the reservoir (defined by reservoir housing 74) to the area between nozzle cap insert 80 and nozzle cap 76 before exiting the center opening 110 of nozzle cap 76. When the pressurized air exits center cap opening 110, the air exists through annulus 81 defined by the outer wall of cannula 78 and inner wall of opening 110. More specifically, when the cap insert 80 is secured to cap 76 and cannula 78 resides within opening 110, the pressurized air exits through annulus 81. See Fig. 43Ia. As such, theDocket No. 0118.014100 -114- diameter of opening 110 is greater than the outer diameter of cannula 78. In particular, opening 110 of nozzle cap 76 has an inner wall with a diameter, defined by Diameter Do(See Fig. 43Ia). Fig. 43Ia also shows cannula 78 that has an outer wall defined by Diameter Dc. Diameter Dois slightly larger than Diameter Dcand the difference is defined by Distance Dd. The resultant difference in diameter, Distance Dd, creates annulus 81 through which pressurized air received from aerosol reservoir housing 74 forms a vortex and flows to the drying chamber 28 to facilitate the formation of small droplets of fluid to be dried. In an embodiment, the outer diameter of cannula 78, Diameter Dc, is between about .030 and about .070 inches (e.g., .030, .040, .050, .060, .070) and the diameter of opening 110 is between 0.075 and 0.100. For ease of use, “Distance Dd” is also referred to as the “radial distance of annulus 81.” In an embodiment, the Diameter Dcis .050+ / - .0005 inches and the Diameter Dois 0.082 + / - .001 inches. The radial distance between the outside surface of cannula 78 and the inner surface of opening 110, Distance Dd, is the space through which the rotating vortex of pressurized aerosol gas flows and assists in creation of small droplets of plasma to mix with the hot drying gas during spray drying. In an embodiment, Ddhas a range between 0.005 and 0.030 inches (e.g., 0.015 and 0.021 inches).

[0487] Along these lines, the data from Example 27 show that that the radial distance of annulus 81, Dd, has an impact on both the yield for dried product from the drying process and on the preservation of vWF. Yield is the ratio of starting solids in the to-be-dried liquid material by weight to dried material recovered by the drying process by weight.

[0488] Before the present invention, one source of loss of yield occurred when dried sprayed material that was not fully dried and retaining residual moisture above about 2.5% contacted and stuck to the interior structures of the drying disposable during drying without being recoverable.

[0489] Example 27 describes the reduction in the amount of material visibly stuck to the underside of baffle plate 8 after completion of the drying cycle. The data described in Example 27 show that overall yield by weight was increased by changing the radial distance of annulus 81 Ddfrom 0.021” to 0.015”. The yield was acceptable with the annulus dimension at 0.021”. However, the yield percentage was improved by more than 2.2% by reducing the annulus width / diameter to 0.015”. Other features of disposable 100 increase yield and include, in part, drying jets 142 that form an air wall within plasma drying chamberDocket No. 0118.014100 -115- 28, as further described herein.

[0490] Example 27 also described in increase in the recovery of vWF as measured by Ristocetan (RCO) assay by changing the radial distance of annual 81 from 0.021” to 0.015.” The vWF recovery was acceptable with the annulus dimension at 0.091”. However, vWF recovery was increased by more than 2.0% by reducing the annulus width to 0.082”. Other features of disposable 100 also increase vWF recovery and include, in part, the angled edge cannula 78, as described herein.

[0491] The space between nozzle cap insert 80 and nozzle cap 76 before exiting the center opening 110 of nozzle cap 76 is generally referred to herein as the “vortex generator” which includes a series of channels and curved pads, as further described below. Pressurized air passes through recesses 118 that act as openings in nozzle assembly 20 to allow air to enter and travel down the channels and between the curved pads. See Figs. 43G, 43H, 43I and 43Ic.

[0492] Referring to Fig. 43H, the bottom of surface of cap insert 80 not occupied by 122 on the underside of projections 119 / recesses 118 and will act as walls for the channels in the vortex generator. Pads 122 form a kidney-like shape that assists in locating recesses 118 for the vortex air flow pattern. The nozzle cap, cap 76, shown in Fig. 43I, has complementary receivers 112 to receive the pads from the cap insert 80. The complementary fit between nozzle insert 80 and nozzle cap 76 is shown in Fig. 43Ic (as a cross-section). Nozzle cap 76 also has nozzle cap channels 106 extending from bulbous head 108 and ending at opening 110. The surface of the bottom of cap insert 80 that is not occupied by pads 122 and further complemented by channels 106 of cap 76, is the space through which the pressurized air flows. The vortex generator includes the recesses 118 and the surface of the bottom of cap insert 80 that is not occupied by pads 122, bulbous heads 108 and channels 106 of cap 76, the shape and position of each cause the pressurized air to form a vortex air flow pattern. Bulbous head 108 receives the pressurized air flow through recess 118 and the curved ramp like surface of channel 106 provides a curved boundary for the air to flow. In other words, recesses 118 is an entrance port and feeds the air flow and channels 106 feeds the vortex. Channels 106 are arched and further accentuates the curved air flow and directs the tangential air flow toward opening 110 in which cannula 78 resides. These channels, channels 106, guide the air in a circular fashion from bulbous head 108 to nozzle cap openingDocket No. 0118.014100 -116- 110, all working in concert to expel pressurized air as a vortex through opening 110. The design provides tangential momentum to provide an efficient generation of a vortex. Channels 106 are in the form of an arc or curve, and the radius of the curvature ranges from about 0.10 inches to about 0.25 inches, and in an embodiment about .140 + / - .010 inches radius. The vortex generated includes 4 channels but can have between about 2 and 12 channels (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 channels). Other types of shaped channels can be used. Fig. 43I shows the design molded into nozzle cap 76. Designs A and B both show more linear channels but Design A have no equivalent of a bulbous head and Design B shows a bell-shaped head. Design C is similar to the design shown in Fig. 43I but with a lobe instead of a bulbous shape at the end. The present invention includes nozzle caps having Designs A and B but found the Design C appears to be a more efficient vortex generator. The various designs demonstrate that any combination of channels, heads and shapes can be used to generate a vortex in the annulus. Other types of channels include conical shaped channels including convert or divergent cone shapes and the like.

[0493] As the vortex is generated, pressure and velocity flow patterns are shown in Fig. and 43P. Fig. 43O shows the static gas pressure in psig at the top and tangential velocity contours in the pressurized aerosol gas flow in m / s at the bottom. As can be seen, there is an inverse relationship between pressure and velocity. In areas where the pressure is increased, the velocity is decreased and vice versa. In particular, at bulbous head 108 where the pressurized gas enters the vortex generator, there is higher relative static gas pressures (e.g., about 2.54 x 101psig) and relatively low velocity flow rates (e.g., about 2.00 x 101m / s). Conversely, at annulus 81, relatively low or negative gas pressures (e.g., about -2.24 psig) and higher velocity (e.g., between about -1.58x102to about -3.75x102m / s). As the pressurized aerosol gas travels along the curved nozzle cap channel 106, the pressure and velocity are at rates in between. As such, the vortex generator of the present invention has gas pressure of between about 2.54 x 101psig to about -2.24 psig and velocity at a rate between about 2.00 x 101m / s and about -3.75 x 102m / s. In an embodiment, any vortex generator can be used with the present invention so long as gas pressure and velocity are produced in these ranges. Similarly, Fig. 43P shows a more detailed velocity pattern that occurs in annulus 81. The pressurized gas moves between exits of channels 106 as it integrates into the vortex. The pressurized gas accelerates when it enters annulus 81 andDocket No. 0118.014100 -117- becomes a vortex.

[0494] The vortex generator in this embodiment includes a curved pad / ramp, a bulbous head to receive the pressurized air flow and curved channels extending to the exit opening. The present invention can include other vortex generator elements such as wings, edges, wedges, vanes and the like. Other shaped channels can also be employed to create a vortex generator. One of skill in the art can utilize other vortex generators of residing within the insert and the cap of the nozzle assembly of the present invention so long as the pressurized air exits between the inner surface of opening 110 and the outer surface of the cannula 78 in a vortex.

[0495] The pressurized air circulates between the outer surface of cannula 78 and the inner surface of opening 110. Specifically, the pressurized gas exits through annulus 81. The plasma is pumped through cannula 78 by peristaltic pump 214 at approximately room temperature. The plasma travels down the inside of stainless-steel cannula 78 and is drawn out of cannula 78 by the pressurized aerosol air flow exiting annulus 81. The high-speed aerosolization air flow atomizes the liquid droplets. In an embodiment, the steady-state plasma feed rate is between about 6 and about 23 mL / min (e.g., 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, or 23). In a preferred embodiment, 13.5 mL / min is the steady- state feed rate after system is warmed-up, in thermal equilibrium.

[0496] In an embodiment, the plasma feed rate is related to or dependent on the outlet temperature. This is done in a closed loop. As the outlet temperature lowers, the system adjusts to lower the plasma feed rate. Conversely, as the outlet temperature increases, the system increases the plasma feed rate. The outlet temperature may be lower when the spray dryer is warming up or when time passes between spray drying runs, for example. In particular, in an embodiment, the plasma feed rate can be modulated as follows:

[0497] Table 7 Dryer Inlet System Dryer Outlet Dryer Outlet Plasma ReductionDocket No. 0118.014100 -118- 100 0.18 59.3 11.8 10.5 22.2% 80 0.18 50 11.8 6.6 51.1%

[0498] ing constant system heat loss, full droplet evaporation, and a dryer outlet relative humidity of <11.8%.

[0499] As such, when the outlet temperature is lower, the plasma feed rate lowers to maintain the target drying chamber outlet temperature needed to dry the plasma to a residual moisture of less than 2%. When the outlet temperature is higher within the range, the plasma feed rate can also be increased, and still maintain a residual moisture content of plasma of less than 2% only if the total gas flow can be increased and / or the drying chamber outlet temperature is allowed to increase to maintain target system relative humidity.

[0500] In an embodiment, the closed loop includes exhaust temperature ranging between about 62oC and about 68oC and plasma feed rates ranging between about 6 and about 23 mL / min.

[0501] Just below the cannula there is negative pressure, while pressurized gas flow is at a high velocity. Generally, the velocity increases along a pathway of decreasing static pressure. The pressurized aerosol gas travels through the series of channels 106 and creates a vortex flow which both atomizes the plasma droplets and directs the initial droplet trajectory. Aerosol flow rate is between about 20 splm and about 60 slpm (e.g., about 20, 25, 30, 35, 40, 45, 50, 55, 60 slpm) and in an embodiment is about 40 slpm. This occurs with a pressure of between about 180 kPa to about 260 kPa (e.g., about 180, 190, 200, 210, 220, 230, 240, 250, 260 kPa) and in an embodiment about 227.5 kPa (33 psig). The aerosol flow acts to draw the liquid feed out of the cannula where it forms a film across the end. The expansion of the aerosol gas as it exits the opening locally cools the near nozzle gas field, which also acts to delay evaporation slightly by cooling the liquid droplets. Also, in an embodiment, the pressure just below the end of cannula is less than that inside the cannula and pressurized gas velocity is accelerating when traveling along the outside surface of the cannula. Upon exit, the spherical plasma droplets hit the pressurized gas to aerosolize and form a spray plume which is surrounded by a ring of drying gas jets, which are further described below. See Fig. 43N. The mixing of aerosol and drying gas sets the initial conditions for the evaporationDocket No. 0118.014100 -119- process.

[0502] Fig. 43Q is a schematic that shows the liquid plasma droplet undergoing the drying process. The plasma droplet is atomized at the nozzle assembly cannula exit into the drying chamber and is generally spherical. The dried plasma particle is formed with heat and mass transfer. Drying takes place in two stages. These are: evaporative drying stage (constant rate) drying which occurs in the initial drying (e.g., in, less than 1 second) and falling rate drying (diffusion limited) which occurs after the evaporative drying stage and continues so long as the dried particle is subject to ambient relative humidity lower than its internal relative humidity.

[0503] The factors involved in the evaporative drying stage of the plasma droplet include the temperature of the plasma and the drying gas, the surface area of the droplet, the humidity in the drying gas and the air circulation within the plasma drying chamber. When initially exiting the nozzle assembly, the temperature of the drying gas is between about 90oC to about 130oC (e.g., between about 100oC to about 114oC) and the temperature of the plasma droplet is between about 20oC and about 65oC in the plume as shown in Fig 43Sa. The heat flows from a point of higher temperature to that of a lower temperature, and in this case the drying gas heat flows to the plasma droplet. With respect to the surface area, the droplet is spherical thereby maximizing its surface area and the droplet size is very small so the mass and heat transfer can happen quickly. The relative humidity in the drying gas is very dry (e.g., about 0.1% RH) and therefore the low humidity of the surrounding drying gas promotes evaporation of the plasma particle. Finally, as described in more detail below, the drying gas is emitted using several drying gas jets in an angled and downward direction into the plasma drying chamber and into the plume of atomized droplets to initiate rapid mixing of the drying air and the atomized droplets, which increases the rate of evaporation of the liquid droplets. The drying rate is constant and as the liquid particle evaporates and loses moisture, the moisture transfers from the liquid plasma droplet to the drying gas, and the heat from the drying gas transfers to the plasma droplet making it into a dried particle. The plasma droplet enters the drying chamber essentially at room temperature and the temperature stays constant the majority of the evaporation period. See Fig. 43S. Once most all of the moisture leaves the particle, the temperature of the particle increases to equilibrate with the dryer chamber exit temperature of 65oC. During evaporation, the droplet is maintained at a lowerDocket No. 0118.014100 -120- temperature thereby protecting heat sensitive proteins such as vWF. See Fig. 43S. During the evaporation process, the temperature of the liquid droplet and the proteins therein experience a lower temperature, the thermodynamic wet bulb temperature, compared to the inlet drying gas temperature, thereby protecting the proteins. See Fig. 43R. Evaporation reduces protein temperature to near the thermodynamic wet bulb value and when the evaporation slows the particle temperature rises. See Fig. 43Sa.

[0504] The starting liquid droplet size produced by the nozzle assembly impacts the residence time in the drying chamber needed to complete evaporation. The larger the liquid droplet, the smaller the ratio of evaporation surface area to droplet mass and the slower the mass transfer rate from the droplet. This slower rate requires a greater distance between nozzle assembly 20 and the lower filter 36 to avoid overly wet particles depositing on the filter membrane of lower filter 36. An overly wet particle causes “plugging or blinding” of the porous filter membrane and the inability to complete the process as the chamber pressure would elevate excessively, preventing production of powder. See Fig 43T.

[0505] Additionally, Fig. 43T shows that in all cases, the majority of the evaporation occurs in the upper portion of disposable 148. This is particularly seen when using a small droplet size, e.g., under 15 microns. As seen from Fig. 43T, the drying chamber can be shortened to that where the majority of the evaporation occurs while still allowing a dried particle to achieve less than 2.5% residual moisture before being deposited on lower filter 36. In other words, in an embodiment, drying chamber 28 can be shortened by an amount between about 8 inches and 1 inch when the droplet size is below about 15 microns and achieves a residual moisture of below about 2.5%.

[0506] Turning to the plenum, the functions of the plenum include 1) allowing for the introduction and flow of the drying gas to the disposable, 2) housing the nozzle assembly, and 3) provide support for the drying chamber during the spray drying process. The underside of plenum 6 is shown in Fig. 43J. Plenum 6 has two openings, opening 96 to receive nozzle assembly 20 and drying gas inlet port 22 to receive the drying gas.

[0507] Nozzle assembly receiver opening 96 is complementary in shape to the top of nozzle reservoir housing 74 and manifold 72. The top of nozzle assembly 20 is secured in opening 96. The length of nozzle assembly coincides with the height of plenum 6 such that the bottom of nozzle assembly 20 extends past baffle plate 8. See Figs. 46A and 46B. InDocket No. 0118.014100 -121- certain embodiments, the cannula is flush with the nozzle assembly and the baffle plate. Nozzle assembly 20 can be secured with adhesive, fasteners, or with an interlocking assembly (e.g., a spring latch, screw fit and the like).

[0508] The other opening of plenum 6 is drying gas inlet port 22 which receives the drying gas. The drying gas source (not shown) flows into the plenum through drying gas inlet deflector 242, which is shown in Fig. 43K. Once the disposable is secured and aligned, and the door to the spray dryer is closed and spray drying begins. Drying gas inlet deflector 242 lowers through drying gas inlet port 22 to provide the drying gas to plenum 6. Drying gas inlet deflector 242 has the shape of an elbow so that the drying gas flows toward the far inside side wall of the plenum creating a tangential mixture, as shown in Fig. 43Ka. The right angle of deflector 242 distributes the drying gas throughout plenum 6, creating a low velocity, highly uniform pressure reservoir. Uniformity is desired to create low velocity, uniform pressure of the drying gas as it exists each of drying jets 142. When the drying gas is not deflected off the side of plenum 6 but instead in a downward direction, the air pressure can be asymmetrical with drying jet closer to the drying gas inlet experiencing high pressures as compared to those farther away from the inlet. Accordingly, the present invention includes a drying gas inlet that is deflected to the side of plenum e.g., with a 90-degree elbow as is the case with deflector 242. Other geometries of deflector 242 can be employed to create a uniform air pressure in plenum 6. For example, the deflector can be angled at degree less than a 90-degree angle, as measured from the top surface of the plenum. For example, the deflector can have angle ranging between about 60 and about 110 degrees relative to the top surface of plenum 6 so that the drying gas pressure across the width of plenum 6 is substantially uniform. Alternatively, more than one drying gas inlet from opposing sides can be used to create a substantially uniform drying gas pressure across the width of plenum 6.

[0509] As shown in Fig. 43J, plenum 6 has concentric ribs 98 and radiating ribs 102. These ridges provide support for the structure of plenum 6. The additional support provided by the concentric and radiating ribs allow the plenum to withstand the pressure and heat of the spray drying process. Since the drying gas is a low velocity, uniform pressure air container, concentric ribs 98 and radiating ribs 102 do not contribute to or affect the drying gas air flow. Similarly, the projections on the inside side wall of the plenum, projections 104, are used in the injection molding process when making the plenum and are not involved inDocket No. 0118.014100 -122- the drying gas flow.

[0510] Referring to Figs. 43L, 43La, and 43M, the baffle plate has several functions as follows: A) serves as a support in securing disposable 100 when the disposable is aligned and inserted into spray dryer 200, B), creates drying gas air flow channels and releases the drying air into plasma drying chamber 28 of disposable 100, and C) supports baffle filter 94.

[0511] Fig. 43L shows the top, inside view of baffle plate 8. The inner surface of baffle plate 8 has baffle plate nozzle opening 140 through which a portion of nozzle assembly 20 resides. Baffle plate 8 also includes raised outer ring 124 with its base 126 and raised inner ring 128 with its base 130. Outer sealing ring 90 is placed around outer ring 124 and inner sealing ring 92 is placed round inner ring 128. The sealing rings prevent drying gas from flow out the edges of filter 94 and instead the drying gas flows through it. The inner side of baffle plate 8 further includes locator 132 for insertion of locator notch 26 of plenum 6. The plenum and baffle plate have locators to align one another. Baffle plate 8 has baffle locator 132 that receives plenum locator 152 on plenum 6.

[0512] Baffle plate ribs, 134, 136 and 138 provide support to baffle plate filter 94 (shown in Fig. 43L) while keeping most of the surface of filter 94 lifted off the baffle plate during use. The baffle plate ribs also act as a guide for the drying gas flow. It has been determined that if the baffle plate filter 94 lies flat on the inner side of baffle plate 8 without ribs, the drying gas flow slows and does not freely flow through the plurality of drying jets 142. To obviate that phenomenon, in particular, baffle plate 8 has radiating ribs 134 which connects inner ring base 130 with outer ring base 126. Each radiating rib 134 has a consistent profile throughout its length and allows the filter to sit in a raised position, as compared to resting directly on inner surface of the baffle plate. Radiating ribs 134 also form pie-shaped air channel 139 leading to drying jet 142. Radiating ribs 134 are the side walls of pie-shaped air channel 139. Baffle plate 8 has two types of ribs that extend from outer ring base 126 but do not connect with or reach inner ring base 130. Of these types of ribs, there is shorter radiating rib 136 and intermediate radiating rib 138. Both shorter radiating ribs 136 and intermediate radiating ribs 138 have a consistent profile in height as it proceeds from outer ring base 126 inward and then quickly tapers at tapered end 137. The tapered end 137 aids in supporting the filter without creating a corner on which the filter could be pierced. In particular, baffle filter 94 rests atop ribs 134, 136 and 138 and is pressed against the ribsDocket No. 0118.014100 -123- during spray dryer operation by air pressure e.g., of about 11.5 psig. The tapered ends on ribs 136 and 138 reduce the stress placed on the baffle filter, prevents damage to the filter and reduces loss of filtration efficiency. See Fig. 43La. The ribs keep baffle filter 94 from adhering to the inner baffle plate surface. Connecting ribs and non-connecting ribs are interspersed on the baffle plate inner surface and in the embodiment shown in Fig. 43L form a pattern within pie-shaped air channel 139 (e.g., connecting rib, shorter non-connecting rib, two intermediate non-connecting ribs, shorter non-connecting rib, connecting rib, etc.). Each pie-shaped air flow channel 139 defined by two connecting ribs on either side. The connecting and non-connecting ribs can be any pattern so long as they provide support for baffle filter 94 while also allowing drying gas to flow through filter 94 and beneath filter 94 and through the air channels 139 to drying jets 142. For example, Fig. 43La shows another arrangement of ribs 136 and 138.

[0513] In an embodiment, the pressure drops when the drying gas passes baffle filter 94. The input drying air pressure in plenum 6 before traveling through the filter into the pie- shaped air flow channel 139, during operation, is between about 8 and about 15 psig, and in an embodiment is approximately 10.4 psig, When the drying gas passes baffle filter 94, the pressure drops by about 40-60% or in an embodiment by approximately 6 psig, The pressure decrease across baffle filter 94 is utilized to aid the uniform distribution of drying gas being injected into the drying chamber as shown in Fig. 43Ma. This feature helps to minimize asymmetric drying within the chamber to enable the overall shorter length. The resultant pressure in the drying chamber 28 in area defined by Dimension Z ranges between about 4 psig and about 7 psig and in an embodiment is approximately 5.5 psig. Although further described later herein, lower filter 36 of disposable 100 causes a pressure drop by about 40- 80% or in an embodiment by approximately 3.0 -5.5 psig that increases as dried plasma accumulates on the filter resulting in exiting air pressure of approximately 1-2.5 psig. In an embodiment, the gas exhaust port 208 of spray dyer 200 is slightly constricted so that the exhaust gas leading to the outside is about 1.5 to about 3 psig. If there are valves, sensors or tubing length past the exhaust exit those will add a small pressure increase.

[0514] In an embodiment, the flow of drying gas passing through and over the baffle plate 8 is not restricted. In an aspect, the height and placement of the ribs 134, 136, 138 of baffle plate 8 are such that the baffle plate filter 96 does distort somewhat under the pressureDocket No. 0118.014100 -124- of the drying air but has no material effect on the pressure drop. Note that baffle plate filter 96 does crea...

Claims

Docket No. 0118.014100 -343- CLAIMS What is claimed is: 1) A spray dried plasma that is spray dried from a donor plasma, wherein the spray dried plasma has a reduced amount of pathogens, as compared to donor plasma. 2) The spray dried plasma of Claim 1, wherein the plasma is reconstituted, to thereby obtain reconstituted spray dried plasma. 3) A reconstituted previously spray dried plasma that is spray dried from a donor plasma, wherein the reconstituted spray dried plasma has a reduced amount of pathogens, as compared to donor plasma and has one or more plasma proteins that induce clot formation. 4) The reconstituted previously spray dried plasma of Claim 3, wherein the one or more plasma proteins having a level within a corresponding clinical reference range. 5) The reconstituted previously spray dried plasma of Claim 3, wherein the one or more plasma proteins that induces clot formation comprises von Willebrand Factor (vWf). 6) The reconstituted previously spray dried plasma of Claim 5, wherein the one or more plasma proteins that induces clot formation is measured by von Willebrand Factor Antigen assay or von Willebrand Factor Ristocetin Cofactor assay. 7) The reconstituted previously spray dried plasma of Claim 6, wherein von Willebrand Factor Antigen or von Willebrand Factor Ristocetin Cofactor is between about 50 IU / dL and about 200 IU / dL. 8) A spray dried plasma having a reduced amount of pathogens, said spray dried plasma is made using a spray drying disposable device, having a spray drying head and a plasma drying chamber, in a spray drying system having a drying gas source, a plasma source and a pressurized aerosol gas source, a) wherein the spray drying disposable device comprising: i) a spray drying head comprising: (1) a spray dry nozzle assembly in fluid communication with the plasma source and the pressurized aerosol gas source, wherein the pressurized aerosol gas flows in a vortex pattern, wherein, when in use, the pressurized aerosol gas atomizes the plasma in the drying chamber to obtain atomized plasma droplets;Docket No. 0118.014100 -344- wherein, when in use, the spray drying head source supplies drying gas to the drying chamber at a temperature of between about 90° C and about 130° C; ii) the drying chamber, attached to a baffle plate, wherein atomized plasma droplets evaporate in the presence of the drying gas emitted from the one or more drying gas jets to thereby obtain dried plasma particles and humid air; iii) a capture filter, residing in the drying chamber, wherein the capture filter captures the dried plasma particles and allows the humid air to pass; iv) a gas outlet, wherein said gas outlet is attached to the exhaust port of the spray drying apparatus, wherein the humid air flows through the gas outlet; wherein an amount of pathogens is reduced in the spray dried plasma, when reconstituted, as compared to spay dried plasma not spray dried using the spray dried disposable or to that present in the donor plasma. 9) The spray dried plasma of Claim 8, wherein the atomized plasma droplets have a temperature between about 20oC and about 65oC. 10) A spray dried plasma having a reduced amount of pathogens, said spray dried plasma is made using a spray drying disposable device, having a spray drying head and a plasma drying chamber, in a spray drying system having a drying gas source, a plasma source and a pressurized aerosol gas source, a) wherein the spray drying disposable device comprising: i) a spray drying head comprising: (1) a spray dry nozzle assembly in fluid communication with the plasma source and the pressurized aerosol gas source, wherein the pressurized aerosol gas flows in a vortex pattern, wherein, when in use, the pressurized aerosol gas atomizes the plasma in the drying chamber to obtain atomized plasma droplets; (2) a plenum having a drying gas inlet in communication with the drying gas source, wherein, when in use, the drying gas resides in the plenum with uniform air pressure, wherein the plenum supports the nozzle assembly; (3) a baffle plate forming the floor of the plenum having one or more drying gas jet, wherein drying gas jet provides drying gas to the drying chamber;Docket No. 0118.014100 -345- ii) the drying chamber, attached to the baffle plate, wherein atomized plasma droplets evaporate in the presence of the drying gas emitted from the one or more drying gas jet to thereby obtain dried plasma particles and humid air; iii) a capture filter, residing in the drying chamber, wherein the capture filter captures the dried plasma particles and allows the humid air to pass; iv) a gas outlet, wherein said gas outlet is attached to the exhaust port of the spray drying apparatus, wherein the humid air flows through the gas outlet; wherein an amount of pathogens is reduced in the spray dried plasma, when reconstituted, as compared to spay dried plasma not spray dried using the spray dried disposable or to that present in the donor plasma. 11) The spray dried plasma of Claim 10, wherein the drying gas has a temperature of between about 90° C and about 130° C. 12) The spray dried plasma of Claim 10, wherein the atomized plasma droplets have a temperature between about 20oC and about 65oC. 13) The spray dried plasma of Claim 10, wherein the spray drying disposable device further comprises a baffle plate filter in communication with the baffle plate. 14) The spray dried plasma of Claim 10, wherein the baffle plate filter comprises 0.2 micron filter. 15)Amethod of spray drying plasma wherein the spray dried plasma, when reconstituted, has a reduced amount of pathogens and has one or more plasma proteins that include clot formation, the method comprises the step of: a) spray drying plasma from a donor plasma; said spray dried plasma is made with a spray drying disposable device, having a spray drying head and a plasma drying chamber, in a spray drying system having a drying gas source, a plasma source and a pressurized aerosol gas source, wherein, when in use, the spray drying head source supplies drying gas to the drying chamber at a temperature of between about 90° C and about 130° C; wherein the reconstituted spray dried plasma has a reduced amount of pathogens, as compared to donor plasma and has one or more plasma proteins that induce clot formation.Docket No. 0118.014100 -346- 16) A method of spray drying plasma of Claim 15, wherein the spray drying disposable device comprises: a) a spray drying head comprising: i) a spray dry nozzle assembly in fluid communication with the plasma source and the pressurized aerosol gas source, wherein the pressurized aerosol gas flows in a vortex pattern, wherein, when in use, the pressurized aerosol gas atomizes the plasma in the drying chamber to obtain atomized plasma droplets; ii) a plenum having a drying gas inlet in communication with the drying gas source, wherein, when in use, the drying gas resides in the plenum with uniform air pressure, wherein the plenum supports the nozzle assembly; iii) a baffle plate forming the floor of the plenum having one or more drying gas jets, wherein the one or more gas jets provide drying gas to the drying chamber; b) the drying chamber, attached to the baffle plate, wherein atomized plasma droplets evaporate in the presence of the drying gas emitted from the one or more drying gas jet to thereby obtain dried plasma particles and humid air; c) a capture filter, residing in the drying chamber, wherein the capture filter captures the dried plasma particles and allows the humid air to pass; d) a gas outlet, wherein said gas outlet is attached to the exhaust port of the spray drying apparatus, wherein the humid air flows through the gas outlet; wherein an amount of pathogens is reduced in the spray dried plasma, when reconstituted, as compared to spay dried plasma not spray dried using the spray dried disposable or to that present in the donor plasma. 17) The method of spray drying plasma of Claim 16, wherein the atomized plasma droplets have a temperature between about 20oC and about 65oC. 18) The method of spray drying plasma of Claim 16, wherein the spray drying disposable device further comprises a baffle plate filter in communication with the baffle plate. 19) The method of spray drying plasma of Claim 16, wherein the baffle plate filter or the capture filter or both comprises 0.2 micron filter. 20) The method of spray drying plasma of Claim 15, wherein the donor plasma is pretreated with an amount of an spray dry stable acidic substance (SDSAS) and one or more amino acids.Docket No. 0118.014100 -347- 21) A method for storing spray dried plasma with reduced pathogens, the method comprises; a) subjecting the dried plasma to a temperature between about -80ºC and about 45oC for a period of time ranging from about 1 day and about 48 months; wherein the amount of pathogens are not increased, as compared to that before step a). 22) The method of Claim 21, wherein the amount of pathogens is decreased, as compared to that before step a). 23) The method of Claim 21, wherein when rehydrated, the level of one or more plasma proteins is within a clinical reference range. 24) The method of Claim 21, wherein when rehydrated, the level of one or more plasma proteins induces clot formation.

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