Plasma products

Abstract

The present invention relates to spray-dried plasma compositions that have one or more of the following properties: upon reconstitution, they are largely amorphous and free of cholesterol crystals; upon reconstitution, they have small average size of large particulates, low residual moisture; they are rapidly reconstituted in less than 4 minutes; they are highly stable when stored under refrigeration, at room temperature, or at elevated temperatures, allowing for extended storage; upon reconstitution, they exhibit retention of the most fragile proteins, including von Willebrand factor; upon reconstitution with sterile water for injection (SWFI), the reconstituted plasma has a pH that approximates normal plasma pH; and they do so without treatment or storage with CO2 or other pH adjusting agents.

Description

[Technical Field]

[0001] Related Applications This application is a continuation-in-part of U.S. Patent Application No. 17,945,125, entitled "Blood Plasma Product," by Qiyong Peter Liu et al., filed September 15, 2022.

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

[0003] government support This invention was made with government support under Contract Nos. HHSO100201200005C and 75A50121C00059 awarded by the Biomedical Advanced Research and Development Authority (BARDA). The government has certain rights in this invention. [Background technology]

[0004] Dried plasma is of great medical importance, and its development has been the subject of decades of development and expense. Known methods for drying plasma include freeze-drying, also known as lyophilization, and spray-drying. Dried plasma has applications in the military, mass casualty events, and the treatment of certain disorders. It also has potential use on a more universal basis as a preventative emergency use system for people at various risks.

[0005] Regarding plasma, transfused human plasma is often important for hemorrhage control and wound treatment in trauma victims and in surgery. Unfortunately, plasma is not readily available in many settings worldwide, including battlefields, first responders, and rural settings away from large hospitals, as well as in the second and third world.

[0006] The primary reason that liquid human plasma is not as widely available as it should be is that plasma is generally frozen for long periods or can only be stored as a liquid for very short periods of time. Therefore, when large amounts of plasma are needed (e.g., in a mass casualty event), it may not be available in such quantities, or when plasma is needed in an emergency, it may not be available in time because it must be thawed, which can take 30-45 minutes or more.

[0007] Freeze-dried plasma, on the other hand, presents a different set of challenges. Upon rehydration, freeze-dried plasma can form cholesterol crystals, which contribute to inflammation and atherosclerosis. The pH of the reconstituted plasma is alkalotic, which can be harmful to the transfusion recipient, especially if transfused in large quantities. Furthermore, in most cases, freeze-dried plasma must be treated with acid to raise its pH and / or stored with CO2 to achieve a near-normal pH. Some known, available freeze-dried plasmas present logistical challenges as well. These plasmas are stored in glass bottles, which are heavy and prone to damage if not handled properly. The reflux system required for the rehydration process using glass bottles can cause problems if the rehydration fluid is added too quickly. Furthermore, the presence of undesirable solids or undesirable particulates is undesirable for any type of plasma.

[0008] Therefore, there is a need for dried plasma that has no or minimal cholesterol crystals upon rehydration. There is also a need for dried plasma to induce clot formation upon reconstitution. There is a further need for dried plasma that can be easily stored at various temperatures and environments (e.g., battlefield environments) without heavy glass bottles. There is also a need for rapid rehydration of dried plasma that has a near-normal pH and can be readily transfused in emergency situations. Summary of the Invention [Means for solving the problem]

[0009] The present invention provides a plasma that has a) dried particles having a size in the range of about 1 micron to about 7 microns, b) upon reconstitution, a reduced mean particle size for particles having a size of about 2 μm to 60 μm as compared to particles in donor plasma as measured using a Coulter Multisizer 4 by the electrical sensing zone method, c) a reduced number of cholesterol crystals upon reconstitution when viewed at 100x magnification as compared to freeze-dried plasma, d) a residual moisture in the range of about 0.5% to about 2.5% (e.g., 2.5%, 2.0%, 1.5%, 1.0%, or 0.5%), e) is stable when stored at temperatures in the range of about -80°C to about 45°C for periods of about 1 day to about 48 months as compared to reconstituted spray-dried plasma before storage, f) upon reconstitution, is suitable for transfusion and / or is stable for transfusion after storage for up to about 26 hours, g) is stable when stored in the range of about 100x magnification when viewed at 100x magnification when compared to freeze-dried plasma before storage, and The present invention relates to spray-dried plasma having several novel properties, including any combination of: (i) having a pH of about 6.5 to about 7.8 upon reconstitution with Spray-Dried Plasma Injection (SWFI); (ii) having protein functionality, including von Willebrand factor (vWF), or inducing clot formation upon reconstitution; and (iii) having C5a and C3a levels comparable to those of apheresis plasma upon reconstitution.

[0010] In one embodiment, the reconstituted spray-dried plasma of the present invention has a reduced mean particle size compared to the particulates in donor plasma, with the particulates having a size of about 2 μm to about 60 μm, as measured using a Coulter Multisizer 4 by the electric sensing zone method. The mean particle size is reduced by about 60%, about 50%, about 40%, about 30%, about 25%, about 20%, about 15%, about 10%, about 5%, about 4%, about 3%, about 2%, or about 1% compared to that found in donor plasma. As measured, the particulates in the reconstituted plasma had a mean size in the range of about 2 μm to about 3.5 μm, compared to donor plasma, which had a mean size in the range of about 2 μm to about 7 μm.

[0011] With respect to storage, spray-dried plasma is stable over extended periods of time compared to liquid plasma or conventional plasma. Stability involves comparing the respective values ​​of one or more of these plasma proteins and / or properties of the spray-dried plasma of the present invention upon reconstitution before and after storage. In one embodiment, the value of the plasma protein / plasma property after storage is within the clinical range for that plasma protein. In another embodiment, the value of the plasma protein / plasma property after storage is within about 25% or less (25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, and 1%) of the value before storage. Such plasma properties include pH, osmolality (mOsm / kg), particle size, particle load, and particle distribution. Plasma proteins and their functions include, for example, activated partial thromboplastin time (aPTT), prothrombin time, international normalized ratio (INR), thrombin time, 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 F1+2 (pmol / L), thrombin-antithrombin complex (Thrombin-Antithrombin Examples include total protein (mg / mL or g / dL), total ATP (μg / ...In one embodiment, the acceptable or clinical range for von Willebrand Factor Ristocetin Cofactor (VWF:RCo) is about 10 to about 200 IU / dL, or about 50 to about 200 IU / dL, and the von Willebrand Factor Antigen (VWF:Ag) value is about 50 to 200 IU / dL.

[0012] In one embodiment, the clinical reference ranges are as follows:

[0013] [Table 1]

[0014] [Table 2]

[0015] The storage period for the spray-dried plasma is in the range of about 3 hours to about 48 hours (e.g., about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 12 months, about 13 months, about 14 months, about 15 months, about 16 months, about 17 months, about 18 months, about 19 months, about 20 months, about 21 months, about 22 months, about 23 months, about 24 months, about 25 months, about 26 months, about 27 months, about 28 months, about 29 months, about 30 months, about 31 months, about 32 months, about 33 months, about 34 months, about 35 months, about 36 months, about 37 months, about 38 months, about 39 months, about 40 months, about 41 months, about 42 months, about 43 months, about 44 months, about 45 months, about 46 months, about 47 months, about 48 months, about 49 months, about 50 months, about 51 months, about 52 months, about 53 months, about 54 months, about 55 months, about 56 months, about 57 months, about 58 months, about 59 months, about 60 months, about 61 months, about 62 months, about 63 months, about 64 months, about 65 months, about 66 months, about 67 months, about 68 months, about 69 months, about 70 months, about 71 months, about 72 months, about 73 months, about 74 months, about 75 months, about 76 months, about 77 months, about 78 months, about 79 months, about 80 months, about 81 months, about 8 The spray-dried plasma of the present invention can be stored at temperatures ranging from -80°C to 45°C. In one embodiment, the spray-dried plasma of the present invention can be stored at a warm temperature (e.g., from about 25°C to about 45°C) for about 1 hour to at least about 3 months. In one embodiment, the spray-dried plasma of the present invention can be stored at room temperature (e.g., from about 20°C to about 25°C) for about 1 hour to at least about 12 months. In one embodiment, the spray-dried plasma of the present invention can be stored at refrigerated temperatures (eg, about 1° C. to about 6° C.) for about 1 hour to at least about 48 months.

[0016] The reconstituted spray-dried plasma of the present invention, in one embodiment, is stable for transfusion for up to 26 hours, and the levels of one or more plasma proteins in the reconstituted spray-dried plasma are within about 25% compared to the levels of one or more plasma proteins measured immediately after spray-drying or compared to the corresponding clinical reference range.

[0017] The dried plasma of the present invention, upon reconstitution with SWFI, has a pH close to normal, for example, about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7 or about 7.8.

[0018] Furthermore, the reconstituted spray-dried plasma of the present invention has reduced amounts of the complement-activating protein fragments C5a and C3a. In particular, the spray-dried plasma has an amount of C5a of about 0.1 to about 74 ng / mL, or within the clinical reference range for apheresis plasma.

[0019] In another embodiment, the amount of vWF is measured by von Willebrand factor ristocetin cofactor and is within about 20% (e.g., 10%) of the amount of vWF in the donor plasma or within the clinical reference range. Similarly, vWF can be measured by von Willebrand factor antigen and is within about 20% (e.g., 10%) of the amount of vWF in the donor plasma or within the clinical reference range. In one embodiment, a von Willebrand factor ristocetin cofactor assay or a von Willebrand factor antigen assay is used to measure the amount of vWF, and the amount of vWF in either assay ranges from about 50 IU / dL to about 200 IU / dL.

[0020] The present invention includes a method for rehydrating a spray-dried plasma unit with spray-dried plasma by combining an amount of SWFI with the spray-dried plasma unit. In one embodiment, the spray-dried plasma is reconstituted in a period ranging from about 2 minutes to about 5 minutes, as measured from initial contact by a user to complete reconstitution without visible clumps.

[0021] The present invention includes spray-dried plasma produced from a disposable spray-drying 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 disposable spray-drying device includes a spray-drying head having a spray-drying nozzle assembly in fluid communication with the plasma source and the pressurized aerosol gas source, where the pressurized aerosol gas flows in a vortex pattern, and in use, the pressurized aerosol gas atomizes the plasma in the drying chamber to obtain atomized plasma droplets. The spray-drying head also includes a plenum having a drying gas inlet in communication with the drying gas source, where in use, the drying gas resides within the plenum at a uniform air pressure, and the plenum further includes a baffle plate supporting the nozzle assembly and forming a floor of the plenum with one or more drying gas jets, where the drying gas jets provide the drying gas to the drying chamber. The disposable described herein includes a drying chamber attached to a baffle plate, in which atomized plasma droplets evaporate in the presence of drying gas emitted from one or more drying gas jets, thereby obtaining dried plasma particles and moist air, and a trapping filter in the drying chamber, which traps the dried plasma particles and allows the moist air to pass through. The disposable also includes a gas outlet attached to an exhaust port of the spray-drying device, through which the moist air flows.

[0022] The present invention further includes a method for spray-drying plasma using a disposable spray-drying device for use in a spray-drying system as described herein. The method includes drying liquid plasma using a disposable spray-drying device of the spray-drying system. The method steps can further include reconstituting the spray-dried plasma, thereby obtaining reconstituted spray-dried plasma. In one embodiment, the reconstituted spray-dried plasma has a reduced number of cholesterol crystals compared to freeze-dried plasma when viewed at 100x magnification.

[0023] Advantageously, the spray drying method of the present invention effectively prevents or minimizes the formation of cholesterol crystals. Additionally, the spray-dried plasma of the present invention is largely amorphous upon reconstitution, yet contains no or minimal cholesterol crystals, and does not increase the average particle size (e.g., as measured by a reliable measurement system such as a Beckman Coulter Multisizer 4 using the electrical sensing zone method in the 2-60 micron range) in the reconstituted plasma when compared to a matched control that was not spray-dried. The dried plasma of the present invention has low residual water content compared to liquid plasma, making it stable at various temperatures and during extended storage. Another advantage of the dried plasma of the present invention is its rapid reconstitution, e.g., in less than four minutes. The dried plasma of the present invention can be reconstituted with sterile water for injection and maintains a near-normal plasma pH without treatment with or storage in CO2, or other post-drying pH adjustments. Furthermore, the reconstituted dried plasma of the present invention further provides protein function for the most vulnerable proteins, including von Willebrand factor, and other active proteins in donor plasma with reduced complement activation, a marker of inflammation. Furthermore, another advantage of the dried plasma of the present invention is the simplified storage, transportation and use options (e.g., refrigerated / ambient temperature storage / hot or a mixture thereof).

[0024] An additional advantage is that recipients at high risk for plasma transfusion (e.g., soldiers, police, travelers) can transport their own spray-dried plasma due to the lightweight nature of these units. In such cases, a healthcare provider (e.g., first responders) or other person assisting such a recipient would carry a reconstitution solution (e.g., sterile water for injection (SWFI)) so that in an emergency situation in which the recipient requires a plasma transfusion, the healthcare provider can reconstitute the recipient's own dried plasma for emergency transfusion, thereby eliminating issues such as incompatibility (e.g., ABO Rh matching and exogenous infection).

[0025] The foregoing and other objects, features, and advantages of the present invention will become apparent from the following more particular description of preferred embodiments of the invention, as illustrated in the accompanying drawings, in which like reference characters 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. [Brief explanation of the drawings]

[0026] [Figure 1] 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™ plasma. [Figure 2] 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 FLYP™ plasma. [Figure 3] 1 is a panel of black and white photographs showing phase contrast microscopy at 100x magnification of cholesterol crystals in freeze-dried plasma using LYOPLAS™ plasma (left) and FLYP™ plasma (right). [Figure 4] 1 is a panel of black and white photographs showing phase contrast microscopy of spray-dried plasma without visible cholesterol crystals observed as follows: donor plasma at 100x magnification before spray drying (top left panel), spray-dried plasma at 100x magnification (top right panel), donor plasma at 400x magnification before spray drying (bottom left panel), and spray-dried plasma at 400x magnification (bottom right panel). [Figure 5] 1 is a composite bar / line graph showing the microparticle concentration (microparticles / mL) and microparticle size (logarithmic scale (μm)) of single unit dried plasma at initial rehydration and after 7.5 months of room temperature storage 4 hours after rehydration compared to paired thawed frozen control plasmas as follows: Control plasma (CP) at time 0 (T=0), spray-dried plasma of the present invention (ODP) at time 0 (T=0), control plasma (CP) at time 4 hours (T=4), spray-dried plasma of the present invention (ODP) at time 4 (T=4). [Figure 6]1 is a composite bar / line graph showing particle concentration (particles / mL) and particle size (logarithmic scale (μm)) of single unit dried plasma at initial rehydration and after 12 months of refrigerated storage 4 hours after rehydration compared to its paired thawed frozen control plasma: Control plasma (CP) at time 0 (T=0), spray-dried plasma of the present invention (ODP) at time 0 (T=0), control plasma (CP) at time 4 hours (T=4), spray-dried plasma of the present invention (ODP) at time 4 (T=4). [Figure 7] 10A-10C are panels of black and white scanning electron microscope (SEM) photographs of spray-dried plasma particles from Run #3, with the top left panel at 2000x, the top right panel at 5000x, the bottom left panel at 5000x, the middle right panel at 1000x, and the bottom left panel at 5000x, with measurements overlaid and showing sizes ranging from 0.99 μm to 7.87 μm, and the bottom right panel at 2000x. These photographs demonstrate the small size and amorphous characteristics of the present invention. [Figure 8A] 10 shows panels of black and white scanning electron microscope (SEM) photographs of spray-dried plasma particles from run #7, with the top left panel at 2000x magnification, the top right panel at 1000x magnification, the bottom left panel at 5000x magnification, the middle right panel at 5000x magnification, the bottom left panel at 1000x magnification, and the bottom right panel at 2000x magnification. [Figure 8B] Panel of black and white scanning electron microscope (SEM) photographs of spray-dried plasma particles from run #7 at 5000x magnification with measurements overlaid on top, showing sizes ranging from 1.46 μm to 6.53 μm. [Figure 9A] 1 is a bar graph showing the results of clotting time (R) in minutes from a thromboelastography TEG study with reconstituted WB and mock resuscitation (abbreviations: WB:FFP = whole blood (WB) reconstituted with fresh frozen plasma (FFP); WB:ODP - whole blood reconstituted with spray-dried plasma (ODP) of the present invention; 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 indicated by dotted lines. [Figure 9B]1 is a bar graph showing the results of clotting rate (angle) from a thromboelastography TEG study with reconstituted WB and mock resuscitation (abbreviations: WB:FFP = whole blood (WB) reconstituted with fresh frozen plasma (FFP); WB:ODP - whole blood reconstituted with spray-dried plasma (ODP) of the present invention; 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 indicated by dotted lines. [Figure 9C] 1 is a bar graph showing results of clot strength (MA) in maximum amplitude (mm) from a thromboelastography TEG study with reconstituted WB and mock resuscitation (abbreviations: WB:FFP = whole blood (WB) reconstituted with fresh frozen plasma (FFP); WB:ODP - whole blood reconstituted with spray-dried plasma (ODP) of the present invention; 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 indicated by dotted lines. [Figure 9D] 1 is a bar graph showing the results of the lysis index (30 min) in LY30 as a percentage (%) from a thromboelastography TEG study with reconstituted WB and mock resuscitation (abbreviations: WB:FFP = whole blood (WB) reconstituted with fresh frozen plasma (FFP); WB:ODP - whole blood reconstituted with spray-dried plasma (ODP) of the present invention; 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 indicated by dotted lines. [Figure 9E] 1 is a bar graph showing the results of the lysis index (60 min) of LY60 in percentage (%) from a thromboelastography TEG study with reconstituted WB and mock resuscitation (abbreviations: WB:FFP = whole blood (WB) reconstituted with fresh frozen plasma (FFP); WB:ODP - whole blood reconstituted with spray-dried plasma (ODP) of the present invention; 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 indicated by dotted lines. [Figure 10A]1 is a bar graph showing the results of clotting time (R) in minutes from a thromboelastography TEG study using plasma only (abbreviations: FFP = fresh frozen plasma (FFP); ODP - spray-dried plasma (ODP) of the present invention; 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 indicated by dotted lines. [Figure 10B] 1 is a bar graph showing the results of clotting rate (angle) from a thromboelastography TEG study using plasma only (abbreviations: FFP = fresh frozen plasma (FFP); ODP - spray-dried plasma (ODP) of the present invention; 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 indicated by dotted lines. [Figure 10C] 1 is a bar graph showing the results of clot strength in maximum amplitude (MA) (mm) from a thromboelastography TEG study using plasma only (abbreviations: FFP = fresh frozen plasma (FFP); ODP - spray-dried plasma (ODP) of the present invention; 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 indicated by dotted lines. [Figure 11] 1 is a bar graph showing the results of vWF (von Willebrand Factor):Ristocetin (Rist) cofactor activity (percent (5)) from a ristocetin cofactor assay. (Abbreviations: WB:FFP = whole blood (WB) reconstituted with fresh frozen plasma (FFP); WB:ODP = whole blood reconstituted with spray-dried plasma (ODP) of the present invention; 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 indicated by dotted lines. [Figure 12] 1 is a bar graph showing the results of ADAMTS-13 (von Willebrand factor-cleaving protease) activity by the ADAMTS-13 assay. (Abbreviations: FFP = fresh frozen plasma (FFP); ODP - spray-dried plasma (ODP) of the present invention.) [Figure 13A]1A-1C are two bar graphs showing the results of a platelet adhesion Bioflux study, showing the percentage (%) of arterial shear at intensity NS(10min) 900 s-1 in fluorescence intensity units (FIU) and area NS(10min) coverage. (Abbreviations: NS = normal shear conditions; WB:FFP = whole blood (WB) reconstituted with fresh frozen plasma (FFP); WB:ODP - whole blood reconstituted with spray-dried plasma (ODP) of the present invention; 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 indicated by dotted lines. [Figure 13B] Included are two bar graphs showing the results of a platelet adhesion Bioflux study, showing the percentage (%) of pathological shear at 4000 s of intensity HS(10 min) in fluorescence intensity units (FIU) and area HS(10 min) coverage. (Abbreviations: HS = high shear conditions; WB:FFP = whole blood (WB) reconstituted with fresh frozen plasma (FFP); WB:ODP - whole blood reconstituted with spray-dried plasma (ODP) of the present invention; 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 indicated by dotted lines. [Figure 14A] Included are two bar graphs showing the results of a CAT (calibrated automated thrombogram) thrombin generation assay, showing the lag time in minutes and the endogenous thrombin potential (ETP) (nM, min). (Abbreviations: WB:FFP = whole blood (WB) reconstituted with fresh frozen plasma (FFP); WB:ODP - whole blood reconstituted with spray-dried plasma (ODP) of the present invention; 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 indicated by dotted lines. ETP reflects coagulation status and is a measure of whether someone is prone to bleeding (low ETP) or clotting (high ETP). [Figure 14B]1 includes two bar graphs showing the results of a CAT (calibrated automated thrombogram) thrombin generation assay, showing peak thrombin (nM) and time to peak (min). (Abbreviations: WB:FFP = whole blood (WB) reconstituted with fresh frozen plasma (FFP); WB:ODP - whole blood reconstituted with spray-dried plasma (ODP) of the present invention; 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 indicated by dotted lines. [Figure 15A] 1 includes three bar graphs showing the flow cytometry results of the remaining cellular material in total events, total labeled events, and CD41A (platelets) events. (Abbreviations: FFP = fresh frozen plasma (FFP); ODP - spray-dried plasma (ODP) of the present invention. [Figure 15B] Contains three bar graphs showing the results of flow cytometry of residual cellular material for CD45 (WBC) and CD235 (RBC). (Abbreviations: FFP = fresh frozen plasma (FFP); ODP - spray-dried plasma (ODP) of the present invention; WBC - white blood cells; RBC - red blood cells). [Figure 16] Panels A-C are schematic diagrams showing the unfolding / refolding model of the vWF A2 domain and proton cleavage by ADAMTS13. (A) Illustration of the vWF A2 domain in its native folded state. (B) The first step of unfolding occurs from the C-terminus of the vWF A2 domain and is influenced by the presence of a proximal disulfide bond (cysteine ​​indicated by C). Initial unfolding occurs up to and including the central b4 sheet, which contains the scissile bond (YM). This intermediate step of unfolding exposes the high-affinity ADAMTS13 spacer-binding site. (C) Once the stabilizing effect of the calcium-binding site (CBS) is overcome, this leads to the complete unfolding of the vWF A2 domain and positions the ADAMTS13 active site for nucleophilic attack of the Y1605-M1606 scissile bond. [Figure 17]This is a bar graph showing that formulations of citrated plasma stabilize during spray drying at approximately 50% von Willebrand factor:ristocetin cofactor (vWF:RCo) activity without any effect on 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 normalized to control plasma (fresh frozen plasma (FFP)) at time 0, the completion of spray drying. CP indicates control plasma; SpDP indicates spray-dried plasma; PreT indicates plasma formulation by SDSAS. [Figure 18] This is a bar graph showing that formulation of plasma with citrate confers 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), and % von Willebrand factor:ristocetin cofactor (vWF:RCo)) during 6 weeks of storage at 4°C. SpDP indicates spray-dried plasma. PreT indicates plasma preparation by SDSAS. CP indicates control plasma, and FFP is fresh frozen plasma. [Figure 19] This bar graph shows that pre-treatment of plasma with citrate confers 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), and % von Willebrand factor:ristocetin cofactor (vWF:RCo)) after 2 weeks of storage at 25°C. SpDP indicates spray-dried plasma. PreT indicates plasma preparation by SDSAS. CP indicates control plasma, and FFP is fresh frozen plasma. [Figure 20]1 is a bar graph showing that formulation of plasma with citrate confers stability to coagulation factors (prothrombin (PT), activated partial thromboplastin time (aPTT), fibrinogen (FGN), factor V (FV), factor VII (FVII), factor VIII (FVIII), factor IX (FIX), and vWF antigen (vWF-Ag)) during 2 weeks of storage at 37°C. SpDP indicates spray-dried plasma. PreT indicates plasma preparation by SDSAS. CP indicates control plasma, and FFP is fresh frozen plasma. [Figure 21] 1 is a photographic image showing that formulation of plasma with citrate stabilizes 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. [Figure 22A] This is a line graph showing the CP / FFP and pH of delivered plasma under a constant plasma delivery 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. [Figure 22B] 1 is a line graph showing the resulting activity (%, IU / dL) of vWF:RCo activity of CP / FFP and fed plasma under a constant plasma feed rate of 10 mL / min but variable aerosol gas flow rates (0, 5, 10, 15, 20 L / min). CP denotes control plasma; FFP denotes fresh frozen plasma. [Figure 23A] A line graph showing the pH of CP / FFP and supplied plasma at an aerosol gas flow rate of 10 L / min; fluid = 2 mL / min, 10 L / min; fluid = 4 mL / min, 10 L / min; fluid = 6 mL / min, 10 L / min; fluid = 8 mL / min, 10 L / min; and 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 ristocetin assay). [Figure 23B]1 is a line graph showing the resulting vWF:RCo activity (%, IU / dl) of donated plasma at CP / FFP and 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, 10 L / min; fluid = 8 mL / min, 10 L / min; fluid = 10 mL / min. CP indicates control plasma; FFP indicates fresh frozen plasma. [Figure 24] 1 is a bar graph showing the effect of different SDSAS formulations on vWF:RCo recovery and pH during nebulization. pH levels before and after nebulization are indicated above the bar graph. CP indicates control plasma; FFP indicates fresh frozen plasma; vWF indicates von Willebrand factor. [Figure 25A] Bar graphs showing the effect of different SDSAS formulations on vWF:RCo recovery and pH during spray drying. (A) Citric acid. pH levels before and after spraying are indicated at the top of the bar graph. vWF stands for von Willebrand factor. vWF:RCo (vWF activity measured by vWF ristocetin assay). [Figure 25B] (B) Bar graphs showing the effect of different SDSAS formulations on vWF:RCo recovery and pH during spray drying. (C) Lactic acid. pH levels before and after spraying are indicated at the top of the bar graph. vWF stands for von Willebrand factor. vWF:RCo (vWF activity measured by vWF ristocetin assay). [Figure 25C] (C) Bar graphs showing the effect of different SDSAS formulations on vWF:RCo recovery and pH during spray drying. (D) pH. pH levels before and after spraying are indicated at the top of the bar graph. vWF stands for von Willebrand factor. vWF:RCo (vWF activity measured by vWF ristocetin assay). [Figure 26] 1 is a color line graph showing the amount of C3 ang / mL and pH for batches 1543, 1542, 1541 and their respective control plasmas (CP) for the experiments performed in Example 16. [Figure 27] 1 is a line graph showing the amount of C5 ang / mL and pH for batches 1543, 1542, 1541 and their respective control plasmas (CP) for the experiments conducted in Example 16. [Figure 28] 1 is a bar graph showing C5a analysis of control plasma (CP) / tube with 400 mM glycine HCl, 400 mM glycine HCl + 1 M glycine, 148 mM citric acid, 148 mM citric acid + 1 M glycine (n=7) and non-frozen plasma (NFP) pretreated rapidly and slowly with control plasma / non-frozen plasma (CP / NFP). [Figure 29] 1 is a bar graph showing C3a measurements 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), not frozen, pre-treated) mean of n=20±1 SD. [Figure 30] 1 is a bar graph showing mean C5a measurements in ng / mL for CP / FFP, CP / FFP / PreT, and ODP / NFP / PreT, n=20±1 SD. [Figure 31] 1 is a bar graph showing antigen measurements normalized to ODP / NFP / PreT activity and CP / FFP. [Figure 32] 1 is a bar graph showing aPTT (activated partial thromboplastin time), PT (prothrombin time), and TT (thrombin time) of ODP / NFP / PreT normalized by CP / FFP. [Figure 33] 1 is a bar graph showing vWF antigen (von Willebrand factor), vWF:RCo (vWF activity measured by vWF ristocetin assay), and vWF activity of ODP / NFP / PreT (on-demand plasma (applicant's inventive spray-dried plasma), not frozen, pre-processed) normalized to CP / FFP (control plasma / fresh frozen plasma). [Figure 34] 1 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) not frozen, pre-processed) normalized with CP / FFP (control plasma / fresh frozen plasma). [Figure 35]1 is a bar graph showing chemistry analyzer results of ODP / NFP / PreT (on-demand plasma (applicant's inventive spray-dried plasma), not frozen and pre-processed) for IgG, IgM, IgA, total protein, albumin, triglycerides, cholesterol, LDL cholesterol, HDL cholesterol and calcium normalized to CP / FFP (control plasma / fresh frozen plasma). [Figure 36] 1 is a bar graph showing thromboelastography 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). [Figure 37A] 1 is a bar graph showing clotting profile results and ELISA assay results for various clotting factors and complement activation for unfrozen plasma pretreated with 400 mM glycine HCl and spray-dried at exhaust gas temperatures of 49, 50, 51, and 52° C. [Figure 37B] 1 is a bar graph showing coagulation activation and complement activation marks (D-dimer, TAT, PF1.2, C3a, and C5a) of fresh frozen plasma normalized to control plasma pretreated with 400 mM glycine HCl and spray-dried at exhaust gas temperatures of 49, 50, 51, and 52° C. [Figure 38A] 1 is a bar graph showing C5a levels in ng / mL at 10 minutes, 1 hour, 2 hours, and 21 hours for rehydrated plasma pretreated with 400 mM, 280 mM, and 140 mM glycine HCl (GlyHCl) and 148 mM, 100 mM, and 50 mM citric acid (CA). [Figure 38B] 1 is a bar graph showing the pH of plasma pretreated with 400 mM, 280 mM, 140 mM glycine HCl (GlyHCl) and 148 mM, 100 mM, and 50 mM citric acid (CA). [Figure 39] 1 is a line graph plotting the pH level of the pretreatment solution and pretreatment plasma sample versus the glycine concentration (mM). [Figure 40]1 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 and 60 minutes using rapid mixing (e.g., when a larger volume of plasma is quickly added to a relatively small volume of pretreatment solution). [Figure 41A] 1 is a line graph showing the pH of plasma (batches 1949 and 1950) pretreated with pretreatment solution and 400 mM lactate and supplemented with increasing concentrations of glycine (0, 400, 600, 800, 1000, 1200, 1400, and 1600 mM). [Figure 41B] 1 is a bar graph showing C5a levels in ng / mL in plasma (batches 1949 and 1950) pretreated with 400 mM lactate and supplemented with increasing concentrations of glycine, where the pretreated plasma had concentrations of 0, 20, 30, 40, 50, 60, 70, and 80 mM glycine. [Figure 42A] Schematic diagram showing a perspective view of a spray drying disposable device including a liquid plasma bag, a spray drying head, and a spray drying chamber, the disposable having alignment elements that allow it to be aligned with the spray drying equipment and finishing equipment. [Figure 42B] 1 is a model representation of the three-dimensional flow geometry of the disposable flow model in operation, which is used to create the computer flow models described herein. [Figure 43A] FIG. 42B is a schematic diagram showing a perspective view of the spray drying head of the spray drying disposable device shown in FIG. 42A. [Figure 43B] 42B is a schematic diagram showing an exploded view of the spray drying nozzle assembly and spray drying head of the spray drying disposable device shown in FIG. 42A. [Figure 43C] FIG. 1 is a schematic diagram showing a perspective view of a spray drying nozzle assembly from the spray drying head of a spray drying disposable device. [Figure 43D] 43D is a schematic diagram showing a perspective view of the spray drying nozzle assembly of FIG. 43C, but with the aerosol reservoir housing transparent to show the internal structure of the assembly. [Figure 43E] FIG. 43D is a schematic diagram showing a perspective view of the spray drying nozzle assembly of FIG. 43C with the aerosol reservoir housing and the nozzle cap and nozzle cap insert removed to show the manifold and cannula. [Figure 43F] FIG. 1 is a schematic diagram showing a front view of one embodiment of a beveled edge cannula that is part of a spray drying nozzle assembly. [Figure 43G] FIG. 10 is a schematic diagram showing a perspective top view of the cannula and the nozzle cap insert that guides the aerosolized air. [Figure 43H] FIG. 10 is a schematic diagram showing a perspective bottom view of the nozzle cap insert with the cannula inserted. [Figure 43I] FIG. 2 is a schematic diagram showing a top view of the nozzle cap. [Figure 43Ia] FIG. 43I is a schematic diagram showing a bottom view of the nozzle cap of FIG. 43I, with the annular member residing within the cap opening. [Figure 43Ib] 43I is a schematic diagram showing three possible vortex generator flow patterns that can be used with the nozzle cap insert of FIG. 43I. [Figure 43Ic] FIG. 43I is a schematic diagram showing a cross-sectional view of the nozzle cap insert of FIG. 43H residing within the nozzle cap of FIG. 43I. [Figure 43J] FIG. 1 is a schematic diagram showing a perspective bottom view of the plenum of the spray drying head. [Figure 43K] FIG. 1 is a schematic diagram illustrating a partial front view of a spray dryer showing a portion of a drying gas deflector. [Figure 43Ka] FIG. 10 is a diagram showing modeled dry gas flow in a plenum chamber using a 15 m / s constant velocity amplitude surface. [Figure 43L] FIG. 2 is a schematic diagram showing a top perspective view of a baffle plate of a spray drying head. [Figure 43La] FIG. 43La is a schematic diagram showing a cut-out portion of the baffle plate rib design shown in FIG. 43L and a cut-out portion of another variation of the baffle plate rib design, and also shows a cross section of one of the ribs. [Figure 43M] FIG. 2 is a schematic diagram showing a perspective bottom view of a baffle plate of a spray drying head. [Figure 43Ma] Model representation showing uniform jet penetration and drying gas distribution at a constant velocity of 25 m / s, thus introducing uniform drying gas circumferentially around the spray plume. [Figure 43N] FIG. 1 is a schematic diagram illustrating droplet plume formation, aerosol gas flow, and drying gas flow to promote rapid mixing in a disposable device of the present invention. [Figure 43Na] FIG. 10 is a model diagram showing gas velocity magnitude contours within the disposable center plane, illustrating the drying jet penetration into the drying chamber and the effect of the baffle plate flow paths and their interaction with the high velocity spray plume, which acts to draw the drying gas jet radially inward to aid in the desired rapid mixing of the droplets with the gas flow. [Figure 43O] FIG. 1 is a model diagram showing the gas pressure flow (psig) (top) and tangential velocity flow (m / s) (bottom) of the vortex generated within the nozzle insert and cap assembly. [Figure 43P] 1 is a model diagram showing gas velocity amplitude flow (m / s) in the components of a vortex generator. [Figure 43Q] FIG. 1 is a schematic diagram showing the transformation of droplets into dry particles using a disposable of the present invention. [Figure 43R] 1 is a line graph showing droplet wet bulb temperature (° C.) and drying gas temperature (° C.) for droplets dried to particles with 0% relative humidity (RH), 10% RH, and 20% RH. This particular graph illustrates the concept but is not specific to plasma. [Figure 43S] 1 is a line graph showing the evaporative mass transfer of droplet temperature over time for all mean droplet trajectories averaged over three simulated drying gas inlet temperatures in the model: 80° C., 100° C., and 114° C. Note that the evaporation process cools the droplets to keep the delicate liquid protein below 30° C. [Figure 43Sa]Line graph of temperature in °C vs. time (seconds) of plasma droplets as they become particles in the model. Once evaporation is complete, the proteins encased in solid particles are more resistant to high temperatures because they equilibrate with the dryer outlet temperature. In this case, evaporation occurs in less than a fraction of a second (e.g., 0.01-0.05 seconds). [Figure 43T] FIG. 1 is a model diagram showing the paths of droplets having sizes of 5 microns, 15 microns, and 25 microns during evaporation. The diagram shows that smaller droplet size allows for more rapid evaporation mass transfer over a shorter path, allowing for a physically smaller drying chamber. [Figure 44A] FIG. 42B is a schematic diagram showing an exploded view of the drying chamber of the spray drying disposable device shown in FIG. 42A. [Figure 44B] FIG. 1 is a schematic diagram showing a front view of a separator or spacer that can be used in the drying chamber of a spray drying disposable. [Figure 45A] FIG. 2 is a schematic diagram showing a front view of the spray drying apparatus with the door closed. [Figure 45B] FIG. 1 is a partial front view of the spray drying apparatus without the door to reveal the drying chamber housing with alignment elements that allow alignment with the spray drying disposable device. [Figure 45C] FIG. 1 is a schematic diagram showing a front view of the spray drying apparatus with the door handle engaged and the door open. [Figure 46A] 1 shows an alignment element for aligning the spray drying disposable device with the spray drying apparatus. [Figure 46B] FIG. 1 is a schematic diagram showing a partial front view of a spray drying apparatus without the door, with the spray drying disposable device installed, and with the deflector engaged. [Figure 46C]2 is a schematic diagram showing the structure 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 - peristaltic pump, PP - pneumatic piston, PR - pressure regulator, PRV - pressure relief valve / rupture disc, PT - pressure transducer, PV - valve (downward arrow is fail closed, upward arrow is fail open), S - scale, SS - solenoid, TC - thermocouple, TS - thermocouple sensor, and TT - temperature transducer. [Figure 46D] 1 is a flow chart showing the steps of a leak detection method using a spray dryer and spray dried disposables. [Figure 46E] 1 is a flowchart showing the steps of a pressure detection method using a spray dryer and spray drying disposables. [Figure 46F] 1 is a flow chart showing the steps of a method for detecting the integrity of a filter and disposable interface using a spray dryer and spray dried disposables. [Figure 46G] 1 is a line graph showing the slope using a method to detect filter integrity and demonstrating that the filter is intact by measuring pressure (psig), elapsed time (min), and rate of pressure change (psi / min). [Figure 46H] 1 is a line graph showing the slope using a method to detect filter integrity and demonstrate that the capture filter failed intact by measuring pressure (psig), elapsed time (minutes), and rate of pressure change (psi / minute). [Figure 46I] 1 is a line graph showing the slope using a method to detect filter integrity and showing that a baffled filter failed intact by measuring pressure (psig), elapsed time (minutes), and rate of pressure change (psi / minute). [Figure 47A] FIG. 1 is a schematic diagram showing a front view of the finishing device in the loading position with no spray drying disposables attached. [Figure 47B]47B is a schematic diagram showing a front view of the finishing device of FIG. 47A without the front cover with the shuttle in the lowered position. FIG. [Figure 47C] FIG. 47C is a schematic diagram showing a front view of the finishing device of FIG. 47B with the shuttle in the raised position and no disposables attached. [Figure 47D] FIG. 10 is a schematic diagram showing a front view of another embodiment of the finishing device in a raised position and without spray drying disposables attached. [Figure 47E] FIG. 47D is a schematic diagram showing a front view of the finishing device of FIG. 47D without the front cover with the shuttle in the raised position. [Figure 47F] FIG. 47D is a schematic diagram showing a front view of the finishing device of FIG. 47D without the front cover with the shuttle in the lowered position. [Figure 47G] FIG. 47D is a schematic diagram showing a front view of the finishing device of FIG. 47D but without the front cover with the shuttle in the raised and inverted position. [Figure 47H] FIG. 47D is a schematic diagram showing a front view of the finishing device of FIG. 47D without the front cover with the shuttle in a lowered and inverted position. [Figure 47I] FIG. 47D is a schematic diagram showing an enlarged perspective view of a portion of the rail system of the finishing device of FIG. 47D. [Figure 48A] FIG. 1 is a schematic diagram showing a front view of the finishing device with the disposable spray drying head in the aligned and mounted position. [Figure 48B] FIG. 1 is a schematic diagram showing a front view of the finishing device with the disposables in an aligned, raised position. [Figure 48C] A schematic diagram showing a front view of the finishing device in a raised position with a portion of the disposable attached after the first sealing and separation step has been completed and the frame has been rotated into place. [Figure 48D] FIG. 10 is a schematic diagram showing a front view of another embodiment of the finishing device in a raised position with spray drying disposables attached. [Figure 48E] FIG. 48E is a schematic diagram showing a front view of the finishing device shown in FIG. 48D in the lowered position with spray dryer disposables attached. [Figure 48F] FIG. 48D is a schematic diagram showing a front view of the finishing device shown in FIG. 48D in the raised and inverted positions with spray dryer disposables attached. [Figure 48G] FIG. 48E is a schematic diagram showing an enlarged top view of the receiver of the finishing device shown in FIG. 48D. [Figure 48H] FIG. 48E is a schematic diagram showing an enlarged perspective view of the receiver of the finishing device shown in FIG. 48D. [Figure 48I] FIG. 48E is a schematic diagram showing an enlarged perspective view of the tensioner of the finishing device shown in FIG. 48D. [Figure 48J] FIG. 48E is a schematic diagram showing an enlarged perspective view of a separator having a roller that engages with a tensioner of the finishing device shown in FIG. 48D. [Figure 48K] FIG. 48E is a schematic diagram showing an enlarged perspective view of the impactor, separator, and sealer of the finishing device shown in FIG. 48D. [Figure 48L] FIG. 48E is a schematic diagram showing a side view of the impactor, separator, and sealer of the finishing device shown in FIG. 48D. [Figure 49A] FIG. 1 is a schematic diagram showing a front view of a spray-dried plasma unit obtained from a spray-drying disposable device after processing by a finishing apparatus. [Figure 49B] 4 is a schematic diagram showing the structure of a 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 (downward arrow is fail closed, upward arrow is fail open), SC - speed controller, SR - sensor, and VG - vacuum generator. [Figure 50] 1 is a bar graph comparing the vWF% ratio of reconstituted plasma (vs. undried control aliquots) dried using disposable devices with composite nozzle assemblies without a bevel, with a bevel and a reference stainless steel nozzle. [Figure 51A] 1 is a flowchart illustrating steps of a pre-processing methodology. [Figure 51B] 1 is a flow chart showing steps of a spray drying method using a spray dryer and spray drying disposables. [Figure 51C] 1 is a flow chart showing the steps of a finishing method using a finisher and spray-dried disposables to create dried plasma units. [Figure 51D] 1 is a flow chart showing steps of a storage method after the spray drying unit has been prepared. [Figure 52] Schematic diagram showing cannula geometry to demonstrate shear on vWF protein as it exits the cannula. The curved arrow indicates the aerosol gas vortex direction within the annulus. To demonstrate the amount of shear impact on the liquid plasma at the cannula exit region, the diagram shows cannula edges at 15, 90, 45 degrees (with chamfers and sharp edges), and 60 degrees, showing how the angled edges reduce the shear contact of the cannula. DETAILED DESCRIPTION OF THE INVENTION

[0027] A description of a preferred embodiment of the present invention follows.

[0028] The present invention relates to spray-dried plasma. The spray-dried plasma is obtained using a spray-drying device and disposables as further described herein. The spray-dried plasma of the present invention has the following properties: 1) It is largely amorphous, as defined as having no distinct shape, e.g., small protein and lipid particles (<20 microns) with high specific surface area and short diffusion lengths, unlike freeze-dried plasma, where the majority of amorphous dried particles typically form a solidified, porous "cake"; 2) upon reconstitution, there is little or no presence of crystals (e.g., cholesterol) created by the drying process; 3) upon reconstitution, the system of the present invention does not create microparticles and the number of larger microparticles is reduced, particularly for microparticles having a size of approximately 2 μm to 60 μm compared to the microparticles in donor plasma as measured using a Coulter Multisizer 4 with an electric detection zone method; 4) Low residual moisture, 5) Rapid reconfiguration, measured from initial contact by the user to complete reconfiguration without visible lumps, in less than 4 minutes; 6) It is stable when stored below refrigeration temperatures, at room temperature and at elevated temperatures, or a combination thereof, compared to liquid plasma, and can be stored for extended periods of time. 7) Stable for at least 4 hours (e.g., up to 26 hours) after reconstitution, and the product remains sterile; 8) Upon reconstitution, the protein functions of most plasma proteins in donor plasma are preserved, including fragile proteins such as von Willebrand factor and other active proteins. 9) Upon reconstitution with sterile water for injection (SWFI), the reconstituted plasma has a near-normal plasma pH (not absolutely high (alkalosis)) that is favorable for use in transfusion and is immediately usable for transfusion without treatment with or storage in CO2, or other post-drying pH adjustment; 10) Upon reconstitution, the plasma does not activate complement (C5a, C3a) compared to apheresis plasma (complement activation is involved in inflammation).

[0029] The spray-dried plasma of the present invention is used for transfusions and the like and is an alternative to liquid, frozen, or freeze-dried plasma. In one embodiment, the dried plasma preparation of the present invention, also referred to herein as a "plasma unit," "dried plasma unit," "plasma preparation," or "unit," is a standard dose of dried plasma intended for rapid rehydration at the point of care. The dried plasma of the present invention allows local blood centers and the like to prepare single-donor units of plasma or pooled plasma in a dried form that can be more easily stored, transported, and transfused than frozen plasma.

[0030] The dried plasma units of the present invention are, in part, a source of plasma protein for patients who are deficient or have a deficient plasma protein for which no specific replacement factor is available. The dried plasma units of the present invention can also be used for humans who require blood volume and / or clotting factor treatment, such as for external or internal bleeding or medical conditions. The dried plasma units of the present invention provide a plasma product when traditional plasma (such as fresh frozen plasma (FFP) or plasma frozen within 24 hours (PF24)) is unavailable or impractical, such as for pre-hospital transfusions, rural / basic hospitals, and military applications.

[0031] Additional indications for using the dried plasma units of the present invention include the management of preoperative or bleeding patients requiring replacement of multiple clotting factors (e.g., liver disease, disseminated intravascular coagulation (DIC)), and transfusion-receiving patients with clinically significant coagulation deficiency. The plasma of the present invention is capable of clot formation upon reconstitution.

[0032] The dried plasma units of the present invention are suitable for use in massive transfusions because they are capable of inducing clot formation and do not expose the recipient to the risks of high pH, ​​cholesterol or other crystals, particulates, or complement activation, compared to other blood products currently available and expected to be available for transfusion.

[0033] To better understand the properties of plasma, a description of spray drying systems, disposables and methods is provided herein.

[0034] Plasma properties: Dried plasma is amorphous with few or no crystals The present invention relates to reconstituted, pre-spray-dried plasma produced by a process that produces few or no crystals (e.g., cholesterol crystals). As used herein, "reconstituted plasma" refers to reconstituted plasma spray-dried using the methods, disposables, and spray-drying systems described herein. In certain embodiments, the reconstituted plasma has few or no crystals visible using a phase-contrast microscope, in one embodiment, set at 100x, 400x, or both. Cholesterol crystals are cholesterol that form a type of lattice structure, often in the form of elongated rods or thin, rectangular plates. In another embodiment, the reconstituted plasma has fewer crystals (e.g., cholesterol crystals) compared to pre-freeze-dried reconstituted plasma. Crystals in reconstituted freeze-dried plasma have been observed in the range of about 1 μm to about 3 μm. The formation of dried plasma with few or no crystals is, in part, a result of the spray-drying process, the nozzle design of the disposable, the rapid mixing of liquid plasma droplets with the drying gas during spray-drying, the size of the droplets, and the parameters under which the plasma is dried. These aspects are further described herein.

[0035] Without being bound by any particular theory, it is believed that spray-drying plasma using the disposable 100 and dryer 200 described herein allows for rapid mass transfer (e.g., less than one second) to convert liquid plasma into an amorphous dry material. As a result, the formation of crystalline solids of many molecules, including cholesterol, is suppressed because there is no minimum time or mobility required to orient the molecules in a low-energy crystalline state during the production of the spray-dried plasma of the present invention. In contrast, freeze-drying has a long time window (e.g., hours or days) during which crystals, including cholesterol crystals, can form and stabilize. See Examples 1 and 2.

[0036] Examples 1 and 2 describe that no crystal formation is observed in the rehydrated spray-dried plasma of the present invention, but is present in the freeze-dried plasma. Crystals can be identified by microscopic examination, i.e., compound, phase-contrast, or electron microscope at various magnifications (e.g., 40x, 80x, 100x, 200x, 400x, 800x, 1000x, and 2000x). In one embodiment and in the Examples, crystals are identified in all of the freeze-dried plasma samples at 100x and 400x magnifications.

[0037] In one aspect, cholesterol crystals are undesirable in plasma. The freeze-drying / lyophilization process creates such crystals in rehydrated freeze-dried plasma preparations. Freeze-dried human plasma has crystal formation upon reconstitution, so recipients receive cholesterol crystals during transfusions with previously freeze-dried reconstituted plasma. Cholesterol crystals are undesirable in the human body because they induce inflammation in atherosclerosis, which can lead to heart attacks and strokes. Furthermore, because cholesterol cannot be easily broken down by mammalian cells, they have a persistent effect. In particular, cholesterol crystals have the following undesirable properties: 1) Excess cholesterol is excreted in the bile, transported in specific transporter molecules (HDL, LDL), and modified by cells but not degraded. In certain cases, cholesterol crystals form in the bile and are involved in the formation of stones; 2) Cholesterol crystals are formed during the formation of atherosclerotic plaques. Excessive crystal growth is involved in plaque rupture, and the severity of symptoms is related to the content of cholesterol crystals; 3) Cholesterol crystals spread throughout the body as a result of plaque rupture. Cholesterol crystals released from atherosclerotic plaques can spread to almost any organ. Effects can vary from relatively benign skin symptoms to multiple organ failure, which manifests within hours to 5 months of exposure; 4) that cholesterol crystals caused by human atheroma, a degeneration of the arterial wall caused by accumulated fatty deposits and scar tissue, can still be found in significant amounts after vascular repair; furthermore, that large crystals have been shown to damage endothelial cells, while smaller crystals are thought to be less damaging; and 5) Cholesterol crystals are known to activate the immune system and induce inflammatory processes.

[0038] The effects of cholesterol crystals may be immediate or delayed and may vary from patient to patient. If cholesterol crystal-related effects (mild or severe) are observed in a patient, these effects may be interpreted as being part of the patient's disease or dietary behavior, and not as being caused by treatment with reconstituted lyophilized plasma.

[0039] Due to the undesirable effects of cholesterol crystals, the dried plasma of the present invention is preferable for transfusion compared to freeze-dried plasma. The reconstituted plasma of the present invention does not contain crystals (e.g., cholesterol crystals) visible at 100x and 400x magnification and is largely amorphous. "Amorphous" refers to a non-crystalline solid in which atoms and molecules are not organized in a lattice pattern and lack long-range order. While freeze-drying promotes the formation of cholesterol crystals due to supercooling, the spray-drying process of the present invention minimizes or avoids crystal formation by promoting rapid mixing and rapid mass transfer, in which plasma droplets are dried into amorphous plasma particles in less than one second. Note that donor-derived crystals may be present in the final formulation. The spray-drying process does not create additional crystals in the plasma. If certain crystals are present in the donor plasma, they may also be present in the reconstituted, previously spray-dried plasma. For example, uric acid crystals or calcium phosphate crystals present in the donor plasma may also be found in the reconstituted plasma unit. Thus, the reconstituted, pre-spray-dried plasma of the present invention is largely amorphous or substantially amorphous (e.g., at least about 99%, about 98%, about 97%, about 96%, about 95%, about 94%, about 93%, about 92%, about 91%, about 90% amorphous). As a result, the formation of crystalline solids of many molecules, including cholesterol, is inhibited during the spray drying of the present invention because the conditions required for crystal formation are not present. In contrast, the freeze-drying process promotes crystal formation, including cholesterol crystal formation, by providing the time and conditions for crystal formation. See Figures 1-4. Figure 7 provides a photograph illustrating the small size and amorphous nature of the present invention.

[0040] As a result, the dried plasma of the present invention is largely substantially or entirely amorphous in structure, with few or no crystals (e.g., cholesterol crystals). Because the spray-drying process described herein does not produce plasma, the reconstituted plasma of the present invention has few or no crystals, except for those present in the corresponding donor plasma.

[0041] The average size of microparticles in the reconstituted plasma of the present invention is reduced Various types of particulates, such as protein aggregates, platelets, and microparticles, are naturally present in plasma, including human plasma. When plasma containing excess or large particulates is transfused into a recipient, these protein aggregates or microparticles react with the recipient's body and cause inflammation or other immunological reactions, even though these microparticles may be benign to the donor.

[0042] As used herein, the term "particulate" is distinct from the term "particles." "Particulate" refers to protein aggregates, platelets, and microparticles in reconstituted plasma. "Particles" refer to the individual, discreet components that make up dried plasma.

[0043] A particulate test was performed on the dried plasma of the present invention and it was determined that there was no increase in particulate size or distribution in 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.

[0044] In other words, upon reconstitution, the rehydrated plasma exhibits a reduced average particle size compared to the particles in the donor plasma for particle sizes between about 2 μm and about 60 μm as measured using an electric detection zone Coulter Multisizer 4. Unlike the spray-dried plasma of the present invention, freeze-dried plasma has not been reported to reduce particle size or quantity in the resulting reconstituted plasma for transfusion.

[0045] The reconstituted plasma of the present invention has a reduced average particle size compared to that found in the original donor plasma. Without being bound by theory, it is believed that the spray drying process of the present invention reduces larger particles in the donor plasma to smaller particle sizes, while proteins, even delicate proteins, maintain their functionality and integrity. See Example 3.

[0046] Particle size analysis characterization studies were conducted to assess the impact of spray-drying manufacturing on protein aggregation and other particulates, in-use stability, and shelf life of the formulation. See Example 3. Particle size analysis data support that the rehydrated dried plasma units of the present invention do not exhibit protein aggregation or other particulates due to manufacturing and storage after combined storage temperatures of up to 7.5 months at room temperature, up to 24 months refrigerated, or 1 year refrigerated + 6 months at room temperature, compared to (1) a matched control and (2) t=4 hours after rehydration.

[0047] Particles in the 2-60 micron range, as measured by electrochemical detection zone analysis, do not increase in average size when compared to non-spray-dried matched controls. As measured, the particles in the reconstituted plasma ranged in size from about 2 μm to about 3.5 μm, compared to donor plasma, which had a size range of about 2 μm to about 7 μm. See Figure 5 (room temperature, 7.5 months) and Figure 6 (refrigerated, 12 months). In one embodiment, the maximum particle size of the reconstituted, pre-spray-dried plasma is about 50%, about 45%, about 40%, about 35%, about 30%, about 25%, about 20%, about 15%, about 10%, about 5%, about 4%, about 3%, about 2%, or about 1% of the maximum particle size found in the donor plasma. See Tables 11 and 12 in Example 3. As further described herein, plasma proteins are involved in and enable clot formation, including fragile proteins such as von Willebrand factor (vWF). In all cases, 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 compared to the matched non-spray-dried control. Furthermore, the mean size of microparticles was reduced compared to donor plasma when measured in the range of about 2 μm to about 60 μm. The proportion of microparticles having a mean size in the range of about 2 μm to about 60 μm in the reconstituted plasma of the present invention is reduced compared to donor plasma. In one embodiment, the number of microparticles having a mean size in the range of about 2 μm to about 60 μm found in the reconstituted, previously spray-dried plasma is about 60%, about 50%, about 40%, about 30%, about 25%, about 20%, about 15%, about 10%, about 5%, about 4%, about 3%, about 2%, or about 1% less than the number in donor plasma.

[0048] Particle analysis to determine particle size, particle quantity and distribution can be obtained using, for example, a Beckman Coulter Multisizer 4 (electrical sensing zone method) operating in the range of 2-60 μm.

[0049] The spray-dried plasma of the present invention has low residual moisture, e.g., less than 2.5%. As described herein, the spray drying system of the present invention is designed to produce dried plasma powder with low residual moisture, which 1) reduces any potential clogging of the lower filter, 2) facilitates more rapid reconstitution, creating plasma units suitable for transfusion, and 3) contributes to stability during storage, including storage time, storage temperature, and for storage in different environments.

[0050] Low residual moisture plasma units are obtained using the disposables described herein and dried using the parameters described herein. In particular, the low residual moisture of the dried plasma is due to several aspects of the spray drying system that promote rapid mixing of the drying gas with the plasma droplets during spray drying. Rapid mixing allows for rapid mass transfer / evaporation, which results in plasma formulations with low residual moisture and contributes to the very compact dimensions of the spray-dried disposables of the present invention, particularly the plasma drying chamber.

[0051] For example, as described herein, the design of the liquid nozzle cap insert 80 and nozzle cap 76 allows the pressurized aerosol gas to flow through the annulus 81 in a vortex pattern, maximizing aerosolization and promoting rapid mixing of the aerosolized plasma droplets with the drying gas, as further described herein.

[0052] As further described herein, the improved droplet size provided by the beveled edge of the cannula promotes rapid mixing, faster evaporation, and reduced drying time, in part providing a plasma formulation with low residual moisture. See Figure 43T, which shows that the larger the droplet size, the longer it takes to evaporate the droplets using a hotter drying gas. Droplet size is also affected by the ratio of pressurized gas flow rate to liquid feed rate and nozzle design.

[0053] Additional factors contributing to the rapid mass transfer and evaporative drying phase of plasma droplets include the temperature of the plasma and drying gas, the surface area of ​​the droplets, humidity in the drying gas, and air circulation within the plasma drying chamber. As described herein, when initially exiting the nozzle assembly, the temperature of the drying gas is about 90°C to about 130°C (e.g., about 100°C to about 114°C), and the temperature of the plasma droplets is about 20°C to about 65°C within the plume, as shown in Figures 43S and 43Sa. The simulated droplet temperatures in Figure 43S, obtained by averaging over the entire droplet trajectory during a period of constant rate evaporation, span the range of drying gas inlet temperatures during evaporation near the nozzle, demonstrating that proteins are not exposed to such high drying gas inlet temperatures. In fact, plasma proteins are protected by energy losses due to evaporation when they are most vulnerable in their liquid state. Figure 43Sa focuses on a single droplet along the aerosol particle path. Figure 43Sa shows a longer time scale, illustrating the high temperatures the proteins are exposed to when they are in a solid state, after evaporation and in the capture filter. Heat flows from higher temperature locations to lower temperature locations, in this case the heat of the drying gas flows to the plasma droplets. In terms of surface area, the droplets are spherical, thereby maximizing their surface area, and because the droplets are very small, mass and heat transfer can occur rapidly. 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 particles. Finally, as described in more detail herein, the drying gas is emitted obliquely downward into the plasma drying chamber and the plume of atomized droplets, using several drying gas jets, initiating rapid mixing of the drying air with the atomized droplets. This increases the evaporation rate of the droplets.

[0054] As described herein, yet another factor contributing to low residual moisture formulations is the size of the starting droplets produced by the nozzle assembly. This affects the residence time in the drying chamber required to complete evaporation, with smaller droplet size allowing for more rapid evaporation to occur. The smaller the droplets, the greater the ratio of evaporative surface area to droplet mass, and the faster the rate of mass transfer from the droplets. As can be seen from Figure 43T, the drying chamber can be shortened to the point where most evaporation occurs and still allow the dried particles to achieve less than 2.5% residual moisture before deposition on the lower filter 36.

[0055] The residual moisture content in plasma dried using the disposables and dryers of the present invention is very low, e.g., about 2.5%, about 2%, less than about 1%, and preferably about 1.46% residual moisture, as measured by a Karl Fischer moisture sensor, Model No. C30S Compact KF Coulometer (Mettler, Toledo Billerica, Massachusetts, USA). This very low moisture content results from the effective and efficient evaporation of plasma droplets that occurs at the top and process conditions of the drying chamber 28. In this embodiment, the powder moisture content is in equilibrium with the relative humidity of the chamber exit airflow.

[0056] Dried plasma with less water content improves reconstitution time and protein stability during storage.

[0057] The dried plasma of the present invention is rapidly reconstituted, e.g., in less than 4 minutes. The primary reason that human plasma is not as widely available as it needs to be is that, prior to the present invention, plasma was typically stored frozen for long periods of time or in liquid form for very short periods of time. Thus, when large amounts of plasma are needed (e.g., in a mass casualty event), these quantities may not be available, and if plasma is needed in an emergency, it may need to be thawed, which can take 30-45 minutes or more and may not be available in time.

[0058] The spray-dried plasma of the present invention can be stored for extended periods as described herein and can be rapidly reconstituted in less than four minutes from initial contact by a user to complete reconstitution. The terms "reconstituted" and "rehydrated" are used interchangeably herein and refer 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.

[0059] Rapid reconstitution is due, in part, to the amorphous nature, low residual water content, small particle size, increased particle surface area of ​​dried plasma, and the amount of air present in the plasma unit bag (e.g., about 3 to about 15 mL of air). The low residual water content of dried plasma, less than about 2.5%, promotes faster reconstitution because the low residual water content does not contribute to the clumping of plasma particles into larger aggregates with smaller surface areas. In the case of dried plasma (e.g., solute), the interaction between the dried plasma particles and the solvent (e.g., SWFI) is strong, so that individual solute particles separate from each other, are surrounded by solvent molecules, and enter solution to form liquid plasma. The drier the particles, the faster they enter solution. Similarly, the smaller the dried plasma particles and the greater the exposed surface area of ​​the particles, the easier it is for the solution (e.g., SWFI) to surround the particles and allow them to enter solution. In one embodiment, the particle size of the dried plasma is about 1 to about 7 microns. Particle analysis to determine particle size, particle quantity and distribution of dried plasma can be obtained using Scanning Electron Microscopy (SEM), see Example 7.

[0060] The method of the present invention further includes reconstituting the dried plasma using a physiologically compatible reconstitution solution. The reconstitution solution can be mixed with the dried plasma using one of the spike ports 42A or 42B of the dried plasma unit 60. The reconstitution solution is connected to the dried plasma attached to one of the spike ports, and the water is manually pushed into the dried plasma bag to mix. Furthermore, the spray-dried compounded plasma of the present invention may be reconstituted with sterile water (e.g., sterile water for injection (SWFI) or the like) or clean, non-sterile water, which may be filtered after reconstitution if desired. Under normal circumstances, the clinician / healthcare provider / end user rehydrates the unit using the SWFI unit provided by the system. In a preferred embodiment, sterile water for injection is used for the reconstitution solution. If SWFI is not available, distilled water may be used in one aspect. In other embodiments, for example, when no pretreatment step is performed, the reconstitution solution further comprises an amino acid (e.g., glycine) or a buffer solution (e.g., an acid such as hydrochloric acid or citric acid). The plasma unit, including the dried plasma and bag, weighs about 45 grams to about 55 grams (e.g., about 50 grams). The dried plasma weighs about 15 grams to 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 ranges from about 175 mL to about 230 mL (e.g., 200 mL and about 225 mL). When the dried plasma is rehydrated with the reconstitution solution, thereby producing a plasma unit ready for transfusion, the rehydrated plasma unit has a final volume of about 180 mL to about 236 mL (e.g., about 205 mL and about 231 mL) or about 197 grams to about 247 mL (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 that accounts for possible evaporative losses during storage.

[0061] It is contemplated that the pH of the reconstituted spray-dried compounded plasma of the present invention will be 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 amount of pretreatment components, as further described herein, can be adjusted to achieve the design pH of the reconstituted spray-dried compounded plasma.

[0062] Furthermore, once the reconstitution solution is added, the method steps include shaking, rocking, and / or stirring the reconstituted plasma unit to ensure mixing and homogeneity of the reconstitution solution and dried plasma. The reconstituted plasma is ready for transfusion into a recipient. Examples of recipients include humans, primates, mammals, animals, etc. In one embodiment, the method for reconstituting a dried plasma unit can be carried out in less than 7 minutes (e.g., 7, 6, 5, 4, 3, and 2 minutes), preferably less than about 3.5 minutes, measured from a starting time of time 0 (the time of initial contact between the reconstitution solution and the dried plasma unit) to a final time point (total reconstitution time). This is the time at which the plasma is reconstituted with the reconstitution solution without visible clumps that can be seen with the naked eye.

[0063] This method could be performed by a variety of individuals, some of whom were medically trained and others who were untrained and time-limited. Rehydration was achieved, on average, in less than about 4 minutes.

[0064] 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-loaded SWFI in the kit, as long as the amount of SWFI used for rehydration is the same as specified. In one embodiment, the single-use container is 200 mL of sterile water for injection (SWFI) packaged in a 250 mL bag in an overwrapped pouch. The kit further includes a rehydration tubing set, such as a commercially approved standard sterile fluid transfer set (e.g., a FENWAL™ Plasma Transfer Set with two spikes 4C2243 or equivalent) for transferring the SWFI to the unit. Furthermore, a transfusion tubing set may also be included for transfusing the rehydrated plasma into the patient. An example of a commercially approved standard sterile transfusion / administration set is the FENWAL™ Blood Component Recipient Set with a Standard Blood Filter and Luer Adapter 4C2160 or equivalent.

[0065] The spray-dried plasma of the present invention is stable when stored below refrigeration, at room temperature, or at elevated temperatures, allowing for extended storage periods. The stability of spray-dried plasma is achieved in part by its low residual moisture content, which allows for storage at various temperatures and for extended periods of time.

[0066] "Stability" or "shelf-life stability" or "unit stability" refers to stored spray-dried plasma that, upon reconstitution, behaves equivalently to dried but unstored plasma. Stability involves comparing the amounts and functions of various plasma proteins (e.g., vWF, Factor V, Factor VIII, etc.) and / or plasma properties (e.g., pH, particle size, etc.) present in reconstituted spray-dried plasma before storage (time = 0 seconds) and after storage (e.g., up to 48 months refrigerated, up to 12 months at room temperature). Stability involves comparing the respective values ​​of one or more of these proteins / properties before and after storage to determine their similarity to each other or whether the values ​​are within a clinical range. In one embodiment, the plasma protein / property value after storage is within about 25% or less (25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, and 1%) of the value before storage. In one embodiment, the plasma protein / property value after storage is within the clinical range for that plasma protein. Such plasma properties include pH, osmolality (mOsm / kg), particle size, particle load, and particle distribution.Plasma proteins and their functions include, for example, total protein (mg / mL), activated partial thromboplastin time (aPTT), prothrombin time, and international normalized ratio (IMRR). Examples of plasma characteristics and plasma proteins include: INR (International Thrombosis Rate), thrombin time, 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 PI (%), antithrombin III (%), von Willebrand factor antigen (% or IU / dL), von Willebrand factor ristocetin cofactor (% or IU / dL), C5a (ng / mL), prothrombin fragment F1+2 (pmol / L), thrombin-antithrombin complex (TAT) (μg / L), and protein (mg / mL). Measuring such plasma characteristics and plasma proteins is known in the art. In another embodiment, the acceptable or clinical range for von Willebrand factor antigen ristocetin cofactor (VWF:RCo) is about 50 to about 200 IU / dL, or about 50 to about 200 IU / dL, and the von Willebrand factor antigen (VWF:Ag) value is about 50 to 200 IU / dL.

[0067] In one embodiment, the clinical reference ranges for determining shelf life or unit stability are as follows:

[0068] [Table 3]

[0069] [Table 4]

[0070] The data show that the dried plasma is stable after storage as dried plasma for periods of up to about 24 hours at refrigerated temperatures (2-8°C), 6 months at room temperature (20-25°C), and 3 months at elevated room temperature (30°C). Example 5 describes experiments that were performed to confirm that the spray-dried plasma units remained stable during storage.

[0071] The spray-dried plasma of the present invention can be stored at elevated temperatures, room temperatures, refrigerated temperatures, frozen temperatures, or combinations thereof, as described herein, after release testing has been completed and the formulation has been determined to be safe for use. In certain embodiments, the spray-dried plasma is refrigerated to allow for extended storage.

[0072] The dried plasma of the present invention may be stored at temperatures between -80°C and 45°C. Room temperature is considered to be about 20°C to about 30°C. In one embodiment, refrigerated temperatures are about 1°C to about 6°C. The terms "chilled," "low temperature," "temperature below room temperature," and "temperature below ambient temperature" refer interchangeably to any temperature between 1°C and 20°C. In any of the embodiments of the invention described herein, the temperature is selected from the group of temperatures consisting of 45°C, 44°C, 43°C, 42°C, 41°C, 40°C, 39°C, 38°C, 37°C, 36°C, 35°C, 34°C, 33°C, 31°C, 30°C, 29°C, 28°C, 27°C, 26°C, 25°C, 24°C, 23°C, 22°C, 21°C, 20°C, 19°C, 18°C, 17°C, 16°C, 15°C, 14°C, 13°C, 12°C, 11°C, 10°C, 9°C, 8°C, 7°C, 6°C, 5°C, 4°C, 3°C, 2°C, 1°C, 0°C, -1°C, -5°C, -10°C, -20°C, -30°C, -40°C, -50°C, -60°C, -70°C and -80°C. In some embodiments, the spray-dried plasma of the present invention is stored at a temperature below about 15°C, preferably below 10°C, and more preferably below 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 a warm temperature, for example, above 23°C. In further embodiments, the spray-dried plasma of the present invention can be stored at freezing temperatures (e.g., 0°C to -80°C). It should be noted that while the dried plasma preparation can be stored at freezing temperatures, it is not necessary to maintain it in a frozen state for transfusion. On the one hand, storage conditions can be carefully monitored, and on the other hand, storage conditions can be a range of temperatures depending on the terrestrial conditions. In yet another embodiment, the dried plasma can be stored at a combination of temperatures.

[0073] As used herein in all aspects and embodiments of the present invention, "period of time" or "time period" refers to the period of time during which the spray-dried plasma of the present invention is stored at any given temperature. The term "period" can range from seconds to minutes, hours, days, weeks, months, or years. In preferred embodiments, the term "period" refers to a period of time ranging from about 3 to about 120 hours, e.g., 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, or the like. , 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 38 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 ​​hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours Between, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours, 73 hours, 74 hours, 75 hours, 76 hours, 77 hours, 78 hours, 79 hours, 80 hours, 81 hours, 82 hours, 83 hours, 84 hours, 85 hours, 86 hours, 87 hours, 88 hours, 89 hours, 90 hours, 91 hours, 92 hours, 93 hours, 94 hours, 9 Refers to several times including 5 hours, 96 hours, 97 hours, 98 hours, 99 hours, 100 hours, 101 hours, 102 hours, 103 hours, 104 hours, 105 hours, 106 hours, 107 hours, 108 hours, 109 hours, 110 hours, 111 hours, 112 hours, 113 hours, 114 hours, 115 hours, 116 hours, 117 hours, 118 hours, 119 hours and 120 hours.In certain embodiments, the period for which the spray-dried plasma of the present invention can be stored includes about 1 to about 30 days (e.g., about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 15 days, about 16 days, about 17 days, about 18 days, about 19 days, about 20 days, about 21 days, about 22 days, about 23 days, about 24 days, about 25 days, about 26 days, about 27 days, about 28 days, about 29 days, and about 30 days). In certain embodiments, the spray-dried plasma of the present invention can be stored for about 1 to about 48 months (about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 12 months, about 13 months, about 14 months, about 15 months, about 16 months, about 17 months, about 18 months, about 19 months, about 20 months, about 21 months, about 22 months, about 23 months, about 24 months, about 25 months, about 26 months, about 27 months, about 28 months, about 29 months, about 30 months, about 31 months, about 32 months, about 33 months, about 34 months, about 35 months, about 36 months, about 37 months, about 38 months, about 39 months, about 40 months, about 41 months, about 42 months, about 43 months, about 44 months, about 45 months, about 46 months, about 47 months, and about 48 months).

[0074] In one embodiment, the spray-dried plasma of the present invention can be stored at room temperature for about 1 hour to at least about 7.5 months. In one embodiment, the spray-dried plasma of the present invention can be stored at refrigerated temperatures for about 1 hour to at least about 48 months. In one aspect, the spray-dried plasma of the present invention can be refrigerated for any day or days during storage.

[0075] In various other embodiments, the spray-dried plasma of the present invention is stable, maintaining plasma protein functionality and / or plasma properties over time when compared to corresponding clinical reference intervals or clinical reference intervals before storage, or when compared to thawed previously frozen plasma over the same period after spray drying. In one embodiment, spray-dried plasma samples stored at temperatures between -80°C and 45°C are suitable for transfusion after extended storage periods, in one embodiment, 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 1.5 years, about 2 years, about 3 years, about 4 years, or more.

[0076] The present invention provides, in certain embodiments, novel methods for storing spray-dried plasma, the steps of which 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 substantial or subclinical loss of one or more plasma protein amounts and / or plasma properties.

[0077] The spray-dried plasma of the present invention is stable after reconstitution for up to 26 hours prior to use Spray-dried plasma stability also refers to the properties of the spray-dried plasma that has been reconstituted and is suitable for transfusion over a period of time. The reconstituted, pre-dried plasma of the present invention remains stable and ready for transfusion for up to 26 hours, and upon reconstitution, the plasma is stored at room temperature, refrigerated temperature, or elevated temperature.

[0078] "Stability" or "in-use stability" also refers to pre-spray-dried plasma that is reconstituted over a period of time (e.g., 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 equivalently to spray-dried plasma immediately or simultaneously (e.g., 0-5 hours, 1-15 minutes) after reconstitution. Stability includes comparing the amounts of various plasma proteins and their functions (e.g., vWF, Factor V, Factor VIII, etc.) and / or the properties (e.g., pH, particle size, etc.) of the reconstituted, pre-spray-dried plasma before (time = 0 seconds) and after (e.g., time = 26 hours) storage of the reconstituted plasma. In another embodiment, stability involves determining the amounts of various plasma proteins, their functions (e.g., vWF, Factor V, Factor VIII, etc.) and / or plasma properties (e.g., pH, particle size, etc.) present in reconstituted, pre-spray-dried plasma after storage to determine whether one or more are within their corresponding clinical reference ranges. Stability involves comparing the respective values ​​of one or more of these proteins / properties before and after reconstitution and storage to determine their similarity to each other or whether each value is within its clinical reference range. In one embodiment, the post-storage plasma protein / property value is within about 25% or less (25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, and 1%) of the pre-storage value after reconstitution. When the post-storage value of one or more plasma proteins is within 25% of the pre-storage value or within the clinical reference range, the reconstituted, pre-dried plasma is suitable for transfusion. Such plasma properties include pH, osmolality (mOsm / kg), particle size, particle load, and particle distribution.Plasma proteins and their functions include, for example, total protein (mg / mL), activated partial thromboplastin time (aPTT), prothrombin time, and international normalized ratio (IMRR). Examples of plasma characteristics and proteins include: INR (International Thrombosis Rate), thrombin time, 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 F1+2 (pmol / L), and thrombin-antithrombin complex (TAT) (μg / L). Measuring such plasma characteristics and proteins is known in the art. In another embodiment, the acceptable or clinical range for von Willebrand factor antigen ristocetin cofactor (VWF:RCo) is about 50 to about 200 IU / dL, or about 50 to about 200 IU / dL, and the von Willebrand factor antigen (VWF:Ag) value is about 50 to 200 IU / dL.

[0079] In one embodiment, the clinical reference ranges for determining in-use stability are as follows:

[0080] [Table 5]

[0081] [Table 6]

[0082] The data indicate that the dried plasma, upon reconstitution, is stable for transfusion after storage as reconstituted plasma for periods ranging from about 1 hour to about 26 hours. Example 4 describes experiments performed that confirm that reconstituted, pre-spray-dried plasma units remained stable during storage and prior to transfusion.

[0083] The reconstituted spray-dried plasma of the present invention can be stored at room temperature for transfusion or can be refrigerated, hi certain embodiments, the reconstituted, pre-spray-dried plasma is refrigerated to allow for extended pre-transfusion storage.

[0084] The reconstituted, pre-dried plasma of the present invention may be stored for transfusion at a temperature between 1°C and 45°C. Room temperature is considered to be about 20°C to about 30°C. In one aspect, refrigerated temperatures are about 1°C to about 6°C, and the terms "chilled," "chilled," "below room temperature," and "below ambient temperature" refer interchangeably to any temperature between 1°C and 20°C. In any of the embodiments of the invention described herein, the temperature is selected from the group of temperatures consisting of 45°C, 44°C, 43°C, 42°C, 41°C, 40°C, 39°C, 38°C, 37°C, 36°C, 35°C, 34°C, 33°C, 31°C, 30°C, 29°C, 28°C, 27°C, 26°C, 25°C, 24°C, 23°C, 22°C, 21°C, 20°C, 19°C, 18°C, 17°C, 16°C, 15°C, 14°C, 13°C, 12°C, 11°C, 10°C, 9°C, 8°C, 7°C, 6°C, 5°C, 4°C, 3°C, 2°C, and 1°C. In some embodiments, the reconstituted, pre-spray-dried plasma of the invention is stored at a temperature below about 15°C, preferably below 10°C, and more preferably below 5°C. In some other embodiments, the reconstituted, pre-spray-dried plasma of the invention is stored at room temperature. In other embodiments, the reconstituted, pre-spray-dried plasma of the present invention is stored at warm temperatures, for example, above 23° C. In yet other embodiments, the dried plasma is stored at a combination of these temperatures.

[0085] As used herein in all aspects and embodiments of the invention, a "period of time" or "time period" refers to the period of time during which the reconstituted, pre-spray-dried plasma of the invention is stored at any given temperature prior to transfusion. The term "period" can range from seconds to minutes, hours, days, weeks, or months. In one embodiment, the term "period" refers to a period of time ranging from about 3 to about 30 hours, including, for example, 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.

[0086] In various other embodiments, the reconstituted, pre-spray-dried plasma of the present invention is stored at room temperature prior to transfusion. When compared to corresponding clinical reference ranges or pre-storage clinical reference ranges, or compared to thawed previously frozen plasma over the same period, the reconstituted, pre-spray-dried plasma of the present invention is stable, maintaining plasma protein functionality and / or plasma properties over time. Thus, reconstituted, pre-spray-dried plasma samples stored at or below room temperature are suitable for transfusion after extended storage periods, in one embodiment, 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.

[0087] In certain aspects, the present invention provides a novel method for storing reconstituted spray-dried plasma, the steps of which 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 substantial (e.g., about 25% or less) or subclinical loss of one or more plasma protein amounts and / or plasma properties. The method further includes transfusing the stored reconstituted plasma into a recipient.

[0088] The spray-dried plasma of the present invention exhibits functionality of most vulnerable proteins, including von Willebrand factor, and other active proteins, absence of complement activation (C5a, C3a), and near-normal pH without buffering or treatment with CO2. The spray-dried plasma of the present invention is capable of preserving the most vulnerable proteins and functions of donor plasma, reducing complement activation, and having a near-normal pH upon reconstitution without additional buffering or treatment with CO. This is achieved by pretreating the liquid plasma with a pretreatment solution (e.g., a spray dry stable acidic substance (SDSAS) and one or more amino acids).

[0089] The present invention provides effective preservation, including functional maintenance of plasma coagulation factors in a manner that is not harmful to the plasma or the transfused patient. During spray drying, some plasma proteins degrade to some extent due to shear stress, surface stress (e.g., air-liquid interface stress), exposure to extremes of pH, heat stress, dehydration stress, and other environmental stresses.

[0090] The methods and compositions of the present invention recognize that pH and associated stresses can be reduced or their effects mitigated by using formulations of liquid plasma prior to or simultaneously with spray drying. By formulating liquid plasma with glycine HCl or a similar spray-drying stable acidic substance (SDSAS) and one or more amino acids (e.g., glycine) at novel concentrations, the pH of the plasma is maintained at a non-alkaline level during the spray-drying process. Adding amino acids to the pretreatment solution leads to higher retention of plasma protein function and better subsequent storage stability when compared to unformulated plasma.

[0091] In certain embodiments, the pretreatment solution is added to donor plasma, and upon addition to the plasma, the pretreatment container contains glycine in an amount ranging from about 10 μmol to about 110 μmol of plasma (e.g., about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110 μmol / mL of plasma) and hydrochloric acid (HCl) in an amount ranging from about 10 μmol / mL to about 30 μmol / mL of plasma (e.g., about 10, about 15, about 20, about 25, and about 30 μmol / mL of plasma), thereby obtaining blended plasma. The pH of the blended plasma is in the range of about 6.0 to about 6.6, which offsets the effects of spray drying on pH, resulting in a final rehydrated formulation whose pH is normal physiological pH and whose pH ranges from about 6.5 to about 7.8. In one embodiment, pretreatment of the plasma is optional.

[0092] In one embodiment, the amounts of SDSAS and amino acids in the pretreatment step range as follows:

[0093] [Table 7]

[0094] Proteins and their functionality are also partially protected from mass transfer occurring at lower temperatures, as described herein. As described above, the drying rate is constant, and as the liquid particles evaporate and lose water, water is transferred from the liquid plasma droplets to the drying gas, and heat from the drying gas is transferred to the plasma droplets, turning them into dry particles. The plasma droplets enter the drying chamber at essentially room temperature, and the temperature remains constant for most of the evaporation period. See Figure 43S. Once most of the water leaves the particles, the particle temperature increases and equilibrates with the dryer chamber outlet temperature of 65°C. During evaporation, the droplets are maintained at a low temperature, thereby protecting heat-sensitive proteins such as vWF. See Figure 43S. During the evaporation process, the temperature of the droplets and the proteins therein are exposed to a low temperature, i.e., the thermodynamic wet-bulb temperature, compared to the inlet drying gas temperature, thereby protecting the proteins. See Figure 43R. Evaporation reduces the protein temperature to near the thermodynamic wet-bulb value, and when evaporation is delayed, the particle temperature increases. See Figure 43Sa.

[0095] The term "retention" is defined herein to refer to the percentage of an analyte that is retained after spray drying compared to the analyte in a sample of plasma of the same nature (the same sample before spray drying), which may be frozen, and the analyte is analyzed for native plasma and / or rehydrated plasma at the same protein concentration. The analyte may be any known plasma substance, such as a plasma protein (e.g., vWF antigen or fibrinogen), as described herein, and may be measured by the concentration or activity of the analyte (e.g., vWF:RCo activity), as further described herein. The amount of the analyte may be compared to its corresponding clinical reference range.

[0096] As used herein, a spray-drying stable acidic substance (SDSAS) is any substance, such as an acid or acid salt, that provides a pH that is physiologically suitable for addition to plasma being spray-dried and that is physiologically suitable for the subject (human or otherwise) to whom the reconstituted plasma is administered (transfused). The SDSAS remains sufficiently stable for the spray-drying process (e.g., does not physically evaporate or chemically degrade). The SDSAS provides the pH adjustment described herein that maintains or improves the retention or functionality of von Willebrand factor, for example, in the reconstituted plasma described herein, compared to non-pretreated spray-dried plasma. Specific examples of spray-drying stable acidic substances include glycine HCl, HCl, citric acid, lactic acid, monosodium citrate, and other SDSAS described herein. Other SDSAS may be known in the art or may be determined by straightforward experimentation.

[0097] In one embodiment, the pretreatment solution of the present invention used to obtain the spray-dried formulation comprises one or more SDSAS and one or more amino acids. The addition of amino acids allows for the protection of plasma proteins during spray drying without lowering the pH of the pretreatment solution. The addition of amino acids increases the pH of the pretreatment solution, but surprisingly, this does not affect the pH of the pretreated spray-dried plasma or the rehydrated spray-dried plasma. Furthermore, in one embodiment, the use of amino acids with SDSAS provides spray-dried plasma that, upon rehydration, has low levels of C5a, i.e., anaphylatoxins, or C5a levels equivalent to never-frozen plasma (NFP) or FDA-approved apheresis plasma products. In one embodiment, complement activation is associated with inflammation and should be kept low within a clinically acceptable range. In particular, the pH of the pretreatment solution ranges from about 2.0 to 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), which, when combined with plasma, results in pretreated plasma having a pH of about 6.0 to about 6.6 (e.g., 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6), and when the previously spray-dried plasma is reconstituted, results in reconstituted plasma having a pH of 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 one or more amino acids in the reconstituted plasma is present in an amount of about 1 mM to about 150 mM. Examples of amino acids that can be combined with SDSAS for the pretreatment solution include glycine, alanine, asparagine, glutamine, isoleucine, leucine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine. In certain embodiments, glycine is the amino acid added to the SDSAS composition.

[0098] Thus, treatment of the spray-dried formulation, i.e., plasma provided prior to or concurrently with spray drying, maintains and enables the recovery or functionality of active coagulation factors in the rehydrated plasma subjected to the spray-drying process, as well as long-term stability during storage after drying. As further described below, these improvements to certain embodiments of spray-drying of plasma involving formulation with SDSAS and / or amino acids improve the ease and reduce the cost of rehydration of plasma products by enabling rehydration of the spray-dried plasma 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 processes to control the pH of the reconstituted plasma.

[0099] In one embodiment, the spray-dried plasma of the present invention has improved functionality or retention of active plasma proteins, long-term stability of plasma proteins, and reduced anaphylatoxins. In one embodiment, a method for obtaining dried plasma of the present invention comprises combining donor plasma containing a pretreatment solution having SDSAS and an amino acid with a spray-drying system. The present invention further contemplates adjusting the pH of the donor plasma to a concentration of SDSAS of about 1 mM to about 50 mM, thereby lowering the plasma pH to about 5.0 to about 6.5, to produce blended plasma. In one embodiment, the present invention further contemplates adjusting the pH of plasma to be spray-dried using a pretreatment solution having SDSAS and an amino acid to a concentration of about 1 mM to about 50 mM, and a concentration of an amino acid compound of about 1 mM to about 150 mM, thereby lowering the plasma pH to about 6.0 to about 6.6, to produce blended plasma.

[0100] In one embodiment, to obtain the dried plasma of the present invention, the method includes producing spray-dried plasma by combining plasma with a pretreatment solution, the pretreatment solution including an amino acid (e.g., glycine) in an amount ranging from about 10 μmol plasma to about 110 μmol rehydrated plasma (e.g., about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, or about 110 μmol / mL plasma) and an SDSAS (e.g., hydrochloric acid (HCl)) in an amount ranging from about 10 μmol / mL plasma to about 30 μmol / mL rehydrated plasma (e.g., about 10, about 15, about 20, about 25, or about 30 μmol / mL plasma), thereby obtaining blended plasma. The method also includes drying the blended plasma using a spray-drying system, as described herein, to produce spray-dried blended plasma. In one embodiment, the pretreatment solution has glycine in the plasma in an amount of about 84 μmol / mL and HCl in the plasma in an amount of about 20 μmol / mL.

[0101] In one embodiment, the pretreatment solution comprises glycine in an amount ranging from about 15 mmol to about 30 mmol (e.g., about 15, about 20, about 25, and about 30 mmol), or from about 43 μmol / mL to about 473 μmol / mL, and HCl in an amount ranging from about 3 mmol to about 7 mmol (e.g., about 3, about 4, about 5, about 6, and about 7 mmol), or from about 43 μmol / mL to 129 μmol / mL, thereby obtaining blended plasma, and drying the blended plasma using a spray-drying system to produce spray-dried blended plasma. In one particular embodiment, the pretreatment solution comprises glycine in an amount of about 22 mmol and HCl in an amount of about 5.3 mmol.

[0102] After drying the formulated plasma in a spray-drying system to produce spray-dried formulated plasma, the spray-dried formulated plasma had a functional von Willebrand factor (vWF) retention rate at least 10% to at least 100% points higher than the functional von Willebrand factor (vWF) retention rate obtained from otherwise identical spray-dried plasma that had not been acid-formulated with the pretreatment solution of the present invention. In another embodiment, the spray-dried formulated plasma has a functional von Willebrand factor (vWF) retention rate that is similar to or within about 20% (e.g., about 15%, about 10%, about 5%) of that of never-frozen plasma or FDA-approved plasma products relative to fresh frozen plasma. While SDSAS may be selected from any known in the art, glycine HCl, HCl, citric acid, and lactic acid are preferred materials for use in the present invention. When an amino acid is added to SDSAS to form a pretreatment solution, in one embodiment, glycine is a preferred material of the present invention (e.g., HCl / glycine or citric acid / glycine combinations). The physiologically compatible pretreatment solution is added to the plasma prior to spray drying, preferably immediately prior to or simultaneously with spray drying. Furthermore, the pH of the plasma may be determined prior to adding SDSAS and amino acids to the plasma to determine the appropriate amount of acid to add. In one embodiment, about 7.4 mM citric acid is added to CPD plasma or WB plasma. In one embodiment, the pH of the combined plasma is about 5.5 to about 6.5. The present invention further contemplates that the retention of functional vWF may be about 40 percentage points higher, by about 10 to about 100 percentage points (e.g., about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 50, about 60, about 70, about 80, about 90, or about 100 percentage points), than the retention of functional von Willebrand factor obtained from otherwise identical spray-dried plasma that has not been pretreated with SDSAS and amino acids. In one embodiment, the data show that levels of ristocetin cofactor (vWF:RCo) were absent or undetectable when plasma was spray dried without pretreatment. See Figures 17-19.In this embodiment, the ristocetin cofactor (vWF:RCo) in the pretreated spray-dried plasma was 2-fold, 3-fold, 4-fold, 5-fold or more higher than that in the non-pretreated spray-dried plasma.

[0103] It has been discovered that desirable C5a levels result from the addition of a pretreatment solution having SDSAS and an amino acid, and optionally rapid mixing / agitation of the pretreatment components. In one embodiment, desirable C5a levels are similar to the C5a levels in NFP or FDA-approved apheresis plasma products. In one embodiment, the C5a levels are reduced compared to the C5a levels from rehydrated plasma pretreated with SDSAS alone and optionally rapid mixing / agitation of the pretreatment components. In particular, the C5a levels of reconstituted plasma obtained from the pretreatment solution of the present invention can be from about 0.1 ng / mL to about 74 ng / mL, particularly 20 ng / mL to 70 ng / mL (e.g., about 30 ng / mL). C5a levels are reduced when treated with a pretreatment solution having at least one SDSAS and at least one amino acid compared to plasma subjected to a pretreatment solution having only SDSAS. In one embodiment, C5a levels are reduced by about 20% or less (e.g., 20%, 15%, 10%, 5%, 1%) compared to plasma subjected to a pretreatment solution of the invention containing only acid. In another embodiment, C5a levels are about the same as that in never-frozen plasma or within about 20% or less (e.g., 20%, 15%, 10%, 5%, 1%) of the C5a in never-frozen plasma or an FDA-approved apheresis plasma product.

[0104] The present invention contemplates a reconstituted spray-dried plasma preparation for human transfusion (administration), where the reconstituted spray-dried plasma preparation is reconstituted, for example, with sterile water for injection, and the reconstituted spray-dried plasma preparation has a pH of about 6.5 to about 7.8. The reconstituted plasma of the present invention has a functional von Willebrand factor retention that is 5 percentage points higher than the retention of functional von Willebrand factor obtained from otherwise identical spray-dried plasma that has not been compounded with SDS-AS and / or amino acids, or has a functional von Willebrand factor retention that is about 5 percentage points to about 40 percentage points (e.g., about 25 percentage points to about 35 percentage points) higher than the retention of functional von Willebrand factor obtained from otherwise identical spray-dried plasma that has not been pretreated with SDS-AS and amino acids. Furthermore, the present invention relates to reconstituted spray-dried plasma having a C5a level of about 0.1 ng / mL to about 74 ng / mL, particularly 20 ng / mL to 40 ng / mL (e.g., about 30 ng / mL). In one embodiment, the present invention relates to reconstituted spray-dried plasma pretreated with SDSAS and amino acids, having C5a levels that are reduced, for example, by about 20% or less (e.g., 20%, 15%, 10%, 5%, 1%) compared to plasma subjected to a pretreatment solution with SDSAS. In another embodiment, the C5a levels of the reconstituted plasma of the present invention are similar to those of approved FDA plasma products.

[0105] Autologous use of dried plasma units of the present invention The dried plasma of the present invention can be derived from a third-party donor or from the intended recipient. The latter is known as autologous plasma. Autologous refers to plasma collected from and donated to the same individual. Autologous spray-dried plasma is desirable because it is compatible with the recipient's plasma, e.g., does not require ABO / Rh matching prior to use, and is less likely to harbor pathogens or allergic immunogens to the recipient.

[0106] The spray-dried plasma for autologous use of the present invention features at least two major components: a dried plasma unit and a reconstitution fluid. In one embodiment, the reconstitution solution is sterile water (SWFI), and the resulting reconstituted plasma has a near-physiological pH. In one embodiment, the sterile water is provided as a pre-measured standard volume in a pouch.

[0107] In one embodiment, an intended recipient (e.g., a soldier, police officer, firefighter, or adventurer) who may be at elevated risk for plasma transfusion can provide / donate their own plasma, which can be dried using the methods described herein for obtaining dried plasma units having one or more of the properties described herein. One or more dried plasma units, derived from dried plasma donated by the intended recipient, can be transported by or on behalf of the intended recipient to be readily available for their use. In this embodiment, the dried plasma units are lightweight, their packaging is sturdy, and they can be stored at various temperatures (room and / or warm), making autologous use encompassed by the present invention. When the intended recipient needs to transfuse plasma, a healthcare provider (e.g., a doctor, nurse, or first responder) can transport a reconstitution solution (e.g., SWFI) and reconstitute the intended recipient's own dried plasma for emergency transfusion. In an emergency, immediate plasma with a large amount of clotting factors is available to the bleeding recipient. Early delivery of plasma, which is needed in large quantities in situations such as trauma, increases the likelihood of a favorable outcome.

[0108] In one embodiment, the reconstitution solution (e.g., SWFI) can be carried by either the healthcare provider or the intended recipient. However, in a battlefield-like situation, it is preferable for the healthcare provider to carry the reconstitution solution, as the reconstitution solution tends to be relatively heavy and the SWFI is readily available to the healthcare provider. Advantages of autologous dried plasma units include elimination of compatibility issues and reduced infection transmission. Autologous dried plasma does not require ABO Rh matching, reducing infection transmission from non-autologous or potentially allogeneic donors.

[0109] The manufacturing system for dried plasma of the present invention is relatively compact and can be deployed in a wide variety of environments. The manufacturing process for spray-dried plasma of the present invention is automated and easy to use by personnel with minimal education and straightforward training.

[0110] Thus, providing an autologous dried plasma unit involves providing liquid plasma by the intended recipient (which may be obtained using methods known in the art and / or the methods described herein) and drying the intended recipient's liquid plasma using the spray drying system described herein. Finally, the method involves reconstituting the recipient-donated dried plasma unit and, when necessary, transfusing the recipient with the reconstituted recipient-donated dried plasma. The recipient-donated dried plasma unit may be reconstituted by a healthcare provider or other qualified individual.

[0111] This autologous dried plasma system can be easily adapted to other situations and is available for use by anyone who desires their own plasma nearby, and is available for future use. Other situations in which autologous dried plasma may be used include rural migrants who are far from plasma sources, or people who live anywhere but prefer to transfuse their own plasma in an emergency. The intended recipient can store their own dried plasma unit at home, in a safe room, in a vehicle, or with a healthcare provider, and the local healthcare provider can deliver the reconstituted solution. Based on the storage data herein, these dried plasma units for autologous use can be replenished every one to four years. Widespread deployment of the dried plasma units of the present invention may be undertaken as a preventative measure against death or permanent damage due to hemorrhagic trauma or disease.

[0112] The recipient can be a human or mammal.

[0113] Pretreatment Formulations and Processes - Detailed Description Embodiments of the present disclosure relate to methods and compositions for spray-drying liquid samples. In certain embodiments, the liquid sample is plasma obtained from a blood donor. In preferred embodiments, the blood donor is a human. However, it can be understood that the disclosed embodiments may be utilized 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 (colloidal suspensions of very small particles).

[0114] The pH control of the reconstituted dried plasma according to the present invention using spray drying as described herein is improved compared to reported pH control of reconstituted plasma produced by freeze-drying or lyophilization. An excessively high or excessively low pH of plasma (including a pH greater than 7.8) is associated with increased morbidity or mortality ("alkalosis"). In this respect, the present invention is superior to prior art freeze-dried formulations and processes. The drying process, unless controlled in some manner, leads to loss of CO2, increasing the pH of the dried product. The present invention does this without additional processing steps, reconstituting using only sterile water, and has been approved in clinical trials by the United States Food and Drug Administration. Reconstitution using only sterile water is highly desirable for dried plasma products. In the event of loss or damage of the pre-packaged sterile water for reconstitution, which is pre-measured and provided as part of an emergency kit, ER, OR, or any other emergency use of dried plasma, a readily available measured amount of sterile water can be used for reconstitution.

[0115] "Human freeze-dried plasma is alkalotic at pH levels around 8." Zaza M, Kalkwarf KJ, Holcomb JB. Dried Plasma. Damage Control Resuscitation. 2019;145-162. Published May 6, 2019. doi:10.1007 / 978-3-030-20820-2_8, page 8, second full paragraph. Zaza et al. defend this by citing only Saillol et al.'s 2013 paper, "The evolving role of lyophilized plasma in remote damage control resuscitation in the French Armed Forces Health Service." Transfusion. 2013;53:65S-71S, and stating, "However, [freeze-dried plasma] is well tolerated clinically in humans." The Saillol paper concerns the French Army's freeze-dried plasma, known as French LYophilized Plasma (FLYP). Saillol et al. acknowledge that "upon reconstitution [of FLYP], the pH is near 8" (Id. at 67S). The Saillol et al. report is limited to combat situations of severe hemorrhage, where the protocol involves tranexamic acid, FLYP containing red blood cells in a 1:1 ratio, and other measures to control the patient's blood pH (Id. at 66S). Saillol et al. acknowledge that "further studies will be required to determine the specific efficacy of FLYP in the therapeutic management of civilian patients with severe hemorrhage" (see the last sentence of the abstract, p. 65S). In contrast, the lower, better-controlled pH of the present invention is expected to be suitable for use in all situations where plasma transfusion is indicated under any circumstances and in any volume.

[0116] The pH of reconstituted dried plasma produced by the Terumo-BCT freeze-drying process has been reported to be high, ranging from 7.66 to 7.94. 7.94 is very close to 8.0. The so-called French Lyophilized Plasma (FLYP) plasma product produced by the French Army has also been reported to 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 increase in pH after freeze-drying, as expected based on other freeze-dried 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 are no reports of Terumo freeze-dried plasma products being approved for clinical trials in the United States or elsewhere. FLYP plasma was not the subject of clinical trials in the United States. The Flaumenhaft authors dismiss the high pH of Terumo-BCT reconstituted freeze-dried plasma material as being "within the Terumo-BCT required range of 7.0 to 8.0" and "aligned" with the FLYP plasma pH, which is closer to 8.0. In contrast, the pH of the spray-dried plasma formulations of the present invention does not exceed 7.8 and generally has a pH range close to physiological pH.

[0117] The reported pH of reconstituted dried plasma produced by Teleflex's freeze-drying process has not been reported. However, because the pH is significantly higher than physiological (more alkaline) at the end of the freeze-drying process, Teleflex, in its agreement with the United States Food and Drug Administration, states that its REPLAS™ freeze-dried plasma requires additional processing and equipment to lower the pH of the Teleflex formulation upon reconstitution. Specifically, an acidic reconstitution fluid must be used to restore the pH of the reconstituted [freeze-dried plasma] to physiological pH before infusion. Van, et al., J Trauma Injury, Infection and Critical Car Vol 71 No 1, p 22 (July 2011). In fact, according to Van, preliminary studies in our laboratory revealed that the pH of reconstituted LP without acid is approximately 9, and its infusion leads to rapid death (ID. at 20). The REPLAS™ freeze-drying process is described as including the following steps: "The vacuum chamber is ruptured with medical-grade carbon dioxide (CO2) gas, which compensates for the loss of dissolved CO2 from the starting plasma material during the freeze-drying process," and "In addition, REPLAS™ is 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 formulation." Jose A. Cancelas, Investigator's Agreement A Phase 1, Single-Center, Partial Double-blind, 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, Apr 19, 2018; pp-24.25.Downloaded December 6, 2021 at https: / / clinicaltrials.gov / ProvidedDocs / 26 / NCT02930226 / Prot_000.pdf. The spray-dried plasma system of the present invention does not require the elaborate and expensive use of CO2 gassing of the dried plasma, or CO2 storage of the dried plasma to control pH in the reconstituted plasma product, or the equipment required to perform these additional pH correction processes.

[0118] plasma Plasma is the fluid that remains after blood has been centrifuged (for example) to remove cellular material such as red blood cells, white blood cells, and platelets. Plasma is generally yellow in color and clear to opaque. It contains proteins (6-8%, e.g., serum albumin, globulins, fibrinogen, etc.), glucose, clotting factors (clotting proteins), electrolytes (Na + , Ca 2+ , Mg 2+ , HCO3 - It contains dissolved components of blood, such as soluble components (e.g., Cl), and hormones. Whole blood (WB) plasma is plasma isolated from whole blood without added medications, except for one or more anticoagulants. Citrate phosphate dextrose (CPD) plasma, as the name suggests, contains citrate, sodium phosphate, and a sugar (usually dextrose) added as an anticoagulant. The citrate level in whole blood-derived CPD plasma is approximately 20–30 mM. Therefore, the final citrate concentration in whole blood-derived CPD plasma formulated with 7.4 mM citrate is approximately 27.4–37.4 mM.

[0119] The plasma of the present invention may be dried in pools or units. Pooling multiple plasma units has several advantages. For example, a lack of factor retention on an equal volume basis can be compensated for by adding volume from the pool to the final product. There are negative aspects as well. Supplementing volume from the pool to improve factor retention is expensive. Importantly, pooled plasma must be constantly tested for pathogens, since any pathogen entering the pool from, for example, a single donor, risks harming hundreds or thousands of patients if undetected. Even if detected, pathogen contamination of the pooled plasma can render the entire pool worthless. Testing can be avoided by inactivating the plasma pathogens by radiation or chemically, such as with organic solvent detergent treatment, but each of these treatments adds cost and complexity to pooled plasma processing. In either case, pooled plasma processing is generally not suitable for blood centers and is generally only practically suitable for industrial, high-volume production environments.

[0120] Conversely, unit-by-unit (unit) collection and processing is well suited to a blood center environment, eliminating the risk of pathogen contamination of pooled plasma by allowing for pre-processing testing for pathogens and unit tracking to ensure each unit leaves the blood center site pathogen-free. The inventors have discovered that effective and efficient storage and recovery of functional clotting factors is the standard by which successful unit plasma processing should be measured. Such efficiency is also highly useful in a pooled plasma environment as well.

[0121] clotting factors There are many plasma factors involved in coagulation. The methods and compositions of the present invention involve recovering amounts of functional fibrinogen, Factor V, Factor VII, Factor IX, and vWF from rehydrated plasma that has undergone a spray-drying process. These plasma factors are important in patient treatment, particularly after traumatic injury, to promote wound clotting. Therefore, rapid administration of plasma is an important factor contributing to positive clinical outcomes. The spray-dried plasma of the present invention can be easily reconstituted within minutes at the site of trauma, without patient transport or time delays. Furthermore, the spray-dried plasma of the present invention has high levels of functional proteins that are stable for extended periods of time without freezing.

[0122] Functional vWF is generally difficult to recover and is an indicator of overall factor conservation. The present invention encompasses recovering a certain amount of functional vWF in rehydrated spray-dried plasma that is at least about 5 percentage points higher (e.g., about 5, about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, or about 60 percentage points higher) than the amount of functional vWF in rehydrated spray-dried plasma that has not undergone the pretreatment process of the present invention. The present invention encompasses recovering a certain amount of functional vWF in rehydrated spray-dried plasma that is about 5 to about 40 percentage points higher, or about 10 to about 35 percentage points higher, than the amount of functional vWF in rehydrated spray-dried plasma that has not undergone the pretreatment process of the present invention. vWF activity is typically assayed using an assay known as von Willebrand factor:ristocetin cofactor (vWF:RCo), which is known to those skilled in the art. The vWF:RCo assay measures the ability of a patient's plasma to aggregate platelets in the presence of the antibiotic ristocetin. The rate of ristocetin-induced aggregation is related to the concentration and functional activity of 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 whether functional coagulation plasma proteins are present in reconstituted, pre-spray-dried plasma is to perform a Bioflux test. See Examples 9 and 10.

[0123] In one embodiment, von Willebrand factor antigen (% or IU / dL) and / or von Willebrand factor ristocetin cofactor (% or IU / dL) are measured before and after spray drying to determine vWF retention. In one embodiment, the acceptable or clinical range for von Willebrand factor antigen ristocetin cofactor (VWF:RCo) is about 50 to about 200 IU / dL, and the von Willebrand factor antigen (VWF:Ag) value is about 50 to 200 IU / dL. In one embodiment, the invention involves determining the amount 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 higher (e.g., about 5, about 10, about 15, about 20, about 25 percentage points higher) than the amount of vWF in rehydrated spray-dried plasma that has not been subjected to the pretreatment steps of the invention.

[0124] A pretreatment solution having one or more spray-dried stable acidic substances (SDSAS) and one or more amino acids. The present invention contemplates the use of a pretreatment solution comprising one or more physiologically compatible spray-drying stable acidic substances (SDSAS) combined with one or more amino acids as a compounding agent for plasma before it is spray-dried. As used herein, phrases such as "SDSAS and amino acids" refer to a pretreatment solution having at least one SDSAS and / or at least one amino acid. Similarly, the use of "SDSAS" or "amino acids" refers to one or more SDSAS or one or more amino acids, respectively. The phrases "compounded plasma" or "pretreated plasma" or "PreT" refer to a mixture of a pretreatment solution (e.g., at least one SDSAS and / or at least one amino acid) and plasma before spray drying. Dry-state blended plasma refers to spray-dried plasma that has been pretreated with a pretreatment solution.

[0125] While the present invention is not limited by theory, the inventors hypothesize that the SDSAS (e.g., citric acid, lactic acid, hydrochloric acid, etc.) of the present invention exerts its effect by preventing or reducing the increase in plasma pH during the spray-drying process. Adding an amino acid to the SDSAS allows the pretreatment solution to still have an acidic pH, but not so low that it is harmful to plasma proteins. Non-limiting examples of suitable SDSAS are hydrochloric acid (HCl), citric acid, and lactic acid. When SDSAS is combined with an amino acid, an example is glycine HCl. SDSAS (e.g., HCl, citric acid, or lactic acid) and an amino acid (e.g., glycine) can be added to 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, bicarbonate and H2O are converted from CO2 and H2O. + The reaction that generates H + The pH shifts from 0 to 1, thereby increasing the pH (the so-called Le Chatelier principle). Human blood / plasma contains carbonate (H2CO3) and bicarbonate anions (HCO3 - ), which is important for maintaining a blood pH of 7.35-7.45, as values ​​higher than 7.8 can lead to death. In this buffer, hydronium (HO) + ) and bicarbonate anion are in equilibrium with carbonic acid (Equation 1). Furthermore, carbonic acid in the first equilibrium can decompose into CO2 gas and water, which leads to a second equilibrium between carbonic acid and water (Equation 2).

[0126] In summary, blood buffers are: H2CO3 + H2O ⇔ H3O + +HCO3 - (Equation 1).

[0127] For the simultaneous equilibrium of H2CO3⇔H2O+CO2 (Equation 2).

[0128] Spray drying generates CO2, which is converted into H2CO3 and H3O + As a result, the pretreatment solution of the present invention serves to safely lower the pH of formulated plasma prior to spray drying to provide spray-dried plasma that, upon reconstitution with sterile water for injection, has a resulting physiologically compatible pH.

[0129] The addition of glycine helps offset this change. The addition of an amino acid prevents the pH from decreasing excessively. Thus, in one embodiment, plasma is pre-treated with SDSAS and an amino acid. Due to the blending / pre-treatment step, vWF activity loss is reduced and / or the amount of native vWF is increased compared to spray-dried plasma not subjected to the blending step of the present invention. SDSAS is present in the pre-treatment solution in an amount of about 1 mM to about 50 mM, which lowers the pH of the blended plasma to about 5.5 to about 6.5 or about 7.2, producing blended plasma. An amino acid, such as glycine, is also present in the pre-treatment solution with SDSAS in an amount of about 1 mM to about 150 mM, which lowers the pH of the blended plasma to about 6.0 to about 6.6.

[0130] Because the SDSAS and amino acids of the present invention are physiologically compatible in this manner, the inventors have further determined that the rehydration step can be performed with water (e.g., SWFI) alone. Alternatively, sodium phosphate or other agents can optionally be added to the rehydration solution. Furthermore, any other suitable rehydration fluid may be used, as can be determined by one of skill in the art.

[0131] Experiments conducted by the present inventors using spray drying have revealed that von Willebrand factor activity in plasma dried by spray drying is partially affected by shear forces and an increase in plasma pH that occurs during the aerosolization process (see Examples below). The present invention demonstrates that utilizing a process in which plasma is combined with at least one SDSAS and at least one amino acid significantly improves the retention and stability of active vWF compared to conditions in which SDSAS and amino acids are not used as blending agents.

[0132] An SDSAS is a substance that does not readily evaporate at room temperature and atmospheric pressure. Typically, the boiling point of an SDSAS is greater than about 150°C at atmospheric pressure. In addition to glycine HCl, non-volatile acids suitable for use as the SDSAS of the present invention include phosphorus-containing acids, such as ortho-phosphoric acid, pyrophosphoric acid, meta-phosphoric acid, polyphosphoric acid, alkyl- and aryl-substituted phosphonic and phosphinic acids, phosphorous acid, and their equivalents, 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, citric acid, lactic acid, gluconic acid, oxalic acid, halogenated acetic acids, arenesulfonic acids, molybdic acid, phosphotungstic acid, tungstic acid, chromic acid, sulfamic acid, and their equivalents.

[0133] In one embodiment, the pretreatment solution of the present invention may comprise one or more SDSAS and one or more amino acids. Adding an amino acid to SDSAS allows for the protection of plasma proteins without lowering the pH too much and causing adverse effects such as protein damage and complement activation. The addition of an amino acid increases the pH of the pretreatment solution, but surprisingly, this does not affect the pH of the rehydrated spray-dried plasma (ODP). In particular, the pH of the pretreatment solution ranges from about 2.0 to about 4.0, resulting in a combined plasma (e.g., before spray drying) having a pH of about 6.0 to about 6.6, and a rehydrated plasma having a pH of about 6.5 to about 7.8. In one embodiment, the pretreatment solution of the present invention may comprise SDSAS and at least one (e.g., one or more) amino acids. The total concentration of the one or more amino acids present in the pretreatment solution is in an amount of about 1 mM to about 150 mM. Examples of amino acids that can be added to the SDSAS pretreatment solution include alanine, asparagine, glutamine, glycine, isoleucine, leucine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine. In a specific embodiment, glycine is added to the SDSAS composition as shown in the examples. The addition of amino acids increases the pH of the pretreatment solution, but surprisingly, this does not affect the pH of the rehydrated spray-dried plasma (ODP). See Example 17. Even more surprisingly, the addition of amino acids mitigates C5a elevation. See Example 16. These amino acids have at least two pKa values, as follows:

[0134] [Table 8]

[0135] SDSAS useful in the process of the present invention are capable of replacing (or compensating for) volatile acids, i.e., CO2, which escapes from plasma during spray drying. As noted above, example or suitable acids include, but are not limited to, glycine HCl, ascorbic acid, citric acid, gluconic acid, and lactic acid.

[0136] Volatile acids, as defined herein, have a pKa of less than about 3 and a boiling point at atmospheric pressure of less than about 150° C. Typically, the pKa of a volatile acid is in the range of about 1 to about 15. Non-limiting examples of volatile acids are hydrogen chloride, hydrogen bromide, hydrogen iodide, acetic acid, formic acid, hydrogen sulfide, hydrogen selenide, sulfur dioxide, fluorosulfonic acid, methanesulfonic acid, trifluoroacetic acid, trifluoromethanesulfonic acid, and the like.

[0137] Volatile strong acids can be immobilized with amino acids or their equivalents to make them nonvolatile and more usable. For example, volatile hydrogen chloride can be converted to glycine hydrochloride (glycine HCl, glycine hydrochloride). To distinguish this from adding an amino acid to a pretreatment solution, this describes how SDSAS is made. The pretreatment solution involves adding an amino acid that is not bound to SDSAS, but to which the acid is already conjugated and stable. Alternatively, glycine and HCl can be added to the pretreatment solution in these amounts to form SDSAS (e.g., glycine HCl) and free amino acid (e.g., glycine) in solution. Because a 1:1 relationship exists between glycine and HCl, more glycine than HCl can be added to obtain a solution containing free glycine and glycine HCl. In one embodiment of the present invention, HCl and glycine are added to a solvent such as SWFI to produce a pretreatment solution with a final concentration in the combined plasma of about 16.8 mM HCl and about 69.6 mM glycine. In other words, in one embodiment of the present invention, 5.2 mM HCl and 21.5 mM glycine are added to 50 mL of a solvent, such as SWFI, to produce a pretreatment solution. The present invention provides a method for producing a pretreatment solution by adding 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) of HCl and about 15 to about 30 mmol of glycine (e.g., about 15, 17, 20, 22, 25, 27, and 30 mmol of glycine) to a solvent, such as SWFI, in 50 mL of solvent to obtain the following blended plasma: 260. In yet another embodiment, about 440 mM of glycine and about 106 mM of HCl are present in the pretreatment solution. In one embodiment, 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 are present in the pretreatment solution.

[0138] 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 about 1 mM to about 50 mM SDSAS and about 1 mM to about 150 mM (e.g., 50 mM to about 100 mM) amino acids. The following table shows how the above calculations were obtained.

[0139] [Table 9]

[0140] In one embodiment, a pretreatment solution of the present invention can have a 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 using 53 mL of pretreatment solution made from 1.61 g of glycine and 0.52 g of HCl, or equivalent. This ratio results in approximately 67 mM glycine and approximately 16 mM HCl in approximately 319 mL of combined plasma before the plasma is spray-dried.

[0141] A study of pooled, ABO-matched, never-frozen plasma (NFP) from whole blood treated with a citrate phosphate dextrose (CPD) anticoagulation regimen and pretreated with the pretreatment solution of the present invention demonstrated that dilution of the pretreatment solution of the present invention with plasma dried at +20%, +10%, +5%, -5%, -10%, and -20% did not significantly affect the properties of the dried plasma upon reconstitution and when assayed by a 25-assay panel. Notably, the effect of the pretreatment dilution range on the sensitive vWFRCo assay showed normalized retention of vWF by the assay within the error range exhibited by the control plasma. pH was also well controlled over a range of 7.32 to 6.84, with the pH of the control plasma being 7.11.

[0142] These results demonstrate that the pretreatment solution of the present invention readily enables the production of spray-dried plasma that, after reconstitution, demonstrates comparable assay panel results to NFP, FFP, and PF24, demonstrating assay characteristics that are essentially the same as NFP, FFP, and PF24.

[0143] The pretreatment solution of the present invention can have a weight ratio of glycine to HCl of about 5 to 3. The formulated plasma treated with the pretreatment solution of the present invention can have a mmol / mL ratio of glycine to HCl of about 5 to 3.

[0144] In one embodiment, the present invention involves adding a volatile acid and an amino acid as separate compounds (e.g., not as a salt) to form a solution. The volatile acid and amino acid should be added in amounts to produce about 1 mM to about 50 mM SDSAS and about 1 mM to about 150 mM amino acid. When a 1:1 relationship exists between the volatile acid and amino acid, such as with HCl and glycine, each is added in equal amounts ranging from about 1 mM to about 50 mM, and additional amino acid is added to achieve a free amino acid concentration in the solution of about 1 mM to about 150 mM. In other embodiments, strong corrosive acids can be converted to acid salts for use in pretreating plasma prior to spray drying. Examples include NaHSO4 and NaH2PO4, i.e., acid salts of sulfuric acid.

[0145] In one embodiment, the pretreatment solution has glycine in an amount ranging from about 10 μmol plasma to about 110 μmol plasma (e.g., about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110 μmol / mL plasma) and hydrochloric acid (HCl) in an amount ranging from about 10 μmol / mL plasma to about 30 μmol / mL plasma (e.g., about 10, about 15, about 20, about 25, and about 30 μmol / mL plasma), thereby obtaining blended plasma. In one embodiment, the pretreatment solution has glycine in an amount of about 84 μmol / mL plasma and HCl in an amount of about 20 μmol / mL plasma.

[0146] In another embodiment, the pretreatment solution has an amount of glycine and an amount of HCl that form a ratio that allows free glycine to be present in the pretreatment solution. In one aspect, the ratio of glycine to HCl is about 1.5 to 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 certain embodiments, the ratio of glycine to HCl is 4.15. In yet another embodiment, the ratio of glycine to HCl affects the pH of the pretreatment solution to be about 2.0 to about 4.0, or results in the combined plasma of step a) having a pH of about 6.0 to about 6.6. When reconstituted with sterile water, combined plasma having the above-described ratio of glycine to HCl produces a pH of about 6.7 to about 7.8.

[0147] The invention further includes methods of producing spray-dried plasma by combining plasma with a pretreatment solution, the pretreatment solution having glycine in an amount ranging from about 15 mmol to about 30 mmol (e.g., about 15, 20, 25, and 30 mmol) and HCl in an amount ranging from about 3 mmol to about 7 mmol (e.g., about 3, 4, 5, 6, and 7 mmol). In certain embodiments, the pretreatment solution has glycine in an amount of about 22 mmol and HCl in an amount of about 5.3 mmol.

[0148] Non-volatile acids and acid salts are collectively defined and included as spray-dried stable acidic substances (SDSAS) in the present invention. In one embodiment, the pretreatment solution of the present invention comprises an SDSAS and at least one amino acid.

[0149] In one embodiment, the SDSAS and / or one or more amino acids of the present invention are 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 one embodiment, the SDSAS of the present invention is added to the plasma simultaneously as the plasma is being pumped through the spray-drying apparatus. The term "simultaneously" is defined herein to mean 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. By adding an amino acid to the SDSAS that increases the pH of the pretreatment solution, in one embodiment, the plasma formulation (e.g., the pretreatment solution and the plasma to be spray-dried) may be allowed to store or stand for up to about 24 hours (e.g., 1 hour, 5 hours, 10 hours, 15 hours, 20 hours, or 24 hours) before spray-drying.

[0150] In one embodiment, the present invention involves mixing the pretreatment solution with the plasma to be spray-dried using a technique called rapid mixing. The rapid mixing step is optional. One of the discoveries of the present invention involves rapid or immediate mixing of the pretreatment composition with the plasma. It has been discovered that slow mixing of the pretreatment solution with the plasma allows localized contact or pockets of unmixed acid to come into contact with plasma proteins, which can affect these proteins, specifically increasing C5a. In contrast, when the pretreatment solution and plasma are rapidly mixed and / or stirred, in one embodiment, the amount of C5a is equivalent to that of fresh frozen plasma or other similar FDA-approved formulations on the market. Rapid mixing and / or stirring allows for immediate, complete, and rapid mixing of the pretreatment solution (e.g., SDSAS and one or more amino acids) with the plasma. See Examples 17 and 18. Rapid mixing is defined as adding a large amount of plasma to a relatively small amount of pretreatment solution prior to spray-drying the plasma. Generally, when a much smaller volume (e.g., about 10 to about 30% (about 10%, about 15%, about 20%, about 25%, about 30%) of the larger volume is added to a larger volume, mixing the two volumes results in rapid and thorough mixing of the two volumes. In a preferred embodiment, 260 mL of plasma is added to 50 mL of pretreatment solution. In one embodiment, once rapid / instant mixing occurs, the operator can gently invert the bag containing both the pretreatment solution and the plasma several times (e.g., 1 to 5 times) to further mix the two together. In contrast, with respect to mixing of the pretreatment solution and plasma, when a small amount of pretreatment solution is injected into a large amount of plasma being spray-dried, the small volume takes longer to fully mix into the larger volume, and small pockets may form within the larger volume. During this time, local contact or pockets of unmixed acid formed within the mixture have been found to cause an increase in the amount of C5a in the resulting reconstituted plasma. Agitation is defined as constant shaking or movement of the components of the pretreatment solution (e.g., SDSAS, amino acids, and plasma). Rapid mixing or agitation results in a homogeneously mixed plasma formulation with little or no local contact or pockets of unmixed acid.

[0151] The present invention relates to the addition of SDSAS and at least one amino acid to spray-dried plasma for a period of time prior to spray drying that is short enough to obtain a formulation ("plasma formulation") having a desired pH and to prevent denaturation or damage of certain plasma proteins, such as von Willebrand factor, or an increase in C5a due to prolonged exposure to low pH conditions. In one embodiment, maintaining a time delay of 30 minutes or less between formulation of plasma with SDSAS and spray drying, as described below, results in improved retention of plasma proteins, including von Willebrand factor, without undesirable protein damage due to prolonged exposure to low pH conditions prior to spray drying.

[0152] The time period between pretreatment blending and spray drying varies depending on the pH / acidity of the plasma blend produced by mixing the SDSAS, amino acids, and plasma. In one embodiment, the time period between contacting the SDSAS and amino acids with the plasma and spray drying the plasma ranges from about 0 seconds (e.g., the time at which aerosolization occurs: time 0) to about 30 minutes. In one embodiment, to minimize protein denaturation, the time between adding the pretreatment solution to the plasma and spray drying should be kept to a minimum. The practical maximum time between blending and spray drying is determined empirically. Blending this close to time 0 is also referred to as "simultaneous blending."

[0153] There are several ways in which co-blending can be performed. In one embodiment, a blending station is provided in conjunction with the spray dryer. In conjunction with the blending station, the weight or volume of the plasma before spray drying is determined, and the SDSAS and amino acid doses are measured to achieve the desired pH of the plasma blend. The doses may be introduced into the plasma by any convenient method, including injection through a port on the plasma bag. In one embodiment, the bag containing the plasma and the bag containing the pretreatment solution are sterilely connected by tubing using a tube sealer that can sterilely heat seal the two ends of the tubing. In such cases, the transfer of the plasma to the pretreatment bag can be performed manually or by using a collection monitor or scale. Gravity can be used, and the transfer can be assisted by suspending the plasma bag higher than the pretreatment bag. The blending station may be operated manually, semi-manually, or automatically. Of course, the timing of administration must be controlled as described above. The timing control may be manual, semi-manual, or automatic.

[0154] In another embodiment, appropriate doses of SDSAS and one or more amino acids are introduced into the plasma flow path of the spray dryer prior to the spray drying head. The introduction of the pretreatment solution is controlled manually, semi-manually, or automatically to produce the desired plasma formulation.

[0155] In a further embodiment, an appropriate amount of pretreatment solution (e.g., SDSAS and one or more amino acids) is introduced into the spray-drying chamber close enough 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. The introduction of the pretreatment solution (e.g., SDSAS, one or more amino acids) is controlled manually, semi-manually, or automatically to produce the desired plasma formulation.

[0156] In yet another embodiment, the pretreatment solution is combined with the donor plasma using a sterile connection device and scale, as further described herein.

[0157] C5a G protein-coupled receptors are widespread throughout the human body, comprising approximately 60% of known cellular receptor types and mediating transmembrane signaling for a wide variety of endogenous ligands. These receptors are involved in a wide variety of physiological and pathophysiological processes, including, but not limited to, those related to the cardiovascular system, central and peripheral nervous system, reproduction, metabolism, digestion, immunological, inflammatory and growth disorders, and other cell regulatory and proliferative disorders. One of the most well-studied G protein-coupled receptors is the human and mammalian complement (C) system, which contains over 20 components involved in the orchestrated reactions that result in complement activation. The blood complement system has diverse functions related to a wide range of host defense mechanisms, including antimicrobial and antiviral effects. Products derived from C component activation include the nonself-recognition molecules C3b, C4b, and C5b, and the anaphylatoxins C3a, C4a, and C5a, which influence various cellular immune responses. These anaphylatoxin molecules are involved in pro-inflammatory effects, both acute and chronic inflammation, and the associated pain and tissue damage.

[0158] This data reveals the surprising result that adding a certain amount of glycine to the acid pretreatment solution increases the pH of the pretreatment solution, providing additional protection to plasma proteins and mitigating C5a elevation. Addition of appropriate levels of glycine does not affect the maximum pH of the rehydrated spray-dried plasma (ODP).

[0159] It is desirable that the pretreatment solutions of the present invention produce rehydrated plasma with reduced C5a levels or levels of FDA-approved apheresis plasma products. In particular, the present invention relates to pretreatment solutions that produce C5a levels similar to those of fresh frozen plasma or never-frozen plasma, or available FDA-approved apheresis plasma products. As shown in Figure 28 and the results of Example 17, when local contact with low-pH acids can be avoided, C5a levels are similar to those of never-frozen plasma. Local contact with low-pH acids can be avoided by raising the pH using an amino acid such as glycine, by using the rapid mixing technique described above, or by a combination of these. As can be seen from Figure 28, when using glycine HCl, which has a pH of 1.32, without the addition of an additional basic amino acid such as glycine, the rapid mixing technique reduces C5a levels from about 64 ng / mL to about 31 ng / mL. When glycine was added to SDSAS to lower the pH (e.g., a glycine HCl / glycine pretreatment solution), rapid mixing techniques resulted in C5a levels of approximately 12 ng / mL, close to the level of never-frozen plasma (NFP), which is approximately 10 ng / mL. Using citric acid, which has a pH of 2.28 without the addition of an amino acid, and rapid mixing techniques also resulted in C5a levels similar to those of NFP (12.66 ng / mL). The pH of citric acid is higher than that of glycine HCl. When a citric acid pretreatment solution is combined with an amino acid (glycine), the pH of the solution is 3.4, and rapid mixing does not actually affect C5a levels, as both are close to the pH of NFP, e.g., approximately 10 ng / mL. It has been discovered that desirable C5a levels result from the addition of a pretreatment solution with SDSAS and an amino acid, rapid mixing / agitation of the pretreatment components, or a combination of both. In particular, the C5a level in the reconstituted plasma obtained from the pretreatment solution of the present invention can be from about 4.7 ng / mL to about 74 ng / mL, particularly from 8 ng / mL to 12 ng / mL (eg, about 10 ng / mL).The C5a level is reduced compared to plasma not subjected to a pretreatment solution having at least one SDSAS and at least one amino acid. In one embodiment, the C5a level is reduced by about 20% or less (e.g., 20%, 15%, 10%, 5%, 1%) compared to plasma not subjected to a pretreatment solution of the present invention. In another embodiment, referring to FIG. 28, rapid mixing and certain pretreatment formulations result in C5a levels that are approximately the same as those in never-frozen plasma, or within a range of about 20% or less (e.g., 20%, 15%, 10%, 5%, 1%) of the C5a in never-frozen plasma or in an FDA-approved apheresis plasma product.

[0160] Similarly, in one embodiment, rapid mixing and / or stirring is not required 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 even though the pH of the pretreatment solution varies, e.g., ranges from about 1 to about 4, the pH of the plasma remains approximately the same, e.g., about 6.2.

[0161] Protein stability of pretreated plasma Proteins can undergo physical degradation (e.g., unfolding, aggregation, formation of insoluble particulates) by several 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 shifts, pH changes and exposure to extremes, shear stress, surface adsorption / interfacial stress, etc. Proteins in solution can be converted to solid formats for improved storage (i.e., powders or other dry formats by significantly reducing or removing water and other volatile components of the protein solution) using several methods.

[0162] Freeze-drying (also known as lyophilization) is the most common processing method for removing water from biopharmaceuticals, enhancing the stability, temperature tolerance, and shelf life of these formulations. It involves freezing a suspension, colloid, or solid and then "drying" it under vacuum via sublimation (phase transition). During this process, proteins can suffer from low-temperature denaturation, interfacial stress (adsorption at the water / ice interface), exposure to increasing alkaline pH (CO2 loss), and dehydration stress. Freeze-drying is well established in the industry; however, it requires expensive equipment that takes up a large amount of space within manufacturing facilities. Freeze-drying can also take days to complete, and manufacturers requiring powdered products must incorporate a granulation step into the process. In environments with limited budgets, time, and space, freeze-drying can be a challenging option for some companies. Due to the space required, drying plasma using freeze-drying technology is limited to plasma manufacturers and cannot be performed in blood centers.

[0163] Due to the inherent difficulties in freeze-drying plasma in terms of time, space and cost, the present invention relates to an improved spray-drying process for plasma that overcomes the known difficulties associated with spray-drying plasma.

[0164] In the spray-drying process, a viscous liquid is pumped through a supply line to a nozzle, and the exiting fluid stream is atomized into a large number of droplets under an aerosol gas. The droplets are then contacted with a drying gas to form dry particles. This is a much shorter and less expensive process than the freeze-drying process, and it can be performed in laboratories and blood centers. However, prior to the present invention, this process can subject plasma proteins to a wide range of shear stresses, interfacial stresses, thermal stresses, dehydration stresses, and pH extremes.

[0165] Aerosolization subjects liquid samples to shear stress, producing extremely rapid and significant expansion of the gas-liquid interface. The combined effect of shear stress and gas-liquid interfacial stress can cause serious and detrimental effects on labile compounds such as proteins. Complex biomolecules are difficult to spray-dry because they are highly sensitive to high shear stress. While some control over the amount of shear stress encountered can be obtained, for example, by selecting the type of atomizer and aerosolization pressure used, applying spray-drying techniques to human plasma is extremely challenging due to the large number of diverse proteins it contains. Different proteins may be susceptible to different stresses, which can make it difficult to determine process conditions suitable for all protein types found in plasma. In particular, vWF, naturally designed for shear sensitivity in its biological function, is the most shear-sensitive human plasma protein. Most other plasma proteins remain largely intact after spray drying, with the exception of vWF. As shown in the Examples section, spray drying reduced vWF activity below detection levels (see Example 27, Figure 17).

[0166] Ionizable amino acid residues have been shown to play important roles in protein binding to other molecules and enzyme mechanisms. They also have a significant impact on protein structure, stability, and solubility. The types of interactions these side chains have with their environment change depending on their protonation state. As a result, their pKa values ​​and the factors that influence them are of intense biochemical interest. Strongly altered pKa values ​​are often found in the active sites of enzymes, enhancing the ability of ionizable residues to act as nucleophiles, electrophiles, or general bases and acids. As a result of changes in the protonation of these residues, protein stability is pH-dependent. Therefore, it is believed that inhibiting alkalinization of plasma during spray drying could potentially improve the processability and storage stability of many plasma proteins.

[0167] As mentioned above, the spray-drying process subjects plasma proteins to forces different from those found in the freeze-drying process. First, spray-drying subjects plasma proteins to high stress during the aerosolization process because the plasma is forced through a narrow orifice exposed to the high-velocity airflow required to generate droplets of a suitable size for drying. Second, the spray-drying process exposes plasma proteins to the high temperatures required to force water out of the aerosolized droplets. Third, the spray-drying process subjects plasma proteins to a dramatic and rapid increase in pH as a result of the rapid release of CO2 during drying. Because freeze-drying does not subject plasma proteins to these forces, especially the unique combination of these forces, those skilled in the art would likely find no suggestion or motivation in the freeze-drying art to improve the spray-drying process for plasma.

[0168] Despite the difficulties associated with spray drying plasma, the spray drying process of the present invention results in high retention and stability of functional plasma proteins, particularly but not limited to vWF, with retention of vWF in the rehydrated spray-dried plasma being at least about 5 percentage points higher (e.g., about 5, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80 percentage points higher) compared to the amount of active / native vWF in rehydrated spray-dried plasma that has not been subjected to the pretreatment step of the present invention.

[0169] The compositions and processes of the present invention relate to the effect of formulating liquid plasma with an SDSAS, e.g., glycine HCl, alone or in combination with an amino acid, on the retention and stability of functional vWF and other coagulation factors through the spray-drying process (during storage of the dried, spray-dried, and rehydrated plasma). This can be accomplished by adding an SDSAS, e.g., glycine HCl, citric acid, or lactic acid, alone or in combination with an amino acid, such as glycine, to the liquid plasma before spray-drying begins or simultaneously with the spray-drying process. During the spray-drying process, CO2 loss occurs, which makes the pH of the plasma composition more alkaline (e.g., increases), and the addition of the SDSAS and amino acids maintains the plasma pH in a range that prevents significant denaturation of coagulation factors, particularly vWF. Therefore, pretreatment of plasma with citric acid, glycine HCl, or other SDSAS, or in conjunction with amino acids, serves at least three purposes: 1) increasing the in-process retention of plasma proteins, 2) increasing the stability of plasma proteins during storage, and 3) allowing the spray-dried plasma to be rehydrated with water (e.g., sterile water, WFI), eliminating the need for a specific rehydration solution.

[0170] When SDSAS and amino acids are combined prior to drying the liquid plasma, the acids are present in the dried plasma product at a level consistent with improved shelf life and reduced degradation of coagulation factors during storage. "Levels consistent with increasing shelf life" herein also refers to levels that result in a physiological pH upon reconstitution of the spray-dried plasma. The use of SDSAS and amino acids also allows for simple rehydration with low-cost, readily available sterile water for injection, or, in emergencies, purified water at physiological pH. The convenience, reduced costs, and improved safety associated with direct rehydration with water are evident. Advantages include savings in the ability to transport the dried plasma product without the weight and bulk of the rehydration fluid, cost savings from not having to specially formulate the rehydration fluid, and reduction or elimination of refrigeration or freezing during storage.

[0171] Thus, the inventors have discovered that plasma formulations with SDSAS and amino acids result in spray-dried plasma with high retention or functionality of plasma proteins, particularly vWF, highly improved storage properties of the dried plasma, and a near-neutral pH when rehydrated with water without a rehydration buffer fluid. Thus, the present invention allows for the production of spray-dried plasma without the added expense and complexity of pretreatment with polyols or other additional stabilizers known in the art. However, the use of stabilizers is not contraindicated and may be beneficial in some cases.

[0172] In a further embodiment, novel compositions of matter for plasma spray drying are produced by administering, by any means, pre-spray dried plasma containing citrate (also known as citric acid) or other suitable SDSAS and amino acids at appropriate concentrations as disclosed herein.

[0173] In further embodiments, the freshly administered citrate-loaded plasma prior to spray drying has a citrate concentration of about 27.5 mM and about 40.4 mM, or about 31.6 mM and 34.2 mM.

[0174] In a further embodiment, the novel spray-dried plasma formulations are produced by spray-drying plasma that has been formulated with appropriate levels of a suitable SDSAS (e.g., citric acid) and a suitable amino acid prior to or concurrently with drying, and then drying the plasma to the desired level of moisture, which is generally less than 2%.

[0175] In various embodiments, citric acid or other SDSAS and amino acids are added to plasma as compounding agents. Experiments relating to the effect of SDSAS and amino acids on protecting the activity of proteins found in plasma are further described in the Exemplification section herein. For example, citric acid is used at concentrations of about 1 to about 15 mM, or about 5 to about 10 mM (e.g., 7.4 mM). Thus, plasma proteins may be better protected when citric acid is added at the indicated concentrations before or simultaneously with spray drying. The activity of vWF is provided in the examples because this factor is particularly sensitive to denaturation and damage due to spray drying (see Example 17 and Figure 18) and is therefore a good indicator protein for the beneficial effect of SDSAS and amino acids on the retention and stability of spray-dried plasma proteins.

[0176] Examples of other physiologically compatible SDSAS and amino acids are known to those of skill in the art and are described herein.

[0177] In one embodiment, single-donor plasma expressed from collected whole blood or apheresis, never frozen, and collected less than 24 hours ago, is preferably utilized in this process. Plasma is collected from blood by standard techniques known to those skilled in the art, as described herein. Plasma is collected through a process called plasmapheresis, which refers to procedures in which plasma is separated from blood by either centrifugation or membrane filtration. The system process can also be utilized with pooled plasma, if desired, as well as starting plasma material produced with any of the currently available anticoagulation systems known as CPD, CP2D, ACD-A, and ACD-B. A sterile, non-pyrogenic, single-use container of SDSAS, e.g., 50 mL of solution glycine and hydrochloric acid, is packaged in a 500 mL container inside an overwrap pouch. In one embodiment, the process of the present invention involves converting a single-donor unit of plasma collected by standard procedures into a single unit of spray-dried plasma.

[0178] The dried plasma of the present invention can be derived from a third-party donor or from the potential recipient himself. The latter is known as autologous plasma. Autologous plasma is highly desirable because it overcomes compatibility issues, e.g., it does not need to be ABO / Rh typed prior to use, and the plasma is less likely to carry any foreign substances, such as pathogens or immunogens, to the recipient.

[0179] In vitro characterization data demonstrate that the system's manufacturing effect is comparable between units manufactured with different starting materials. Units manufactured from apheresized plasma (ACD-A anticoagulated) showed similar percent changes attributable to manufacturing effects in the starting material compared to units manufactured from whole blood-derived plasma (CPD anticoagulated). Statistical analysis (ANOVA) was performed on the percent changes before and after manufacturing between the two starting materials across 20 assays, including clotting time, coagulation function, and activation markers. Of the 20 assays, total protein concentration, PT, TT, and factor VIII and XIII activity were determined to be statistically significantly different; however, the mean percent changes were similar, and all mean values ​​were within the clinical reference range. In summary, the in vitro test results support the conclusion that manufacturing effects in both apheresized and whole blood plasma are comparable, and the coagulation profiles are within ±20% of their paired controls, or within the normal reference range.

[0180] Spray Dryer and Spray Drying Process Generally, a spray dryer system (spray dryer) is provided for spray drying a liquid sample such as plasma.

[0181] The pretreated plasma is dried using components and a system for using a disposable spray-drying device. The spray-drying system includes a spray-drying apparatus (hereinafter referred to as a "drying apparatus," "machine," "spray dryer," or "dryer"), a spray-drying finishing apparatus (hereinafter referred to as a "finishing apparatus," "seal and separator," or "finisher"), and a disposable spray-drying device (hereinafter referred to as a "disposable device" or "disposable"). The present invention includes a system that enables a disposable spray-drying device with a liquid spray nozzle and drying chamber to efficiently dry liquids, including liquid human or animal plasma, while protecting active components such as plasma proteins. The disposable spray-drying device is installed in a spray dryer that controls the plasma flow, pressurized aerosol gas flow, drying gas flow, temperature, pressure, etc., within the disposable. Upon completion of the spray-drying process, the disposable containing the dried plasma powder is aligned and processed by the spray-drying finishing apparatus, where portions of the disposable are sealed and separated to form a dried plasma unit. Furthermore, the present invention advantageously provides an apparatus for performing the functions of spray-drying and finishing products containing dried human plasma.

[0182] The disposable spray dryer of the present invention has a compact drying chamber that produces dry powder (<2% residual moisture) at a high powder production rate. The disposable is small, easy to handle, and provides a high-performance, easy-to-use drying chamber. The drying system of the present invention is a significant improvement, providing a removable, disposable drying chamber for spray drying that is suitable for processing small batch sizes such as individual blood units.

[0183] Certain of Applicant's older disposable drying chambers were very long, measuring 58 inches to 66 inches or more in length, allowing sufficient time (flight path) to dry plasma to acceptable residual moisture levels. See Applicant's Patent Nos. 8,533,971, 8,595,950, 8,434,242, 8,601,712, 8,533,972, and 10,843,100. However, their lengths made these prior art disposables unacceptable for practical use because they were difficult and inefficient to handle during installation in spray dryer equipment. As further described herein, the shorter disposables of the present invention are easier to handle than these prior art disposables, which required user reach and bending distances of more than 6' and less than 5', respectively. The shorter disposables make spray drying of human plasma practical for real-world applications by real-world people. Additionally, the disposable drying chamber of the present invention is a removable, disposable drying chamber that maintains the quality and integrity of the plasma while improving processing time and product quality at a low cost.

[0184] Several challenges have been overcome by shortening the drying chamber of the present invention. For example, drying any product to a predetermined degree of dryness involves exposing the material being dried to sufficient heat energy to achieve the desired level of dryness while maintaining the functionality of the substance being dried. However, shortening the drying chamber also shortens the drying path.

[0185] The disposable drying chamber of the present invention is improved by: more efficiently produced plasma; Fairly short; Easy to use for people of a wide range of status; Drying materials in less time; Decrease in inlet air temperature; A nozzle assembly and dry environment are achieved to obtain rapid mixing of the atomized droplets with the drying gas and rapid evaporation; Achieve a lower level of residual dryness, e.g., less than 2.5% residual moisture; and As further described herein, a specially designed, cost-effective composite spray drying nozzle is utilized.

[0186] Overview of spray-dried disposables In particular, the disposable 100 has two general regions: a spray drying head 2 and a plasma drying chamber 28 .

[0187] Overview of the spray drying head 42A and 43A show a disposable 100 spray-drying head 2 having a guide 4 and a baffle plate 8 with a ridge 9 offset so as to be disposed in a plenum 6. The plenum 6 has a guide 4 at the top of the spray-drying head 2. Within the guide 4 is a spray-drying nozzle assembly 20 having a plasma flow inlet 18 connected to liquid plasma via a plasma tube 16 and a pressurized aerosol gas inlet 14 connected to pressurized gas via an aerosol tube 10 and an aerosol filter 12. Also shown is a drying gas inlet port 22, which communicates with a drying gas source (not shown), which may be air, nitrogen, or other drying gas source. Optionally, the drying gas inlet port 22 may be covered by a removable cover, such as a self-adhesive paper label or equivalent. This cover must be removed immediately prior to installation of the disposable 100 into the spray dryer 200. The drying case source may 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) coupled to an Atlas-Copco CD45 desiccant drying system, which supplies clean dry air (CDA) to the spray dryer, heating the air to the appropriate temperature for spray drying. In one embodiment, the drying gas flows through a CDA filter, such as a Millipore Series 3000 0.2 micron filter CTGB71TP3 manufactured by Millipore Sigma of Danvers, MA, USA. The CDA source, in one embodiment, is used as a source for the drying gas and pressurized gas. In certain embodiments, the spray drying nozzle assembly 20 includes a "manifold" that regulates the plasma line and the aerosol line. When the plasma source, pressurized gas source, and drying gas source are combined, droplets of liquid plasma are formed and dried into dry plasma (e.g., fine amorphous plasma powder). The plenum 6 has a notch that is a locator, also referred to herein as a locator 26 or a second locator, as described further herein.

[0188] Briefly, guide 4 fits into receiver 204 of spray dryer 200, which also ensures proper alignment of disposable 100 and dryer 200 (FIGS. 45B and 45C). Guide 4 also aligns spray dryer head 2 with spray dryer 200 in a specific orientation such that drying gas inlet 22 receives a drying gas source (not shown). Ridge 9 fits into and supports ridge receiver 207 of spray dryer 200. Guide 4, along with ridge 9, allows for lateral alignment of disposable 100 and spray dryer 200 (e.g., in the plane defined by the top and bottom surfaces of the spray dryer) that keeps disposable 100 stationary, thereby eliminating vertical movement of the dryer within the spray dryer chamber housing. Additionally, ridges 9 on disposable 100 fit into receptacles 404 on finisher 400 to secure disposable 100 to finisher 400 while finisher 400 displaces the plasma, seals and separates the disposable, and converts it into dried plasma units 60. See Figures 46A-46C. This alignment configuration also provides for easy and consistent mounting of the disposable to both the dryer and finisher.

[0189] A first locator, i.e., locator 206 (FIGS. 45B, 45C, and 46A), and a second locator, i.e., locator 26 (FIGS. 42A and 43A), are positioned on the spray-drying apparatus 200 and the second locator is positioned on the spray-drying disposable 100 to engage the spray-drying apparatus 200 during installation of the disposable 100 to align the disposable with the spray-drying apparatus. The same locator, locator 26 (second locator) on the disposable is also used to align the disposable with a third locator, i.e., locator 452 (see FIGS. 47A-47C), on the spray-drying finishing apparatus 400, which directs the dried plasma to specific compartments of the disposable and seals and separates the dried plasma into plasma units with dried plasma. This arrangement aligns the disposable and the spray dryer coaxially, for example, about the axis defined by the center of the receiver in guide 4 (see axis A in Figure 43A). This arrangement also allows for easy and consistent mounting of the disposable to both the dryer and the finisher.

[0190] As part of the disposable, the spray-drying head 2 includes a nozzle assembly 20. This nozzle assembly allows the spray drying of the plasma to occur within the disposable. Overall, the system design includes a spray dryer and a disposable modified to have a nozzle as part of the disposable in place of the spray dryer, so that the spray drying occurs entirely within the disposable. This design helps maintain the plasma within the disposable throughout the drying and finishing processes, and away from dryer or finisher parts that require decontamination between uses. The design also minimizes contamination from external pathogens by keeping the plasma within the disposable throughout the process. The nozzle assembly coordinates the plasma flow and the pressurized / aerosol gas flow so that both are emitted at the appropriate velocity and airflow to atomize the liquid plasma at the nozzle tip, ready for rapid mixing with the drying gas. The disposable 100 spray drying head 2 further includes a plenum 6 and a baffle plate 8 that directs drying air for rapid mixing with the aerosolized plasma and creates an air curtain to minimize the accumulation of dried plasma on the drying chamber walls.

[0191] Overview of the plasma drying chamber The drying chamber 28 is a disposable area where the plasma dries. The drying chamber is designed to capture the dried plasma while allowing humid air to escape. The drying chamber design also allows the drying chamber to be sealed and separated in a manner that forms a commercially available dried plasma unit.

[0192] Drying chamber 28 has three general regions: a top defined by dimension X (see FIGS. 44 and 46A ); a middle defined by dimension U, which is the region between positions 44A and 44B; and a bottom defined by dimension V, which is the portion below position 44B and includes filter 36 and separator 38. The top is the space where atomized liquid plasma impinges on the drying gas, evaporating the liquid within the droplets and drying them. In particular, the atomized plasma rapidly mixes with the drying gas and dries, as described further herein. As the plasma rapidly mixes and dries, it circulates and moves downward toward the filter. While most evaporation occurs in the top of drying chamber 28 (dimension X), drying continues as the plasma flows into the middle (dimension U) and bottom of drying chamber 28 (dimension V).

[0193] Drying chamber 28 also includes an intermediate section 46, defined by dimension U, having "seal and separate" positions 44A and 44B, label 40, spike ports 42A and 42B, and hang slot 34. Intermediate section 46 also includes locator pin opening 32C. The intermediate section is subsequently processed by a spray dryer finishing device, which is responsible for moving the dried plasma to a specific position in the plasma drying chamber and sealing and separating it at or near cut positions 44A and 44B. The portion between positions 44A and 44B becomes dried plasma unit 60, which is ultimately rehydrated and transfused into the patient.

[0194] The disposable 100 further includes a locating feature for reversibly attaching the outer wall of the disposable 100 to the finisher 400. Locating openings 32A, 32B, and 32C are present on the outer edge of the wall of the spray drying disposable device 100 (FIGS. 42A, 48A). Locating pins 432A, 432B, and 432C are positioned on the finisher 400 so that the drying chamber 28 of the disposable 100 is aligned on the finisher when the locating openings 32A, 32B, and 32C are positioned around the locating pins 432A, 432B, and 432C of the finisher 400. See FIGS. 47C, 48B, and 48C.

[0195] The lower section of the drying chamber 28 includes a lower filter 36 (also referred to herein as the "trap filter"), a lower filter separator 38, a drying gas outlet port 30, and locator pin openings 32A and 32B. Optionally, the gas outlet 30 may be covered by a removable cover, such as a self-adhesive paper label or its equivalent. In one embodiment, this cover must be removed immediately 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 moist air, and the separator acts as a spacer between the drying chamber wall and the filter, allowing air to pass more easily and preventing pressure buildup. Moist air refers to the air moving through the drying chamber and includes a combination of drying gas, aerosolized gas, and moisture removed from the plasma droplets. During drying of the plasma, the moist air passes through the air flow path through the lower filter 36, the lower filter separator 38, and exits through the gas outlet 30, leaving the dried plasma in the lower filter 36.

[0196] The disposable 100 further includes another alignment configuration relating the gas outlet 30 of the disposable 100 and the gas exhaust port 208 of the dryer 200. The spray dryer has a gas exhaust port 208 for exhausting the drying gas, and the bottom of the disposable 100 has a gas outlet 30 that mates with the exhaust port 208 of the dryer 200 (FIGS. 45B and 46A). Furthermore, the spray dryer finisher 400 has a receiver 414 for the drying gas outlet 30 to secure the bottom of the disposable 100 to the finisher 400 (FIGS. 48B and 48C). Additionally, this drying gas configuration allows for common mounting of the disposable to both the dryer and the finisher.

[0197] Furthermore, the overall length of the disposable (as measured from the spray drying head to the bottom of the drying chamber) is limited to about 40 inches or less (e.g., about 40, about 39, about 38, about 37, about 36, about 35, about 34, about 33, about 32, about 31, about 30, about 29, about 28, about 27, about 26, about 25, or about 24 inches or less), preferably about 34.8 inches. Disposables having lengths of about 40 inches or less have been difficult to achieve because drying of the plasma occurs in a smaller space and smaller volume, yet is gentler and without degrading plasma proteins. The length of the disposable, as measured from the bottom of the spray drying head 2 or the bottom of the baffle plate 8 to the bottom of the filter 36, as shown as dimension Y in Figure 46A, is about 31 inches or less (e.g., about 31, about 30, about 29, about 28, about 27, about 26, about 25, about 24, about 23, about 22, about 21, about 20, about 19 inches or less), and in one embodiment, is preferably about 25.9 inches. In another aspect, the area of ​​the disposable 100 encompassed by dimension Z, i.e., the length from the bottom of the spray drying head 2 to the top of the filter 36, is about 22 inches or less (e.g., about 22, about 21, about 20, about 19, about 18, about 17, about 16, about 15, about 14 inches), and preferably about 19.11 inches. In yet another aspect, the length of dimension X, i.e., the length between the bottom and top 46 of the spray-drying head 2, is less than about 16 inches (e.g., about 16, about 15, about 14, about 13, about 12, about 11, about 10, about 9, or about 8 inches), and preferably about 12.14 inches. In one embodiment, the length of a disposable can be varied or shortened. For example, the length of a 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, a disposable can be shortened anywhere along dimension Y and dimension Z by the same amount.

[0198] Computational model For some of the figures, computational models were used to show the flow paths, particle evaporation, etc. Figure 42B shows the three-dimensional fluid geometry of the disposable in operation used in the model.

[0199] The three-dimensional model shown in Figure 42B is based on the disposable shown in Figure 42A and the dryer shown in Figures 45A-45C. These computer simulations demonstrate that the flow and mixing processes were generated by first constructing the geometry of the three-dimensional fluid domain. See Figure 42B. This geometry was extracted from a computer-aided design (CAD) model of the system hardware to refine a high-fidelity representation of the fluid domain inside the ODP system. Volume: 0.0195 m 3 The fluid domain was discretized into 3.3M spatial cells, and the computational grid was generated using the commercially available Ansys-Gambit grid mesh generation software. The fluid model was calculated using a commercially available computer code, Ansys-Fluent version 2019-R1, running on an HPZ840 multiprocessor workstation.

[0200] The simulation uses ideal gas properties, the KE turbulence model and the following: Drying gas inlet temperature = 114°C Drying chamber exhaust temperature = 65℃ System heat loss = 0.18kW Drying gas flow = 750 slpm Atomizer aerosol gas flow = 40 slpm Supply rate = 13.5 mL / min and varies depending on exhaust gas temperature Liquid water droplets with a non-volatile mass of 8.5% and a diameter of 5 microns (monodisperse size) A steady-state segregated solver was used, assuming an exhaust port pressure of 2.76 kPa (0.4 psig).

[0201] The inlet and product capture filters are modeled using a "porous zone" function with fluid resistance values ​​set to match the measured operating pressures in the drying chamber of 71.7 kPa (10.4 psig) and the drying gas manifold of 27.6 kPa (4 psig) at the start of the batch.

[0202] Two customized C programs, "prsc_udf_multi_2017.c" and "processdata_multi_2017.c," were developed at PARSEC to obtain averaged droplet drying paths from a converged Fluent-coupled DPM solution with the goal of calculating the average droplet diameter and temperature during a constant rate drying period for a given set of process conditions. The program "prsc_udf_multi_2017.c" is used to export droplet tracking data step-by-step for information of interest. The program "processdata_multi_2017.c" reads the data file exported from the first program and obtains the average path from all tracked particles. The output file can be imported into an Excel file.

[0203] The data shown in Figure 42B, Figure 43Ka, Figure 43Ma, Figure 43Na, Figure 43O, Figure 43P, Figure 43S, Figure 43Sa and Figure 43T were generated using this model.

[0204] Overview of spray dryers The 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, a disposable compressed air line, a disposable exhaust line, a housing exhaust line, and a leak detection line.

[0205] Along with these flow lines, the spray dryer 200 of the present invention further includes a leak detection method 1000 that utilizes a series of pressure transducers, flow sensors and valves to assess whether the disposable 100 has a leak during spray drying.

[0206] The spray dryer 200 further includes a pressure detection method 1200 that enables a pressure transducer present on the outside of the wall of the disposable 100 to measure the pressure inside the disposable 100. This process involves heating the wall of the disposable, allowing it to soften. During use, the wall of the disposable exerts a force on force sensors PT08 224A and PT09 224B, which measure such force. The amount of force is used to calculate the amount of pressure within the disposable. If the pressure within the disposable 100 exceeds a set amount (e.g., greater than 7.02 psi), the computer system causes the dryer 200 to enter fail-safe mode. If the pressure is within a set acceptable amount (e.g., between about 6.7 psi and less than about 7.02 psi), the computer system determines whether enough plasma has been dried by determining the amount of donor plasma remaining in the donor plasma bag. If a sufficient amount of plasma has been dried, the computer system communicates that the drying run was successful and the finishing process can begin. If an insufficient amount is dried, the drying run fails and the disposable 100 is discarded.

[0207] Additionally, the dryer 200 includes a method 1400 for determining the integrity of the bottom filter 36, the baffle filter 94, and / or the plenum interface. This methodology utilizes pressure transducers in the inlet and housing to determine the pressure gradient during spray drying. The gradient is compared to a model, and if the gradient deviates from the model, the computer system determines that the disposable 100 has failed.

[0208] Finishing machine overview Once spray drying is completed on dryer 200, a finisher 400 or 400' of the present invention moves the plasma to the desired compartment, then seals the disposable walls and cuts the disposable walls to form the dried plasma units. The finisher 400 or 400' provides an impactor, sealer, separator, and air extraction.

[0209] Workflow Overview A summary of the process using the disposables, spray dryer, and finisher described herein is as follows: The spray-drying plasma methodology of the present invention includes pre-treating a provided liquid plasma unit or a thawed previously frozen liquid plasma unit, drying the liquid plasma using a spray-drying apparatus equipped with a spray-drying disposable device to obtain a disposable with dried plasma, and finishing the disposable using a finishing apparatus designed to seal and separate the disposable, converting it into a dried plasma unit. The unit can be used or stored. Once ready for use, the plasma unit is rehydrated and ready for transfusion to a recipient.

[0210] Regarding pretreatment, the pretreatment process involves adding a biocompatible component (e.g., a spray-drying stable acidic substance) to liquid plasma (or thawed fresh frozen plasma) that protects plasma proteins during the spray-drying process, which involves high temperature and pressure.

[0211] In one embodiment, creating the pretreatment solution involves adding to a solvent such as SWFI 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 of HCl and about 15 to about 30 mmol of glycine (e.g., about 15, 17, 20, 22, 25, 27, and 30 mmol of glycine) in 50 mL of solvent to obtain 260 mL of formulated plasma, i.e., about 440 mM glycine and about 106 mM HCl are present in the pretreatment solution. In one embodiment, 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 are present in the pretreatment solution. Pretreatment vessels are commercially available and can be formulated, filled, and finished, for example, by Berkshire Sterile Manufacturing (Lee, Massachusetts, USA). In one embodiment, the pretreatment solution has about 440 mM / 50 ml glycine and 106 mM / 50 ml hydrochloric acid. (United States Pharmacopeial Convention ("USP") Monographs (12601 Twinbrook Parkway, Rockville, MD 20852-1790, USA). The pretreatment solution, when combined with liquid plasma to form formulated plasma, protects plasma proteins during the drying process. The formulated plasma has a pH in the range of about 5.5 to about 7.2, which offsets the effects of spray drying on pH, and is generally closer to normal physiological pH, about 6.5 to 7.8 (e.g., about 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4). The resulting rehydration product is in the pH range of 7.5, 7.6, 7.7, and 7.8 (7.5, 7.6, 7.7, and 7.8), although in some cases, a pH lower than 6.5 or higher than 7.8 may be harmful to the recipient. The dried plasma product obtained from this invention retains its von Willebrand factor and other blood proteins, and is plasma with low cholesterol crystals, low particulates, low pathogens, and well-controlled pH due to the pretreatment steps described above. Furthermore, the resulting dried plasma has certain properties that are distinct from and superior to freeze-dried plasma.

[0212] As known in the art, a sterile connection device (SCD) is used to connect the plasma unit to the pretreatment container and the liquid plasma; in one embodiment, a volume of plasma is transferred using, for example, a blood collection monitor / mixer. After the liquid plasma is transferred to the pretreatment container, in one embodiment, it is gently mixed in the pretreatment container by inversion. Other mixing methods, such as rocking, shaking, and stirring, can be used. Additionally, mixing can be performed by an operator or by devices known in the art. The bag containing the liquid plasma is tubing sealed, separated, and discarded. The pretreatment container 64 with the pretreatment solution and liquid plasma (i.e., formulated plasma 66) is then connected to the disposable device with the plasma tubing 16 using the SCD, resulting in the improved spray-drying disposable device shown in FIG. 42A.

[0213] The spray dryer disposable device 100 is a sterile, non-pyrogenic, single-user container (e.g., approximately 35 inches long) that utilizes a pathogen-retaining filter to filter the air before it enters the drying chamber and as it exits the drying chamber. See FIG. 42A. The spray dryer disposable device is sterilely connected to liquid plasma with plasma tubing, tubing 16.

[0214] Briefly, the drying process is as follows. See Figures 45A-4C, 46A-5B. In one embodiment, pre-treated plasma is aseptically spray-dried in a disposable spray dryer device 100. See Figures 45A-4C. During the process, in one embodiment, a positive air flow is maintained. The pre-treated plasma is atomized using a nozzle contained within the single-use disposable spray dryer device to generate 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 disposable spray dryer device is then undocked from the spray dryer and transported to a finishing device.

[0215] The finishing process is outlined below. See Figures 47A-47C, 48A-48C, and 34. Once undocked from the spray-drying apparatus 200, the disposable device 100 with dried plasma is transferred to the finishing apparatus 400. The finishing apparatus 400 mechanically, acoustically, or otherwise impacts or agitates the spray-drying disposable device containing dried plasma to consolidate the dried plasma powder into what will eventually become the spray-dried plasma unit of the spray-drying disposable device. The finishing apparatus utilizes an impactor to assist the dried plasma in moving first to the bottom of the disposable and then to the compartment that will become the spray-dried plasma unit 60 in the second example. The spray-dried plasma unit 60 is sealed and separated from the rest of the disposable device using an impulse seal. This is the final closure step to produce the plasma unit 60. In one embodiment, the seal is visually inspected, the excess portion of the disposable device is discarded, and the dried plasma unit 60 is produced (see Figure 49).

[0216] For an overview of the dried plasma storage process, dried plasma units 60 are removed from the finishing equipment and stored in resealable, moisture-barrier foil pouches containing desiccant. See Patent No. 9,561,184. In one embodiment, the dried plasma units are quarantined and stored refrigerated until all required blood screening tests are completed. Once final release criteria are met, the pouches are opened and the dried plasma units are relabeled for release. The dried plasma units are then placed in resealable or other pouches, sealed, and stored according to the storage protocol.

[0217] In one embodiment, the dried plasma unit is compatible with commercially available fluids or other transfer sets for rehydration with sterile water for injection (SWFI). Once rehydrated, the dried plasma unit 60 is also compatible with blood administration sets for transfusion. In certain embodiments, the spray-dried plasma unit 60 is rehydrated in its existing container using an appropriate amount of sterile water for injection (e.g., 200 mL, 208 mL) prior to transfusion.

[0218] Detailed explanation of spray dried disposables Detailed description of the spray drying head 42A, there is shown a perspective view of a spray drying disposable device 100. As noted above, the disposable generally has two parts: a spray drying head 2 and a drying chamber 28. The spray drying head includes a plenum 6, a guide 4, a baffle plate 8, a baffle filter 94, a nozzle 20, and a locator notch 26 (also referred to herein as a "second locator").

[0219] In one aspect, the purpose of spray drying head 2 is, in part, to A) assist in securing disposable 100 to dryer 200, B) regulate the flow of drying air, aerosolized pressurized gas, and plasma flow, C) house the nozzle assembly, and D) house the baffle filter.

[0220] With regard to securing the disposable 100 to the dryer 200, the system of the present invention includes an integrated universal alignment system. In one embodiment, the locator notch 26 on the plenum 6 is shown in FIG. 43A. FIG. 43A also better illustrates the plenum 6, guide 4, baffle plate 8, and ridge 9. The locator notch 26, also referred to as the second locator, aligns with the locator protrusion 206, also referred to as the first locator (shown in FIGS. 43K, 45B, 45C, and 46A), on the spray dryer 200. This arrangement allows the spray dryer head 2 of the disposable 100 to be coaxially aligned with the spray dryer 200. The arrangement can include any arrangement that attaches, mates with, complements, or otherwise communicates with a locator on the dryer to a locator on the disposable. Examples of alignment configurations may include recess / protrusion configurations, complementary configurations, hook / receptacle configurations, channel / groove configurations, latch and catch configurations, magnetic configurations, etc. In FIG. 46A, a male locator is on the spray dryer and a complementary female locator is on the disposable, but the configurations can be reversed. The complementary nature of the configurations allows for easy matching and alignment by the operator and prevents the door from closing unless the disposable is aligned with the spray dryer. In one embodiment, the alignment configuration allows for alignment of the locator on the disposable with the locator on the dryer and can include any configuration that allows for alignment of the disposable with the finisher. In another embodiment, the dryer and finisher have identical locators that mate with the locators on the disposable to create a common alignment. Having a universal configuration reduces training requirements and increases muscle memory because operators will insert disposables into the spray dryer and finisher in a similar manner.

[0221] In one embodiment, the system of the present invention provides positive feedback to the operator when the first locator of the spray dryer and the second locator of the disposable are aligned. In one embodiment, the spray dryer 200 has a spring clip 232 on the top of the drying chamber housing that engages with guide 4 when the disposable is aligned and secured to the spray dryer. See FIG. 46A. The 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 that provides communication to a display indicating alignment). The retaining clip 232 is also an alignment element because it aligns with ridge 9, as described further below.

[0222] FIG. 43A also shows the guide 4 being offset from the center of the baffle plate 8. The offset design of the guide on the plenum 6 allows the disposable 100 to be attached to the receiver 204 (shown in FIGS. 45B and 46A) of the spray-drying machine 200 in a specific orientation. Before inserting the disposable device into the dryer, the operator removes and discards the adhesive covers, if present, from the top exposing the drying gas inlet port 22 and the bottom exposing the gas outlet 30. The use of these covers is optional. In a preferred embodiment, the operator removes and discards the adhesive cover from only the drying gas inlet port 22 and inserts the spray-drying head 2 into the spray-drying head receiver 404. The cover for the drying gas outlet 30 on 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 typically aligns the ridge 9 formed by the baffle plate 8 on the spray-drying head 2 of the disposable 100 and inserts it into the groove 207 of the spray dryer 200. See FIG. 46A. Once engaged, the operator uses their hand to push the spray-drying head 2 further inward, aligning it with the groove receiver 207 as long as the notch locator 26 on the spray-drying head 2 is within about 30 degrees (e.g., about 30 degrees, about 25 degrees, about 20 degrees, about 15 degrees, about 10 degrees, about 5 degrees) of alignment with the protrusion locator 206 on the dryer 200. Insertion and alignment of the spray-drying head can be accomplished quickly, for example, within 10 seconds (2-5 seconds). The receiver 204 of the guide 4 also functions as a drying gas inlet on the spray dryer and provides a drying gas source (not shown). The ridges 9 of the spray drying head 2 also support and complement the grooves 207 of the receiver 210, allowing the spray drying head 2 of the disposable 100 to be aligned laterally relative to the dryer 200.

[0223] In one embodiment, the receiver 210 has a groove 207 as shown in Figure 45B. The ridges and grooves between the spray drying head and the dryer can be any configuration that provides support and lateral alignment for fitting the spray drying head within the drying chamber housing 202. In addition to the groove 207, the receiver can be a ledge, ledge, arm, stop, base, or other structure that can engage with a baffle plate to stabilize the spray drying head throughout the spray drying process.

[0224] The operator then inserts the disposable device into the receptacle 204 of the spray drying apparatus 200 by positioning the guide 4. Once inserted and aligned, the spray drying disposable cannot move up or down any further. When using this guide and the above arrangement, the disposable is aligned so that it cannot move up or down, but rather coaxially around the axis defined by the center of the guide 4. The guide is then fitted into the receptacle 204, as shown in Figures 45B and 46A, so that the fit is snug or tight. In this embodiment, with the spray drying end aligned and in the engaged position, the operator can remove the bottom adhesive cover at the drying chamber gas outlet 30 and attach it to the gas exhaust port 208, as described further herein.

[0225] When the arrangement (locators 26 and 206) is aligned, guide 4 is inserted into receiver 204, ridge 9 is inserted into groove receiver 207, and retaining clip 232 is engaged, so that, in one embodiment, the spray drying head is inserted, secured, and aligned. Specifically, in one embodiment, retaining clip 232 engages with ridge 9 and holds spray drying head 2 in place. Retaining clip 232 provides an audible indicator that the spray drying head is properly aligned and inserted. In the embodiment shown in FIGS. 45B, 45C, and 31A, the retaining clip is a spring clip. The retaining clip can be any type of retainer that engages with ridge 9, including, for example, a snap, pin, clasp, slide, etc. The retainer can be made of metal, plastic, rubber, etc. The retainer that engages the spray drying head is optional.

[0226] In the embodiment shown in Figures 45B, 45C, and 46A, a spring clip is used as an audible indicator to allow the operator to know that the spray drying head 2 of the disposable 100 is properly inserted and aligned with the dryer 200, although any type of indicator can be provided. The indicator can be audible, visual, or tactile. In one embodiment, an audio indicator provides audible feedback that the loading of the spray drying head 2 of the disposable 100 has been completed correctly, mechanically or otherwise. The audible indicator can be mechanical, such as a sound that the spring clip has locked into place, or it can be generated by a sensor (mechanical or pressure / contact sensor) in communication with an actuator or processor that receives a signal of correct positioning of the spray drying head 2 of the dryer 200 and provides a visual indicator to the operator, for example, on the display 212 or on the indicator light 234. In an alternative embodiment, the sensor sends feedback to the processor, which provides an audio indicator to a speaker to notify the operator of proper alignment. In yet another embodiment, the feedback can be in the form of a tactile response, such as a vibration to alert the operator to incorrect or correct placement. The feedback can include an audible indicator (e.g., an audible click) or a visual indicator (e.g., a sensor providing a communication to a display indicating alignment).

[0227] 43B is an exploded view of the spray drying head 2, showing a portion of the spray drying nozzle assembly 20 along with the plenum 6, outer filter seal ring 90, inner filter seal ring 92, plenum filter 94, and baffle plate 8 having ridges 9. From the top to the bottom of FIG. 43B, the nozzle assembly 20 includes a strain relief valve 75, a plasma and pressurized aerosol gas manifold 72, an aerosol reservoir 74, a cannula 78 having an opening 79, a liquid nozzle cap insert 80, and a nozzle cap 76. The nozzle cap 76 has an inner wall having a diameter D o43Ia also has an opening 110 with a diameter defined by a diameter D c 1 shows a cannula 78 having an outer wall (outer diameter) defined by a diameter D o is slightly larger than the diameter Dc, and the difference is the distance D d The resulting difference in diameter is the distance D d forms an annulus 81 in which pressurized air received from aerosol reservoir housing 74 forms a vortex, promoting the formation of small droplets of fluid that flow into drying chamber 28 and are dried. See Example 27.

[0228] Thus, the cannula length is about 2 to about 5 inches, and in one embodiment, 3.500 inches + / - 0.005 inches.

[0229] 43A and 43B, the spray-drying nozzle assembly 20 has a plasma flow inlet 18 (shown in FIG. 42A) connected to pre-treated liquid plasma 66 via plasma tubing 16, and a pressurized aerosol gas inlet 14 connected to a pressurized gas source (not shown) via aerosol tubing 10 and aerosol filter 12. Additionally, a 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 are combined, liquid plasma particles are formed under the pressurized (aerosolized) gas and dried into finely divided dried plasma (e.g., plasma powder).

[0230] Figures 43C and 43D show detailed perspective views of the spray drying nozzle assembly 20. In particular, Figures 43B and 43J show where and how the spray drying nozzle assembly fits within assembly opening 96 of plenum 6 of spray drying head 2. Plasma and pressurized aerosol gas manifold 72 conditions and directs the plasma source via inlet 18 and the pressurized aerosol gas source via inlet 14. Strain relief 75 fits into and communicates with manifold 72 to support tubes 10 and 16 and prevent them from collapsing under pressure during packaging, shipping, and spray drying. Strain relief 75 also prevents the tubes from collapsing during packaging and shipping. The spray-drying nozzle assembly 20 includes an aerosol gas reservoir housing 74 in which pressurized aerosol gas is held and accumulated before being released through a liquid nozzle cap insert 80 and a nozzle cap opening 110 (shown in FIGS. 43G, 43H, 43I, 43Ia, and 43Ic). The nozzle assembly 20 is housed by the aerosol gas reservoir housing 74 and secured by a nozzle cap 76. The liquid nozzle cap insert 80 guides the cannula 78 and holds it in place during use. An annulus 81 is disposed between the outer surface of the cannula 78 and the inner surface of the opening 110. The design of the liquid nozzle cap insert 80 and the nozzle cap 76 allows the pressurized aerosol gas to flow through the annulus 81 in a vortex pattern, maximizing aerosolization and promoting rapid mixing of the aerosolized plasma droplets with the drying gas, as described further herein. The entire nozzle assembly 20 is secured to an opening 96 in the plenum 6, which includes a baffle plate 8 having ridges 9, with a filter 94 therebetween, sealed by an inner filter seal ring 92 and an outer filter seal ring 90. See Figure 43B.

[0231] Figure 43D shows the aerosol gas reservoir housing 74 as transparent so that the attachment of the cannula 78 and the liquid nozzle cap insert 80 and nozzle cap 76 can be seen, and Figure 43E shows the manifold 72 and cannula 78 with the aerosol gas reservoir housing 74, liquid nozzle cap insert 80, and nozzle cap 76 removed. Figure 43F shows the bottom tip of the cannula 78, the end facing the manifold, with an outer wall surface 84, an inner wall surface 86, a flat edge 88, and a beveled or angled edge 82 (e.g., a chamfer) on the bottom surface of the cannula.

[0232] It has been discovered that cannulas having beveled edges (e.g., chamfers) on the inner diameter, when used in spray drying to produce atomized plasma particles, help or allow many proteins in the plasma to remain intact, functional, or both. Thus, the beveled edge cannulas of the present invention reduce the amount of protein degradation during spray drying because the beveled edge cannula reduces shear on the liquid plasma film.

[0233] In a specific embodiment, the blood protein, vWF, was measured. vWF is a more fragile and easily degraded protein, as further described herein. In one embodiment, vFW retention is maintained when using a spray-drying nozzle with a beveled cannula of the present invention compared to a nozzle with a non-beveled cannula. Indeed, based on the data described in Example 28, the use of a composite nozzle with a beveled cannula resulted in increased vFW retention compared to a composite nozzle with a non-beveled cannula and a reference stainless steel nozzle (Buchi Model no. 4244 Buchi Corporation of New Castle, Delaware, United States). In one embodiment, the use of a nozzle with a beveled cannula resulted in an increase in functional vFW retention by an amount ranging from at least about 1% to about 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%) compared to a nozzle with a non-beveled cannula. In particular, as described in Example 28, the data show that compared to the same system operated with a composite nozzle having a cannula that did not include a beveled edge, spray drying with a beveled cannula having a 45-degree angle and a length of 0.005 inches increased vWF RCO assay results by about 9% to 22%, a surprisingly favorable increase of 3.7% compared to the reference Buchi nozzle.

[0234] Plasma proteins that are preserved throughout the beveled-edge cannula spray-drying process include von Willebrand factor (vWF), which is involved in coagulation, repair of vascular injury, and platelet adhesion. In particular, vWF is a large adhesive glycoprotein with established functions in hemostasis. It functions as a carrier of factor VIII and acts as a vascular injury sensor by attracting platelets to sites of vascular injury. Regulation of vWF multimer size and platelet tethering function is mediated by ADAMTS13, a constitutively active plasma metalloprotease. It is secreted into the blood and degrades large vWF multimers, reducing their activity. Unusually, the protease activity of ADAMTS13 is not controlled by a natural inhibitor but by a conformational change in its matrix induced when vWF is subjected to elevated rheological shear forces. This results in the conversion of vWF from a globular to an elongated protein. This conformational change unfolds the vWF A2 domain, exposing cryptic exosites and scissile bonds. To enable vWF proteolysis, ADAMTS13 creates multiple interactions that, when exposed by shear forces, guide the protease to the substrate and position it to engage the cleavage site. ADAMTS13 (a disintegrin and metalloproteinase with thrombospondin type 1 motifs, member 13), also known as von Willebrand factor-cleaving protease (vWFCP), is a zinc-containing metalloprotease enzyme.

[0235] Without being bound by theory of operation, it is believed that during spray drying, plasma proteins are subjected to substantial shear forces due to the atomization mechanism as the solution fluidizes from the end of a fine nozzle to form droplets that come into contact with the drying air. The process of unfolding multimeric vWF is expected to be driven by the hydrodynamic forces of high shear stress during spray drying combined with air-liquid interfacial stress. Shear-induced structural changes in vWF, when combined with other physical factors associated with spray drying, such as high temperature and / or unfavorable pH, and air-liquid interfacial stress, can lead to protein denaturation (if denatured vWF cannot properly refold after spray drying) and proteolysis (if denatured vWF exposes proteolytic sites of ADMATS13), potentially damaging vWF activity and other proteins in the spray-dried plasma.

[0236] The spray drying system of the present invention is optimized to reduce protein damage caused by shear forces and temperature, and the specially designed cannula of the present invention helps minimize shear and damage to proteins, including vWF.

[0237] In one embodiment, the cannula of the present invention has a bottom edge where at least a portion of the bottom edge is beveled, also referred to as a beveled edge cannula. In one example, the entire bottom edge can be beveled, or a portion of the bottom edge can be a flat edge (e.g., at an angle of about 90° from the outer or inner wall surface). In another embodiment, a portion of the cannula's bottom edge is a flat edge (e.g., at an angle of about 90° from the outer or inner wall surface), such as flat edge 88, as shown in FIG. 43F, and a portion of the cannula's bottom edge is beveled (e.g., at an angle of 45° from the outer wall surface or an angle of 135° from the inner wall surface), such as beveled edge 82. This embodiment shown in FIG. 43F can have a flat edge (90° from the outer wall) that is angled at a 45° angle, and is referred to as having a "chamfer" or "beveled edge."

[0238] When the cannula has a bottom edge and the entire bottom edge is sloped from the outer wall to the inner wall, the angle measured from the outer wall surface is in the range of about 30° to about 60° (e.g., about 30°, about 35°, about 40°, about 45°, about 50°, about 55°, about 60°), and the angle measured from the inner wall surface is in the range of about 120° to about 150° (e.g., about 120°, about 125°, about 130°, about 135°, about 140°, about 145°, about 150°). The length of the beveled bottom edge ranges from 0.001 inches to about 0.010 inches (e.g., about 0.001, about 0.002, about 0.003, about 0.004, about 0.005, about 0.006, about 0.007, about 0.007, about 0.008, about 0.009, about 0.010 inches).

[0239] When the cannula has a bottom edge that is flat and beveled, the flat edge is at an angle of about 90° (e.g., about 85% to about 95%) from the outer wall surface. The beveled edge, when measured from the outer wall surface (assuming the beveled edge crosses the outer wall surface), is at an angle ranging from about 30° to about 60° (e.g., about 30°, about 35°, about 40°, about 45°, about 50°, about 55°, about 60°), and in one embodiment, is 45° + / - 5°. When measured from the inner wall surface, the beveled edge is at an angle ranging from about 120° to about 150° (e.g., about 120°, about 125°, about 130°, about 135°, about 140°, about 145°, about 150°), and in one embodiment, is 135° + / - 5°. See, for example, Figure 52 for 45° and 30° beveled edges. The length of the flat edge portion ranges from 0.001 inch to about 0.009 inch (e.g., about 0.001, about 0.002, about 0.003, about 0.004, about 0.005, about 0.006, about 0.007, about 0.008, about 0.009 inch), and the length of the beveled edge portion ranges from about 0.001 inch to about 0.009 inch (e.g., about 0.001, about 0.002, about 0.003, about 0.004, about 0.005, about 0.006, about 0.007, about 0.007, about 0.008, about 0.009 inch), and in one embodiment, is 0.005 + / - 0.003. The ratio between the length of the flat edge and the length of the beveled edge ranges from about 5 percent to about 500 percent. In one embodiment, the flat edge is adjacent to the outer wall surface and the beveled edge is adjacent to the inner wall surface.

[0240] Compared to a non-beveled cannula with a 90° angle, a beveled edge cannula with or without a flat edge results in less stress / shear on the plasma droplets exiting the cannula. Without being bound by any particular theory, it is believed that when a plasma droplet exits the edge of a non-beveled 90° cannula, a portion of the plasma droplet or plasma membrane is subjected to a shearing effect, which in the process degrades a high percentage of the plasma proteins within. In this case, a non-beveled 90° cannula exerts a shear force on the droplet, thereby degrading the proteins in the plasma. As in the present invention, when a plasma droplet exits a cannula with a beveled edge, the plasma droplet experiences little shear. As the plasma is drawn by the airflow of the beveled edge cannula, it accelerates based on the plasma supply, causing the plasma to be pulled around the cannula edge. Unlike a cannula with a non-beveled 90° edge, the plasma is not rotated 90 degrees. Beveling the edges of the cannula reduces the rotation of the plasma as it exits the cannula, which results in less shear on the liquid film as the plasma is withdrawn. The film of liquid plasma exiting a beveled cannula is thicker and accelerates more slowly, reducing the shear on the liquid.

[0241] The inner diameter of the cannula ranges from about 0.010 inches to about 0.040 inches, and in one embodiment, is 0.030 inches + / - 0.002 inches. The outer diameter ranges from about 0.030 inches to about 0.060 inches, and in one embodiment, is 0.050 inches + / - 0.0005 inches. The beveled edge of the cannula affects the size of the atomized droplets. Upon exiting the beveled cannula, droplets in this range range in size from about 5 microns to about 35 microns, and in one embodiment, the droplet size is about 10 microns. The small droplet size, defined in part by the beveled edge of the cannula, promotes rapid mixing, faster evaporation, and reduced drying time. See Figure 43T, which shows that the larger the droplet size, the longer it takes to evaporate the droplets using a hotter drying gas. The droplet shape is driven by its surface tension, which dominates, producing a sphere after exiting the cannula. Droplet size is also primarily influenced by the pressurized gas velocity as a ratio to the liquid rate of delivery (ALR) and the nozzle design.

[0242] The cannula of the present invention can be made from stainless steel materials suitable for medical devices. Examples of stainless steel grades that can be used include grades 304 and 316 stainless steel. The stainless steel used in the cannula of the present invention is commercially available, for example, from Bergsen Metals (Santa Fe Springs, California, USA) or Fort Wayne Metals (Fort Wayne, Indiana, USA). The nozzle assembly (excluding the cannula), nozzle insert, nozzle cap, plenum and baffle plate, outer filtering, inner filtering, etc. can be made from plastics used in medical devices, such as polycarbonate, polypropylene, polysulfone, or combinations thereof. Each of the above parts can be made from the same material, different materials, or combinations thereof. Such plastics are commercially available and can be purchased from, for example, 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 can be molded from, for example, Egli Machine Co. (Sidney, NY, USA) and Southwest Mold, Inc. (Tempe, AZ, USA). Other materials now known or later developed can be used for the cannula and / or nozzle, so long as, when combined, they result in increased control or vWF retention in the spray-dried plasma.

[0243] Stainless steel nozzles, such as the Buchi Model no. 4244 (Buchi Corporation of New Castle, Delaware, United States), are often used in spray drying, but they are expensive to manufacture or purchase, especially for disposable devices that are discarded after each spray drying run. For example, a typical Buchi stainless steel nozzle body (part no. 4244) costs between $1,000 and $2,000. The nozzle assembly of the present invention is a composite nozzle for use in spray drying, particularly spray drying of delicate materials such as human plasma, costing 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 composite nozzles, as it is used by the applicant to produce dried human plasma that preserves plasma proteins at controllably acceptable levels.

[0244] As shown, most of the nozzle assembly, except for the cannula, which is made from stainless steel, is made from less expensive plastic materials, as described above. Therefore, the nozzle assembly is also referred to as a "composite nozzle" or "composite nozzle assembly," which refers to two or more different types of materials used to make the nozzle assembly (e.g., the stainless steel cannula and the polycarbonate nozzle insert and nozzle cap). Example 28 shows that the beveled cannula of the composite nozzle assembly has improved vWF retention compared to those with non-beveled cannulas, and provides vWF retention that is as good as that of expensive stainless steel nozzles.

[0245] As the plasma exits the tip of the cannula, it is exposed to the pressurized aerosol gas in the nozzle cap 76. More specifically, the pressurized aerosol gas exits in a vortex pattern through the annulus 81 and impinges on the liquid plasma droplets flowing from the chamfered / beveled edge 82 of the cannula 78, atomizing the plasma into a plume. As the atomized plasma exits the spray-drying nozzle assembly, it is exposed to the drying gas and is dried into plasma powder in the drying chamber. In one embodiment, the tip of the cannula 78 is flush with the distal end of the opening 110.

[0246] The liquid nozzle cap insert 80 secures the bottom of the cannula 78 and directs the pressurized aerosol gas flow. FIG. 43G shows a perspective top view of the liquid nozzle cap insert 80. As can be seen, the cap insert 80 has an inner wall 116 and a cannula anchor 120 with an opening (not shown) through which the cannula 78 extends. The top of the inner wall 116 forms a ridge, and the side of the inner wall 116 defines a series of recesses 118 and protrusions 119. The cannula anchor 120 supports the tip of the cannula 78 during plasma flow. The beveled edge 82 of the cannula 78 from which the plasma exits and the annular portion 81 from which the pressurized aerosol gas is released form the actual nozzle. The cannula anchor 120 is a hollow, cylindrical substrate, but can be any shape as long as the cannula is supported and its position is maintained during spray drying. A recess 118 in the wall allows pressurized air to pass from the reservoir (defined as reservoir housing 74) through the region between nozzle cap insert 80 and nozzle cap 76 before exiting central opening 110 in nozzle cap 76. As the pressurized air exits central cap opening 110, it exits an annulus 81 defined by the outer wall of cannula 78 and the inner wall of opening 110. More specifically, when cap insert 80 is secured to cap 76 and cannula 78 is present within opening 110, the pressurized air exits through annulus 81. See FIG. 43Ia. Thus, the diameter of opening 110 is larger than the outer diameter of cannula 78. Specifically, opening 110 in nozzle cap 76 has a diameter D o(See FIG. 43Ia.) FIG. 43Ia also shows the inner wall having a diameter D c 1 shows a cannula 78 having an outer wall defined by a diameter D o is the diameter D c is slightly larger than the distance D d The resulting difference in diameter is the distance D d The cannula 78 has an outer diameter, D c is about 0.030 to about 0.070 inches (e.g., 0.030, 0.040, 0.050, 0.060, 0.070), and the diameter of the opening 110 is 0.075 to 0.100. d " is also referred to as "the radial distance of the annular portion 81." In one embodiment, the diameter D c is 0.050+ / -0.0005 inches, and the diameter D o is 0.082 + / - 0.001 inches. The distance D, which is the radial distance between the outer surface of the cannula 78 and the inner surface of the opening 110 d is a space where a rotating vortex of pressurized aerosol gas flows and assists in the generation of small droplets of plasma that are mixed with the hot drying gas during spray drying. d has a range of 0.005 to 0.030 inches (e.g., 0.015 inches and 0.021 inches).

[0247] Along these lines, the data from Example 27 shows that the radial distance of the annulus 81, D d It has been shown that the drying process affects both the yield of the dry product and the preservation of vWF. The yield is the ratio of the starting solids (weight) in the liquid material being dried to the dry material (weight) recovered by the drying process.

[0248] Prior to the present invention, one cause of yield loss occurred when dried spray material that was not completely dried and retained more than about 2.5% residual moisture came into contact with and adhered to the internal structure of the dried disposable during drying, making it impossible to recover.

[0249] Example 27 demonstrates that there was a reduction in the amount of material visibly adhering to the underside of the baffle plate 8 after the drying cycle was completed. The data presented in Example 27 demonstrates that the overall yield (weight) was 1.0 times lower than that of the annular portion 81D. d The graph shows that the yield increased by changing the radial distance of the annulus from 0.021 inches to 0.015 inches. The yield with the 0.021 inch annulus size was acceptable. However, by decreasing the annulus width / diameter to 0.015 inches, the yield percentage improved by over 2.2%. Other features of the disposable 100 that increase yield include, in part, the drying jets 142 that form an air wall inside the plasma drying chamber 28, as further described herein.

[0250] Example 27 also describes an increase in vWF retention as measured by a Ristocetan (RCO) assay by changing the radial distance of the annular 81 from 0.021 inches to 0.015 inches. The vWF retention with the annular dimension of 0.091 inches was acceptable. However, by decreasing the annular width to 0.082 inches, the vWF retention increased by more than 2.0%. Other features of the disposable 100 also increase vWF retention, including, in part, the beveled edge cannula 78, as described herein.

[0251] The space between the nozzle cap insert 80 and the nozzle cap 76 before exiting the central opening 110 in the nozzle cap 76, also commonly referred to as the "vortex generator," contains a series of channels and curved pads, as described further below. Pressurized air passes through recesses 118, which act as openings in the nozzle assembly 20, allowing the air to enter and travel down the channels and between the curved pads. See Figures 43G, 43H, 43I, and 43Ic.

[0252] Referring to FIG. 43H, the bottom surface of the cap insert 80, unoccupied by the pads 122 on the underside of the protrusions 119 / recesses 118, serves as the walls of the vortex generator channels. The pads 122 form a kidney-like shape to aid in the placement of the recesses 118 for the vortex airflow pattern. The nozzle cap shown in FIG. 43I, cap 76, has a complementary receiving portion 112 for receiving the pads from the cap insert 80. The complementary fit between the nozzle insert 80 and nozzle cap 76 is shown (in cross section) in FIG. 43Ic. The nozzle cap 76 also has a nozzle cap channel 106 extending from the spherical head 108 and terminating in an opening 110. The bottom surface of the cap insert 80 not occupied by the pads 122 and further complemented by the channel 106 of the cap 76, is the space through which pressurized air flows. The vortex generator includes the bottom surface of the cap insert 80 not occupied by the recess 118 and pad 122, the spherical head 108, and the channel 106 in the cap 76, the shapes and positions of which cause the pressurized air to form a vortex airflow pattern. The spherical head 108 receives the pressurized airflow through the recess 118, and the curved ramp-like surface of the channel 106 provides a curved boundary for the flowing air. In other words, the recess 118 is the inlet port, providing the airflow, and the channel 106 provides the vortex. The channel 106 is arched, further enhancing the curved airflow and directing the tangential airflow toward the opening 110 where the cannula 78 resides. These channels, i.e., the channel 106, guide the air in a circular motion from the spherical head 108 to the opening 110 in the nozzle cap, all working in concert to discharge the pressurized air as a vortex through the opening 110. The design provides tangential momentum to provide efficient vortex generation. The channels 106 are in the form of an arc or curve, with a radius of curvature ranging from about 0.10 inches to about 0.25 inches, and in one embodiment, a radius of about 0.140 + / - 0.10 inches. The generated vortex includes four channels, but may have about 2 to 12 channels (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 channels). Other types of shaped channels may be used.FIG. 43I shows the designs molded into the nozzle cap 76. Designs A and B show more linear channels, but Design A does not have a portion equivalent to a spherical head, and Design B shows a bell-shaped head. Design C is similar to the design shown in FIG. 43I, but has lobes at the ends instead of spherical shapes. The present invention includes nozzle caps with Designs A and B, but it has been found that 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 vortices in the annulus. Other types of channels include conical channels, including diverging or diverging conical shapes.

[0253] Once the vortex is generated, the pressure and velocity flow patterns are shown in Figures 43O and 43P. Figure 43O shows the static gas pressure (psig) at the top and the tangential velocity profile (m / s) for the pressurized aerosol gas at the bottom. As can be seen, there is an inverse relationship between pressure and velocity. In regions where pressure increases, velocity decreases, and vice versa. In particular, at the spherical head 108 where the pressurized gas enters the vortex generator, there is a higher relative static gas pressure (e.g., 2.54 x 10 1 psig) and a relatively low velocity flow rate (e.g., about 2.00 x 10 1 Conversely, in annulus 81, there is a relatively low or negative gas pressure (e.g., about -2.24 psig) and a higher velocity (e.g., about -1.58 x 10 2 ~Approx.-3.75×10 2 m / s). As the pressurized aerosol gas travels along the curved nozzle cap channel 106, the pressure and velocity are in the ratio between them. Thus, the vortex generator of the present invention generates a vortex flow of approximately 2.54×10 1 psig to approximately -2.24 psig gas pressure and approximately 2.00 x 10 1 m / s~approx.-3.75×10 243P shows a more detailed view of the velocity pattern that occurs in annulus 81. The pressurized gas travels between the outlets of channel 106 as the gas becomes entrained in vortices. The pressurized gas accelerates as it enters annulus 81, forming vortices.

[0254] The vortex generator in this embodiment includes a curved pad / ramp, a spherical head for receiving the pressurized airflow, and a curved channel extending to the exit opening. The present invention may include other vortex generator elements, such as wings, edges, wedges, vanes, etc. Other shaped channels may also be utilized to create the vortex generator. One skilled in the art may utilize other vortex generators present within the insert and cap of the nozzle assembly of the present invention, as long as the pressurized air in the vortex exits between the inner surface of the opening 110 and the outer surface of the cannula 78.

[0255] Pressurized air circulates between the exterior surface of cannula 78 and the interior surface of opening 110. Specifically, the pressurized gas exits through annulus 81. Plasma is pumped through cannula 78 by peristaltic pump 214 at approximately room temperature. The plasma travels down the interior of stainless steel cannula 78 and is drawn out of cannula 78 by the pressurized aerosol air stream exiting annulus 81. The high-velocity aerosolizing air stream atomizes the droplets. In one embodiment, the steady-state plasma delivery rate is about 6 to about 23 (e.g., 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, or 23) mL / min. In a preferred embodiment, 13.5 mL / min is the steady-state delivery rate after the system has warmed to thermal equilibrium.

[0256] In one embodiment, the plasma supply rate is related to or dependent on the outlet temperature. This is done in a closed loop. As the outlet temperature decreases, the system adjusts to decrease the plasma supply rate. Conversely, as the outlet temperature increases, the system increases the plasma supply rate. The outlet temperature may decrease, for example, as the spray dryer warms up or as time passes between spray drying runs. In particular, in one embodiment, the plasma supply rate may be adjusted as follows:

[0257] [Table 10]

[0258] The chart values ​​are calculated based on thermodynamic principles and assume constant system heat loss, complete droplet evaporation, and a dryer outlet relative humidity of less than 11.8%.

[0259] Thus, when the outlet temperature is decreased, the plasma feed rate is decreased to maintain the target drying chamber outlet temperature required to dry the plasma to less than 2% residual moisture. At higher outlet temperatures within the range, the plasma feed rate can also be increased, and a plasma residual moisture content of less than 2% can still be maintained only if the total gas flow is increased and / or the drying chamber outlet temperature is increased to maintain the target system relative humidity.

[0260] In one embodiment, the closed loop comprises an exhaust temperature in the range of about 62° C. to about 68° C. and a plasma delivery rate in the range of about 6 to about 23 mL / min.

[0261] Although negative pressure exists directly below the cannula, the pressurized gas flow is high velocity. Generally, velocity increases along paths of decreasing static pressure. The pressurized aerosol gas travels through a series of channels 106, atomizing the plasma droplets and forming vortices that guide the initial droplet trajectories. The aerosol flow rate is about 20 slpm to about 60 slpm (e.g., about 20, 25, 30, 35, 40, 45, 50, 55, 60 slpm), and in one embodiment, about 40 slpm. This occurs at pressures of about 180 kPa to about 260 kPa (e.g., about 180, 190, 200, 210, 220, 230, 240, 250, 260 kPa), and in one embodiment, about 227.5 kPa (33 psig). The aerosol flow acts to draw the liquid supply from the cannula, forming a membrane across its end. The expansion of the aerosol gas as it exits the orifice causes localized cooling near the nozzle gas field, which also acts to cool the droplets and slightly delay evaporation. Additionally, in one embodiment, the pressure just below the end of the cannula is less than the pressure inside the cannula, and the pressurized gas velocity accelerates as it travels along the outer surface of the cannula. Upon ejection, the spherical plasma droplets collide with the pressurized gas and aerosolize, forming a spray plume surrounding the ring of the drying gas jet, as described further below. See Figure 43N. The mixing of the aerosol and drying gas sets the initial conditions for the evaporation process.

[0262] Figure 43Q is a schematic diagram showing droplets of liquid plasma undergoing a drying process. Plasma droplets are atomized by the nozzle assembly cannula and exit the drying chamber, and are generally spherical. Dried plasma particles are formed through heat and mass transfer. Drying occurs in two stages: an evaporative drying stage (constant rate) that occurs during initial drying (e.g., less than 1 second), and a drop-rate drying (diffusion-limited) that occurs after the evaporative drying stage and continues as long as the dried particles are subjected to an ambient relative humidity lower than their internal relative humidity.

[0263] Factors contributing to the evaporative drying phase of plasma droplets include the temperature of the plasma and drying gas, the surface area of ​​the droplets, the humidity of the drying gas, and the circulation of air within the plasma drying chamber. When initially exiting the nozzle assembly, the temperature of the drying gas is about 90°C to about 130°C (e.g., about 100°C to about 114°C), and the temperature of the plasma droplets is about 20°C to about 65°C within the plume, as shown in Figure 43Sa. Heat flows from higher temperature locations to lower temperature locations, and in this case, the heat of the drying gas flows to the plasma droplets. Regarding surface area, the droplets are spherical, thereby maximizing their surface area, and because the droplets are very small, mass and heat transfer can occur rapidly. 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 facilitates evaporation of the plasma particles. Finally, as described in more detail below, drying gas is emitted obliquely downward into the plasma drying chamber and into the plume of atomized droplets using several drying gas jets, initiating rapid mixing of the drying air with the atomized droplets. This increases the evaporation rate of the droplets. The drying rate is constant; as the liquid particles evaporate and lose water, water is transferred from the liquid plasma droplets to the drying gas, and heat from the drying gas is transferred to the plasma droplets, turning them into dry particles. The plasma droplets enter the drying chamber at essentially room temperature, and the temperature remains constant for most of the evaporation period. See Figure 43S. Once most of the water leaves the particles, the particle temperature increases and equilibrates with the dryer chamber exit temperature of 65°C. During evaporation, the droplets are maintained at a low temperature, thereby protecting heat-sensitive proteins such as vWF. See Figure 43S. During the evaporation process, the temperature of the droplets and the proteins therein are exposed to a low temperature, i.e., the thermodynamic wet-bulb temperature, compared to the inlet drying gas temperature, thereby protecting the proteins. See Figure 43R. Evaporation reduces the protein temperature to near the thermodynamic wet-bulb value, and when evaporation is delayed the particle temperature increases. See Figure 43Sa.

[0264] The size of the starting droplets produced by the nozzle assembly affects the residence time in the drying chamber required to complete evaporation. The larger the droplets, the smaller the ratio of evaporation surface area to droplet mass, and the slower the rate of mass transfer from the droplets. This slower rate requires a greater distance between the nozzle assembly 20 and the lower filter 36 to avoid deposition of overly wet particles on the filter membrane of the lower filter 36. Overly wet particles can cause "clogging or plugging" of the porous filter membrane, causing excessive chamber pressure buildup and preventing the process from completing, preventing powder production. See Figure 43T.

[0265] Furthermore, Figure 43T shows that in all cases, the majority of evaporation occurs at the top of the disposable 148. This is particularly evident when using small droplet sizes, such as less than 15 microns. As can be seen from Figure 43T, the drying chamber can be shortened to the point where most evaporation occurs while still allowing the dried particles to achieve less than 2.5% residual moisture before deposition on the lower filter 36. In other words, in one embodiment, the drying chamber 28 can be shortened by an amount of approximately 8 inches to 1 inch when the droplet size is less than about 15 microns and less than about 2.5% residual moisture is achieved.

[0266] Turning to the plenum, its functions include: 1) allowing the introduction and flow of drying gas to the disposable, 2) housing the nozzle assembly, and 3) providing support for the drying chamber during the spray drying process. The underside of plenum 6 is shown in Figure 43J. Plenum 6 has two openings: opening 96 for receiving nozzle assembly 20 and drying gas inlet port 22 for receiving drying gas.

[0267] The nozzle assembly receiver opening 96 is complementary in shape to the top of the nozzle reservoir housing 74 and manifold 72. The top of the nozzle assembly 20 is secured in the opening 96. The length of the nozzle assembly matches the height of the plenum 6 so that the bottom of the nozzle assembly 20 extends past the baffle plate 8. See Figures 46A-46B. In certain embodiments, the cannula is flush with the nozzle assembly and baffle plate. The nozzle assembly 20 may be secured using adhesive, fasteners, or a connecting assembly (e.g., spring latch, threaded fit, etc.).

[0268] The other opening in plenum 6 is drying gas inlet port 22, which receives drying gas. A drying gas source (not shown) flows into the plenum through drying gas inlet deflector 242, shown in FIG. 43K. Once the disposables are secured and aligned, the door to the spray dryer closes and spray drying begins. Drying gas inlet deflector 242 lowers through drying gas inlet port 22 to provide drying gas to plenum 6. Drying gas inlet deflector 242 has an elbow shape, as shown in FIG. 43Ka, so that the drying gas flows toward the far inner wall of the plenum, creating a tangential mixture. The right angle of deflector 242 distributes the drying gas throughout plenum 6, creating a low-velocity, very uniform pressure buildup. Uniformity is desirable to create that low-velocity, uniform pressure as the drying gas exits each of the drying jets 142. When the drying gas does not bounce off the sides of the plenum 6 and change direction, but instead deflects downward, the air pressure can be asymmetric with drying jets closer to the drying gas inlet experiencing higher pressure compared to drying jets further from the inlet. Thus, the present invention includes, for example, drying gas inlets deflected against the sides of the plenum with a 90-degree elbow, as with deflector 242. Other geometries for deflector 242 can be utilized to generate uniform air pressure in the plenum 6. For example, the deflector can be angled at an angle less than 90 degrees, as measured from the top of the plenum. For example, the deflector can have an angle ranging from about 60 degrees to about 110 degrees relative to the top of the plenum 6, such that the drying gas pressure across the width of the plenum 6 is substantially uniform. Alternatively, two or more drying gas inlets from opposite sides can be used to generate substantially uniform drying gas pressure across the width of the plenum 6.

[0269] As shown in FIG. 43J, plenum 6 has concentric ribs 98 and radial ribs 102. These ridges support the structure of plenum 6. The additional support provided by the concentric and radial ribs allows the plenum to withstand the pressure and heat of the spray drying process. Because the drying gas is a low-velocity, uniform pressure air vessel, concentric ribs 98 and radial ribs 102 do not contribute to or affect the drying gas airflow. Similarly, protrusions on the inner wall of the plenum, i.e., protrusion 104, are used in the injection molding process when manufacturing the plenum and do not contribute to the drying gas flow.

[0270] Referring to Figures 43L, 43La and 43M, the baffle plate has several functions, such as: A) acting as a support to secure the disposable 100 when it is aligned and inserted into the spray dryer 200; B) creating a drying gas airflow channel to release drying air into the plasma drying chamber 28 of the disposable 100; and C) supporting the baffle filter 94.

[0271] FIG. 43L shows a top inside view of the baffle plate 8. The inner surface of the baffle plate 8 has a baffle plate nozzle opening 140 through which a portion of the nozzle assembly 20 resides. The baffle plate 8 also includes a raised outer ring 124 having a base 126 and a raised inner ring 128 having a base 130. The outer seal ring 90 is disposed around the outer ring 124, and the inner seal ring 92 is disposed around the inner ring 128. The seal rings prevent dry gas from escaping the edge of the filter 94; instead, the dry gas flows through them. The inside of the baffle plate 8 further includes a locator 132 for inserting the locator notch 26 of the plenum 6. The plenum and the baffle plate have locators for aligning them with one another. The baffle plate 8 has a baffle locator 132 that receives a plenum locator 152 on the plenum 6.

[0272] The baffle plate ribs 134, 136, and 138 support the baffle plate filter 94 (shown in FIG. 43L) during use, while maintaining the majority of the filter's surface elevated from the baffle plate. The baffle plate ribs also function as guides for the drying gas flow. It has been demonstrated that if the baffle plate filter 94 were to lie flat on the inside of a baffle plate 8 without ribs, the drying gas flow would be retarded and would not flow freely through the multiple drying jets 142. To prevent this from happening, the baffle plate 8, among other features, includes radial ribs 134 connecting the inner ring base 130 and the outer ring base 126. Each radial rib 134 has a consistent profile throughout its length compared to if it were directly on the inside surface of the baffle plate, allowing the filter to be elevated. The radial ribs 134 also form pie-shaped air channels 139 that lead to the drying jets 142. The radial ribs 134 are the sidewalls of the pie-shaped air channels 139. The baffle plate 8 has two types of ribs that extend from the outer ring base 126 but do not connect to or reach the inner ring base 130. Among these types of ribs are shorter radial ribs 136 and intermediate radial ribs 138. Both the shorter radial ribs 136 and the intermediate radial ribs 138 have a consistent height profile because they proceed inward from the outer ring base 126 and taper rapidly to tapered ends 137. The tapered ends 137 help support the filter without creating corners that could allow the filter to penetrate. In particular, the baffle filter 94 rests on the ribs 134, 136, and 138 and is pressed against them during operation of the spray dryer, e.g., with an air pressure of approximately 11.5 psig. The tapered ends on the ribs 136 and 138 reduce stress on the baffle filter, preventing damage to the filter and reducing loss of filtration efficiency. See FIG. 43La. The ribs prevent the baffle filter 94 from contacting the inner baffle plate surface.The connecting and non-connecting ribs are interspersed on the inner surface of the baffle plate and, in the embodiment shown in FIG. 43L, form a pattern within the pie-shaped air channels 139 (e.g., a connecting rib, a shorter non-connecting rib, two middle non-connecting ribs, a shorter non-connecting rib, a connecting rib, etc.). Each pie-shaped air flow channel 139 is defined by two connecting ribs on either side. The connecting and non-connecting ribs can be in any pattern as long as they support the baffle filter 94 while allowing drying gas to flow through and beneath the filter 94 and through the air channels 139 to the drying jets 142. For example, FIG. 43La shows another configuration of ribs 136 and 138.

[0273] In one embodiment, the pressure drops as the drying gas passes through the baffle filter 94. During operation, the input drying air pressure in the plenum 6 before passing through the filter and into the pie-shaped air flow channels 139 is about 8 to about 15 psig, and in one embodiment, about 10.4 psig. When the drying gas passes through the baffle filter 94, the pressure drops by about 40 to 60%, or in one embodiment, by about 6 psig. The pressure drop across the baffle filter 94 is utilized to aid in the even distribution of the drying gas injected into the drying chamber, as shown in FIG. 43Ma. This feature helps minimize asymmetric drying within the chamber, allowing for a shorter overall length. The resulting pressure within the drying chamber 28 in the area defined by dimension Z ranges from about 4 psig to about 7 psig, and in one embodiment, it is about 5.5 psig. As described further herein, the lower filter 36 of the disposable 100 creates a pressure drop of about 40-80%, or in one embodiment, about 3.0-5.5 psig, which increases as dried plasma accumulates on the filter, resulting in an outlet air pressure of about 1-2.5 psig. In one embodiment, the gas exhaust port 208 of the spray dryer 200 is slightly tapered so that the exhaust gas leaving the outside is at about 1.5 to about 3 psig. If there are valves, sensors, or lengths of tubing past the exhaust outlet, these will apply a slight pressure increase.

[0274] In one embodiment, the flow of dry gas through and across the baffle plate 8 is unrestricted. In one aspect, the height and arrangement of the ribs 134, 136, 138 of the baffle plate 8 are such that the baffle plate filter 96 deflects somewhat under the pressure of the dry air, but does not have a material effect on the pressure drop. Note that, as noted above, the baffle plate filter 96 creates a pressure drop as the dry gas passes through, but the baffle plate 8 itself does not. The cross-sectional area of ​​the pie-shaped channel 139 is equal to or greater than the cross-sectional area of ​​the opening of the dry gas inlet deflector 242. Alternatively, the cross-sectional area of ​​the pie-shaped channel 139 is equal to or greater than the sum of the cross-sectional areas of all the openings of the dry gas jets 142.

[0275] The data presented in Example 8 indicates that the airflow channels 139 of the baffle plate 8 shown in Figure 43L do not interfere with the bacterial filtration efficiency (BFE) of the baffle filter 94, enable a greater than log 6 pathogen reduction when challenged by S. aureus in BFE under American Society for Testing and Materials (ASTM) F2101-14, and do not damage the baffle filter 94 during operation of the spray dryer 200. See Examples 8 and 9.

[0276] In one embodiment, the baffle filter 94 is a 0.2 micron filter (e.g., a 0.22 micron filter) that prevents pathogens that may be introduced into the drying gas from entering the drying chamber. The filter is at least a 0.2 micron filter, and can be, for example, a 0.1 micron filter or smaller, so long as it allows the drying gas to flow as described herein. The filter is strong enough to withstand the heat and pressure of the spray drying process, but flexible enough to allow air to pass through without being deflected when pressed against the ribs. The baffle filter is such a filter, and can be a 4 micron depth filter or membrane filter. Filters are commercially manufactured by Gore (231 East Oak Street, Bozeman, Montana, USA), Lydal (Rochester, New Hampshire, USA), Teijin (Chiyoda City, Tokyo, Japan), or Sabeu (Northeim, Germany). Any type of commercially available filter can be used as long as it prevents pathogens from being introduced into the drying chamber, but is porous enough to allow the drying gas to pass through the filter at the specified flow rate. The baffle filter and lower filter can be made, for example, from a polyethylene filter matrix. In one embodiment, the baffle filter 94 is commercially available from Sabeu (Northeim, Germany) under art. No. 063090.

[0277] Referring to Figure 43M, the underside of the baffle plate can be seen. After the drying gas passes through the baffle filter 94, it is discharged through a plurality of drying gas jets 142. In one embodiment, the drying gas jets 142 are parallel to one another and have centerlines along their lengths that are perpendicular to the baffle plate 8. Despite the described mechanical structure of the jets 142, the drying gas air exiting them is discharged in an inwardly angled manner toward the centerline of the drying chamber 28. Figures 43N and 43Na show the flow pattern of the drying gas jets 142.

[0278] The drying gas jet 142 effectively creates a "wall of dry gas air" within the plasma drying chamber 28, as shown in FIG. 43Ma, while promoting rapid mixing with the atomized plasma particles. The dry jet air flow is directed, in part, toward the plume of atomized liquid plasma droplets for rapid mixing. The wall of dry gas air minimizes accumulation of dried plasma on the interior walls of the plasma drying chamber 28.

[0279] A plurality of indentations 144 are present within the plurality of drying gas jets 142. The indentations 144 are used to further support the structure so that the plenum 6 and baffle plate 8 do not twist due to heat or pressure during spray drying and contribute to the airflow. The drying gas jets 142 are positioned concentrically with the indentations 144. In the embodiment shown in FIG. 43M, there are 16 drying jets 142. The present invention can have more or fewer drying jets, ranging from 2 to 32.

[0280] Furthermore, the drying gas jet 142, like the nozzle assembly, is not flush with the baffle plate but extends through the plane of the baffle plate. By extending the nozzle assembly 20 and drying gas jet 142 through the plane of the baffle plate, drying of the plasma can occur away from the baffle plate surface, reducing the accumulation of dried plasma on the outer surface of the baffle plate and / or the bottom surface of the nozzle assembly during the drying process.

[0281] In light of the above structure, the heated drying gas dries the atomized plasma droplets as follows: The heated drying gas is supplied to the top of the plenum through deflector 242 at a flow rate of about 500 slpm to about 1000 slpm (e.g., about 500, about 550, about 600, about 650, about 700, about 750, about 800, about 850, about 900, about 950, or about 1000 slpm), and in one embodiment, at about 750 slpm. The heated drying gas enters the plenum at a temperature of about 100°C to about 130°C (e.g., about 100°C, about 105°C, about 110°C, about 115°C, about 120°C, about 125°C, or about 130°C), and in one embodiment, at about 114°C. Deflector 242 redirects the gas by 90 degrees to promote uniformity of airflow within the plenum. The drying gas is forced through a baffle filter 94 (e.g., a 0.2 micron, sterilization-rated filter) located at the top of the baffle plate. As described above, the baffle plate 8 is designed with channels forming pie-shaped air channels 139, with the filter providing the upper surface of the channels. The pie-shaped ducts 139 direct the drying gas into 16 individual drying gas jets 142. This flow structure forms jets that are directed inward toward the atomizer to promote plume containment. The mixture of heated drying gas, aerosol gas, droplets, and water vapor drives evaporation, converting the plasma into dry powder. This process is largely completed in less than one second under the powder drying process conditions of the present invention, with individual powders forming in the upper portion of the drying chamber 28, defined by the dimension X.

[0282] An inner concentric ridge 146 on the outside of the baffle plate 8 is a base for attaching the walls of the plasma drying chamber 28. The plasma drying chamber 28 can be attached to the baffle plate 8 with a collar or ring, adhesive, fasteners, etc. The plasma drying chamber 28 can also be attached to the baffle plate 8 at the ridge 146 by heat bonding the chamber to the baffle plate 8. The attachment points can also be molded as part of the baffle plate. The drying chamber can be attached to the baffle plate by several commercially available methods.

[0283] Detailed description of the drying chamber As referred to in this invention, the purpose of drying chamber 28 is to A) allow drying of nebulized plasma while preserving proteins and their function, B) capture dried plasma while allowing gases to escape, and C) for subsequent conversion to commercially available dried plasma units without a filter. In one embodiment, the drying chamber is a dual-purpose, sterile, apyrogenic, single-use chamber in which plasma is dried and collected and stored in a portion of the chamber for use.

[0284] 44 and 46A show the drying chamber 28 including an upper section defined by a length X, a middle section defined by a length U, and a lower section defined by a length V.

[0285] The upper portion 148 is attached to the baffle plate 8 via the baffle plate ring 156 at the outer concentric ring 146. The nozzle assembly 20 protrudes through the baffle plate 8. As described above, the nozzle assembly 20 and the drying gas jet 142 extend through the plane defined by the baffle plate 8. Thus, convergence of atomized plasma occurs at the upper portion 148 of the plasma drying chamber 28. Although the plasma continues to dry as it travels along the length of the drying chamber 28, most of the atomized plasma particles dry at the upper portion 148. When pressurized air exits the nozzle assembly, this is a vortex configuration. When a plasma film exits the cannula and the pressurized air exits the annulus as a vortex, the droplets aerosolize or atomize to form a plume, which decays and expands as the vortex configuration moves downward, as shown in FIG. 43N. Meanwhile, the drying gas jet 142 directs gas such that the flow is angled inward toward the plume to contain the plume and rapidly mix with the aerosolized plasma droplets. The combination of the decaying vortex and the flow from the inclined drying gas stream dilutes the spray plume of plasma droplets, providing more drying gas around the plasma droplets and promoting rapid mixing of the drying gas and droplets. This action promotes efficient evaporation of the plasma droplets, which occurs almost entirely at the top of the drying chamber. When rapid mixing occurs as in the present invention, the droplets evaporate relatively quickly and at a lower temperature than the drying gas, preserving plasma proteins. See Figure 43S. In contrast to freeze-dried plasma, the rapid drying of plasma in the present invention largely obviates the formation of crystals, particularly undesirable cholesterol crystals in dried plasma.

[0286] The gas flow of the drying air jet 142 forms an air curtain to prevent the dried plasma particles from accumulating on the inner walls of the drying chamber. Furthermore, the sloped air wall formed by the drying jet 142 also helps to guide the dried plasma particles downward toward the lower filter 36.

[0287] While most of the plasma undergoes evaporation and dries in the upper compartment 148, drying of the plasma continues in the middle compartment 46, defined by dimension U and containing "seal and separate" positions 44A and 44B, label 40, spike ports 42A and 42B, and hang slot 34. The "seal and separate" positions 44A and 44B are where the drying chamber 28 is severed to form the dried plasma unit 60 (shown in FIG. 49). As described herein, the finishing device 400 moves the plasma within the disposable, seals and separates at positions 44A and 44B, and cuts off the middle compartment 46 to remove the upper and lower disposable sections 148 and 150, forming the dried plasma unit. The spike ports 42A and 42B are for use with the dried plasma unit. The spike ports may be used to allow reconstitution of the dried plasma with a reconstitution solution or sterile water for injection (SWFI). The spike port is a sealing and / or connection device and may be configured to "twist off" to expose a connection port for use in a sterile environment. Other commercially available connectors and adapters for spike ports may be used as long as they are suitable for a sterile environment. The hanger slot 34 is an opening used to attach the plasma bag 64 to an IV (intravenous) pole. The spike ports 42A and 42B and the hanger slot 34 are made and used in the same manner as IV medical bags. The middle section 46 also includes a locator pin opening 32C. As described further herein, the locator pin opening 32C is used to secure the disposable 100 to the finishing device 400 so that the disposable 100 remains in place during sealing and separation.

[0288] While most of the plasma undergoes evaporation and dries in the upper compartment 148, drying continues in the lower compartment 150, defined by dimension V. Referring to the disposable exploded view of FIG. 44, the lower compartment of the drying chamber 28 includes the lower filter 36, the lower filter separator 38, the drying gas outlet port 30, and the locator pin openings 32A and 32B. Moist air (e.g., drying gas, aerosolized gas, and moisture removed from aerosolized plasma droplets) passes through the lower filter 36, the lower filter separator 38, and the lower compartment 150 and exits through the gas outlet 30, which is secured to the gas exhaust port 208. The moist air travels through a channel or space between the filter 36 and the outer wall of the drying chamber 28, exits through the gas exhaust port 208, and is filtered and released to the outside air. When the air is released to the outside air, a filter is used to prevent contamination of the plasma in the spray dryer in the event of a breakage. Such a filter may be a HEPA filter, a UPLA filter, or the like. HEPA filters for filtering the released air can be purchased commercially.

[0289] The lower / trapping filter 36 separates the dried plasma from the moist air. Specifically, the lower filter 36 traps the dried plasma particles / powder while allowing the moist air to pass through. The dried plasma accumulates on the filter throughout the drying process. The goal of the drying process is to complete the majority of the evaporation of the plasma droplets (i.e., complete the mass transfer process) before the dried particles impact the filter surface. Effective evaporation occurs with a size distribution of atomized plasma droplets ranging from about 1 micron to about 35 microns when rapidly mixing with dry air. As described herein, rapid mixing is enhanced by the vortex flow of the pressurized air, the droplet size of the atomized plasma droplets, and the dry air flow. The length of the drying chamber varies depending on the size of the atomized plasma droplets. A shorter drying chamber reduces the time for the droplets to complete evaporation / mass transfer, while a longer drying chamber allows for larger droplets to be used. The completion of the evaporation process for a particular droplet size depends, in part, on the length of the drying chamber. As the initial plasma particles collect on the filter, subsequent dried plasma particles create a depth of powder through which the airflow penetrates, increasing the pressure in the system, but still allowing moist air to pass through effectively. When the dried plasma particles have less than 2% residual moisture, moist air can pass through the dried plasma on the lower filter 36 and pass through the lower filter 36 and out the gas outlet 30 / gas exhaust port 208.

[0290] The dried plasma produced in the present invention is a fine, highly amorphous, and extremely dry (e.g., less than 2% residual moisture) powder to ensure little or no clogging of the lower filter 36.

[0291] The initial powder entering the lower filter is exposed to the chamber outlet temperature for the duration of the batch, while subsequent powders have less residence time in the filter. Plasma dried using the disposables and dryers of the present invention has a very low residual moisture content, e.g., about 2.5%, about 2%, less than about 1%, and preferably about 1.46%, as measured by a Karl Fischer moisture sensor, Model No. C30S Compact KF Coulometer (Mettler, Toledo Billerica, Massachusetts, USA). This very low moisture content results from the effective and efficient evaporation of plasma droplets occurring at the top of the drying chamber 28 and at the process conditions. In this embodiment, the powder moisture content is in equilibrium with the relative humidity of the chamber outlet airflow. Plasma particles with higher moisture content accumulate on the lower filter 36, forcing moist air to pass through the filter at a slower rate, thereby increasing pressure within the chamber. Essentially, excessively moist plasma particles and inefficient evaporation clog the filter, preventing or significantly reducing the flow of moist air. However, the evaporation of the present invention is efficient, allowing moist air to pass through the trapped dried plasma particles. Dried plasma with less water content improves protein stability during storage.

[0292] In one embodiment, lower filter 36 is a 0.2 micron filter whose pore size is small enough to prevent the passage of plasma particles while allowing moist air to pass through with minimal pressure rise. The filter is at least a 0.2 micron filter, and can be, for example, a 0.1 micron or smaller filter, so long as moist air can flow through it as described herein. Lower filter 36 is commercially available from Lydall Inc. (Rochester, New Hampshire, USA) under model number 70L02A.

[0293] The lower filter 36 is supported by a filter frame built into or attached to the filter 36, which may also be attached to the inner wall of the plasma drying chamber 28. The filter 36 is attached around the entire periphery of the inner wall. In other words, the filter frame or the filter itself is attached entirely around the inner wall of the drying chamber 28, with no openings between the inner wall and the attachment point of the filter 36. Attaching the filter to the inner wall in this manner forms a barrier against dried plasma particles and moist air, which allows the moist air to pass through while forcing the plasma and moist air to move downward toward the gas outlet 30, with the filter 36 capturing the dried plasma. The filter frame 37 is attached to the inner surface of the drying chamber 28 by thermal bonding. In other embodiments, the filter frame may be attached to the inner surface of the drying chamber 28 by a combination of adhesive (e.g., UV adhesive) and RF bonding (e.g., by an insulating unit from UFP, Inc., Chicopee, Massachusetts, USA).

[0294] As shown in FIGS. 44A and 44B, the lower filter separator 38 is positioned between the filter 36 and the inner wall of the drying chamber 28. The separator 38 acts similarly to the ribs of a baffle plate, lifting the filter away from the inner wall of the drying chamber 28. This separating / lifting action prevents the filter from adhering to the inner wall of the drying chamber 28, allowing moist air to pass through more easily and preventing pressure buildup. The lower filter separator 38 may be textured or ribbed to maintain a space between the filter 36 and the inner wall of the drying chamber 28. In one embodiment, the filter separator 38 is ribbed with a plurality of spacers. See FIGS. 42A and 44A. In another embodiment, the filter separator 38′ has a weave of porous, coarse circular filaments. See FIG. 44B. In the embodiment shown in FIG. 44B, the separator is two-ply. The separator can be made from one or more sheets (e.g., 1, 2, 3, 4, 5, 6). Any type of spacer or standoff can be used to maintain separation between the filter 36 and the inner wall of the drying chamber 28. Another example of a separator includes a flexible, three-dimensional matrix of polymer filaments. In the embodiment shown in FIG. 44, the separator 38 surrounds most of the filter 36. In other embodiments, the separator 38 need only surround the filter 36 sufficiently to maintain a space between the filter 36 and the inner wall of the drying chamber 28. The spacer / separator 38 is made from a material that can withstand the heat and pressure of the spray-drying process and does not affect the plasma. In one embodiment, the separator is injection molded and can be made from an olefin-based or thermoplastic elastomer, such as polyester or polypropylene. In another embodiment, the separator can be made from nylon with a thermoplastic polyurethane frame. In the embodiment shown in Figures 42A and 44A, separator 38 is Baltex NPD 88 grade with a width of 8.750 inches + / - 0.65, a height of 13.000 inches + / - 0.65, and a thickness of approximately 0.197 inches.The material used in this embodiment is 100% polyester spacer mesh fabric. In the embodiment shown in Figure 44B, separators 38' are two pieces, model 02257 (Freudenberg Filtration Technologies Weinheim, Baden-Wuerttemberg, Germany), approximately 8.80 inches + / - 0.06 inches wide, 6.40 inches + / - 0.06 inches high, and 0.197 inches + / - 0.030 / 0.010 inches thick. During plasma drying, moist air passes through lower filter 36 and lower filter separator 38 or 38' and exits gas outlet 30, leaving the dried plasma on lower filter 36.

[0295] Another important aspect relates to the length of the disposable 100. In previous versions, the disposable was approximately 66 inches long. It is envisioned that longer disposables require more time, space, and heat to dry the plasma particles. However, longer disposables are difficult for operators to install and use, and are complex, inefficient, and difficult to handle. See Examples 30 and 31. In fact, a 66-inch long disposable is longer than the height of many operators, such as American women who are 5'66" tall, which is the 79th percentile according to CDC MHANES 2015-2016 data. Men 5'66" and shorter remain a significant number of potential operators of the present invention, and according to the same data, comprise the 14th percentile for the United States.

[0296] Shortening the disposable to that shown in Figure 42A posed several challenges. A shorter disposable meant there was a shorter drying chamber. A shorter drying chamber meant that the plasma particles had to be evaporated and dried over a shorter distance in a smaller volume and in a shorter time without damaging all of the proteins in the plasma. In other words, the plasma had to be dried gently, yet quickly, and in a small space.

[0297] Despite these obstacles, the present invention includes disposables 100 having a length of about 40 inches or less (e.g., about 40, about 39, about 38, about 37, about 36, about 35, about 34, about 33, about 32, about 31, about 30, about 29, about 28, about 27, about 26, about 25, or about 24 inches or less), preferably about 34.8 inches. At 34.8 inches in length, disposables 100 are easily handled by minimally trained personnel and easily installed and removed from dryers and other pieces of processing equipment for heights ranging from the 5th percentile (4'11") to the 99th percentile (6'6") for men and women, according to data for the United States. See Examples 30-33.

[0298] The length of the disposable, as measured from the bottom of the spray drying head 2 or the bottom of the baffle plate 8 to the top of the bottom filter 36, is about 31 inches or less (e.g., about 31, about 30, about 29, about 28, about 27, about 26, about 25, about 24, about 23, about 22, about 21, about 20, about 19 inches or less), as shown as dimension Y in Figure 44, and in one embodiment is preferably about 25.90 inches. In another aspect, the area of ​​the disposable 100 encompassed by dimension Z, i.e., the length from the bottom of the spray drying head 2 to the top of the filter 36, is about 22 inches or less (e.g., about 22, about 21, about 20, about 19, about 18, about 17, about 16, about 15, about 14 inches), and preferably about 19.11 inches. In yet another embodiment, the length of dimension X, i.e., the length between the bottom and top 46 of the spray drying head 2, is less than about 16 inches (e.g., about 16, about 15, about 14, about 13, about 12, about 11, about 10, about 9, about 8 inches), and preferably about 12.14 inches.

[0299] In fact, when comparing a disposable of the present invention to an earlier development version of a disposable in which the length of the disposable is about 66 inches and dimension Z is about 48 inches, the difference in overall length is about 30 inches, or a 46% reduction, and the difference in dimension Z is about 29 inches, or a 60% reduction. A significant difference can be achieved by shortening the overall length of the disposable along dimensions X, Y, and / or Z. In another embodiment, the length of the present invention can be further shortened by about 1 inch to about 8 inches (e.g., 1 inch, 2 inches, 3 inches, 4 inches, 5 inches, 6 inches, 7 inches, or 8 inches) along dimension X, thereby reducing the overall length by the same amount. In one embodiment, length X ranges from about 30 inches to about 37 inches. In other embodiments, the disposable can be shortened anywhere along dimensions Y and Z by the same amount.

[0300] This shorter disposable 100 dries liquid plasma while preserving protein function, even for the most fragile proteins, such as von Willebrand factor and other proteins. The spray drying system of the present invention, shown in the figure, meets FDA vWF requirements.

[0301] The reduced length of the disposable 100, including the spray drying head 2 and plasma drying chamber 28 of the present invention, results in a system that is significantly more usable by operators of various heights than the prior art. For example, the height of the upper loading slot of a prior art dryer using the described 66" inch disposable was 72.5 inches (more than 6') compared to only 54" height for the dryer 200 shown in Figures 45 and 46. While the bent or folded down (exhaust) loading height of prior art dryers using the 66" disposables described herein was only 16" above the floor, the disposables 100 of dryer 200 described herein have a much more comfortable height of 27.5". Multiple ergonomic validation episodes (see Examples 30-33) demonstrated that shorter disposables allow for unhindered installation of disposables 100 into spray dryer 200 by operators ranging from the 5th (4'11") to the 95th (6'6") percentile of height in the United States. Shorter disposables make it easier for the operator to reach and bend or flex to install the disposable into the spray dryer, and to safely and effectively install and remove the disposable before and after operation of the spray dryer.

[0302] The plasma drying chamber's tubing, baffle plate ring 156, exhaust ports, etc. are made from blow-molded or flat polyvinyl chloride (PVC) and heat-bonded to form the drying chamber. The disposable heat-bonded baffle plate ring 156 can be attached to the baffle plate using, for example, an adhesive. In one embodiment, the drying chamber expands to take the shape of the enclosure, i.e., the dryer's spray-drying chamber housing, during use. Other commercially available materials and other vinyl materials can be used to fabricate the plasma drying chamber of the present invention. Sheets of PVC material are injection molded and / or heat-bonded to form the spray-drying chamber and then sterilized.

[0303] The plasma drying chamber of the present invention, in one embodiment, functions as a single-use chamber in which spray drying occurs. Sterilization by gamma or X-ray irradiation provides sterility of the plasma drying chamber. Development and manufacturing of the drying chamber are performed under ISO 13485 design control. Certification of materials that contact the drying air or donor plasma inside the drying chamber assembly provides lot traceability. Certification provides for toxicity testing and certification for human use.

[0304] The overall size of the spray dryer is generally much smaller than other production-type spray dryers, which are often prohibitively large and unsuitable for use in blood centers, battlefield medical units, or similar locations. The spray drying system of the present invention is adapted for and designed for use in blood component laboratories, whereas other production spray dryers are typically used in large industrial facilities.

[0305] As noted above and in co-pending application No. (Attorney Docket No. 0118.0168-000, filed on the same day, entitled "Usability Of A Disposable For A Spray Drying Plasma System," the entire teachings of which are incorporated herein by reference), the spray dryer of the present invention is largely automated and usable by personnel with limited training. This contrasts with the high level of training and skill required for industrial or laboratory-scale spray drying, such as that sold by Buchi Corporation, 19 Lukens Drive, Suite 400, New Castle, DE 19720, United States, Model No. 4244.

[0306] The liquid plasma was pretreated before undergoing the spray-drying process, as further described in a co-pending related patent application (Application No. 17945126). The pretreatment solution protects plasma coagulation factors during the spray-drying process. A quantity of never-frozen or frozen plasma (e.g., approximately 260 mL) is transferred to a plasma pretreatment vessel containing, for example, 50 mL of a spray-drying stable acidic substance (SDSAS), such as a glycine and hydrochloric acid solution. In one embodiment, single-donor plasma expressed from collected whole blood or obtained by apheresis, which has never been frozen and is less than 24 hours old, is preferably utilized in this process. Plasma is collected from the blood by standard techniques known to those skilled in the art, as described herein. Plasma is collected through a process called plasmapheresis. Plasmapheresis refers to a procedure in which plasma is separated from blood by either centrifugation or membrane filtration. The system process can also be utilized with pooled plasma, if desired, as well as with starting plasma material made with any of the currently available anticoagulation systems known as CPD, CP2D, ACD-A, and ACD-B. A sterile, non-pyrogenic, single-use container containing SDSAS, e.g., 50 mL of solution glycine and hydrochloric acid, is packaged in a 500 mL container inside an overwrap pouch. In one embodiment, the process of the present invention involves converting a single donor unit of plasma collected by standard procedures into a single unit of spray-dried plasma.

[0307] In vitro characterization data demonstrate that the impact of the system's spray-drying process is comparable between units spray-dried using different starting materials. Units manufactured from apheresized plasma (ACD-A anticoagulated) showed similar percent changes attributable to manufacturing effects in the starting material compared to units spray-dried from whole blood-derived plasma (CPD anticoagulated). Statistical analysis (ANOVA) was performed on the percent changes before and after manufacturing between the two starting materials across 20 assays, including clotting time, coagulation function, and activation markers. Of the 20 assays, total protein concentration, PT, TT, and factor VIII and XIII activity were determined to be statistically significantly different, but the mean percent changes were similar, and all mean values ​​were within the clinical reference range. In summary, the results of the in vitro testing support the conclusion that manufacturing effects in both apheresized plasma and whole blood plasma are comparable, and that coagulation profiles are within ±20% of their paired controls, or within the normal reference range.

[0308] Detailed description of spray dryer The spray dryer 200 provides pressurized aerosol gas, plasma, and drying gas to the disposable 100 and exhaust to the moist air. The disposable 100 is placed inside the dryer, which provides the pressurized gas, plasma, and drying gas so that drying can occur inside the disposable.

[0309] Figure 45A is a front view of the spray drying apparatus 200 with the door closed, and Figure 45B shows the spray drying apparatus 200 without the door to allow a view into the dryer. Disposables are placed into the dryer for spray drying. Figure 45B shows the projected locator 206, which is a first locator that receives the notch locator 26 of the disposable device 100. Additionally, a receiver 204 (see Figures 45B and 46A) above the projected locator 206 allows the disposable to be easily received so that it is aligned. Figures 45B and 45C also show the spray drying head receiver 210 for receiving the spray drying head 2, including the baffle 6.

[0310] At the top, spray dryer 200 includes an aerosol line 216 that provides a pressurized atomizing gas source (not shown). The gas source provides clean, dry air with a dew point of minus 40° C., such as an Atlas-Copco SF22 oil-free scroll compressor combined with an Atlas-Copco CD45 desiccant dryer (Atlas Copco Manufacturing company, Nacka, Sweden). The pressurized gas source need not be located near spray dryer 200, but can optionally be located at a distance and in a different space. Such a device is intended to be easily connected to or in communication with a device that receives pressurized air. The spray dryer apparatus 200 heats the air from the source to an appropriate temperature (e.g., about 100 to about 120°C (e.g., about 100, 105, 110, 115, 120°C), in one embodiment at about 114°C. See Figures 45A-45C. In one embodiment, there are redundant in-line filters (e.g., commercially available filters of 0.2 μm or less) in the drying gas line and aerosolization gas line in addition to the filters in the spray dryer disposable device, as further described herein.

[0311] A display 212 provides instructions and information to the operator. An aerosol line 216 is adjacent to the spray dryer head 2 installed in the dryer 200. The aerosol line 216 has a luer lock that attaches to the aerosol filter 12 (which may be a luer lock). In one embodiment, they thread together. The aerosol line 216 is adjacent to the aerosol filter 12 and the aerosol tube 10 when the spray dryer head 100 is installed in the dryer 200. In one embodiment, the aerosol line 216 exits the dryer head 2 between about 4 inches and 10 inches, with about 6 inches being the preferred distance, measured from the top of the spray dryer head 2 to the attachment point of the aerosol filter 12. Furthermore, the aerosol line 216 is directed downward with the luer lock filter at the bottom so that an operator can easily reach and attach the aerosol filter 12 to the aerosol line 216 about 4'6" and 5'6" above the floor, with about 5' being the preferred distance.

[0312] Indicator lights 234 (see FIG. 45A) are located above the display 212 and provide color / visual information (e.g., green = proceed, red = problem, yellow = assistance needed) to the operator. Below the display 212 is a peristaltic pump 214 that pumps liquid plasma through guides 226 into the nozzles of the spray-drying head 2. The peristaltic pump 214 has a plunger latch 214A that is used to secure the plasma tubing 16. The peristaltic pump 214 disposably dispenses plasma at the rates described herein. The dryer 200 also includes a hook 222 for hanging the plasma bag, an emergency shut-off switch 218, and a circuit breaker 220. See FIGS. 45A, 45B, and 45C. The tube guides 226 allow the user to easily position and align the plasma tubing 16 that leads to the pretreated liquid plasma bag 64 and the aerosol tubing 10 that attaches to the aerosol line 216. The aerosol tube 10 is attached to an aerosol line 216 via an aerosol filter 12 with a thread lock (eg, luer lock or luer taper) and provides a source of pressurized gas (not shown).

[0313] FIG. 46B shows the disposable spray dryer 100 installed in the dryer 200. This figure illustrates the position of the aerosol filter 12 in the aerosol line 216 of the disposable 100. A plasma / aerosol guide, guide 226, is provided to protect the path of the plasma tubing 16 and aerosol tube 10. The plasma tubing provides the flow of liquid (to be dried) plasma 66, which travels from the liquid plasma bag 64 through the pump 214 to the disposable 100 during spray drying. The aerosol tube 10 provides a continuous flow of pressurized air from the aerosol line 216 to the disposable 100. A continuous flow of plasma and pressurized air is necessary to ensure continuous spray drying, making this guide an important aspect of the present invention. The plasma / aerosol guide 226 allows the tubing to be properly positioned to ensure it does not kink or buckle during operation. The plasma / aerosol guide 226 is positioned sufficiently between the plasma bag hook 222 and the plasma flow inlet 18 on the installed disposable, and between the aerosol tube 10, the aerosol line 216, and the aerosol inlet 14 on the installed disposable. The placement of the plasma / aerosol guide 226 allows for easy threading of both the plasma tube 16 and the aerosol tube 10. Once the disposable 100 is aligned, the operator threads the tubes 10 and 16 through the plasma / aerosol guide 226. In one embodiment, the plasma / aerosol guide 226 has a retention notch for retaining the plasma tube and / or the aerosol tube within the plasma / aerosol guide during operation of the spray dryer. In one embodiment, the plasma / aerosol guide 226 is readily visible to an operator with a height between the 5th and 95th percentiles when standing in front of the dryer (e.g., approximately 2 feet from the dryer) under load.

[0314] 46B also shows the placement of the aerosol line 216, which provides the source of pressurized gas. Once the operator threads the aerosol tube 10 through the plasma / aerosol guide 266, the operator attaches the aerosol filter 12 to the aerosol line 216 by connecting a luer lock or thread lock. In this embodiment, the connection is easy to make and involves only aligning and rotating the luer lock / thread lock. The proximity of the spray drying head 2, plasma / aerosol guide 226, and aerosol line 216 allows the operator to quickly and easily screw in and attach the aerosol line.

[0315] Similarly, once threaded through the plasma / aerosol guide 226, the plasma tube 16 is threaded through the peristaltic pump 214 and a latch 214A is closed onto the plasma tube to hold the plasma tube 16 in place during spray drying. Again, the proximity of the spray drying head 2, plasma / aerosol guide 226 and peristaltic pump 214 allows for quick and easy screwing and fastening.

[0316] The operator controls (e.g., display 212, pump latch 214A, tube guide 226, aerosol line 216, door handle 230) are positioned to be easily visible and operable by a wide range of stator operators. Operators of shorter stature could not easily see the display 212 in previous versions. This problem has been addressed with the present invention, and the display 212 is now easily visible and accessible to 99% of all people with a variety of conditions. These controls are within approximately 12, 13, 14, 15, 16, 17, 18, 19, or 20 inches of each other, and in one embodiment, they are within approximately 15 inches of each other. In one embodiment, the controls are readily visible to operators with heights between the 5th and 99th percentiles when standing in front of the dryer (e.g., about 2 feet from the dryer) and under load. The controls are positioned not only in close proximity to each other, but also in close proximity to the components on which they are attached or used. Furthermore, the controls are oriented in the direction of attachment of their respective components. Additionally, the layout of the plasma / aerosol guide 226, pump latch 214A, and aerosol line 216 are logically arranged according to the air / plasma flow.

[0317] Dryer 200 has a gasket 203. Gasket 203 resides around the perimeter of housing 202 and / or along the inside of door 228. Gasket 203 has a partial hourglass shape when dryer 200 is viewed from the front. Gasket 203 helps form a seal between door 228 and housing 202 to help retain gas between housing 202 and disposable 100 during spray drying operations. Gasket 203 acts as an insulator to prevent gas from escaping dryer 200. Gasket 203 can be made from rubber or a similar material that can be molded into the slot or recess where it resides.

[0318] Similarly, emergency shutoff 218 is located so that it can be easily located by the operator, specifically, it is located lower and further away from the operator controls mentioned above. The idea is that by locating emergency shutoff 218 away from the main controls, it encourages the operator to make a deliberate decision to use it. Next to emergency shutoff 218 is circuit breaker 220. Emergency shutoff 218 and circuit breaker 220 provide two ways to turn off dryer 200 in the event of an emergency.

[0319] 45A-45C also show the spray drying apparatus 200 including an exhaust port 208 that accepts the gas outlet port 30 of the disposable device 100. The alignment apparatus of the present invention, in one embodiment, includes attaching the gas outlet 30 of the disposable 100 to the gas exhaust port 208 of the spray dryer 200 or to the gas outlet receiver 414 of the finisher 400. (See FIGS. 46A-46C and 47A-47C.) By attaching the gas outlet 30 of the disposable 100 to the gas exhaust port 208 of the spray dryer 200 during spray drying, the bottom of the disposable 200 can be secured to the dryer, thereby holding the disposable in place during the plasma drying process. Similarly, the finisher 400 is designed to accept the gas outlet 30 of the disposable 100 via the gas outlet receiver 414 to keep the disposable in place during the plasma drying process by shaking / impinging the plasma in place, removing air, sealing, and separating the plasma. The gas outlet 30 of the disposable device 100 is a cylindrical outlet with a lip made from a strong, rigid plastic material. The exhaust port 208 of the spray dryer 200 has an O-ring and a gaske...

Claims

1. 1. Spray-dried plasma made from donor plasma, the spray-dried plasma comprising: a) the dry particles have a size ranging from about 1 micron to about 7 microns; b) upon reconstitution, there is a reduction in the average size of the microparticles, as measured using a Coulter Multisizer 4 by electrical sensing zone method, compared to the microparticles in said donor plasma, for microparticles having a size between about 2 μm and 60 μm; c) having some cholesterol crystals upon reconstitution when viewed at 100x magnification compared to freeze-dried plasma; d) a residual moisture content ranging from about 0.5% to about 2.5%; e) stable when stored at temperatures ranging from about -80°C to about 45°C for periods ranging from about 1 day to about 48 months compared to reconstituted spray-dried plasma before storage; f) upon reconstitution, is stable for transfusion after storage for up to about 26 hours; g) having a pH of about 6.5 to about 7.8 upon reconstitution with sterile water for injection (SWFI); h) having an amount of von Willebrand factor (vWF) that induces clot formation upon reconstitution; i) upon reconstitution, have C5a levels, C3a levels, or both, compared to apheresed plasma; and j) any combination thereof; 2. Spray-dried plasma comprising:

2. 2. The spray-dried plasma of claim 1, wherein upon reconstitution, the average size of the microparticles is reduced by about 60%, about 50%, about 40%, about 30%, about 25%, about 20%, about 15%, about 10%, about 5%, about 4%, about 3%, about 2%, or about 1% for microparticles having a size of about 2 μm to about 60 μm compared to the average size found in the donor plasma as measured using a Coulter Multisizer 4 by electrical sensing zone method.

3. 2. The spray-dried plasma of claim 1, wherein the spray-dried plasma is stable upon storage and the level of one or more plasma proteins in the spray-dried plasma upon reconstitution is within about 20% of the level of one or more plasma proteins in the reconstituted spray-dried plasma before storage.

4. 2. The spray-dried plasma of claim 1, wherein the spray-dried plasma is stable upon storage and the levels of one or more plasma proteins of the spray-dried plasma upon reconstitution are within the corresponding clinical reference ranges.

5. 2. The spray-dried plasma of claim 1, wherein upon reconstitution, the reconstituted spray-dried plasma is suitable for transfusion for up to about 26 hours and the level of one or more plasma proteins in the reconstituted spray-dried plasma is within about 20% of the level of one or more plasma proteins in spray-dried plasma reconstituted at the same time after spray-drying.

6. 2. The spray-dried plasma of claim 1, having a pH of about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7 or about 7.8 upon reconstitution with SWFI.

7. 2. The spray-dried plasma of claim 1, wherein the amount of C5a levels is from about 0.1 to about 74 ng / mL.

8. 2. The spray-dried plasma of claim 1, wherein the functionality of vWF is measured by a Bioflux assay.

9. 2. The spray-dried plasma of claim 1, wherein the amount of vWF is measured by a von Willebrand factor ristocetin cofactor assay.

10. 10. The spray-dried plasma of claim 9, wherein the amount of vWF is measured by the von Willebrand factor ristocetin cofactor assay and the amount of vWF is in the range of about 50 IU / dL to about 200 IU / dL.

11. 2. The spray-dried plasma of claim 1, wherein the amount of vWF is measured by a von Willebrand factor antigen assay.

12. 12. The spray-dried plasma of claim 11, wherein the amount of vWF is measured by the von Willebrand factor antigen assay and the amount of vWF is in the range of about 50 IU / dL to about 200 IU / dL.

13. 2. The spray-dried plasma of claim 1, wherein the residual moisture is about 2.5%, about 2.0%, about 1.5%, about 1.0%, or about 0.5%.

14. The spray-dried plasma of claim 1 , wherein the recipient is a mammal.

15. The spray-dried plasma of claim 1 , wherein the recipient is a human.

16. 1. A rehydrated, pre-spray-dried plasma, wherein the pre-spray-dried plasma is dried from donor plasma, and wherein the rehydrated, pre-spray-dried plasma has a reduced average size of the microparticles, as compared to the microparticles in the donor plasma, for microparticles having a size of from about 2 μm to about 60 μm, as measured using a Coulter Multisizer 4 by an electric sensing zone method.

17. 17. The rehydrated pre-spray-dried plasma of claim 16, wherein for microparticles having a size of about 2 μm to about 60 μm, the average size of the microparticles is reduced by about 60%, about 50%, about 40%, about 30%, about 25%, about 20%, about 15%, about 10%, about 5%, about 4%, about 3%, about 2%, or about 1% compared to the average size found in the donor plasma as measured using a Coulter Multisizer 4 by electrical sensing zone method.

18. 1. A method for rehydrating spray-dried plasma units having spray-dried plasma, comprising: a) combining said spray-dried plasma units with an amount of SWFI; the spray-dried plasma reconstitutes in a period ranging from about 2 minutes to about 5 minutes, measured from initial contact by a user to complete reconstitution without visible clumps; method.

19. 1. A method for storing spray-dried plasma, said method comprising: a) subjecting the dried plasma to a temperature of about 20°C to about 25°C for a period of time ranging from about 1 day to about 6 months; A method comprising:

20. 20. The method of claim 19, wherein upon reconstitution, clot formation is induced by a level of one or more plasma proteins in the spray-dried plasma.

21. 1. A method for storing spray-dried plasma, said method comprising: a) subjecting the dried plasma to a temperature of about 1°C to about 6°C for a period of time ranging from about 1 day to about 48 months; A method comprising:

22. 22. The method of claim 21, wherein upon reconstitution, clot formation is induced by a level of one or more plasma proteins in the spray-dried plasma.

23. 1. Spray-dried plasma dried from a disposable spray-drying 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, the disposable spray-drying device comprising: i) a spray drying head, (1) a spray-drying nozzle assembly in fluid communication with the plasma source and the pressurized aerosol gas source, the pressurized aerosol gas flowing in a vortex pattern, and in use, the pressurized aerosol gas atomizing the plasma in the drying chamber to obtain atomized plasma droplets; (2) a plenum having a dry gas inlet in communication with the dry gas source, wherein, in use, the dry gas resides in the plenum having a constant air pressure, and the plenum supports the nozzle assembly; and (3) a spray drying head comprising: a baffle plate forming a floor of the plenum having one or more drying gas jets, the drying gas jets supplying drying gas to the drying chamber; ii) the drying chamber attached to the baffle plate, wherein the atomized plasma droplets evaporate in the presence of the drying gas emitted from one or more of the drying gas jets, thereby obtaining dried plasma particles and moist air; iii) a trapping filter present in the drying chamber, the trapping filter trapping the dried plasma particles and allowing the moist air to pass through; iv) a gas outlet, the gas outlet being attached to an exhaust port of the spray drying apparatus and the moist air flowing through the gas outlet; A disposable spray drying device comprising:

24. 1. A method of spray-drying plasma using a disposable spray-drying device having a spray-drying head and a plasma drying chamber for use in a spray-drying system having a drying gas source, a plasma source, and a pressurized aerosol gas source, the disposable spray-drying device comprising: i) a spray drying head, (1) a spray-drying nozzle assembly in fluid communication with the plasma source and the pressurized aerosol gas source, the pressurized aerosol gas flowing in a vortex pattern, and in use, the pressurized aerosol gas atomizing the plasma in the drying chamber to obtain atomized plasma droplets; (2) a plenum having a dry gas inlet in communication with the dry gas source, wherein, in use, the dry gas resides in the plenum having a constant air pressure, and the plenum supports the nozzle assembly; and (3) a spray drying head comprising: a baffle plate forming a floor of the plenum having one or more drying gas jets, the drying gas jets supplying drying gas to the drying chamber; ii) the drying chamber attached to the baffle plate, wherein the atomized plasma droplets evaporate in the presence of the drying gas emitted from one or more of the drying gas jets, thereby obtaining dried plasma particles and moist air; iii) a trapping filter present in the drying chamber, the trapping filter trapping the dried plasma particles and allowing the moist air to pass through; iv) a gas outlet, the gas outlet being attached to an exhaust port of the spray drying apparatus and the moist air flowing through the gas outlet; Equipped with the method comprising drying liquid plasma using the spray drying disposable device of the spray drying system, thereby obtaining spray-dried plasma; method.

25. 25. The method of claim 24, further comprising reconstituting the spray-dried plasma, thereby obtaining reconstituted spray-dried plasma, wherein the reconstituted spray-dried plasma has a reduced number of cholesterol crystals when viewed at 100x magnification compared to freeze-dried plasma.

26. 25. Dried spray-dried plasma produced by the steps of the method of claim 24.

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