Methods and kits for preparing pathogen inactivated whole blood

Abstract

The present invention provides methods for preparing pathogen inactivated whole blood and other pathogen inactivated blood product compositions, as well as kits and compositions related thereto.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application is a divisional application of Chinese invention patent application No. 201980074126.4, filed on September 20, 2019, and entitled “Method and Kit for Preparing Pathogen-Inactivated Whole Blood.” This application claims priority to U.S. Provisional Application No. 62 / 734,117, filed on September 20, 2018, the disclosure of which is incorporated herein by reference in its entirety. Technical Field

[0003] The present disclosure relates to pathogen-inactivated whole blood and improved methods and kits for preparing pathogen-inactivated whole blood and other blood product compositions. Background Art

[0004] Pathogen inactivation (PI) technologies are increasingly being used to treat blood products to help reduce the risk of transfusion-transmitted infections and complications caused by donor leukocytes. At least three PI technologies commercially available in certain countries utilize ultraviolet light to treat one or more blood products, such as plasma, platelets, red blood cells, and / or whole blood, in the presence or absence of pathogen-inactivating compounds (e.g., photosensitizers) (see, e.g., Schubert, 2018; Schlenke, 2014; Prowse 2013). Such PI technologies include, for example, aminostalin / UV-based System (Cerus), Riboflavin / UV-based system (TerumoBCT) and UVC light-only System (Macopharma). At least one other PI technology in late clinical development for treating red blood cells (RBCs) also targets nucleic acids by using a fragile pathogen-inactivating compound (e.g., S-303) but without the need for UV light (Henschler 2011).

[0005] Exemplary breakable PI compounds are described in more detail in U.S. Patents 6,093,725 and 6,514,987. The blood system utilizes the fragile nucleic acid-targeting alkylating agent compound S-303 (e.g., amustaline) for pathogen inactivation. Amustaline is a modular compound that contains an acridine anchor (e.g., an intercalator targeting nucleic acids), an effector (e.g., a dialkylating agent group that reacts with nucleophiles such as DNA bases), and a linker (e.g., a small flexible carbon chain containing an unstable ester bond that can be hydrolyzed at neutral pH). The anchor targets the nucleic acids of pathogens and cells by embedding into helical regions, and the effector reacts rapidly (e.g., within a few minutes) with the nucleic acid bases to form a monoadduct that is then cross-linked, thereby blocking replication. Within a few hours, the linker can hydrolyze, resulting in the dissociation of the anchor from the effector. For RBCs The blood system also includes a quencher, such as glutathione (GSH), to reduce nonspecific binding of amatarin (Henschler 2011; U.S. Patent 8,900,805). After mixing the donor RBC units, amatarin, GSH, and treatment solution, the mixture is transferred to an incubation bag and incubated at room temperature for 18-24 hours for pathogen inactivation and amatarin hydrolysis, followed by centrifugation. The solution containing residual unhydrolyzed amatarin, GSH, and treatment solution is then removed and replaced with red blood cell additive solution before storage.

[0006] PI treatment using the pathogen inactivating compounds amatalin and GSH has also been demonstrated for whole blood units. However, for whole blood PI treatment, a post-incubation step of centrifugation and solution replacement with fresh additive solution similar to that used for treating red blood cells cannot be performed, and therefore residual unhydrolyzed amatalin and GSH are not removed or reduced by such a solution replacement step, which may not allow the PI-treated whole blood units to be used for transfusion into subjects if the residual amatalin remains above certain concentrations in the treated unit. In addition, compared to red blood cell processing processes, volume limitations of amatalin and GSH added to whole blood for pathogen inactivation may affect dosing and benefit from improvements in processing kits that better facilitate compound transfer. Finally, the use of current pathogen inactivation technologies (including amatalin / GSH systems) may be subject to certain limitations, particularly in geographic areas where suitable facilities may not be available and in war zones where blood components are not readily available or difficult to process. PI techniques typically require a suitable power source and equipment for one or more processing steps, including, for example, lighting devices to provide ultraviolet light exposure during processing (e.g., aminostalin / UV and riboflavin / UV systems) and / or devices for other processing steps, such as using a sterile docking (e.g., tube welding) device to connect a bag containing a blood product to a PI system (e.g., to a kit, a processing device) and / or a centrifuge, incubator, rocker, stirrer, or other device. Therefore, improvements to PI techniques and related processing kits are desired that can further improve pathogen inactivation of blood products (e.g., whole blood), such as by reducing residual levels of PI compounds in treated products, improving dosing of PI compounds and quenchers in blood products, and / or providing the opportunity to perform PI processing without the need for equipment that may require a power source or only a modest power supply (such as from a portable battery power source). Summary of the Invention

[0007] To meet these and other needs, the methods, kits, and compositions described herein are particularly useful for preparing pathogen-inactivated whole blood and other pathogen-inactivated blood product (e.g., RBC, plasma, cryoprecipitate) compositions, including, for example, pathogen-inactivated whole blood compositions having reduced residual levels of pathogen-inactivating compounds in the compositions, which can be particularly beneficial for subsequent transfusion (e.g., transfusion of PI-treated whole blood compositions).

[0008] In one aspect, the present disclosure provides a method for preparing a pathogen-inactivated whole blood composition, comprising: (a) mixing (e.g., combining, adding) (i) a whole blood composition, (ii) an effective amount of a pathogen-inactivating compound (PIC), and optionally (iii) an effective amount of a quencher in a first container; (b) incubating the mixture (e.g., admixture) of the whole blood composition, the PIC, and the optional quencher in the first container under conditions and for a duration sufficient to inactivate the pathogens, if present; (c) aseptically transferring the mixture to a second container; and (d) incubating the mixture in the second container under conditions and for a duration sufficient such that at least one of (i)-(iii) applies: (i) the concentration of the PIC after incubation in the second container is at least 25% less than the concentration of the PIC in the mixture at the time of transferring step (b); (ii) the concentration of the PIC after incubation in the second container is at least 10% less (e.g., lower) than the concentration of the PIC in the mixture of step (a) (e.g., the initial concentration, i.e., the concentration prior to incubating step (b)). 5 times; or (iii) the concentration of the PIC after incubation in the second container is about 1 nM or less, thereby producing a pathogen-inactivated whole blood composition. In some embodiments, the method comprises: (a) mixing (e.g., combining, adding) (i) a whole blood composition and (ii) an effective amount of a pathogen-inactivating compound (PIC) in a first container; (b) incubating the mixture of the whole blood composition and the PIC in the first container under conditions and for a duration sufficient to inactivate the pathogen, if present; (c) transferring the mixture to a second container under aseptic conditions; and (d) incubating the mixture in the second container under conditions and for a duration sufficient such that at least one of (i)-(iii) applies: (i) the concentration of the PIC after incubation in the second container is at least 25% less than the concentration of the PIC in the mixture at the time of the transfer step (b); (ii) the concentration of the PIC after incubation in the second container is at least 10% less than the concentration of the PIC in the mixture of step (a); 5times; or (iii) the concentration of the PIC after incubation in the second container is about 1 nM or less, thereby producing a pathogen-inactivated whole blood composition. In some embodiments, the method comprises: (a) mixing (e.g., combining, adding) (i) a whole blood composition, (ii) an effective amount of a pathogen-inactivating compound (PIC), and (iii) an effective amount of a quencher in a first container; (b) incubating the mixture of the whole blood composition, the PIC, and the quencher in the first container under conditions and for a duration sufficient to inactivate the pathogen, if present; (c) aseptically transferring the mixture to a second container; and (d) incubating the mixture in the second container under conditions and for a duration sufficient to ensure that at least one of (i)-(iii) applies: (i) the concentration of the PIC after incubation in the second container is at least 25% less than the concentration of the PIC in the mixture at the time of the transfer step (b); (ii) the concentration of the PIC after incubation in the second container is at least 10% less than the concentration of the PIC in the mixture of step (a); 5 times; or (iii) the concentration of PIC after incubation in the second container is about 1 nM or less, thereby producing a pathogen-inactivated whole blood composition. In some embodiments, the method further comprises (e.g., after incubation step (d)) storing the pathogen-inactivated whole blood composition under refrigerated conditions (e.g., refrigerated storage, storage at between about 2° C. and about 6° C.).

[0009] In some embodiments, the concentration of PIC in the mixture (e.g., blend) of step (a) (e.g., initial concentration, i.e., concentration prior to incubation step (b)) is at least about 0.1 mM. In some embodiments, the concentration of PIC in the mixture of step (a) is about 0.1 mM to about 10 mM. In some embodiments, the concentration of PIC in the mixture of step (a) is about 0.1 mM to about 1.0 mM. In some embodiments, the concentration of PIC in the mixture of step (a) is about 0.2 mM to about 0.5 mM. In some embodiments, the concentration of PIC in the mixture of step (a) is about 0.2 mM. In some embodiments, the concentration of PIC after incubation in the second container is at least 25% less than the concentration of PIC in the mixture at the time of transfer step (b). In some embodiments, the concentration of PIC after incubation in the second container is at least 50% less than the concentration of PIC in the mixture at the time of transfer step (b). In some embodiments, the concentration of PIC after step (d) is at least 25% less than the concentration of PIC in the mixture at the time of transfer step (b). In some embodiments, the concentration of PIC after step (d) is at least 50% less than the concentration of PIC in the mixture at the time of transfer step (b). In some embodiments, the concentration of PIC after incubation in the second container is at least 10% less than the concentration of PIC in the mixture at the time of step (a). 5In some embodiments, the concentration of PIC after step (d) is at least 10 times less than the concentration of PIC in the mixture of step (a). 5 times. In some embodiments, the concentration of the PIC after incubation in the second container is about 1 nM or less. In some embodiments, the concentration of the PIC after incubation in the second container is less than about 1 nM. In some embodiments, the concentration of the PIC after incubation in the second container is about 0.75 nM or less. In some embodiments, the concentration of the PIC after incubation in the second container is less than about 0.75 nM. In some embodiments, the concentration of the PIC after step (d) is less than about 2 nM. In some embodiments, the concentration of the PIC after step (d) is less than about 1 nM. In some embodiments, the concentration of the PIC after step (d) is less than about 0.75 nM. In some embodiments, the PIC comprises a functional group that serves as or forms a reactive electrophilic group. In some embodiments, the functional group is selected from the group consisting of: mustard, mustard intermediate, and mustard equivalent. In some embodiments, the functional group is or is capable of forming an aziridinium ion. In some embodiments, the reactive electrophilic group is capable of reacting with nucleic acids. In some embodiments, the PIC comprises a nucleic acid binding ligand. In some embodiments, the nucleic acid binding ligand is an intercalator. In some embodiments, the intercalator is an acridine. In some embodiments, the PIC comprises a frangible linker connecting the functional group and the nucleic acid binding ligand. In some embodiments, the PIC is β-alanine, N-(acridin-9-yl), 2-[bis(2-chloroethyl)amino]ethyl ester.

[0010] In some embodiments, step (a) comprises mixing (e.g., combining, adding) a whole blood composition, an effective amount of PIC, and an effective amount of a quencher in a first container. In some embodiments, the concentration of the quencher in the mixture (e.g., admixture) of step (a) (e.g., initial concentration, i.e., concentration prior to incubation step (b)) is at least about 1 mM. In some embodiments, the concentration of the quencher in the mixture of step (a) is about 1 mM to about 10 mM. In some embodiments, the concentration of the quencher in the mixture of step (a) is about 2 mM. In some embodiments, the concentration of the PIC in the mixture of step (a) is about 0.2 mM and the concentration of the quencher in the mixture of step (a) is about 2 mM. In some embodiments, the molar ratio of the quencher concentration to the PIC concentration in the mixture of step (a) is about 5:1 to about 20:1 quencher to PIC. In some embodiments, the molar ratio of the quencher concentration to the PIC concentration in the mixture of step (a) is about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, or about 20:1 quencher to PIC. In some embodiments, the molar ratio of the quencher concentration to the PIC concentration in the mixture of step (a) is about 10:1 quencher to PIC. In some embodiments, the quencher comprises a sulfhydryl group, wherein the sulfhydryl group is capable of reacting with a reactive electrophilic group of the pathogen-inactivating compound. In some embodiments, the quencher comprises cysteine ​​or a derivative of cysteine. In some embodiments, the quencher is glutathione or a pharmaceutically acceptable salt thereof. In some embodiments, the quencher is glutathione monosodium salt.

[0011] In some embodiments, the mixture of step (a) further comprises an anticoagulant. In some embodiments, the whole blood composition further comprises an anticoagulant (e.g., an anticoagulant solution). In some embodiments, the first container comprises an anticoagulant, and the whole blood composition is mixed with the anticoagulant in the first container (e.g., before the whole blood composition is mixed with the PIC). In some embodiments, the whole blood composition (e.g., including a coagulant) has a volume of about 250 mL to about 700 mL. In some embodiments, the whole blood composition has a volume of about 400 mL to about 600 mL. In some embodiments, the ratio of anticoagulant to whole blood (e.g., in a whole blood composition) is about 1:5 to about 1:9. In some embodiments, the ratio of anticoagulant to whole blood is about 1:7.

[0012] In some embodiments, the whole blood composition is added (e.g., introduced, transferred) to the first container through the first inlet of the first container. In some embodiments, the whole blood composition is added to the first container from a donor collection container (e.g., a whole blood collection container) containing a whole blood composition. In some embodiments, the whole blood composition is added to the first container from a donor collection container containing a whole blood composition that is coupled to the first container (e.g., a first inlet in the first container). In some embodiments, the whole blood composition is added to the first container from a donor collection container containing a whole blood composition through a conduit (e.g., one or more segments of tubing extending therebetween, connected to provide a fluid communication path) between the donor collection container and the first container (e.g., the first inlet of the first container). In some embodiments, the whole blood composition is added to the first container by collecting the whole blood composition from a donor (e.g., a whole blood donor) into the first container (e.g., directly into the first container without first collecting it into an intermediate container, without first collecting it into the donor collection container).

[0013] In some embodiments, a whole blood composition is collected from a whole blood donor into a first container. In some embodiments, the whole blood composition is a whole blood donation collected from a donor into a first container (e.g., directly into the first container without first collecting into a different container). In some embodiments, the whole blood composition is collected from a donor into the first container through a first conduit (e.g., one or more segments of tubing). In some embodiments, the whole blood composition is a whole blood donation collected from a donor into the first container using a vascular access device (e.g., a needle, a venipuncture needle), such as a vascular access device connected to a container (e.g., a first container) by a conduit (e.g., a tube, one or more segments of tubing) extending between the vascular access device and the first container (e.g., and providing a fluid communication path from or through the vascular access device to the first container). In some embodiments, a whole blood composition is collected from a whole blood donor into the first container via a first conduit extending between a vascular access device (e.g., a needle, a venipuncture needle) and a first container (e.g., a first inlet of the first container) and providing a fluid communication path for transferring the whole blood composition from the donor (e.g., from the vascular access device, through the vascular access device) into the first container. In some embodiments, the vascular access device (e.g., a needle) and the conduit extending therefrom constitute a needle assembly.

[0014] In some embodiments, the PIC and optional quencher (e.g., PIC and quencher) are added to the first container through an inlet of the first container (e.g., first inlet, second inlet). In some embodiments, the PIC and optional quencher (e.g., PIC and quencher) are added to the first container after the whole blood composition is added to the first container. In some embodiments, the PIC and quencher are added to the first container sequentially (e.g., added to the whole blood composition). In some embodiments, the PIC is added to the first container before the quencher (e.g., added to the whole blood composition). In some embodiments, the quencher is added to the first container before the PIC (e.g., added to the whole blood composition). In some embodiments, the PIC and optional quencher (e.g., PIC and quencher) are added to the first container through the first inlet of the first container after the whole blood composition is added to the first container through the first inlet of the first container. In some embodiments, the PIC and optional quencher (e.g., PIC and quencher) are added to the first container through the second inlet of the first container. In some embodiments, the PIC and optional quencher (e.g., PIC and quencher) are added to the first container via a conduit (e.g., one or more tubing segments) that extends between a container containing the PIC (PIC container) and an optional container containing the quencher (quencher container) and an inlet of the first container and provides a fluid communication path for transferring the PIC from the PIC container to the first container and, optionally, for transferring the quencher from the optional quencher container to the first container. In some embodiments, the PIC is added to the first container via a conduit (e.g., a tube, one or more tubing segments) that extends between the PIC container and an inlet (e.g., first inlet, second inlet) of the first container (e.g., and provides a fluid communication path for transferring the PIC from the PIC container to the first container). In some embodiments, the conduit comprises, at one end (e.g., opposite the end connected to the first container), (a) a first adapter for coupling (e.g., connecting) the PIC container to the conduit, and optionally (b) a displacement device (e.g., an air displacement device) adapted to increase transfer of the PIC into the first container (e.g., through the conduit and into the first container). In some embodiments, the PIC and quencher are added to the first vessel via a conduit (e.g., a tube, one or more tube segments) that extends between the PIC vessel and the quencher vessel and an inlet (e.g., a first inlet, a second inlet) of the first vessel and provides a fluid communication path for transferring the PIC from the PIC vessel to the first vessel and the quencher from the quencher vessel to the first vessel.In some embodiments, the PIC and quencher are added to the first container via a conduit extending between the PIC container and the quencher container and an inlet of the first container, and wherein the conduit comprises at one end (e.g., opposite the first container connection end): (a) a first adapter for coupling (e.g., connecting) the PIC container to the conduit, (b) a second adapter for coupling (e.g., connecting) the quencher container to the conduit, and optionally (c) a displacement device (e.g., an air displacement device) adapted to increase transfer of the PIC and quencher into the first container (e.g., through the conduit and into the first container). In some embodiments, the PIC and quencher are added to the first container via a conduit extending between the PIC container and the quencher container and an inlet of the first container, and wherein the conduit comprises at one end (e.g., opposite the first container connection end): (a) a first adapter for coupling (e.g., connecting) the PIC container to the conduit, (b) a second adapter for coupling (e.g., connecting) the quencher container to the conduit, and (c) a displacement device (e.g., an air displacement device) adapted to increase transfer of the PIC and quencher into the first container (e.g., through the conduit and into the first container). In some embodiments, the PIC container and the quencher container are the same container (e.g., a container having separate chambers for the PIC and the quencher).

[0015] In some embodiments, incubation step (b) is from about 2 hours to about 24 hours. In some embodiments, incubation step (b) is from about 18 hours to about 22 hours. In some embodiments, incubation step (b) is from about 20 hours. In some embodiments, incubation step (b) is at room temperature (e.g., 20°C-25°C, 22°C). In some embodiments, transfer step (c) is carried out by a conduit extending (e.g., connected) between the first container and the second container. In some embodiments, the conduit connects the first outlet of the first container to the first inlet of the second container (e.g., providing a fluid communication path therebetween). In some embodiments, incubation step (d) is from about 1 hour to about 8 hours. In some embodiments, incubation step (d) is from about 2 hours to about 4 hours. In some embodiments, incubation step (d) is at a temperature substantially the same as incubation step (b). In some embodiments, incubation step (d) is at room temperature (e.g., 20°C-25°C, 22°C).

[0016] In some embodiments, the method further comprises transferring the pathogen-inactivated whole blood composition from the second container to a third container. In some embodiments, the method further comprises: (e) storing the pathogen-inactivated whole blood composition at about 2-6°C (e.g., after incubation step (d), storing in the second container, the third container). In some embodiments, the pathogen-inactivated whole blood composition is suitable for transfusion (e.g., transfusion into a subject). In some embodiments, the method further comprises preparing (e.g., separating the pathogen-inactivated whole blood composition into) one or more of a red blood cell composition, a plasma composition, or a platelet composition from the pathogen-inactivated whole blood composition. In some embodiments, the method further comprises preparing a cryoprecipitate composition from the pathogen-inactivated whole blood composition. In some embodiments, the method further comprises preparing (e.g., separating the pathogen-inactivated whole blood composition into) a plasma composition from the pathogen-inactivated whole blood composition, and preparing a cryoprecipitate composition from the plasma composition.

[0017] In some embodiments, the first container (e.g., a container sheet) comprises polyvinyl chloride (e.g., plasticized polyvinyl chloride). In some embodiments, the first container comprises polyethylene (e.g., ethylene-vinyl acetate, ethylene-vinyl acetate copolymer). In some embodiments, the first container comprises a polyolefin (e.g., a polyolefin blend). In some embodiments, the second container (e.g., a container sheet) comprises polyvinyl chloride (e.g., plasticized polyvinyl chloride). In some embodiments, the second container comprises polyethylene (e.g., ethylene-vinyl acetate, ethylene-vinyl acetate copolymer). In some embodiments, the second container comprises a polyolefin (e.g., a polyolefin blend). In some embodiments, the first container and / or the second container does not comprise diethylhexyl phthalate.

[0018] On the other hand, the present disclosure provides a pathogen-inactivated whole blood composition prepared by any of the aforementioned methods. In some embodiments, the pathogen-inactivated whole blood composition is suitable for transfusion. In some embodiments, the pathogen-inactivated whole blood composition is a pathogen-inactivated whole blood unit suitable for transfusion. Another aspect of the present disclosure provides a pathogen-inactivated red blood cell composition prepared by any of the aforementioned methods (e.g., a pathogen-inactivated red blood cell composition suitable for transfusion). Another aspect of the present disclosure provides a pathogen-inactivated plasma composition prepared by any of the aforementioned methods (e.g., a pathogen-inactivated plasma composition suitable for transfusion). Another aspect of the present disclosure provides a pathogen-inactivated platelet composition prepared by any of the aforementioned methods (e.g., a pathogen-inactivated platelet composition suitable for transfusion). Another aspect of the present disclosure provides a pathogen-inactivated cold precipitate composition prepared by any of the aforementioned methods (e.g., a pathogen-inactivated cold precipitate composition suitable for transfusion).

[0019] On the other hand, the present disclosure provides a method for infusing a pathogen-inactivated whole blood composition into a subject in need thereof, the method comprising infusing a pathogen-inactivated whole blood composition prepared by any of the aforementioned methods or the aforementioned pathogen-inactivated whole blood composition into the subject. In some embodiments, the method for infusing a pathogen-inactivated whole blood composition is achieved by infusing the pathogen-inactivated whole blood composition through a catheter (e.g., an infusion line, an infusion line assembly) connected (e.g., connected) to an outlet (e.g., a first outlet, a spike port) of a container (e.g., a second container, a third container) containing the pathogen-inactivated whole blood composition. In some embodiments, the method for infusing a pathogen-inactivated whole blood composition comprises infusing the pathogen-inactivated whole blood composition through an infusion line assembly connected to a second container. In some embodiments, the method for infusing a pathogen-inactivated whole blood composition comprises coupling an infusion line (e.g., an infusion line assembly) to an outlet (e.g., a first outlet, a spike port) of a second container and infusing the pathogen-inactivated whole blood composition into the subject. In some embodiments, a method of transfusing a pathogen-inactivated whole blood composition comprises coupling an infusion line assembly to a second container and infusing the pathogen-inactivated whole blood composition into a subject. In some embodiments, a method of transfusing a pathogen-inactivated whole blood composition comprises coupling an infusion line assembly to a third container and infusing the pathogen-inactivated whole blood composition into a subject. In some embodiments, a method of transfusing a pathogen-inactivated whole blood composition comprises infusing a pathogen-inactivated whole blood composition prepared by any of the methods and / or kits disclosed herein into a subject suffering from trauma (e.g., for trauma resuscitation). In some embodiments, the subject is acutely bleeding due to trauma. In some embodiments, the subject is a trauma patient exhibiting a systolic blood pressure of <100 mmHg, <90 mmHg, or less than 80 mmHg. In some embodiments, the pathogen-inactivated whole blood composition is ABO compatible with the subject. In some embodiments, the pathogen-inactivated whole blood composition is RhD compatible with the subject. In some embodiments, the pathogen-inactivated whole blood composition is from a type O donor. In some embodiments, the transfusion method comprises administering two or more transfusions (e.g., two or more units) of a pathogen-inactivated whole blood composition prepared by any method of the present disclosure and / or using any kit of the present disclosure. In some embodiments, the pathogen-inactivated whole blood composition is transfused into a subject in need thereof (such as a soldier on the battlefield) at the site of injury.

[0020] In another aspect, the present disclosure provides a kit for preparing a pathogen-inactivated whole blood composition, comprising: (a) a vascular access device (e.g., a needle) for withdrawing whole blood from a donor (e.g., by venipuncture), (b) a first container suitable for mixing (e.g., combining, adding) the whole blood and a pathogen-inactivating compound (PIC), wherein the first container comprises (e.g., includes) at least a first inlet and a first outlet; (c) a first conduit (e.g., a tube, one or more tube segments) extending between the vascular access device and the first container (e.g., the first inlet of the first container) (e.g., and providing a fluid communication path for transferring whole blood from the donor (e.g., through the vascular access device) to the first container); and (d) a container (PIC container) containing the pathogen-inactivating compound. In some embodiments, the vascular access device is a needle (e.g., a venipuncture needle) for achieving vascular access in the donor. In some embodiments, the vascular access device and the first conduit constitute a needle assembly.

[0021] In some embodiments, the PIC container is configured to be coupled (e.g., via a fluid communication path (e.g., a conduit, a tube)) to a first container (e.g., a first inlet of the first container). In some embodiments, the PIC container is configured to be coupled to a first conduit. In some embodiments, the PIC container is coupled to the first container via a fluid communication path (e.g., a conduit, a tube). In some embodiments, the first container further comprises a second inlet. In some embodiments, the first container further comprises a second inlet and the PIC container is configured to be coupled (e.g., via a fluid communication path) to the second inlet of the first container. In some embodiments, the PIC container is coupled (e.g., via a fluid communication path (e.g., a conduit, a tube)) to the second inlet of the first container. In some embodiments, the kit further comprises a second PIC container. In some embodiments, the second PIC container is suitable for storing PIC (e.g., lyophilized PIC). In some embodiments, the second PIC container is suitable for reconstitution of lyophilized PIC. In some embodiments, the PIC container coupled to or configured to be coupled to the first container of the kit is suitable for storing PIC. In some embodiments, the PIC container coupled to or configured to be coupled to the first container of the kit is suitable for reconstitution of lyophilized PIC. In some embodiments, the second PIC container is suitable for transferring the PIC to a PIC container that is coupled to or configured to be coupled to the first container of the kit provided herein. In some embodiments, the second PIC container is suitable for transferring the PIC to a catheter that is coupled to or configured to be coupled to the first container of the kit provided herein. In some embodiments, the kit further comprises a container comprising a quencher (quencher container). In some embodiments, the quencher container is configured to be coupled (e.g., via a fluid communication path) to the first container (e.g., the first inlet of the first container). In some embodiments, the quencher container is configured to be coupled to the first catheter. In some embodiments, the quencher container is coupled to the first container via a fluid communication path (e.g., a catheter, a tube). In some embodiments, the quencher container is configured to be coupled (e.g., via a fluid communication path) to the second inlet of the first container. In some embodiments, the quencher container is coupled (e.g., via a fluid communication path (e.g., a catheter, a tube)) to the second inlet of the first container. In some embodiments, the kit further comprises a second quencher container. In some embodiments, the second quencher container is suitable for storing the quencher (e.g., a lyophilized quencher). In some embodiments, the second quencher container is suitable for reconstituting a lyophilized quencher. In some embodiments, the quencher container coupled to or configured to be coupled to the first container of the kit is suitable for storing the quencher. In some embodiments, the quencher container coupled to or configured to be coupled to the first container of the kit is suitable for reconstituting a lyophilized quencher.In some embodiments, the second PIC container is suitable for transferring the quencher to a quencher container that is coupled to or configured to be coupled to the first container of the kit provided herein. In some embodiments, the second quencher container is suitable for transferring the quencher to a conduit that is coupled to or configured to be coupled to the first container of the kit provided herein. In some embodiments, the PIC container and the quencher container are the same container (e.g., portion of the same container, separate chamber of the same container). In some embodiments, the PIC container and the quencher container are separate chambers within the same container.

[0022] In some embodiments, the kit further comprises a second conduit, wherein the second conduit comprises at least one adapter at one end for coupling the PIC container and / or quencher container to the second conduit. In some embodiments, the kit further comprises a second conduit, wherein the second conduit comprises at least one adapter at one end for coupling the PIC container to the second conduit. In some embodiments, the kit further comprises a second conduit, wherein the second conduit comprises at least one adapter at one end for coupling the quencher container to the second conduit. In some embodiments, the second conduit comprises at one end a first adapter for coupling the PIC container to the second conduit and a second adapter for coupling the quencher container to the second conduit. In some embodiments, the kit further comprises a second conduit, wherein the second conduit is coupled to the PIC container and / or quencher container at one end. In some embodiments, the second conduit is configured to couple to the first container (e.g., to provide a fluid communication path between the PIC container and / or quencher container and the first container). In some embodiments, the second conduit is configured to couple to the first inlet of the first container. In some embodiments, the second conduit is configured to couple to the second inlet of the first container. In some embodiments, the second conduit is coupled to the first container (e.g., providing a fluid communication path between the PIC container and / or quencher container and the first container). In some embodiments, the second conduit is coupled to the first inlet of the first container. In some embodiments, the second conduit is coupled to the second inlet of the first container. In some embodiments, the second conduit is configured to couple to the first conduit (e.g., providing a fluid communication path between the PIC container and / or quencher container and the first container). In some embodiments, the fluid communication path is an openable fluid communication path.

[0023] In some embodiments, the kit further comprises a displacement device (e.g., an air displacement device) adapted to increase the transfer of the PIC and / or quencher into the first container (e.g., through the second conduit and into the first container). In some embodiments, the kit further comprises a displacement device adapted to increase the transfer of the PIC into the first container. In some embodiments, the kit further comprises a displacement device adapted to increase the transfer of the quencher into the first container. In some embodiments, the kit further comprises a displacement device adapted to increase the transfer of the PIC and quencher into the first container. In some embodiments, the displacement device is an air displacement device. In some embodiments, the air displacement device is a sample diversion pouch. In some embodiments, the displacement device (e.g., an air displacement device) is coupled to the second conduit. In some embodiments, the first container comprises an anticoagulant (e.g., an anticoagulant solution).

[0024] In some embodiments, the PIC comprises a functional group that acts as or forms a reactive electrophilic group. In some embodiments, the functional group is selected from the group consisting of a mustard, a mustard intermediate, and a mustard equivalent. In some embodiments, the functional group is or is capable of forming an aziridinium ion. In some embodiments, the reactive electrophilic group is capable of reacting with a nucleic acid. In some embodiments, the PIC comprises a nucleic acid-binding ligand. In some embodiments, the nucleic acid-binding ligand is an intercalator. In some embodiments, the intercalator is an acridine. In some embodiments, the PIC comprises a frangible linker connecting the functional group and the nucleic acid-binding ligand. In some embodiments, the PIC is β-alanine, N-(acridin-9-yl), 2-[bis(2-chloroethyl)amino]ethyl ester.

[0025] In some embodiments, the quencher comprises a sulfhydryl group, wherein the sulfhydryl group is capable of reacting with a reactive electrophilic group of the pathogen-inactivating compound. In some embodiments, the quencher comprises cysteine ​​or a derivative of cysteine. In some embodiments, the quencher is glutathione or a pharmaceutically acceptable salt thereof. In some embodiments, the quencher is glutathione monosodium salt.

[0026] In some embodiments, the kit further comprises a second container suitable for storing the pathogen-inactivated whole blood composition. In some embodiments, the kit further comprises a second container suitable for storing the pathogen-inactivated whole blood composition, wherein the second container comprises (e.g., includes) at least a first inlet and a first outlet. In some embodiments, the second container is coupled to the first container. In some embodiments, the kit further comprises a third conduit extending between the first container (e.g., the first outlet of the first container) and the second container (e.g., the first inlet of the second container) (e.g., and providing a fluid communication path for transferring (e.g., under sterile conditions) a mixture comprising the whole blood composition and the PIC (e.g., the pathogen-inactivated whole blood composition) from the first container to the second container). In some embodiments, the kit further comprises a third container suitable for storing the pathogen-inactivated whole blood composition. In some embodiments, the kit further comprises a third container suitable for storing a pathogen-inactivated whole blood composition, wherein the third container comprises (e.g., includes) at least a first inlet and a first outlet, and a fourth conduit extending between the second container (e.g., the first outlet of the second container) and the third container (e.g., the first inlet of the third container) and providing a fluid communication path for transferring (e.g., under sterile conditions) a mixture comprising the whole blood composition and the PIC (e.g., the pathogen-inactivated whole blood composition) from the second container to the third container. In some embodiments, the fluid communication path is an openable fluid communication path.

[0027] In some embodiments, the kit further comprises an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject (e.g., administering pathogen-inactivated whole blood). In some embodiments, the infusion line assembly is configured to be coupled (e.g., via a nosepiece port) to a first container, a second container, or a third container. In some embodiments, the kit further comprises an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject, wherein the infusion line assembly comprises a second vascular access device (e.g., a needle, an infusion needle) and an infusion line conduit extending between the first container, the second container, or the third container (e.g., an outlet of the first container, the second container, or the third container) and the second vascular access device (e.g., wherein the infusion line conduit provides a fluid communication path for aseptically transferring the pathogen-inactivated whole blood composition from the container (e.g., the first container, the second container, the third container) to the vascular access device). In some embodiments, the kit further comprises an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject, wherein the infusion line assembly comprises a second vascular access device and an infusion line conduit extending between the first outlet of the second container and the second vascular access device (e.g., wherein the infusion line conduit provides a fluid communication path for aseptically transferring the pathogen-inactivated whole blood composition from the second container to the vascular access device). In some embodiments, the infusion line assembly comprises a filter.

[0028] In some embodiments, the first container (e.g., a container sheet) comprises polyvinyl chloride (e.g., plasticized polyvinyl chloride). In some embodiments, the first container comprises polyethylene (e.g., ethylene-vinyl acetate, ethylene-vinyl acetate copolymer). In some embodiments, the first container comprises a polyolefin (e.g., a polyolefin blend). In some embodiments, the second container (e.g., a container sheet) comprises polyvinyl chloride (e.g., plasticized polyvinyl chloride). In some embodiments, the second container comprises polyethylene (e.g., ethylene-vinyl acetate, ethylene-vinyl acetate copolymer). In some embodiments, the second container comprises a polyolefin (e.g., a polyolefin blend). In some embodiments, the first container and / or the second container does not comprise diethylhexyl phthalate.

[0029] In another aspect, the present disclosure provides a kit for preparing a pathogen-inactivated whole blood composition, comprising: (a) a first container suitable for mixing (e.g., combining, adding) a whole blood composition, a pathogen-inactivating compound (PIC), and a quencher, wherein the first container comprises (e.g., includes) at least a first inlet and a first outlet, (b) a second container suitable for storing the pathogen-inactivated whole blood composition, wherein the second container comprises (e.g., includes) at least a first inlet and a first outlet, and wherein the second container is coupled to the first container; (c) a container comprising the pathogen-inactivating compound (PIC container), (d) a container comprising the quencher (quencher container), (e) a first conduit, wherein the first conduit is coupled to or configured to be coupled to the PIC container and the quencher container at one end, and wherein the first conduit further comprises a displacement device (e.g., an air displacement device) adapted to increase transfer of the PIC and quencher into the first container (e.g., through the first conduit and into the first container). In some embodiments, the first outlet of the first container is coupled to the first inlet of the second container. In some embodiments, the first container is coupled to the second container by a second conduit (e.g., a tube) that provides a fluid communication path for transferring the whole blood composition (e.g., a pathogen-inactivated whole blood composition) from the first container to the second container. In some embodiments, the present disclosure provides a kit for preparing a pathogen-inactivated whole blood composition, comprising: (a) a first container suitable for mixing (e.g., combining, adding) a whole blood composition, a pathogen-inactivating compound (PIC), and a quencher, wherein the first container comprises (e.g., includes) at least a first inlet and a first outlet, (b) a second container suitable for storing the pathogen-inactivated whole blood composition, wherein the second container comprises (e.g., includes) at least the first inlet and the first outlet; (c) a container comprising the pathogen-inactivating compound (PIC container), (d) a container comprising the quencher (quencher container), (e) a first conduit, wherein the first conduit is coupled to or configured to be coupled to the PIC container and the quencher container at one end, and wherein the first conduit further comprises a displacement device (e.g., an air displacement device) adapted to increase transfer of the PIC and quencher into the first container (e.g., through the first conduit and into the first container); and (f) a second conduit extending between the first container and the second container (e.g., and providing a fluid communication path for transferring the whole blood composition (e.g., pathogen-inactivated whole blood composition) from the first container to the second container). In some embodiments, the first conduit provides a fluid communication path from the PIC container and the quencher container. In some embodiments, the fluid communication path is an openable fluid communication path. In some embodiments, the first container further comprises (e.g., includes) a second inlet.

[0030] In some embodiments, the first catheter further comprises at least one adapter for coupling the first catheter to the PIC container and the quencher container. In some embodiments, the first catheter comprises a first adapter for coupling the PIC container to the first catheter and a second adapter for coupling the quencher container to the first catheter. In some embodiments, the first catheter is coupled to the PIC container and the quencher container. In some embodiments, the PIC container and the quencher container are the same container. In some embodiments, the first catheter is coupled or configured to be coupled to the first container, thereby providing a fluid communication path between the PIC container and the quencher container and the first container. In some embodiments, the first catheter is coupled or configured to be coupled to an inlet (e.g., a first inlet, a second inlet) of the first container. In some embodiments, the kit further comprises a vascular access device (e.g., a needle) for withdrawing whole blood from a donor (e.g., by venipuncture). In some embodiments, the kit further includes a vascular access device for withdrawing whole blood from a donor and a third conduit extending (e.g., connected) between the vascular access device and the first container (e.g., the first inlet of the first container) and providing a fluid communication path for transferring whole blood from the donor (e.g., from the vascular access device, through the vascular access device) into the first container.

[0031] In some embodiments, the PIC container is configured to be coupled (e.g., via a fluid communication path (e.g., a conduit, a tube)) to a first conduit. In some embodiments, the PIC container is configured to be coupled to a first conduit. In some embodiments, the PIC container is coupled to the first conduit via a fluid communication path. In some embodiments, the kit further comprises a second PIC container. In some embodiments, the second PIC container is suitable for storing PIC (e.g., lyophilized PIC). In some embodiments, the second PIC container is suitable for reconstitution of lyophilized PIC. In some embodiments, the PIC container coupled or configured to be coupled to the first conduit is suitable for storing PIC. In some embodiments, the PIC container coupled or configured to be coupled to the first conduit is suitable for reconstitution of lyophilized PIC. In some embodiments, the second PIC container is suitable for transferring the PIC to a PIC container coupled or configured to be coupled to the first conduit. In some embodiments, the second PIC container is suitable for transferring the PIC to a conduit coupled or configured to be coupled to the first container of the kit provided herein. In some embodiments, the kit further comprises a container comprising a quencher (quencher container). In some embodiments, the quencher container is configured to be coupled (e.g., via a fluid communication path (e.g., a conduit, a tube)) to the first conduit. In some embodiments, the quencher container is configured to be coupled to the first conduit. In some embodiments, the quencher container is coupled to the first conduit via a fluid communication path. In some embodiments, the kit further comprises a second quencher container. In some embodiments, the second quencher container is suitable for storing a quencher (e.g., a freeze-dried quencher). In some embodiments, the second quencher container is suitable for reconstituted freeze-dried quencher. In some embodiments, the quencher container coupled or configured to be coupled to the first conduit is suitable for storing a quencher. In some embodiments, the quencher container coupled or configured to be coupled to the first conduit is suitable for reconstituted freeze-dried quencher. In some embodiments, the second PIC container is suitable for transferring the quencher to the quencher container coupled or configured to be coupled to the first conduit. In some embodiments, the second quencher container is suitable for transferring the quencher to a conduit coupled or configured to be coupled to the first container of the kit provided herein. In some embodiments, the PIC container and the quencher container are the same container (e.g., part of the same container, a separate chamber of the same container). In some embodiments, the PIC container and the quencher container are separate chambers within the same container.

[0032] In some embodiments, the first container further comprises (e.g., includes) a second inlet, wherein the first conduit is configured to couple to the second inlet of the first container. In some embodiments, the first container further comprises (e.g., includes) a second inlet and the first conduit is coupled to the second inlet of the first container. In some embodiments, the first conduit provides a fluid communication path from the PIC container and the quencher container to the first container. In some embodiments, the first conduit is configured to couple to a third conduit, thereby providing a fluid communication path between the PIC container and the quencher container and the first container. In some embodiments, the fluid communication path is an openable fluid communication path. In some embodiments, the first container contains an anticoagulant.

[0033] In some embodiments, the PIC comprises a functional group that acts as or forms a reactive electrophilic group. In some embodiments, the functional group is selected from the group consisting of a mustard, a mustard intermediate, and a mustard equivalent. In some embodiments, the functional group is or is capable of forming an aziridinium ion. In some embodiments, the reactive electrophilic group is capable of reacting with a nucleic acid. In some embodiments, the PIC comprises a nucleic acid-binding ligand. In some embodiments, the nucleic acid-binding ligand is an intercalator. In some embodiments, the intercalator is an acridine. In some embodiments, the PIC comprises a frangible linker connecting the functional group and the nucleic acid-binding ligand. In some embodiments, the PIC is β-alanine, N-(acridin-9-yl), 2-[bis(2-chloroethyl)amino]ethyl ester.

[0034] In some embodiments, the quencher comprises a sulfhydryl group, wherein the sulfhydryl group is capable of reacting with a reactive electrophilic group of the pathogen-inactivating compound. In some embodiments, the quencher comprises cysteine ​​or a derivative of cysteine. In some embodiments, the quencher is glutathione or a pharmaceutically acceptable salt thereof. In some embodiments, the quencher is glutathione monosodium salt.

[0035] In some embodiments, the kit includes a third container suitable for storing a pathogen-inactivated whole blood composition, wherein the third container includes (e.g., including) at least a first inlet and a first outlet, and wherein the third container is coupled to the second container. In some embodiments, the kit includes a third container suitable for storing a pathogen-inactivated whole blood composition, wherein the third container includes (e.g., including) at least a first inlet and a first outlet, and wherein the kit also includes a fourth conduit extending between the second container (e.g., the first outlet of the second container) and the third container (e.g., the first inlet of the third container) (e.g., and providing a fluid communication path for transferring the whole blood composition (e.g., pathogen-inactivated whole blood composition) from the second container (e.g., under aseptic conditions) to the third container). In some embodiments, the fluid communication path is an openable fluid communication path. In some embodiments, the kit also includes an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject (e.g., administering pathogen-inactivated whole blood). In some embodiments, the infusion line assembly is configured to be coupled (e.g., through a sharp mouth port) to the second container or the third container. In some embodiments, the kit further comprises an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject, wherein the infusion line assembly is coupled to a second container. In some embodiments, the kit further comprises an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject, wherein the infusion line assembly is coupled to a third container. In some embodiments, the kit further comprises an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject, wherein the infusion line assembly comprises a second vascular access device (e.g., a needle, an infusion needle) and an infusion line conduit extending between the second container or the third container (e.g., an outlet of the first container, the second container, or the third container) and the second vascular access device (e.g., wherein the infusion line conduit provides a fluid communication path for aseptically transferring the pathogen-inactivated whole blood composition from the container (e.g., the second container, the third container) to the vascular access device). In some embodiments, the kit further comprises an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject, wherein the infusion line assembly comprises a second vascular access device and an infusion line conduit extending between a first outlet (e.g., a nose port) of the second container and the second vascular access device (e.g., wherein the infusion line conduit provides a fluid communication path for aseptically transferring the pathogen-inactivated whole blood composition from the second container to the vascular access device). In some embodiments, the infusion line assembly comprises a filter.

[0036] In some embodiments, the first container (e.g., a container sheet) comprises polyvinyl chloride (e.g., plasticized polyvinyl chloride). In some embodiments, the first container comprises polyethylene (e.g., ethylene-vinyl acetate, ethylene-vinyl acetate copolymer). In some embodiments, the first container comprises a polyolefin (e.g., a polyolefin blend). In some embodiments, the second container (e.g., a container sheet) comprises polyvinyl chloride (e.g., plasticized polyvinyl chloride). In some embodiments, the second container comprises polyethylene (e.g., ethylene-vinyl acetate, ethylene-vinyl acetate copolymer). In some embodiments, the second container comprises a polyolefin (e.g., a polyolefin blend). In some embodiments, the first container and / or the second container does not comprise diethylhexyl phthalate.

[0037] It will be appreciated that one, some, or all of the features of the various embodiments described herein may be combined to form other embodiments. These and other aspects will become apparent to those skilled in the art. These and other embodiments will be further described by the detailed description that follows. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1a An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. The dashed line portion illustrates a source container containing whole blood (e.g., a whole blood composition) for connection to the kit. An optional infusion line assembly ("infusion") is also shown. Abbreviations: PIC, pathogen-inactivating compound; WB, whole blood. Drawing not to scale.

[0039] Figure 1b An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. The dashed line portion illustrates a source container containing whole blood (e.g., a whole blood composition) for connection to the kit. An optional infusion line assembly ("infusion") is also shown. Abbreviations: PIC, pathogen-inactivating compound; WB, whole blood. Drawing not to scale.

[0040] Figure 1c An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. The dashed line portion illustrates a source container containing whole blood (e.g., a whole blood composition) for connection to the kit. An optional infusion line assembly ("infusion") is also shown. Abbreviations: PIC, pathogen-inactivating compound; WB, whole blood. Drawing not to scale.

[0041] Figure 2aAn exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. The dashed line portion illustrates a source container containing whole blood (e.g., a whole blood composition) for connection to the kit. An optional infusion line assembly ("infusion") is also shown. Abbreviations: PIC, pathogen-inactivating compound; WB, whole blood. Drawing not to scale.

[0042] Figure 2b An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. The dashed line portion illustrates a source container containing whole blood (e.g., a whole blood composition) for connection to the kit. An optional infusion line assembly ("infusion") is also shown. Abbreviations: PIC, pathogen-inactivating compound; WB, whole blood. Drawing not to scale.

[0043] Figure 2c An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. The dashed line portion illustrates a source container containing whole blood (e.g., a whole blood composition) for connection to the kit. An optional infusion line assembly ("infusion") is also shown. Abbreviations: PIC, pathogen-inactivating compound; WB, whole blood. Drawing not to scale.

[0044] Figure 2d An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. The dashed line depicts a source container containing whole blood (e.g., a whole blood composition) for connection to the kit. Abbreviations: PIC, pathogen-inactivating compound; WB, whole blood. Drawing not to scale.

[0045] Figure 2e An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. Abbreviation: PIC, pathogen-inactivating compound. Drawing not to scale.

[0046] Figure 2f An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. Abbreviations: PIC, pathogen-inactivating compound; WB, whole blood. Drawing not to scale.

[0047] Figure 2g An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. Abbreviation: PIC, pathogen-inactivating compound. Drawing not to scale.

[0048] Figure 3a An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. The dashed line portion illustrates a source container containing whole blood (e.g., a whole blood composition) for connection to the kit. An optional infusion line assembly ("infusion") is also shown. Abbreviations: PIC, pathogen-inactivating compound; WB, whole blood. Drawing not to scale.

[0049] Figure 3b An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. The dashed line portion illustrates a source container containing whole blood (e.g., a whole blood composition) for connection to the kit. An optional infusion line assembly ("infusion") is also shown. Abbreviations: PIC, pathogen-inactivating compound; WB, whole blood. Drawing not to scale.

[0050] Figure 3c An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. The dashed line portion illustrates a source container containing whole blood (e.g., a whole blood composition) for connection to the kit. An optional infusion line assembly ("infusion") is also shown. Abbreviations: PIC, pathogen-inactivating compound; WB, whole blood. Drawing not to scale.

[0051] Figure 4a An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. The dashed line portion illustrates a source container containing whole blood (e.g., a whole blood composition) for connection to the kit. An optional infusion line assembly ("infusion") is also shown. Abbreviations: PIC, pathogen-inactivating compound; WB, whole blood. Drawing not to scale.

[0052] Figure 4b An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. The dashed line portion illustrates a source container containing whole blood (e.g., a whole blood composition) for connection to the kit. An optional infusion line assembly ("infusion") is also shown. Abbreviations: PIC, pathogen-inactivating compound; WB, whole blood. Drawing not to scale.

[0053] Figure 4c An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. The dashed line portion illustrates a source container containing whole blood (e.g., a whole blood composition) for connection to the kit. An optional infusion line assembly ("infusion") is also shown. Abbreviations: PIC, pathogen-inactivating compound; WB, whole blood. Drawing not to scale.

[0054] Figure 4d An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. The dashed line depicts a source container containing whole blood (e.g., a whole blood composition) for connection to the kit. Abbreviations: PIC, pathogen-inactivating compound; WB, whole blood. Drawing not to scale.

[0055] Figure 5aAn exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. An optional infusion line assembly ("infusion") is also shown. Abbreviation: PIC, pathogen-inactivating compound. Drawing not to scale.

[0056] Figure 5b An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. An optional infusion line assembly ("infusion") is also shown. Abbreviation: PIC, pathogen-inactivating compound. Drawing not to scale.

[0057] Figure 5c An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. An optional infusion line assembly ("infusion") is also shown. Abbreviation: PIC, pathogen-inactivating compound. Drawing not to scale.

[0058] Figure 5d An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. Abbreviation: PIC, pathogen-inactivating compound. Drawing not to scale.

[0059] Figure 5e An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. Abbreviation: PIC, pathogen-inactivating compound. Drawing not to scale.

[0060] Figure 5f An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. Abbreviation: PIC, pathogen-inactivating compound. Drawing not to scale.

[0061] Figure 5g An exemplary kit for preparing a pathogen-inactivated whole blood composition according to some embodiments is shown. Abbreviation: PIC, pathogen-inactivating compound. Drawing not to scale. DETAILED DESCRIPTION

[0062] In some aspects, the present disclosure provides for the preparation of pathogen-inactivated whole blood and other blood product compositions, including improved methods and kits for the inactivated whole blood and other blood product compositions suitable for transfusion. It has been found that the kits disclosed herein and the related methods using such kits provide improved pathogen inactivation processes, including, for example, processes for producing pathogen-inactivated whole blood and other blood product compositions with reduced residual levels of pathogen-inactivating compounds in the compositions, which can be particularly beneficial for subsequent transfusions of whole blood compositions. Improved kits and related methods also provide new opportunities for preparing pathogen-inactivated whole blood compositions under conditions where suitable facilities and / or power sources may not be readily available, such as in war zones.

[0063] The terms "pathogen inactivation" and "pathogen inactivation process" refer to processes that can be used to inactivate pathogens that may be present in a whole blood composition (such as a whole blood donation), wherein it is understood that the process does not necessarily completely inactivate all pathogens that may be present, but greatly reduces the amount of pathogens, thereby significantly reducing the risk of transfusion-related diseases (e.g., transfusion-transmitted infections, TTIs). Inactivation of pathogens can be determined by measuring the number of infectious pathogens (e.g., virus particles, bacteria) in a certain volume before and after the pathogen inactivation process, and the level of inactivation is typically expressed as a log reduction in pathogen infectivity, or a log reduction in titer. Methods for determining the log reduction in titer and their measurement for pathogen inactivation are known in the art. When the inactivation process is tested for multiple pathogens, the reduction in specific active pathogens is at least about 1 log reduction, at least about 2 log reductions, at least about 3 log reductions, at least about 4 log reductions, or at least about 5 log reductions in titer. Multiple pathogen inactivation processes are known in the art and are used to demonstrate the use of various commercially available pathogen inactivation processes, such as Methods for achieving pathogen activation levels using the HER2® blood system (Cerus Corp) are known in the art. In certain embodiments, the pathogen inactivation process may include treatment with a pathogen inactivating compound.

[0064] The term "suitable for transfusion" refers to a whole blood composition (e.g., a pathogen-inactivated whole blood composition) or other blood product (e.g., a blood product prepared from a whole blood composition) that can be used for transfusion (e.g., transfusion) into a subject (e.g., a human patient) according to medical judgment. Non-limiting examples of other blood products include red blood cell compositions, plasma compositions, platelet compositions, cryoprecipitate compositions, and cryo-reduced plasma compositions. In some embodiments, suitability refers to having sufficient biological activity for its intended use, i.e., for a use in which transfusion of whole blood or other blood products is indicated (including but not limited to prophylactic and therapeutic transfusions). In some embodiments, suitability refers to having sufficient safety, such as meeting applicable regulatory and / or certification requirements for the safety of administering treated whole blood or blood product compositions. In some embodiments, suitability refers to a pathogen-inactivated whole blood composition or other blood product in which the concentration (e.g., residual concentration) of the pathogen-inactivating compound and / or one or more of its byproducts is below a certain standard (e.g., below the maximum acceptable concentration). In some embodiments, suitability refers to meeting one or more criteria established by an accrediting or regulatory body governing infusion practices, such as the AABB (e.g., having suitable characteristics, having a certain level of biological activity or function, having at least a minimum dose).

[0065] Pathogen-inactivating compounds

[0066] The term "pathogen inactivation compound" or "pathogen inactivating compound" refers to any suitable compound that can be used to inactivate pathogens that may be present in a whole blood composition. Inactivation of pathogens in a whole blood composition is achieved by contacting the pathogens in the whole blood composition with a pathogen inactivating compound (PIC). In any of the embodiments described herein, the PIC (e.g., amatarin, S-303 as described herein) can be present in an effective amount (e.g., an amount effective to inactivate the pathogen, such as an amount sufficient to inactivate, for example, at least about 1 log, at least about 2 log, at least about 3 log, at least about 4 log, at least about 5 log, or at least about 6 log or more of the pathogen (if present) in the whole blood composition). Pathogen inactivating compounds that can be used with the methods of the present disclosure include, for example, compounds that contain a functional group that is or is capable of forming and has formed (e.g., in situ) a reactive group such as an electrophilic group. In some embodiments, the PICs of the present disclosure do not require photoactivation to become reactive. For example, the functional group can be a mustard group, a mustard group intermediate, a mustard group equivalent, an epoxide, formaldehyde, or a formaldehyde synthon. Such functional groups can form reactive groups in situ, such as electrophilic aziridine, aziridinium, thioethane or thioethane (thiiranium) ions. Mustard groups can be single- or double- (haloethyl) amine groups or single (haloethyl) sulfide groups. Mustard equivalents are groups that react by a mechanism similar to mustard, such as by forming reactive intermediates, such as aziridine and aziridine groups or thiirane and thiirane groups. Examples include aziridine derivatives, single or double- (methylsulfonylethyl) amine groups, single (methylsulfonylethyl) sulfide groups, single or double- (tosylethyl) amine groups and single- (tosylethyl) sulfide groups. Formaldehyde synthons are any compounds that decompose into formaldehyde, including hydroxylamines, such as hydroxymethylglycine. The reactive groups of PICs can react with nucleic acids of pathogens, such as with nucleophilic groups on nucleic acids. The reactive group is also capable of reacting with the nucleophilic group of the quencher. The PIC may also include a component that targets the compound to a nucleic acid, such as an anchoring moiety. The anchoring moiety comprises a portion that is capable of non-covalently binding to a nucleic acid biopolymer, such as DNA or RNA, and is also referred to as a nucleic acid binding ligand, nucleic acid binding group, or nucleic acid binding moiety. Examples of such compounds are described in U.S. Patents 5,691,132, 6,410,219, 6,136,586, 6,617,157, and 6,709,810, each of which is incorporated herein by reference. Another class of PICs that can be used in the methods of the present disclosure comprises the above-mentioned reactive groups connected to the nucleic acid binding group via a hydrolyzable linker, such as described in U.S. Patent 6,514,987, which is incorporated herein by reference.The anchoring portion of the PIC has an affinity for nucleic acids. This affinity can result from any of several modes of non-covalent binding to nucleic acids, including but not limited to intercalation, minor groove binding, major groove binding, and electrostatic binding (e.g., phosphate backbone binding). Affinity can also result from mixed binding modes (e.g., intercalation and minor groove binding). Binding can be sequence-specific (i.e., having a higher affinity for one or more specific nucleic acid sequences than for other nucleic acid sequences) or non-sequence-specific. Detailed examples of such nucleic acid-binding moieties can be found in the aforementioned patents.

[0067] In some embodiments of the methods, kits, and compositions described herein, the PIC may comprise a functional group that acts as or forms a reactive electrophilic group that reacts with the nucleophile of the selected quencher. In some embodiments, the pathogen-inactivating group comprises a nucleic acid-binding ligand and a functional group that acts as or forms an electrophilic group. A specific example of a suitable PIC for use in the present disclosure is β-alanine, N-(acridin-9-yl), 2-[bis(2-chloroethylamino]ethyl ester (alternatively referred to herein as "S-303" or "amatalene"), the structure of which is as follows, including its salts.

[0068]

[0069] The quenchers used in the methods of the present disclosure are intended to reduce unwanted side reactions of reactive electrophilic species used to inactivate pathogens (e.g., binding of pathogen-inactivating compounds to the surface of RBCs in the whole blood composition, which may lead to an undesirable immune response). In any of the embodiments described herein, the quencher (e.g., glutathione) may be present in an effective amount (e.g., an amount effective to reduce unwanted side reactions, such as the amounts described herein). Suitable quenchers may comprise nucleophilic groups capable of reacting with the electrophilic groups of the PIC. Non-limiting examples are described in detail in U.S. Patent No. 6,709,810, which is incorporated herein by reference in its entirety. In some embodiments, the quencher is capable of significantly reducing unwanted side reactions in the whole blood composition while allowing the PIC to fully inactivate pathogens that may contaminate the whole blood composition. In some embodiments, the methods of the present disclosure provide a combination of an effective amount of a quencher and an effective amount of a PIC, while providing a combination of optimal reduction of unwanted side reactions (e.g., binding of PICs) and sufficient inactivation of pathogens, without significantly altering the properties of the whole blood composition or the blood components of the whole blood (e.g., without reducing red blood cell osmotic fragility, without significantly increasing red blood cell dehydration, without significantly reducing the levels of coagulation factors required for hemostasis, without significantly reducing one or more platelet functions). Various unwanted side reactions can be reduced, such as reactions of PICs with proteins and / or red blood cells in the whole blood composition. In some embodiments, the quencher provides optimal reduction of modifications of the whole blood composition or the blood components therein (such as binding of IgG to red blood cells or binding of PICs to red blood cells). Although the methods of the present disclosure involve ex vivo processing (e.g., pathogen inactivation) of the whole blood composition, some quencher may remain in the composition upon introduction into a subject. Thus, in some embodiments, the quenchers of the present disclosure are suitable for transfusion. Suitable quenchers include, but are not limited to, compounds containing a sulfhydryl group, such as quenchers containing the amino acid cysteine ​​or a suitable cysteine ​​derivative such as N-acetylcysteine. Examples of such quenchers include cysteine ​​and peptides containing at least one cysteine, such as glutathione. In some embodiments, suitable quenchers include cysteine ​​derivatives that can form sulfhydryl groups in situ with or without the use of additional chemicals or added enzymes, such as S-acetylcysteine ​​or other suitable sulfhydryl-derivatized cysteine ​​prodrugs, or peptides containing S-acetylcysteine ​​or other suitable sulfhydryl-derivatized cysteine ​​prodrugs. Suitable cysteine ​​derivatives are those that contain or are capable of forming in situ a cysteinylsulfhydryl group that is capable of reacting with an electrophilic group of the PIC.

[0070] In some embodiments, the quencher is a peptide of 2 to 10 amino acids, wherein at least one amino acid is cysteine, N-acetylcysteine, S-acetylcysteine ​​or other suitable cysteine ​​derivatives. In some embodiments, the quencher is a peptide of at least 3 amino acids, such as about 3-10 amino acids, and about 3-6 amino acids, wherein at least one amino acid is cysteine, N-acetylcysteine, S-acetylcysteine ​​or other suitable cysteine ​​derivatives. In some embodiments, the quencher is a peptide of at least 3 amino acids, such as about 3-10 amino acids, and about 3-6 amino acids, wherein at least one amino acid is cysteine, N-acetylcysteine, S-acetylcysteine ​​or other suitable cysteine ​​derivatives, and further wherein at least 2 or at least 3 amino acids are cysteine, N-acetylcysteine, S-acetylcysteine ​​or other suitable cysteine ​​derivatives.

[0071] In a preferred embodiment, the quencher is neutralized glutathione (also referred to as L-glutathione and γ-L-glutamyl-L-cysteinyl-glycine). Glutathione has many characteristics that can make it particularly useful as a quencher. It is generally present in all cell types. It is believed that it cannot, such as by passing through the cell membrane or lipid shell of bacteria and lipid envelope viruses, or by passing through the viral capsid of non-enveloped viruses and passively penetrate into pathogens. When approaching neutral pH, glutathione is loaded and in the absence of active transport, it cannot penetrate the lipid bilayer to any significant extent. This is consistent with the results of the inactivation of lipid envelope viruses such as HIV and VSV that are not substantially affected by glutathione, including the use of a neutralized glutathione concentration greater than 2mM. The use of glutathione does have some effect on the inactivation of, for example, Yersinia enterocolitica, Staphylococcus epidermidis, and Serratia marcescens. However, this can be managed by using an effective amount of neutralized glutathione and PIC. Thus, preferred quenching methods are provided in which contamination of whole blood compositions by viral or bacterial pathogens is inactivated by at least about 2 log, at least about 3 log, at least about 4 log, at least about 5 log, or at least 6 log or more. Under appropriate conditions as described in the present disclosure, glutathione is also compatible with in vitro storage of whole blood and the resulting pathogen-inactivated whole blood composition is suitable for introduction into the body.

[0072] In some embodiments, the quencher is glutathione in its reduced form. Glutathione disulfide, oxidized forms of glutathione may also be used, as long as the glutathione disulfide is sufficiently reduced in the solution before the solution is added to the mixture comprising the whole blood composition or is sufficiently reduced after the solution is added to the mixture comprising the whole blood composition.

[0073] In some embodiments, quencher is a derivative of glutathione, such as glutathione monoalkyl ester or dialkyl ester, wherein alkyl is a straight or branched group with 1 to 10 carbon atoms. The instantiation of alkyl includes but is not limited to methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, amyl, isopentyl, neopentyl, tert-amyl, 1-methylbutyl, hexyl, isohexyl, 2-methylpentyl, 1-ethylbutyl, heptyl, octyl, nonyl and decyl. For example, the limiting examples of glutathione derivatives include glutathione methyl ester, glutathione monoethyl ester and glutathione monoisopropyl ester. In some embodiments, the diethyl ester (GSSG-(glycyl)-diethyl-ester) of glutathione oxidation is used. In some embodiments, the thioester of glutathione is hydrolyzed to form sulfhydryl after being added to the whole blood composition.

[0074] It will be understood that in some embodiments, the quencher will be provided in the form of a free acid or base, while in other embodiments, the quencher will be provided in the form of a salt. If the quencher is in the form of a salt, then the salt is preferably a pharmaceutically acceptable salt. Pharmaceutically acceptable salts of the compounds (in the form of water- or oil-soluble or dispersible products) include, for example, conventional non-toxic salts or quaternary ammonium salts formed from inorganic or organic acids or bases. Examples of such acid addition salts include acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, gluconoheptate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, toluenesulfonate, and undecanoate. Basic salts include ammonium salts, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts formed with organic bases such as dicyclohexylamine salts, N-methyl-D-glucamine, and salts formed with amino acids such as arginine, lysine, etc. In addition, basic nitrogen-containing groups can be quaternized by reagents such as lower alkyl halides such as methyl, ethyl, propyl and butyl chlorides, bromides and iodides; dialkyl sulfates such as dimethyl, diethyl, dibutyl and diamyl sulfate; long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides; aralkyl halides such as benzyl and phenethyl bromides, etc. Other pharmaceutically acceptable salts include sulfates, glycolates and sulfates.

[0075] For example, in some embodiments, the quencher is in the form of a pharmaceutically acceptable salt formed by glutathione. In some embodiments, the quencher is in the form of a pharmaceutically acceptable salt formed by glutathione and one or more cations, such as sodium, aluminum, calcium, lithium, magnesium, zinc or tetramethylammonium. In some embodiments, the quencher is glutathione (reduced) and is provided in the form of glutathione monosodium salt (for example, available from Biomedica Foscama, Italy). In some other embodiments, glutathione (reduced) is provided in the form of glutathione hydrochloride. In some other embodiments, glutathione is provided in the form of glutathione (reduced) disodium salt. In other embodiments, glutathione monoalkyl ester sulfate is used. In some embodiments, glutathione is provided in the form of glutathione oxidized disodium salt.

[0076] alkali

[0077] In some embodiments, the methods of the present disclosure further comprise providing a suitable base, such as by mixing (e.g., combining) a base with the whole blood composition, the PIC, and a quencher (e.g., step (a)). In some embodiments, the amount of base is sufficient to reduce the level of unwanted reaction of the PIC with red blood cells in the mixture relative to a mixture without the base. In some embodiments, the unwanted reaction of the PIC with red blood cells (RBCs) is modification of the surface of the RBCs by the PIC (see, e.g., U.S. Patent 8,900,805). In some embodiments, step (a) further comprises mixing a suitable base with the whole blood composition, the PIC, and a quencher, and the amount of base is sufficient to reduce the level of binding of the anti-pathogen inactivating compound antibodies to the RBCs in the treated whole blood composition in the resulting mixture relative to a mixture without the base by at least about 5% (or at least about 10%, at least about 25%, at least about 50%, at least about 75%, or at least about 90%). In some embodiments, the base and quencher are mixed with the whole blood composition before, simultaneously with, or no more than about 30 minutes after mixing the PIC with the whole blood composition. In some embodiments, the base and the quencher are mixed together before the base or quencher is mixed with the whole blood composition. In some embodiments, the base is NaOH. In some embodiments, the base is an alkaline buffer. In some embodiments, the base comprises about 0.5 to 1.5 equivalents of base, where an equivalent refers to a molar amount equal to the molar amount of the quencher in the mixture. In some embodiments, the base comprises about 0.75 to 1.25 equivalents of base. In some embodiments, the base comprises about 1 equivalent of base. In some embodiments, the pH of the resulting mixture of step (a) at 37° C. is about 6.0 to 7.5. In some embodiments, the pH is about 6.5 to 7.1. In some embodiments, the pH is about 6.8 or 6.9.

[0078] In some embodiments, when the PIC and quencher are mixed (e.g., combined) with the whole blood composition, the pH of the mixture is at a suitable level to reduce unwanted side reactions of the PIC (e.g., binding of the PIC to the RBC surface, which may lead to an undesirable immune response) and to substantially reduce cellular dehydration of the RBCs during pathogen inactivation of the whole blood. In some embodiments, the unwanted side reaction is modification of the RBC surface by the PIC. In some embodiments, the modification is covalent binding of the PIC to the RBC surface. In other embodiments, the modification is non-covalent binding of the PIC to the RBC surface.

[0079] As described herein, in some embodiments of each method, undesirable (also referred to herein as "unwanted") side reactions of the PIC with RBCs in whole blood are reduced. In some embodiments, the undesirable side reaction that is reduced is modification of the RBC surface by the PIC. In some embodiments, the level of the side reaction is reduced by at least about 5%, at least about 10%, at least about 25%, at least about 50%, at least about 75%, or at least about 90%. The reduction in side reactions can be demonstrated, for example, by measuring the amount of binding of an antibody specific for the PIC to the treated RBCs and / or measuring the in vivo lifespan of the treated RBCs in whole blood and comparing these measurements with RBCs treated by a second, different method (e.g., a method that does not add sufficient quencher and / or base to the reaction mixture, a method that does not add quencher and / or base to the reaction mixture, and / or a treatment performed at a lower pH).

[0080] In some embodiments, when the PIC and quencher are mixed (e.g., combined) with the whole blood composition, the pH of the mixture is in the range of about 6.0 to 8.5, about 6.0 to 7.5, about 6.5 to 7.1, about 6.5 to 7.0, about 6.6 to 6.8, or about 6.6, 6.7, 6.8, or 6.9. Although the pH in the whole blood composition may vary over time, it is desirable that the pH be within a desired range when the quencher is added to the whole blood composition, whether or not it already contains the PIC. In some embodiments, the methods of the present disclosure comprise adding the PIC and the quencher to the whole blood composition. Regardless of the order in which the PIC and / or the quencher are added to the whole blood composition, a desired pH range is required when both the PIC and the quencher are added. In other words, once all three components have been mixed, the pH is within the desired range. In some embodiments, the quencher is added before the PIC. In some embodiments, the PIC is added before the quencher. In some embodiments, the quencher and the PIC are added substantially simultaneously. Thus, "at the time of addition of the PIC and quencher" refers to the point in time when both the quencher and the PIC have been mixed with the whole blood composition. The desired pH can be achieved in several ways and is not limited to adjusting the pH of the whole blood composition, or in some embodiments, without significantly adjusting from the natural pH of the blood product. For example, the desired pH of the whole blood composition can be achieved by adjusting the pH. For example, the pH adjustment can be performed by adding a suitable solution, such as a buffer solution, before adding the PIC and quencher. Alternatively, the pH of the red blood cell composition can be adjusted simultaneously with the addition of the PIC, the quencher, or both. In some embodiments, the pH is adjusted simultaneously with the addition of the quencher. In some embodiments, the quencher is neutralized such that the addition of the neutralized quencher provides the desired pH range in the whole blood composition. For example, neutralization of glutathione can be used to achieve the necessary pH adjustment. In some embodiments, an appropriate level of neutralization of the glutathione can be used, such as by adding one equivalent of base, to provide a quencher that, when added to the whole blood composition, will provide the necessary pH adjustment of the composition. Appropriate neutralization will depend on the quencher used. For example, when using peptides, the pH of the quencher may depend on the amino acid composition of the peptide. In some embodiments, a quencher that does not significantly affect the pH of the red blood cell composition may be used. For example, the use of a peptide containing cysteine ​​results in a solution of the naturally isolated peptide having a more neutral pH. The peptide may also contain one or more amino acids. In some embodiments, the peptide further contains at least one basic amino acid, such as arginine or lysine.

[0081] In some embodiments of the methods described herein, a base is mixed with the whole blood composition along with the PIC and a quencher to increase the pH of the mixture to a desired level and / or improve the quenching of undesirable side reactions, and the base is a basic salt. The basic salt can be dissolved in an aqueous solution before being mixed (e.g., combined) with the whole blood composition. In other embodiments, the salt can be added directly to the whole blood composition in solid form. In some embodiments, the basic salt comprises a quencher and provides the quencher and base to the mixture. In some embodiments, the base used in the methods is a strong base, such as NaOH. Typically, a strong base such as NaOH is first dissolved in an aqueous solution and then mixed with the whole blood composition. In some embodiments, the strong base (e.g., a solution or solid form) is mixed with the quencher, and the quencher is then mixed with the whole blood composition. In some embodiments, the base is an alkaline buffer (added in sufficient amount and having an appropriate pKa to bring the mixture to the desired pH range). If an alkaline buffer is used, then in some embodiments, the buffer will be a pharmaceutically acceptable buffer. In some embodiments, the buffer will have a titratable proton with a pKa in the range of about 7 to 8. Examples of buffers that can be used as alkaline buffers include, but are not limited to, N-(2-hydroxyethyl)-piperazine-N'-2-ethanesulfonic acid (HEPES), phosphate buffered saline (PBS), and sodium phosphate buffer. Other suitable alkaline buffers will be readily identified by one of ordinary skill in the art.

[0082] In some embodiments of the methods and compositions described herein, the pH of the mixture of whole blood, quencher, PIC, and any added base is greater than about 5.5, greater than about 5.7, greater than about 6.0, greater than about 6.3, greater than about 6.5, greater than about 6.7, greater than about 7.0, or greater than about 7.2. In some embodiments of the methods and compositions described herein, the pH of the mixture of whole blood, quencher, PIC, and base (if added) is in the range of about 6.0 to 8.5, about 6.0 to 7.5, about 6.5 to 7.1, about 6.5 to 7.0, or about 6.6 to 6.8, or about 6.6, 6.7, 6.8, or 6.9. In some embodiments, the indicated pH is the pH at room temperature. In some embodiments, the indicated pH is the pH at 37°C. For example, in some embodiments, a composition comprising whole blood is treated with PIC in the presence of a quencher and any added base, wherein the pH of the mixture is in the range of about 6.5 to about 7.0 (or 7.1) at 37°C.

[0083] In some embodiments, the pH of the mixture of whole blood, quencher, and base (if base is added as part of the method) is in the range of about 6.0 to 8.5, about 6.0 to 7.5, about 6.5 to 7.1, about 6.5 to 7.0, or about 6.6 to 6.8, or about 6.5, 6.7, 6.8, or 6.9 before the PIC is mixed (e.g., combined) with the whole blood composition. In some other embodiments, the pH is achieved simultaneously with, within about 1 hour, about 30 minutes, about 20 minutes, about 10 minutes, about 5 minutes, or about 2 minutes of mixing the PIC with the composition comprising whole blood. In some embodiments of those methods in which the pH is adjusted, the pH is adjusted to the desired pH range prior to, simultaneously with, within about 1 hour, about 30 minutes, about 20 minutes, about 10 minutes, about 5 minutes, or about 2 minutes of mixing the PIC with the composition comprising whole blood. In those embodiments where the quencher is glutathione and the PIC is amataline (e.g., S-303), the pH of the mixture comprising the whole blood composition and the quencher is preferably adjusted to a desired pH range (e.g., pH 6.5 to 7.0) prior to mixing S-303 with the whole blood composition.

[0084] In some embodiments, the resulting pH of the composition after mixing (e.g., combining) whole blood, quencher, and base is not necessarily adjusted to the pH of the starting whole blood composition. For example, the whole blood composition may have a pH within a desired range of 6.0-7.5, and the pH of the composition does not change significantly upon addition of the quencher and subsequent addition of the PIC. In such embodiments, the quencher naturally provides the desired pH, or is neutralized to thereby provide the desired pH. The combination of adding a high initial amount of quencher, such as about 5 mM to about 40 mM, and obtaining a pH within the desired range is important. Thus, for peptides, regardless of the peptide quencher, it can be effectively neutralized as needed to provide a suitable pH range when added to the whole blood composition, and can further be selected so as to provide an appropriate amount of buffering within the desired pH range. Thus, a neutralized quencher refers to a quencher that is appropriately titrated with an acid or base as needed so that, when added to a whole blood composition, the resulting mixture has a pH that provides better quenching of unwanted side reactions (e.g., binding of PICs to the RBC surface, which can lead to undesirable immune responses) while avoiding cellular dehydration of the RBCs during inactivation, such as a pH in the range of about 6.0 to 8.5, about 6.0 to 7.5, about 6.5 to 7.0, about 6.5 to 7.1, or about 6.6 to 6.8, or about 6.6, 6.7, 6.8, or 6.9. In some embodiments, the peptides are appropriately neutralized when isolated naturally, i.e., no acid or base is required to provide the desired pH in the final mixture. In addition, preferred quenchers will provide a buffering capacity to maintain the pH within the desired range for the time necessary to quench unwanted side reactions.

[0085] In some embodiments of the methods and compositions described herein, the quencher is neutralized. A quencher is considered to be "neutralized" by a base if a sufficient amount of base has been combined with the quencher such that quenching of unwanted side reactions between the PIC and the RBCs of the whole blood in a mixture comprising a composition comprising whole blood, the PIC, and the quencher is improved. A "neutralized quencher" does not necessarily have a neutral pH and is not necessarily uncharged. In some embodiments, the neutralized quencher is neither in its most protonated form nor in its most deprotonated form. In some embodiments, when the quencher is very acidic, the pH of the neutralized quencher may still be below 7.0 (e.g., about 6.6, 6.7, 6.8, or 6.9). In some embodiments, the pH of a solution of the neutralized quencher may be greater than 7.0. In some embodiments, the pH of a solution of the neutralized quencher will be detectably higher than the quencher before the addition of the base. In some embodiments, the quencher is neutralized with at least about 0.25 equivalents, at least about 0.5 equivalents, at least about 0.75 equivalents, at least about 1 equivalent, at least about 1.25 equivalents, at least about 1.5 equivalents, or at least about 2 equivalents of a base. In some embodiments, the quencher is neutralized with less than about 2 equivalents, less than about 1.5 equivalents, less than about 1.25 equivalents, less than about 1 equivalent, or less than about 0.75 equivalents of a base. In some embodiments, the quencher is neutralized with about 0.25 to about 2 equivalents, about 0.5 to about 1.5 equivalents, or about 0.75 to about 1.25 equivalents of a base. In some embodiments, the quencher is neutralized with about 0.75 equivalents of a base. In other embodiments, the quencher is neutralized with about 1 equivalent of a base. In other embodiments, the quencher is neutralized with about 1.25 equivalents of a base. For example, in some embodiments of the present disclosure, glutathione is neutralized with about 1 equivalent of a suitable base such as sodium hydroxide. In this case, the protonated glutathione solution has a pH of approximately 3, the solution neutralized with 1 equivalent of sodium hydroxide has a pH of approximately 4.5, and the solution neutralized with 2 equivalents of sodium hydroxide has a pH of approximately 9.5. Any suitable peptide quencher comprising at least one cysteine ​​can be appropriately adjusted to provide the desired pH when added to the red blood cell composition.

[0086] Suitable methods for neutralizing glutathione and other quenchers will be apparent to those of ordinary skill in the art. In some embodiments, sodium hydroxide is used to neutralize or partially neutralize the quencher. In some embodiments, solid NaOH pellets are first dissolved in water to produce a concentrated base solution, such as a 1N, 5N, 10N, or 20N NaOH solution. In some embodiments, an appropriate amount of the NaOH solution is then added to the quencher before, simultaneously with, or after the quencher is added to the mixture. Alternatively, NaOH is added to the whole blood composition or the PIC, or a combination of the two, before the quencher is added to the mixture.

[0087] In addition to providing quenchers that are appropriately pH adjusted or neutralized, in some embodiments, preferred quenchers are unable to enter the pathogen in significant amounts once the PIC has penetrated the interior of the pathogen, such that they optimally quench unwanted reactions in the extracellular environment but do not interfere with pathogen inactivation.

[0088] In some embodiments of the methods described herein, the quencher is an acidic compound. In some embodiments, the quencher is provided in the form of a free acid. In some embodiments, the quencher is acidic and at least about 1 equivalent of base is added to neutralize the quencher. In some cases, the solution containing this neutralized quencher can be alkaline, neutral, or even acidic. In some embodiments, about 1 equivalent of base is added to neutralize or partially neutralize the quencher. In some embodiments, about 2 equivalents of base are added. In some embodiments, the quencher is acidic and about 0.5 to about 1.5 equivalents of base are used to neutralize the quencher. In some embodiments, about 0.75 to about 1.25 equivalents of base are used. In some embodiments, about 1 equivalent of base is used.

[0089] In some embodiments, the quencher is neutralized prior to addition to the whole blood composition and / or PIC. In other embodiments, the quencher is neutralized after combining the quencher with the whole blood composition and / or PIC. In some embodiments, the pH of the quencher neutralized prior to addition to the whole blood composition and / or PIC is in the range of about 2.5 to 7.5, about 3.0 to 6.5, about 3.5 to 5.5, about 4.0 to 5.0, or about 4.3 to 4.5, or about 4.4.

[0090] In some embodiments, the quencher is glutathione and is provided in the form of glutathione monosodium salt and neutralized with about 1 equivalent of base, or without base neutralization. In some other embodiments, the quencher is glutathione and is provided in the form of glutathione hydrochloride and neutralized with about 1 equivalent of base.

[0091] In some embodiments of the methods described herein, the initial quencher concentration in the mixture comprising the whole blood composition, the quencher, the PIC, and any added base is increased during the inactivation period and then reduced to a reduced concentration after the inactivation period. In some embodiments, the initial quencher concentration is sufficient to substantially reduce unwanted side reactions of the PIC (e.g., binding of pathogen-inactivating compounds to the RBC surface) and is then reduced to a reduced concentration to substantially reduce adverse effects on the viability (e.g., osmotic fragility and dehydration) and / or lifespan of the RBCs during storage of the treated whole blood composition.

[0092] Whole blood compositions and methods for pathogen inactivation

[0093] Certain aspects of the present disclosure provide pathogen-inactivated whole blood compositions and methods (e.g., methods of preparation) and kits related thereto. A particular benefit provided by the improvements disclosed herein is the opportunity to prepare (e.g., generate) pathogen-inactivated whole blood compositions suitable for transfusion having reduced levels of residual pathogen-inactivating compounds (PICs) in the composition. Reduced residual PIC levels in the treated whole blood compositions can provide improved safety characteristics and / or improved pathogen inactivation process consistency, such as achieving residual PIC levels below a certain threshold. The improved kits and related methods also provide the opportunity to prepare pathogen-inactivated whole blood compositions under conditions where suitable facilities and / or power sources may not be readily available, such as in war zones.

[0094] Whole blood compositions, such as whole blood compositions for use in the pathogen inactivation methods of the present invention, include but are not limited to any whole blood compositions (whole blood products) suitable for humans, mammals, and / or vertebrates, such as for transfusion. In some embodiments, the whole blood composition comprises whole blood from a human donor. In some embodiments, such whole blood cell compositions may include buffers, salts, nutrients, and other solutions, such as anticoagulants. In some embodiments, the whole blood composition of pathogen inactivation may include chemicals, such as pathogen inactivation compounds and quenchers. Suitable anticoagulants and their use are well known in the art and may include, for example, CPD (citric acid-phosphate-dextrose), citric acid-phosphate-dextrose (CP2D), or citric acid-phosphate-dextrose adenine (CPDA-1). The anticoagulant and whole blood composition may be mixed when collected from the donor, for example, the whole blood composition and the anticoagulant are mixed in a blood collection container (e.g., independent of the kit provided herein), or alternatively, the whole blood composition and the anticoagulant are mixed in the kit of the present invention. For example, a first container of a kit provided herein can contain an anticoagulant (eg, an anticoagulant solution) prior to adding (eg, collecting, transferring) the whole blood composition to the first container.

[0095] The whole blood compositions of the present disclosure (e.g., for pathogen inactivation treatment) can generally include any whole blood collection volume known in the art. In some embodiments, the whole blood volume in the whole blood composition is about 200mL to about 650mL, about 250mL to about 600mL, about 300mL to about 600mL, about 300mL to about 500mL, about 400mL to about 550mL, about 400mL to about 500mL, or about 450mL to about 500mL of whole blood (e.g., about 200mL, about 250mL, about 300mL, about 325mL, about 350mL, about 375mL, about 400mL, about 425mL, about 450mL, about 475mL, about 500mL, about 525mL, about 550mL, about 575mL, about 600mL or about 650mL). In some embodiments, the volume of whole blood is a certain change of the above volume, such as ±10% volume or ±50mL.

[0096] The whole blood composition may also include an anticoagulant solution before being transferred to the kit of the present invention (e.g., from a donor collection bag), or alternatively, may be mixed with an anticoagulant in the kit of the present invention. In some embodiments, the first container of the kit of the present invention may include an anticoagulant for mixing (e.g., merging) with the whole blood composition when the whole blood composition is added (e.g., transferred) to the container. Suitable volumes of anticoagulants for mixing with whole blood (e.g., in whole blood compositions) are well known in the art, and in some embodiments may include, for example, about 30 mL to about 120 mL, about 50 mL to about 120 mL, about 60 mL to about 120 mL, about 50 mL to about 100 mL, about 50 mL to about 75 mL, about 60 mL to about 100 mL, about 60 mL to about 90 mL, about 60 mL to about 80 mL, or about 60 mL to about 70 mL. In some embodiments, the anticoagulant of the present disclosure comprises about 30 mL, about 40 mL, about 45 mL, about 50 mL, about 55 mL, about 60 mL, about 65 mL, about 70 mL, about 75 mL, about 80 mL, about 85 mL, about 90 mL, about 95 mL, about 100 mL, about 105 mL, about 110 mL, or about 120 mL.

[0097] The volume of anticoagulant can be any volume provided herein, and when mixing whole blood, the volume results in a suitable anticoagulant to whole blood ratio (e.g., volume ratio). For example, in some embodiments, the ratio of anticoagulant to whole blood (e.g., in whole blood compositions) is about 1:3 to about 1:10, about 1:4 to about 1:10, about 1:5 to about 1:9, about 1:6 to about 1:8. In some embodiments, the ratio of anticoagulant to whole blood is about 1:3, about 1:4, about 1:5, or about 1:6. In some embodiments, the ratio of anticoagulant to whole blood is about 1:8, about 1:9, or about 1:10. In some embodiments, the ratio of anticoagulant to whole blood is about 1:7.

[0098] In some embodiments, a whole blood composition (e.g., comprising whole blood and an anticoagulant) for use in the pathogen inactivation methods of the present disclosure has a volume of about 200 mL to about 700 mL, about 250 mL to about 700 mL, about 300 mL to about 700 mL, about 200 mL to about 650 mL, about 250 mL to about 650 mL, about 250 mL to about 600 mL, about 300 mL to about 650 mL, about 300 mL to about 600 mL, about 300 mL to about 550 mL, about 300 mL to about 500 mL, about 400 mL to about 650 mL, about 400 mL to about 600 mL, about 400 mL to about 550 mL, about 450 mL to about 650 mL, about 450 mL to about 600 mL, about 450 mL to about 550 mL, about 450 mL to about 50 ... In some embodiments, the whole blood composition comprises about 400 mL to 500 mL of whole blood and about 50 mL to about 100 mL of anticoagulant. In some embodiments, the whole blood composition comprises about 450 mL to about 500 mL of whole blood and about 60 mL to about 70 mL of anticoagulant.

[0099] A whole blood composition (e.g., whole blood with an anticoagulant) can be transferred from a blood collection container to a kit of the present disclosure. In some embodiments, the blood collection container containing the whole blood composition can be coupled (e.g., connected) to the kit via a conduit compatible with blood products (e.g., hemocompatible), wherein the conduit provides a fluid communication path for transferring the whole blood composition (e.g., aseptically) into a container of the kit, such as a hemocompatible container (e.g., a blood product bag) of sufficient size for the combined volume of the whole blood composition, the PIC, and any quencher, anticoagulant, and / or other desired additives. For example, flexible tubing is commonly used in the art to couple (e.g., connect) one or more blood product containers (e.g., of a processing device or kit). In some embodiments, a conduit can be used to couple the blood collection container containing the whole blood composition to an inlet (e.g., a first inlet) of a container (e.g., a first container) of the kit, such as by aseptically docking two tubing segments (e.g., one tubing segment connected to the blood collection container and one tubing segment connected to a container of the kit) using a sterile connection device (e.g., an SCD, a tube welder) to provide the fluid communication path. After the whole blood composition is transferred to the container of the kit, the tubing can be sealed and disconnected, thereby disconnecting the blood collection container from the kit.

[0100] Alternatively, the whole blood composition can be collected directly from the donor into the kit of the present disclosure. For example, in some embodiments, the kit of the present disclosure includes a vascular access device (e.g., a needle) for extracting whole blood from the donor (e.g., by venipuncture), and a conduit (e.g., a tube, a flexible tube) extending between the vascular access device and the container of the kit (such as the first inlet of the first container). The conduit thereby provides a fluid communication path for transferring (e.g., aseptically) the whole blood being extracted from the donor using the vascular access device to the container of the kit. Typically, for embodiments in which whole blood is collected directly into a container of the kit (e.g., a first container), the container contains an anticoagulant for mixing (e.g., combining) with the whole blood (e.g., before mixing the whole blood and PIC). After the whole blood composition is collected into the container of the kit, in some embodiments, the conduit can be sealed and disconnected, thereby disconnecting the blood access device from the kit.

[0101] In some embodiments, prior to use in the treatment methods for pathogen inactivation described herein, whole blood compositions as described herein have been subjected to leukopenia (leukopenia). For example, a closed (e.g., functionally closed) system can be used to reduce the leukocytes of whole blood, such as by filtering through an online filter that is integral with or aseptically connected to a whole blood collection device or kit disclosed herein. Suitable leukopenia filters are known in the art. In some embodiments, whole blood compositions have not yet been subjected to leukopenia. Any whole blood compositions that will be contacted with or introduced into a living human, mammal, or vertebrate as disclosed herein can be processed, where such contact has the risk of spreading disease due to contaminated pathogens. Similarly, any whole blood compositions that will be used to prepare one or more other blood products (e.g., red blood cells, plasma, platelets, cryoprecipitate) that will be contacted with or introduced into a living human, mammal, or vertebrate as disclosed herein can be processed, where such contact has the risk of spreading disease due to contaminated pathogens.

[0102] In some embodiments, the disclosed methods for preparing a pathogen-inactivated whole blood composition comprise: (a) mixing (e.g., combining, adding) (i) a whole blood composition; (ii) an effective amount of a pathogen-inactivating compound (PIC); and (iii) optionally, an effective amount of a quencher in a first container; (b) incubating the mixture (e.g., admixture) of the whole blood composition, the PIC, and the optional quencher in the first container under conditions and for a duration sufficient to inactivate the pathogen, if present; (c) aseptically transferring the mixture to a second container; and (d) incubating the mixture in the second container under conditions and for a duration sufficient to reduce the concentration of the PIC to a concentration that is (i) at least 25% less than the concentration of the PIC in the mixture at the time of transferring step (b); or (ii) at least 10% less (e.g., lower) than the concentration of the PIC in the mixture of step (a) (e.g., the initial concentration, i.e., the concentration prior to incubating step (b)). 5 times the concentration; and / or (iii) is about 1 nM or less, thereby producing a pathogen-inactivated whole blood composition. Also provided herein is a pathogen-inactivated whole blood composition prepared by any method of the present disclosure.

[0103] In some embodiments of the methods provided herein, the concentration of PIC, such as amatarin, in the mixture (e.g., admixture) of step (a) (e.g., a mixture of a whole blood composition, an anticoagulant, a PIC, and an optional quencher) is at least about 0.05 mM, at least about 0.1 mM, or at least about 0.2 mM or greater. In some embodiments, the concentration of PIC in the mixture of step (a) is about 0.1 mM to about 10 mM. In some embodiments, the concentration of PIC in the mixture of step (a) is less than about 10 mM, less than about 5 mM, less than about 4 mM, less than about 3 mM, less than about 2 mM, less than about 1.0 mM, or less than about 0.5 mM. The concentration of PIC in the mixture of step (a) generally refers to the initial PIC concentration at the time the components of step (a) (e.g., the whole blood composition, the PIC, any anticoagulant, and any quencher) are mixed (e.g., immediately after combining, mixing) and prior to incubation step (b). In some embodiments, the concentration of PIC in the mixture of step (a) is about 0.05 mM to about 10.0 mM, about 0.05 mM to about 5.0 mM, about 0.05 mM to about 4.0 mM, about 0.05 mM to about 2.0 mM, about 0.05 mM to about 1.0 mM, about 0.05 mM to about 0.5 mM, about 0.1 mM to about 10 mM, about 0.1 mM to about 5 mM, about 0.1 mM to about 4 mM, about 0.1 mM to about 3 mM, about 0.1 mM to about 2 mM, about 0.1 mM to about 1.0 mM, about 0.1 mM to about 0.5 mM, or about 0.1 mM to about 0.3 mM. In some embodiments, the concentration of the PIC in the mixture of step (a) is about 0.05 mM, about 0.07 mM, about 0.08 mM, about 0.09 mM, 0.1 mM, 0.2 mM, 0.3 mM, 0.4 mM, 0.5 mM, 0.6 mM, 0.7 mM, 0.8 mM, 0.9 mM, 1.0 mM, about 2.0 mM, about 3.0 mM, about 4.0 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, or about 10 mM. In some embodiments, the concentration of the PIC in the mixture of step (a) is about 0.2 mM. In some embodiments, the PIC comprises a functional group that acts as or forms a reactive electrophilic group. In some embodiments, the functional group is selected from the group consisting of: mustard, mustard intermediate, and mustard equivalent. In some embodiments, the functional group is or is capable of forming an aziridinium ion. In some embodiments, the reactive electrophilic group is capable of reacting with nucleic acids. In some embodiments, the PIC comprises a nucleic acid-binding ligand. In some embodiments, the nucleic acid-binding ligand is an intercalator. In some embodiments, the intercalator is acridine. In some embodiments, the PIC comprises a frangible linker connecting the functional group and the nucleic acid-binding ligand. In some embodiments, the PIC is β-alanine, N-(acridin-9-yl), 2-[bis(2-chloroethyl)amino]ethyl ester.

[0104] In some embodiments of the methods provided herein, step (a) comprises mixing (e.g., combining, adding) a whole blood composition, an effective amount of PIC, and an effective amount of a quencher in a first container. In some embodiments, the concentration of the quencher in the mixture (e.g., admixture) of step (a) (e.g., initial concentration, i.e., concentration before incubation step (b)) is at least about 1 mM, at least about 2 mM, at least about 3 mM, at least about 4 mM, or at least about 5 mM. In some embodiments, the concentration of the quencher in the mixture of step (a) is less than about 25 mM, less than about 20 mM, less than about 15 mM, or less than about 10 mM. In some embodiments, the concentration of the quencher in the mixture of step (a) is from about 1 mM to about 25 mM. In some embodiments, the concentration of the quencher in the mixture of step (a) is from about 1 mM to about 20 mM. In some embodiments, the concentration of the quencher in the mixture of step (a) is from about 1 mM to about 15 mM. In some embodiments, the concentration of the quencher in the mixture of step (a) is about 1 mM to about 10 mM. In some embodiments, the concentration of the quencher in the mixture of step (a) is about 1 mM to about 5 mM. In some embodiments, the concentration of the quencher in the resulting mixture of step (a) is about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, or about 10 mM. In some embodiments, the concentration of the PIC in the mixture of step (a) is about 0.1 mM, and the concentration of the quencher in the admixture of step (a) is about 1 mM, about 2 mM, about 2 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, or about 10 mM. In some embodiments, the concentration of the PIC in the mixture of step (a) is about 0.2 mM, and the concentration of the quencher in the mixture of step (a) is about 1 mM, about 2 mM, about 2 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, or about 10 mM. In some embodiments, the concentration of the PIC in the mixture of step (a) is about 0.2 mM, and the concentration of the quencher in the mixture of step (a) is about 1 mM, about 2 mM, about 2 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, or about 10 mM. In some embodiments, the concentration of the PIC in the mixture of step (a) is about 0.2 mM and the concentration of the quencher in the mixture of step (a) is about 1 mM to about 5 mM. In some embodiments, the concentration of PIC in the mixture of step (a) is about 0.2 mM and the concentration of the quencher in the admixture of step (a) is about 2 mM. In some embodiments, the quencher comprises a sulfhydryl group, wherein the sulfhydryl group is capable of reacting with a reactive electrophilic group of the pathogen-inactivating compound. In some embodiments, the quencher comprises cysteine ​​or a derivative of cysteine.In some embodiments, the quencher is glutathione or a pharmaceutically acceptable salt thereof. In some embodiments, the quencher is glutathione monosodium salt.

[0105] In some embodiments, once the two components, the quencher and the pathogen-inactivating compound, have been mixed with the whole blood composition, the molar ratio of the quencher to the pathogen-inactivating compound is about 5:1 to about 100:1, about 5:1 to about 50:1, about 5:1 to about 40:1, about 5:1 to about 30:1, about 5:1 to about 20:1, about 5:1 to about 10:1, about 10:1 to about 100:1, about 10:1 to about 50:1, about 10:1 to about 40:1, about 10:1 to about 30:1, or about 10:1 to about 20:1. In some embodiments, once the two components, the quencher and the pathogen-inactivating compound, have been mixed with the whole blood composition, the molar ratio of the quencher to the pathogen-inactivating compound is about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, or about 20:1.

[0106] Any of the methods and kits disclosed herein can be used to add the PIC and optional quencher to a container (e.g., a first container) of the kit. In some embodiments, the PIC and optional quencher can be added to the first container of the kit disclosed herein. In some embodiments, the PIC and optional quencher are added to the first container through the first inlet of the first container. In some embodiments, both the PIC and quencher are added to the first container through the first inlet of the first container. In some embodiments, the PIC is added to the first container from a container containing the PIC (PIC container), wherein the PIC container is coupled to the first container. In some embodiments, the PIC container is coupled to the first inlet of the first container. In some embodiments, the PIC container is coupled to the second inlet of the first container. In some embodiments, the quencher is added to the first container from a container containing the quencher (quencher container), wherein the quencher container is coupled to the first container. In some embodiments, the quencher container is coupled to the first inlet of the first container. In some embodiments, the quencher container is coupled to the second inlet of the first container. In some embodiments, both the PIC container and the quencher container are coupled to the first inlet of the first container. In some embodiments, both the PIC container and the quencher container are coupled to the second inlet of the first container.

[0107] In some embodiments, the PIC and optional quencher are added to the first container via a conduit (e.g., a flexible tube) that extends (e.g., is coupled) between the PIC container and the optional quencher container and the first container and provides a fluid communication path for transferring the PIC from the PIC container to the first container and, optionally, for transferring the quencher from the optional quencher container to the first container. In some embodiments, the PIC and quencher are added to the first container via a conduit that extends between the PIC container and the quencher container and an inlet of the first container and provides a fluid communication path for transferring the PIC from the PIC container to the first container and transferring the quencher from the optional quencher container to the first container. In some embodiments, the PIC and quencher are added to the first container via a conduit that extends between the PIC container and the quencher container and a first inlet of the first container. In some embodiments, the PIC and quencher are added to the first container via a conduit that extends between the PIC container and the quencher container and a second inlet of the first container. In some embodiments, the PIC container and the quencher container are the same container. In some embodiments, the PIC and quencher are added to the first container via a conduit extending between the PIC container and the quencher container and an inlet of the first container, and the conduit comprises, at one end, (a) a first adapter for coupling the PIC container to the conduit, (b) a second adapter for coupling the quencher container to the conduit, and optionally (c) a displacement device (e.g., an air displacement device) adapted to increase transfer of the PIC and quencher through the conduit and into the first container. In some embodiments, the PIC and quencher are added to the first container via a conduit extending between the PIC container and the quencher container and an inlet of the first container, and the conduit comprises, at one end, (a) a first adapter for coupling the PIC container to the conduit, (b) a second adapter for coupling the quencher container to the conduit, and (c) a displacement device (e.g., an air displacement device) adapted to increase transfer of the PIC and quencher through the conduit and into the first container. In some embodiments, air is transferred from the displacement device (e.g., the air displacement device) into the conduit, thereby increasing transfer of the PIC and quencher into the first container (e.g., the percentage of fluid volume transferred). In some embodiments, fluid (e.g., saline) is transferred from the replacement device into the catheter, thereby increasing the transfer of the PIC and quencher into the first container (e.g., the percentage of fluid volume transferred). In some embodiments, the PIC container is coupled to the first adapter of the catheter after the catheter including the adapter is coupled to the first container. In some embodiments, the quencher container is coupled to the second adapter of the catheter after the catheter including the adapter is coupled to the first container.

[0108] In some embodiments, after the whole blood composition is added to the first container, a conduit extending between the PIC container and the optional quencher container and the inlet of the first container is coupled to the first container. In some embodiments, after the whole blood composition is added to the first container, a conduit extending between the PIC container and the quencher container and the inlet of the first container is coupled to the first container. In some embodiments, after the whole blood composition is added to the first container through the first inlet of the first container, a conduit extending between the PIC container and the optional quencher container and the inlet of the first container is coupled to the first inlet of the first container. In some embodiments, after the whole blood composition is added to the first container through the first inlet of the first container, a conduit extending between the PIC container and the optional quencher container and the inlet of the first container is coupled to the first inlet of the first container. In some embodiments, after the whole blood composition is added to the first container through the first inlet of the first container, a conduit extending between the PIC container and the quencher container and the inlet of the first container is coupled to the first inlet of the first container. For example, after transferring the whole blood composition from the blood collection container or vascular access device to the first container via a catheter that couples the blood collection container or vascular access device to the first inlet of the first container, the catheter can be sealed and disconnected, thereby disconnecting the blood collection container or vascular access device from the kit (e.g., from the first container). The PIC source (e.g., a PIC container) and the quencher source (e.g., a quencher container) can then be coupled to the remaining sealed and disconnected catheters extending from the first inlet of the first container of the kit. In some embodiments, both the PIC container and the quencher container are coupled to the catheter extending from the first container of the kit. In some embodiments, after adding the whole blood composition to the first container (e.g., and after coupling the catheter to the first container), the PIC container is coupled to the first adapter of the catheter and the quencher container is coupled to the second adapter of the catheter.

[0109] In some embodiments, a whole blood composition is added to the first container via a first inlet of the first container, and a PIC and optional quencher are added to the first container via a second inlet of the first container. In some embodiments, a conduit extending between the PIC container and optional quencher container and the inlet of the first container is coupled to the second inlet of the first container. For example, in some embodiments, a whole blood composition can be transferred from a blood collection container or vascular access device coupled (e.g., via a first conduit) to the first inlet of the first container to the first container. After transferring the whole blood composition, the conduit can optionally be sealed and disconnected, thereby disconnecting the blood collection container or vascular access device from the kit, and the PIC and optional quencher can then be transferred from a PIC source (e.g., a PIC container) and an optional quencher source (e.g., a quencher container) coupled (e.g., via a second conduit) to the second inlet of the first container to the first container. In some embodiments, the second conduit includes at least one adapter at one end for coupling the PIC container and / or quencher container to the second conduit. In some embodiments, the second conduit includes at one end a first adapter for coupling the PIC container to the conduit and a second adapter for coupling the quencher container to the conduit. In some embodiments, the second conduit is coupled to the second inlet of the first container prior to coupling the PIC container to the first adapter of the conduit or coupling the quencher container to the second adapter of the conduit.

[0110] After the whole blood composition, an effective amount of PIC, and optionally an effective amount of a quencher are added (e.g., transferred) to a container (e.g., a first container) of the kit (e.g., and mixed therein), the mixture (e.g., admixture) is incubated in the first container under conditions and for a duration sufficient to inactivate the pathogen (if present). In some embodiments, the incubation step (e.g., step (b)) is from about 2 hours to about 24 hours. In some embodiments, the incubation step (e.g., step (b)) is from about 6 hours to about 24 hours, from about 12 hours to about 24 hours, or from about 18 to about 24 hours. In some embodiments, the incubation step (e.g., step (b)) is from about 18 hours to about 22 hours (e.g., about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours). In some embodiments, the incubation step (e.g., step (b)) is about 20 hours. In some embodiments, the incubation step (b) is at about 18°C ​​to 25°C. In some embodiments, the incubation step (eg, step (b)) is at room temperature (eg, 22°C).

[0111] In some embodiments, after the incubation step in the first container (e.g., step (b)), the mixture comprising the whole blood composition, PIC, and an optional quencher is transferred (e.g., under sterile conditions) to a second container. In some embodiments, the second container is coupled to the first container. In some embodiments, the mixture is transferred (e.g., transfer step (c)) by a conduit (such as a flexible tube) extending (e.g., coupled, connected) between the first and second containers. In some embodiments, the conduit extends (e.g., connected, providing a fluid communication path) between the first outlet of the first container and the first inlet of the second container. In some embodiments, the method further comprises incubating the mixture in the second container (e.g., for a time sufficient to reduce the PIC). In some embodiments, the incubation step in the second container (e.g., step (d)) is about 1 hour to about 12 hours, about 1 hour to about 10 hours, about 1 hour to about 8 hours, about 1 hour to about 6 hours. In some embodiments, the incubation step (e.g., step (d)) is about 2 hours to about 4 hours. In some embodiments, the incubation step (e.g., step (d)) is about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, or about 12 hours or more. In some embodiments, the incubation step is at about 18°C ​​to 25°C. In some embodiments, the incubation step is at room temperature (e.g., 22°C). In some embodiments, after the incubation step, the pathogen-inactivated whole blood composition is stored in a second container. In some embodiments, the pathogen-inactivated whole blood composition is transferred from the second container to a third container. In some embodiments, the third container is coupled to the second container. In some embodiments, the pathogen-inactivated whole blood composition is transferred from the second container to the third container by a conduit (e.g., such as a flexible tube) extending (e.g., coupling, connection) between the second and third containers. In some embodiments, the conduit extends (e.g., connects, provides a fluid communication path) between the first outlet of the second container and the first inlet of the third container. In some embodiments, the pathogen-inactivated whole blood composition is stored in the third container. In some embodiments, the pathogen-inactivated whole blood composition is stored at about 2-6° C. (eg, stored in the second container, stored in the third container). In some embodiments, the pathogen-inactivated whole blood composition is suitable for transfusion (eg, transfusion of a human subject).

[0112] In some embodiments, after incubating the mixture in the second container (e.g., incubation step (d)), the concentration of the PIC decreases. In some embodiments, after incubating the mixture in the second container (e.g., incubation step (d)), the concentration of the PIC decreases to: (i) a concentration that is at least 25% less than the concentration of the PIC in the mixture at the time of transferring step (b); (ii) a concentration that is at least 10% less (e.g., lower) than the concentration of the PIC in the mixture in step (a) (e.g., the initial concentration, i.e., the concentration before incubation step (b)). 5 times the concentration; and / or (iii) is about 1 nM or less, thereby producing a pathogen-inactivated whole blood composition.

[0113] In some embodiments, after incubating the mixture in the second container (e.g., incubation step (d)), the concentration of the PIC is reduced to a concentration that is at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% less than the concentration of the PIC in the mixture at the time of transfer to the second container (e.g., transfer step (b)). In some embodiments, after incubating the mixture in the second container (e.g., incubation step (d)), the concentration of the PIC is reduced to a concentration that is at least 10% less than the concentration of the PIC in the mixture of step (a) (e.g., the initial concentration, i.e., the concentration before incubation step (b)). 4 times, at least 10 4.5 times, at least 10 5 times, at least 10 5.5 times, at least 10 6 times, at least 10 6.5 times, or at least 10 7 In some embodiments, after incubating the mixture in the second container (e.g., incubation step (d)), the concentration of the PIC decreases to a concentration of less than about 2.0 nM, less than about 1.5 nM, less than about 1.25 nM, less than about 1.0 nM, less than about 0.9 nM, less than about 0.8 nM, less than about 0.75 nM, or less than about 0.7 nM. In some embodiments, after incubating the mixture in the second container (e.g., incubation step (d)), the concentration of the PIC decreases to a concentration less than the limit of quantitation obtained using LCMS.

[0114] The pathogen-inactivated whole blood compositions of the present disclosure and blood components derived therefrom can be characterized for a variety of parameters (e.g., functional characterization, performed using analytical methods known in the art). Non-limiting examples include percent hemolysis, pH (e.g., at 37° C.), pO 2 , pCO 2 , HCO - 3. Total ATP, extracellular K + / Na +, extracellular glucose and lactate, hematocrit, hemoglobin content, albumin level, coagulation factor activity, calibrated automated thrombogram (CAT), thrombin generation capacity, prothrombin time (PT), activated partial thromboplastin time (aPTT), thrombin time (TT), rotational thromboelastometry (ROTEM), P-selectin, GPIIb / IIIa, and platelet aggregation.

[0115] In some embodiments, the method further comprises separating the whole blood composition of pathogen inactivation into one or more (for example, the red blood cell composition of pathogen inactivation, the plasma composition of pathogen inactivation, the platelet composition of pathogen inactivation). In some embodiments, the method further comprises separating the whole blood composition of pathogen inactivation into red blood cell composition and plasma composition. In some embodiments, the method further comprises separating the whole blood composition of pathogen inactivation into at least plasma composition, and preparing cold precipitate composition (for example, the cold precipitate composition of pathogen inactivation) by plasma composition. In some embodiments, preparing cold precipitate composition by plasma composition includes preparing cold precipitate reduced (for example, lacking cold precipitate (cryo-poor)) plasma composition (for example, the cold precipitate reduced plasma composition of pathogen inactivation). The method of whole blood being separated into red blood cell composition, plasma composition and / or platelet composition is well known in the art. In some embodiments, the whole blood of pathogen inactivation prepared according to the present disclosure is separated into one or more (for example, red blood cell composition, plasma composition and / or platelet composition) using methods known in the art. Likewise, methods for preparing cryoprecipitate compositions and cryoprecipitate-reduced (e.g., cryoprecipitate-lacking) plasma compositions from plasma are well known in the art. In some embodiments, cryoprecipitate compositions and / or cryoprecipitate-reduced plasma compositions are prepared from plasma obtained by separating pathogen-inactivated whole blood prepared according to the present disclosure into a plasma composition using methods known in the art.

[0116] In another aspect, the present disclosure provides a pathogen-inactivated whole blood composition prepared by any of the aforementioned methods. In some embodiments, the pathogen-inactivated whole blood composition is suitable for transfusion. In some embodiments, the pathogen-inactivated whole blood composition is a pathogen-inactivated whole blood unit suitable for transfusion. In some embodiments, the pathogen-inactivated whole blood composition suitable for transfusion comprises a PIC concentration of less than about 2.0 nM, less than about 1.5 nM, less than about 1.25 nM, less than about 1.0 nM, less than about 0.9 nM, less than about 0.8 nM, less than about 0.75 nM, or less than about 0.7 nM. Another aspect of the present disclosure provides a pathogen-inactivated red blood cell composition prepared by any of the aforementioned methods. Another aspect of the present disclosure provides a pathogen-inactivated plasma composition prepared by any of the aforementioned methods. Another aspect of the present disclosure provides a pathogen-inactivated platelet composition prepared by any of the aforementioned methods. Another aspect of the present disclosure provides a pathogen-inactivated cryoprecipitate composition prepared by any of the aforementioned methods. Another aspect of the present disclosure provides a pathogen-inactivated cryoprecipitate-reduced plasma composition prepared by any of the aforementioned methods.

[0117] In some embodiments, the methods of the present disclosure include ex vivo use of pathogen-inactivating compounds and quenchers. Ex vivo use includes using the compound to treat a whole blood composition externally on a living human, mammal, or vertebrate, where the treated biological material is intended for use internally on the living human, mammal, or vertebrate. For example, if the blood is intended to be reintroduced into a human or another human, then removing blood from the human and introducing the compound into the blood to inactivate pathogens is defined as ex vivo use of the compound. Reintroducing the human's blood into the human or another human would be an in vivo use of the blood, as opposed to ex vivo use of the compound. If the compound is still present in the blood when the blood is reintroduced into the human, then in addition to its ex vivo use, the compound is also introduced into the body. Some embodiments of the present disclosure relate to ex vivo use of the quencher, where the whole blood composition is intended for use in vivo. In some cases, a certain level of quencher is retained in the whole blood composition so that the quencher is also introduced into the body. Ex vivo use of a material or compound involves using the material or compound externally on a living human, mammal, or vertebrate, where the material or compound is not intended to be reintroduced into the living human, mammal, or vertebrate. An example of in vitro use would be diagnostic analysis of components of a whole blood sample. The methods of the present disclosure can be applied to in vitro use of whole blood compositions, as modifications of whole blood or other components can affect in vitro analysis of components of a blood sample. Therefore, the methods of the present disclosure can provide safety in processing such in vitro samples by substantially quenching modifications in the sample that might otherwise interfere with diagnostic testing of the sample.

[0118] Processed whole blood composition

[0119] In some embodiments, the present disclosure also provides whole blood compositions obtained by each of the processing methods described herein. In some embodiments, the present disclosure also provides whole blood compositions preparable by each of the processing methods described herein.

[0120] In some embodiments of each method and composition described herein, the whole blood in the whole blood composition is mammalian blood. For example, whole blood can be the whole blood of a rodent (e.g., mouse or rat), a canine, a lagomorph (e.g., rabbit), a non-human primate (e.g., chimpanzee) or a human. For example, in some embodiments, whole blood is human whole blood. In some embodiments, whole blood has been subjected to leukopenia. In some other embodiments, whole blood has not yet been subjected to leukopenia. In some embodiments, the composition comprising whole blood may be contaminated by pathogens. In some embodiments, the whole blood composition is contaminated by pathogens. In some embodiments, if present, at least 1 log, or at least 2 log, or at least 3 log, at least 4 log, at least 5 log, at least 6 log or more of the pathogen is inactivated in the composition.

[0121] The kits of the present disclosure are particularly useful for preparing one or more pathogen-inactivated blood components, such as plasma compositions, red blood cell compositions, and / or platelet compositions, from pathogen-inactivated whole blood compositions, e.g., as described herein. Additional pathogen-inactivated blood products, including, for example, cryoprecipitate and / or cryoprecipitate-reduced (e.g., "cryoprecipitate-deficient") plasma, can be prepared from such blood components.

[0122] The pathogen-inactivated whole blood compositions of the present disclosure can be used in a variety of applications known in the art. In certain aspects, provided herein are methods for transfusing the pathogen-inactivated whole blood compositions of the present disclosure into a subject in need thereof. In some embodiments, the subject is a human subject. Any whole blood composition of the present disclosure can be used in a transfusion method.

[0123] Whole Blood Processing and Kits

[0124] Certain aspects of the present disclosure relate to kits (e.g., pathogen inactivation kits). The kits and associated processing devices of the present disclosure are particularly useful for preparing pathogen-inactivated whole blood compositions and other pathogen-inactivated blood products, for example, as described herein. Any of the exemplary components described below, such as containers, bags, and conduits (e.g., tubing), can be used in the kits of the present disclosure.

[0125] In one aspect, a kit for preparing a pathogen-inactivated whole blood composition is provided, the kit comprising: a vascular access device (e.g., a needle, a venipuncture needle) for withdrawing whole blood from a donor; a first container suitable for mixing (e.g., combining, adding) the whole blood and a pathogen-inactivating compound (PIC), wherein the first container comprises (e.g., includes) at least a first inlet and a first outlet, and wherein the vascular access device is coupled to the first inlet of the first container; and a container comprising the pathogen-inactivating compound (PIC container). In some embodiments, a kit for preparing a pathogen-inactivated whole blood composition is provided, the kit comprising: (a) a vascular access device for withdrawing whole blood from a donor; (b) a first container suitable for mixing (e.g., combining, adding) the whole blood and a pathogen-inactivating compound (PIC), wherein the first container comprises at least a first inlet and a first outlet; (c) a first conduit extending between the vascular access device and the first inlet of the first container and providing a fluid communication path for transferring the whole blood from the vascular access device to the first container; and (d) a container (PIC container) containing the pathogen-inactivating compound, wherein the PIC container is configured to be coupled to the first container via a fluid communication path.

[0126] The container for use with whole blood compositions in the kit of the present disclosure is compatible (for example, blood compatibility) for the processing and / or storage of whole blood and / or other blood product compositions. Such container can include any sheet material (for example, PVC, EVA) known in the art for blood product containers, and its size is enough to include the whole blood of desired volume and any other fluid volume (for example, anticoagulant, PIC, quencher) mixed with the whole blood composition. The container can include one or more inlets (for example, ports, access ports) and one or more outlets (for example, ports, access ports) for access, so that fluid such as whole blood and / or pathogen inactivation compounds are transferred into or out of the container. Many suitable ports used in blood product containers are known in the art. In some embodiments, the inlet (for example, port, access port, entry port) can be "open", so as to allow fluid to enter the container, such as when a conduit (for example, tube) is coupled to the inlet (for example, so as to provide aseptic connection and fluid path). Alternatively or in addition, the inlet can be "openable", such as a fragile or pierceable port that allows fluid to enter (e.g., a sterile fluid path) container after the barrier is opened. Similarly, in some embodiments, the outlet (e.g., port, access port, nozzle port) can be open or openable. In some embodiments, the inlet and outlet of the container may have a substantially similar construction. In some embodiments, one or more containers of the present disclosure (e.g., the first container, the second container) include polyvinyl chloride (e.g., PVC, plasticized polyvinyl chloride). In some embodiments, one or more containers of the present disclosure (e.g., the first container, the second container) include polyethylene (e.g., ethylene-vinyl acetate, ethylene-vinyl acetate copolymer). In some embodiments, one or more containers of the present disclosure (e.g., the first container, the second container) include polyolefins (e.g., polyolefin blends). In some embodiments, one or more containers of the present disclosure (e.g., the first container, the second container) include polyvinylidene fluoride (PVDF). The conduits (e.g., tubes, flexible tubing) used with the whole blood compositions in the kits of the present disclosure are also compatible (e.g., blood compatible) with whole blood and / or other blood product compositions. In some embodiments, the containers and / or conduits of the present disclosure do not contain diethylhexyl phthalate (e.g., do not contain DEHP).

[0127] A vascular access device, such as a needle (e.g., a vascular access needle), is typically used to withdraw whole blood from a donor (e.g., by venipuncture) for collection into a container, such as a blood collection container. In some embodiments, the vascular access device is used to collect whole blood from the donor into a container of the present disclosure (e.g., transferred from the vascular access device to a first container of the kit). In some embodiments, the donor whole blood is transferred from the vascular access device to the first container of the kit of the present disclosure via a conduit (e.g., flexible hemocompatible tubing) extending between the vascular access device and the first container to provide a fluid communication path for aseptic transfer of the whole blood. In some embodiments, the vascular access device and the conduit constitute a needle assembly. The whole blood can be transferred (e.g., aseptically transferred) into the first container via an inlet (e.g., opening, port, access port) of the container coupled (e.g., connected, attached) to the conduit. The conduit can include a single continuous tubing segment, or more than one tubing segment, such as a tubing segment having one or more intervening components (e.g., connectors, filters, leukoreduction filters) that maintain the fluid communication path. The catheter may also include an internal frangible seal or closure (e.g., a frangible sleeve) that can be broken and opened by external manipulation of the tube (such as by bending the tube). Such frangible closures are well known in the art.

[0128] In some embodiments, the PIC container is configured to be coupled to the first inlet of the first container via a fluid communication path. In some embodiments, the PIC container is configured to be coupled to the first conduit. In some embodiments, the first container includes a second inlet and the PIC container is configured to be coupled to the second inlet of the first container via a fluid communication path. In some embodiments, the kit further includes a container containing a quencher (quencher container), wherein the quencher container is configured to be coupled to the first container via a fluid communication path. In some embodiments, the quencher container is configured to be coupled to the first inlet of the first container via a fluid communication path. In some embodiments, the quencher container is configured to be coupled to the first conduit. In some embodiments, the quencher container is configured to be coupled to the second inlet of the first container via a fluid communication path. In some embodiments, the PIC container and the quencher container are the same container (e.g., a container having more than one chamber). The PIC and quencher containers of the present disclosure may include any container of the kit suitable for storing the PIC or quencher, including, for example, a glass container (e.g., a vial), a plastic container (e.g., a syringe), a flexible bag, etc. In some embodiments, such PIC and / or quencher containers can be configured to couple (e.g., connect) to components of the kit (e.g., containers, conduits, processing devices) to allow fluid communication and transfer of the PIC and / or quencher. In some embodiments, the kits of the present disclosure may include a first PIC container and a second PIC container, wherein either or both of the first PIC container and the second PIC container are configured to couple to components of the kit (e.g., containers, conduits, processing devices). In some embodiments, the kits of the present disclosure may include a first quencher container and a second quencher container, wherein either or both of the first quencher container and the second quencher container are configured to couple to components of the kit (e.g., containers, conduits, processing devices). For example, in some embodiments, a kit (e.g., a processing device) can include a first PIC container and a first quencher container for reconstitution of a lyophilized PIC and a lyophilized quencher as glass vials, and a second PIC container and a second quencher container as syringes, which are configured to be coupled to a conduit or container of the kit (e.g., wherein the reconstituted PIC and quencher are transferred (e.g., aseptically) from their respective first containers to their respective second containers, which are configured to be coupled to a conduit or container of the kit). In some embodiments, coupling of the PIC and / or quencher container to a component of the kit (e.g., a conduit, a container) can be performed, for example, using an adapter (e.g., a Luer adapter). In some embodiments, coupling of the PIC and / or quencher container to a component of the kit (e.g., a container, a conduit, a processing device) can be performed, for example, using a conduit (e.g., aseptically docked using a tube welder).In some embodiments, the PIC and / or quencher container are coupled to components of the kit (e.g., a container, a conduit, a processing device) during the manufacturing process so as to be integrally connected within the kit. In some embodiments, the PIC container and the quencher container are the same container. For example, the PIC container and the quencher container can be portions (e.g., chambers, subcontainers) within a single container, e.g., to provide a configuration that requires only a single coupling (e.g., to a container, a conduit) to transfer the PIC and quencher to a container (e.g., a first container).

[0129] In some embodiments, the kit further comprises a second conduit, wherein the second conduit comprises at least one adapter at one end for coupling a PIC container (e.g., a second PIC container) and / or a quencher container (e.g., a second quencher container) to the second conduit. In some embodiments, the second conduit comprises at one end a first adapter for coupling the PIC container to the second conduit and a second adapter for coupling the quencher container to the second conduit. In some embodiments, the second conduit comprises at an end a 2-way branch having a first adapter and a second adapter. In some embodiments, the kit further comprises a second conduit, wherein the second conduit is coupled to the PIC container and / or the quencher container at one end. In some embodiments, the kit further comprises a second conduit, wherein the second conduit is coupled to the PIC container and / or the quencher container at one end (e.g., using a 2-way branch). In some embodiments, the second conduit (e.g., the other end of the second conduit) is configured to couple to the first container, thereby providing a fluid communication path between the PIC container and / or the quencher container and the first container. In some embodiments, the second conduit is configured to couple to the first inlet of the first container. In some embodiments, the second conduit is coupled to the second inlet of the first container. In some embodiments, the second conduit is configured to couple to the first conduit (eg, via a sterile docking) thereby providing a fluid communication path between the PIC container and / or quencher container and the first container.

[0130] In some embodiments, the kit further comprises a displacement device (e.g., an air displacement device) adapted to increase the transfer of the PIC and / or quencher into a container (e.g., the first container) of the present disclosure (e.g., through the second conduit and into the first container). Generally, a displacement device, such as an air displacement device, of the present disclosure is any device or component of the kit that can displace air and / or fluid from the displacement device in a manner that facilitates the movement or transfer of a fluid (e.g., PIC and / or quencher) through or from a conduit, such as, in the case of an air displacement device, by compressing the air displacement device so that air (e.g., or a suitable gas) pushes the fluid through a conduit coupled thereto to increase or maximize the transfer (e.g., expel the fluid) and reduce or minimize the volume of residual fluid in the conduit. A displacement device (e.g., an air displacement device) can be particularly useful when transferring a relatively small volume of fluid (e.g., PIC and / or quencher), such as from a conduit into a container, where residual fluid in the conduit (e.g., fluid that has not been drained by gravity) may affect the final concentration of the PIC and / or quencher when mixed with the whole blood composition. Air displacement devices contemplated for use with the kits of the present disclosure may include, but are not limited to, sample bypass transfer bags, syringes, rubber bulbs, flexible containers, cartridges (e.g., gas cartridges), and the like. Typically, the interior of the air displacement device and any gas (e.g., air) contained therein are sterile, e.g., sterilized as part of kit manufacture. In some preferred embodiments, the air displacement device is a sample bypass transfer bag (e.g., an air pillow, a sampling bag), examples of which are well known in the art of blood collection. Other displacement devices contemplated for use with the present disclosure may include, for example, a displacement device containing a fluid, such as a sterile saline solution in a container, which may be used to facilitate transfer of the PIC and / or quencher. In some embodiments, the displacement device (e.g., an air displacement device) is coupled to one end of a conduit (e.g., a second conduit), and the end of the conduit may also be coupled or configured to be coupled to a PIC container and / or a quencher container, such as using a tube branch connector (e.g., a 3-way branch).

[0131] In some embodiments, a PIC (e.g., in a PIC container) comprises any of the pathogen-inactivating compounds provided herein. In some embodiments, the PIC comprises a functional group that acts as or forms a reactive electrophilic group. In some embodiments, the functional group is selected from the group consisting of a mustard, a mustard intermediate, and a mustard equivalent. In some embodiments, the functional group is or is capable of forming an aziridinium ion. In some embodiments, the reactive electrophilic group is capable of reacting with a nucleic acid. In some embodiments, the PIC comprises a nucleic acid-binding ligand. In some embodiments, the nucleic acid-binding ligand is an intercalator. In some embodiments, the intercalator is an acridine. In some embodiments, the PIC comprises a frangible linker connecting the functional group and the nucleic acid-binding ligand. In some embodiments, the PIC is β-alanine, N-(acridin-9-yl), 2-[bis(2-chloroethyl)amino]ethyl ester.

[0132] In some embodiments, the quencher (e.g., in a quencher container) comprises any quencher provided herein. In some embodiments, the quencher comprises a sulfhydryl group, wherein the sulfhydryl group is capable of reacting with a reactive electrophilic group of the pathogen inactivating compound. In some embodiments, the quencher comprises cysteine ​​or a derivative of cysteine. In some embodiments, the quencher is glutathione or a pharmaceutically acceptable salt thereof. In some embodiments, the quencher is glutathione monosodium salt.

[0133] In some embodiments, the first container of the kit contains an anticoagulant (e.g., an anticoagulant solution), such as any suitable anticoagulant known in the art (e.g., and approved by one or more regulatory agencies and / or certification organizations) for use with whole blood. In some embodiments, the first container of the kit contains the anticoagulant before the whole blood composition is transferred to the container (e.g., when the whole blood is collected from the donor into the first container). In some embodiments, the anticoagulant is citrate-phosphate-dextrose (CPD), citrate-phosphate-dextrose (CP2D), or citrate-phosphate-dextrose adenine (CPDA-1). For example, the suitable volume of anticoagulant for mixing with whole blood (e.g., in whole blood compositions) is well known in the art, and in some embodiments may include, for example, about 30 mL to about 120 mL, about 50 mL to about 120 mL, about 60 mL to about 120 mL, about 50 mL to about 100 mL, about 50 mL to about 75 mL, about 60 mL to about 100 mL, about 60 mL to about 90 mL, about 60 mL to about 80 mL, or about 60 mL to about 70 mL. In some embodiments, the anticoagulant of the present disclosure comprises about 30 mL, about 40 mL, about 45 mL, about 50 mL, about 55 mL, about 60 mL, about 65 mL, about 70 mL, about 75 mL, about 80 mL, about 85 mL, about 90 mL, about 95 mL, about 100 mL, about 105 mL, about 110 mL or about 120 mL.

[0134] In some embodiments, the kit further comprises a second container suitable for storing a pathogen-inactivated whole blood composition, wherein the second container comprises (e.g., includes) at least a first inlet and a first outlet. In some embodiments, the kit further comprises a conduit (e.g., a third conduit) extending between the first outlet of the first container and the first inlet of the second container (e.g., and providing a fluid communication path for transferring (e.g., under sterile conditions) a mixture comprising the whole blood composition and the PIC (and a quencher, if present) from the first container to the second container). In some embodiments, the kit comprises a third container suitable for storing a pathogen-inactivated whole blood composition, wherein the third container comprises (e.g., includes) at least a first inlet and a first outlet, and the kit further comprises a conduit (e.g., a fourth conduit) extending between the first outlet of the second container and the first inlet of the third container (e.g., and providing a fluid communication path for transferring (e.g., under sterile conditions) a mixture comprising the whole blood composition and the PIC (and a quencher, if present) from the second container to the third container).

[0135] The kit of the present disclosure can be used in combination with an infusion line (e.g., an infusion line assembly, an intravenous administration device) to infuse the whole blood composition of pathogen inactivation into a subject. In some embodiments, the kit includes an infusion line assembly configured to be coupled (e.g., connected) to the outlet (e.g., a sharp mouth port) of the kit container. In some embodiments, the kit includes an infusion line assembly coupled (e.g., connected) to the outlet (e.g., a sharp mouth port) of the kit container. In some embodiments, after the whole blood composition of pathogen inactivation is prepared and before infusion, the infusion line assembly is coupled (e.g., connected) to the outlet (e.g., a sharp mouth port) of the kit container. In some embodiments, the infusion line is connected to the outlet of the container (e.g., the second container, the third container) of the kit (e.g., before preparing the whole blood composition of pathogen inactivation). In some embodiments, the infusion line assembly is integrally connected to the outlet of the kit container (e.g., before processing, during the manufacture of the kit). In some embodiments, the kit further comprises an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject, wherein the infusion line comprises a second vascular access device (e.g., a needle, an infusion needle) and an infusion line conduit extending between the first container, the second container, or the third container and the second vascular access device, wherein the infusion line conduit provides a fluid communication path for aseptically transferring the pathogen-inactivated whole blood composition from the container (e.g., the first container, the second container, the third container) to the vascular access device (e.g., and aseptically transferring to a donor accessed through the vascular access device). In some embodiments, the kit further comprises an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject, wherein the infusion line assembly comprises a second vascular access device and an infusion line conduit extending between the first outlet of the second container and the second vascular access device, wherein the infusion line conduit provides a fluid communication path for aseptically transferring the pathogen-inactivated whole blood composition from the second container to the vascular access device. In some embodiments, the infusion line assembly provides an openable fluid communication path between a container (e.g., first, second, third) of the kit and a second vascular access device. The infusion line assembly as provided herein is generally suitable for administering or infusing (e.g., aseptically transferring) a pathogen-inactivated whole blood composition from a container of the kit to a subject requiring whole blood transfusion.

[0136] In another aspect, the present disclosure provides a kit for preparing a pathogen-inactivated whole blood composition, comprising: a first container suitable for mixing a whole blood composition, a pathogen-inactivating compound (PIC), and a quencher, wherein the first container comprises (e.g., includes) at least a first inlet and a first outlet; a second container suitable for storing the pathogen-inactivated whole blood composition, wherein the second container comprises (e.g., includes) at least the first inlet and the first outlet, and wherein the second container is coupled to the first container; a container comprising the pathogen-inactivating compound (PIC container); a container comprising the quencher (quencher container); a conduit, wherein the conduit is coupled or configured to be coupled to the PIC container and the quencher container at one end, and wherein the conduit further comprises a displacement device (e.g., an air displacement device) adapted to increase transfer of the PIC and quencher through the first conduit. In some embodiments, the present disclosure provides a kit for preparing a pathogen-inactivated whole blood composition, comprising: (a) a first container suitable for mixing a whole blood composition, a pathogen-inactivating compound (PIC), and a quencher, wherein the first container comprises (e.g., includes) at least a first inlet and a first outlet, (b) a second container suitable for storing the pathogen-inactivated whole blood composition, wherein the second container comprises (e.g., includes) at least a first inlet and a first outlet; (c) a container comprising a pathogen-inactivating compound (PIC container), (d) a container comprising a quencher (quencher container), (e) a first conduit, wherein the first conduit is coupled to or configured to be coupled to the PIC container and the quencher container at one end, and wherein the first conduit further comprises a displacement device (e.g., an air displacement device) adapted to increase transfer of the PIC and quencher through the first conduit; and (f) a second conduit extending between the first container and the second container and providing a fluid communication path (e.g., an openable fluid communication path) for transferring the whole blood composition from the first container to the second container.

[0137] In some embodiments, the first conduit further comprises at least one adapter for coupling the second conduit to the PIC container and the quencher container. In some embodiments, the first conduit comprises a first adapter for coupling the PIC container to the first conduit and a second adapter for coupling the quencher container to the first conduit. In some embodiments, the first conduit is coupled to the PIC container and the quencher container. In some embodiments, the PIC container and the quencher container are the same container. In some embodiments, the first conduit is configured to couple to the first container (e.g., providing a fluid communication path between the PIC container and the quencher container and the first container). In some embodiments, the first conduit is configured to couple to an inlet of the first container. In some embodiments, the kit further comprises a vascular access device (e.g., a needle) for withdrawing whole blood from a donor (e.g., by venipuncture), wherein the vascular access device is coupled to the first container (e.g., via a conduit extending between the vascular access device and the container). In some embodiments, the kit comprises a conduit (e.g., a third conduit) extending between the vascular access device and a first container, such as a first inlet of the first container (e.g., and providing a fluid communication path for transferring whole blood from the vascular access device to the first container). In some embodiments, the first container further comprises (e.g., includes) a second inlet, wherein the first conduit is configured to be coupled to the second inlet of the first container. In some embodiments, the first container further comprises (e.g., includes) a second inlet and the first conduit is coupled to the second inlet of the first container. In some embodiments, the first conduit is configured to be coupled to a third conduit (e.g., so as to provide a fluid communication path between the PIC container and the quencher container and the first container). In some embodiments, the first container of the kit comprises an anticoagulant (e.g., an anticoagulant solution), such as any suitable anticoagulant for use with whole blood known in the art (e.g., and approved by one or more regulatory agencies and / or certification organizations). In some embodiments, before the whole blood composition is transferred to the container (e.g., when whole blood is collected from the donor into the first container), the first container of the kit comprises an anticoagulant. In some embodiments, the anticoagulant is citrate-phosphate-dextrose (CPD), citrate-phosphate-dextrose (CP2D) or citrate-phosphate-dextrose adenine (CPDA-1). Suitable volumes of anticoagulant for mixing with whole blood (e.g., in a whole blood composition) are well known in the art and, in some embodiments, may include, for example, about 30 mL to about 120 mL, about 50 mL to about 120 mL, about 60 mL to about 120 mL, about 50 mL to about 100 mL, about 50 mL to about 75 mL, about 60 mL to about 100 mL, about 60 mL to about 90 mL, about 60 mL to about 80 mL, or about 60 mL to about 70 mL.In some embodiments, the anticoagulant of the present disclosure comprises about 30 mL, about 40 mL, about 45 mL, about 50 mL, about 55 mL, about 60 mL, about 65 mL, about 70 mL, about 75 mL, about 80 mL, about 85 mL, about 90 mL, about 95 mL, about 100 mL, about 105 mL, about 110 mL, or about 120 mL.

[0138] In some embodiments, the PIC comprises a functional group that acts as or forms a reactive electrophilic group. In some embodiments, the functional group is selected from the group consisting of a mustard, a mustard intermediate, and a mustard equivalent. In some embodiments, the functional group is or is capable of forming an aziridinium ion. In some embodiments, the reactive electrophilic group is capable of reacting with a nucleic acid. In some embodiments, the PIC comprises a nucleic acid-binding ligand. In some embodiments, the nucleic acid-binding ligand is an intercalator. In some embodiments, the intercalator is an acridine. In some embodiments, the PIC comprises a frangible linker connecting the functional group and the nucleic acid-binding ligand. In some embodiments, the PIC is β-alanine, N-(acridin-9-yl), 2-[bis(2-chloroethyl)amino]ethyl ester.

[0139] In some embodiments, the quencher comprises a sulfhydryl group, wherein the sulfhydryl group is capable of reacting with a reactive electrophilic group of the pathogen-inactivating compound. In some embodiments, the quencher comprises cysteine ​​or a derivative of cysteine. In some embodiments, the quencher is glutathione or a pharmaceutically acceptable salt thereof. In some embodiments, the quencher is glutathione monosodium salt.

[0140] In some embodiments, the kit includes a third container suitable for storing a pathogen-inactivated whole blood composition, wherein the third container comprises (e.g., includes) at least a first inlet and a first outlet, and wherein the third container is coupled to the second container. In some embodiments, the kit also includes a fourth conduit extending between the first outlet of the second container and the first inlet of the third container (e.g., and providing a fluid communication path for transferring the whole blood composition from the second container (e.g., under sterile conditions) to the third container). In some embodiments, the kit also includes an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject, wherein the infusion line assembly includes a second vascular access device and an infusion line conduit extending between the first outlet (e.g., a nosepiece port) of the second container and the second vascular access device (e.g., wherein the infusion line conduit provides a fluid communication path for aseptically transferring the pathogen-inactivated whole blood composition from the second container to the vascular access device). In some embodiments, the kit further includes an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject, wherein the infusion line assembly includes a second vascular access device and an infusion line conduit extending between a first outlet (e.g., a nose port) of the third container and the second vascular access device (e.g., wherein the infusion line conduit provides a fluid communication path for aseptically transferring the pathogen-inactivated whole blood composition from the third container to the vascular access device).

[0141] In some embodiments, the kits of the present disclosure further include any one or more of a removable clamp, a frangible connector, an in-line filter, and or a sampling container (eg, a sampling bag, a bypass transfer bag).

[0142] A non-limiting example of an exemplary kit for preparing a pathogen-inactivated whole blood composition according to the methods disclosed herein is provided in Figures 1a-1c , 2a-2g, 3a-3c, 4a-4d and 5a-5g.

[0143] Figure 1aThe exemplary kit 100 shown comprises (e.g., includes): a first container 105 suitable for mixing a whole blood composition, a pathogen-inactivating compound (PIC), and an optional quencher, wherein the first container includes a first inlet 110 and a first outlet 115; a container 120 containing the pathogen-inactivating compound (PIC container); a container 125 containing the quencher (quencher container); a conduit 130 configured to couple the PIC container 120 and the quencher container 125 to the first container 105; and an optional infusion line assembly 145 (e.g., configured to couple to the first outlet 115 of the first container 105) for infusing the pathogen-inactivated whole blood composition into a subject. In some embodiments, the conduit 130 includes at one end (e.g., via a 2-way conduit branch): (a) a first adapter 135 for coupling (e.g., configured to couple) the PIC container to the conduit, and (b) a second adapter 140 for coupling (e.g., configured to couple) the quencher container to the conduit, and wherein the conduit is configured to couple to the first inlet 110 of the first container. In some embodiments, the conduit 130 is configured to couple directly to the first inlet 110. In some embodiments, the conduit 130 is configured to couple to the first inlet 110 by coupling to an optional conduit 142 (e.g., a conduit 142 extending from the first inlet 110). In some embodiments, the optional infusion line assembly 145 is configured to couple to the first container 105 (e.g., the first outlet 115 of the first container; via a tip port). The dashed lines indicate that, in some embodiments, a whole blood (WB) composition can be provided to the kit 100 from or in a blood collection container 150 and can be added to the first container 105 of the kit 100 by coupling (e.g., aseptically docking) the blood collection container (e.g., tubing 152 extending from the blood collection container) to the first inlet 110 of the first container 105 (e.g., directly to the first inlet 110, through a conduit 142 extending from the first inlet 110). In some embodiments, the blood collection container (150) can be coupled to the first container (105) (e.g., directly or to the conduit 142) to allow the whole blood to be transferred to the first container, and then the blood collection container (150) can be disconnected from the first container (105), such as using a tube welder, and the conduit (130) can be coupled to the first container (105) (e.g., directly or to the conduit 142) to allow the pathogen inactivating compound and quencher to be transferred to the first container (105).

[0144] An alternative configuration of the exemplary kit of the present disclosure is Figure 1b Similar to Figure 1a, an exemplary kit 100 includes: a first container 105 including a first inlet 110 and a first outlet 115; a PIC container 120; a quenchant container 125; a conduit 130 configured to couple the PIC container 120 and the quenchant container 125 to the first container 105; an optional infusion line assembly 145; and optionally including at one end of the conduit 130 (e.g., via a conduit branch): (a) a first adapter 135 for coupling the PIC container to the conduit, and (b) a second adapter 140 for coupling the quenchant container to the conduit. This configuration also includes a displacement device (e.g., an air displacement device) 155 adapted to improve (e.g., increase) the transfer of the PIC and quenchant into the container 105 (e.g., through the conduit 130 and into the container 105). In some embodiments, a displacement device (e.g., an air displacement device) 155 is coupled to the conduit 130 (e.g., via a 3-way conduit branch), which includes a first adapter 135 and a second adapter 140 at the ends of the conduit for coupling to the PIC container 120 and the quencher container 125, respectively. Non-limiting examples of displacement devices useful in the kits and methods of the present disclosure include sampling bags, bypass transfer bags, air cushions, balls, syringe barrels, containers containing fluid (e.g., saline), etc. The dotted line indicates that a whole blood (WB) composition can be provided to the kit 100 via the blood collection container 150.

[0145] An alternative configuration of the exemplary kit of the present disclosure is Figure 1c Similar to Figure 1b , an exemplary kit 100 includes: a first container 105 including a first inlet 110 and a first outlet 115; a PIC container 120; a quencher container 125; a displacement device (e.g., an air displacement device) 155; a conduit 130 configured to couple the PIC container 120 and the quencher container 125 to the first container 105; and an optional infusion line assembly 145. This configuration includes, at one end of the conduit 130 (e.g., via a conduit branch), the PIC container 120 coupled to the conduit, the quencher container 125 coupled to the conduit, and the displacement device (e.g., an air displacement device) 155 coupled to the conduit, which is suitable for improving (e.g., increasing) the transfer of the PIC and quencher into the container 105 (e.g., through the conduit 130 and into the container 105). The dotted line indicates that a whole blood (WB) composition can be provided to the kit 100 via a blood collection container 150.

[0146] Figure 2aThe exemplary kit 200 shown includes (e.g., comprises): a first container 205 suitable for mixing a whole blood composition, a pathogen-inactivating compound (PIC), and an optional quencher, wherein the first container 205 includes a first inlet 210, a second inlet 212, and a first outlet 215; a container 220 containing the pathogen-inactivating compound (PIC container); a container 225 containing the quencher (quencher container); a conduit 230 coupled to the first container 205 at one end (e.g., the second inlet 212 of the first container 205) and configured at the other end to couple the PIC container 220 and the quencher container 225 (e.g., to provide a fluid communication path between the first container and the PIC container 220 and the quencher container 225, respectively); and an optional infusion line assembly 245 for infusing the pathogen-inactivated whole blood composition into a subject. In some embodiments, the conduit 230 includes at one end (e.g., via a 2-way conduit branch): (a) a first adapter 235 for coupling (e.g., configured to couple) a PIC container to the conduit 230, and (b) a second adapter 240 for coupling (e.g., configured to couple) a quencher container to the conduit 230, and wherein the conduit is coupled (e.g., at the other end) to the second inlet 212 of the first container 205. In some embodiments, the infusion line assembly 245 is configured to couple to the first container 205 (e.g., the first outlet 215 of the first container; via a nosepiece port). The dashed lines indicate that a whole blood (WB) composition can be provided to the kit 200 from or therein a blood collection container 250 and can be added to the first container 205 of the kit 200 by coupling (e.g., aseptically docking) the blood collection container (e.g., a tube 252 extending from the blood collection container) to the first inlet 210 of the first container 205 (e.g., directly coupling to the first inlet 210, coupling through a conduit 242 extending from the first inlet 210).

[0147] An alternative configuration of the exemplary kit of the present disclosure is Figure 2b Similar to Figure 2a, the exemplary kit 200 includes: a first container 205 including a first inlet 210, a second inlet 212, and a first outlet 215; a PIC container 220; a quenchant container 225; a conduit 230 coupled to the first container 205 at one end and configured to couple the PIC container 220 and the quenchant container 225 at the other end; an optional infusion line assembly 245; and optionally including at one end of the conduit 230 (e.g., via a conduit branch): (a) a first adapter 235 for coupling the PIC container to the conduit; and (b) a second adapter 240 for coupling the quenchant container to the conduit. This configuration also includes a displacement device (e.g., an air displacement device) 255 adapted to improve (e.g., increase) the transfer of the PIC and quenchant into the container 205 (e.g., through the conduit 230 and into the container 205). In some embodiments, a displacement device (e.g., an air displacement device) 255 is coupled to a conduit 230 (e.g., via a 3-way conduit branch), which includes a first adapter 235 and a second adapter 240 at the ends of the conduit for coupling to the PIC container 220 and the quencher container 225, respectively. The dotted lines indicate that a whole blood (WB) composition can be provided to the kit 200 via the blood collection container 250.

[0148] An alternative configuration of the exemplary kit of the present disclosure is Figure 2c Similar to Figure 2b , an exemplary kit 200 includes: a first container 205 including a first inlet 210, a second inlet 212, and a first outlet 215; a PIC container 220; a quencher container 225; a displacement device (e.g., an air displacement device) 255; a conduit 230 coupling the first container 205 to the PIC container 220 and the quencher container 225 (e.g., and providing a fluid communication path therebetween); and an optional infusion line assembly 245. This configuration includes, at one end of the conduit 230 (e.g., via a conduit branch), the PIC container 220 coupled to the conduit, the quencher container 225 coupled to the conduit, and the displacement device (e.g., an air displacement device) 255 coupled to the conduit, which is adapted to improve (e.g., increase) the transfer of the PIC and quencher into the container 205 (e.g., through the conduit 230 and into the container 205). The dashed line indicates that a whole blood (WB) composition can be provided to the kit 200 via the blood collection container 250.

[0149] An alternative configuration of the exemplary kit of the present disclosure is Figure 2d Similar to Figure 2c, an exemplary kit 200 includes: a first container 205 including a first inlet 210, a second inlet 212, and a first outlet 215; a PIC container 220; a quencher container 225; a displacement device 255; and a conduit 230 that couples the first container 205 with the PIC container 220 and the quencher container 225 (e.g., and provides a fluid communication path therebetween). This configuration also includes an infusion line assembly 245, wherein the infusion line assembly is coupled to the first container 205 (e.g., to the first outlet 215). The dashed line indicates that a whole blood (WB) composition can be provided to the kit 200 via a blood collection container 250.

[0150] An alternative configuration of the exemplary kit of the present disclosure is Figure 2e Similar to Figure 2d , an exemplary kit 200 includes: a first container 205 including a first inlet 210, a second inlet 212, and a first outlet 215; a PIC container 220; a quencher container 225; a displacement device 255; a conduit 230 coupling the first container 205 with the PIC container 220 and the quencher container 225 (e.g., and providing a fluid communication path therebetween); and an infusion line assembly 245. This configuration also includes a vascular access device 260 for withdrawing whole blood from a donor (e.g., into the container 205); and a conduit 261 extending between the vascular access device 260 and the first inlet 210 of the first container. In some embodiments, the vascular access device 260 and the conduit 261 together comprise a needle assembly. In some embodiments, the container 205 contains an anticoagulant (e.g., an anticoagulant solution) 265.

[0151] An alternative configuration of the exemplary kit of the present disclosure is Figure 2f Similar to Figure 2e , exemplary kit 200 includes: a first container 205 including a first inlet 210, a second inlet 212, and a first outlet 215; a PIC container 220; a replacement set 255; a catheter 230 coupling the first container 205 with the PIC container 220 (e.g., and providing a fluid communication path therebetween); an infusion line assembly 245; a vascular access device 260; and a catheter 261. In some embodiments, container 205 contains an anticoagulant 265.

[0152] An alternative configuration of the exemplary kit of the present disclosure is Figure 2g Similar to Figure 2f, an exemplary kit 200 includes: a first container 205 including a first inlet 210, a second inlet 212, and a first outlet 215; a PIC container 220; a catheter 230 coupling the first container 205 with the PIC container 220 (e.g., and providing a fluid communication path therebetween); an infusion line assembly 245; a vascular access device 260; and a catheter 261. In some embodiments, the container 205 contains an anticoagulant 265.

[0153] Figure 3aThe exemplary kit 300 shown comprises (e.g., includes): a first container 305 suitable for mixing a whole blood composition, a pathogen inactivating compound (PIC), and an optional quencher, wherein the first container includes a first inlet 310 and a first outlet 315; a second container 307 suitable for storing the pathogen-inactivated whole blood composition, wherein the second container 307 includes a first inlet 316 and a first outlet 317, and an optional second outlet 319 (e.g., coupled to or configured to be coupled to a bypass transfer bag); a container 320 containing a pathogen-inactivating compound (PIC container); a container 325 containing a quencher (quencher container); a conduit 330 configured to couple the PIC container 320 and the quencher container 325 to the first container 305; a conduit 332 coupling the first container 305 and the second container 307; and an optional infusion line assembly 345 for infusing the pathogen-inactivated whole blood composition into a subject. In some embodiments, the conduit 330 includes at one end (e.g., by a 2-way conduit branch): (a) a first adapter 335 for coupling (e.g., configured to couple) the PIC container to the conduit, (b) a second adapter 340 for coupling (e.g., configured to couple) the quencher container to the conduit, and wherein the conduit is configured to couple to the first inlet 310 of the first container. In some embodiments, the conduit 330 is configured to couple directly to the first inlet 310 of the container 305. In some embodiments, the conduit 330 is configured to couple to the first inlet 310 by coupling to an optional conduit 342 (e.g., a conduit 342 extending from the first inlet 310 of the container 305). In some embodiments, the optional infusion line assembly 345 is configured to couple to the second container 307 (e.g., the first outlet 317 of the second container; via a tip port). The dotted lines indicate that a whole blood (WB) composition can be provided to the kit 300 from or in a blood collection container 350 and can be added to the first container 305 of the kit 300 by coupling (e.g., aseptically docking) the blood collection container (e.g., tubing 352 extending from the blood collection container) to the first inlet 310 of the first container 305 (e.g., directly to the first inlet 310, through a conduit 342 extending from the first inlet 310). In some embodiments, the blood collection container 350 can be coupled to the first container 305 to allow the whole blood to be transferred into the first container 305, and then the blood collection container 350 can be disconnected from the first container 305, such as using a tube welder, and the conduit 330 can be coupled to the first container 305 (e.g., directly to the first inlet 310, through a conduit 342 extending from the first inlet 310) to allow the pathogen inactivating compound and quencher to be transferred to the first container 305.

[0154] An alternative configuration of the exemplary kit of the present disclosure is Figure 3bSimilar to Figure 3a , an exemplary kit 300 includes: a first container 305 including a first inlet 310 and a first outlet 315; a PIC container 320; a quenchant container 325; a conduit 330 configured to couple the PIC container 320 and the quenchant container 325 to the first container 305; a second container 307 including a first inlet 316, a first outlet 317, and an optional second outlet 319; a conduit 332; an optional infusion line assembly 345; and optionally including at one end of the conduit 330 (e.g., via a conduit branch): (a) a first adapter 335 for coupling the PIC container to the conduit; and (b) a second adapter 340 for coupling the quenchant container to the conduit. This configuration also includes a displacement device (e.g., an air displacement device) 355 adapted to improve (e.g., increase) the transfer of the PIC and quenchant into the container 305 (e.g., through the conduit 330 and into the container 305). In some embodiments, a displacement device (e.g., an air displacement device) 355 is coupled to a conduit 330 (e.g., via a 3-way conduit branch), which includes a first adapter 335 and a second adapter 340 at the ends of the conduit for coupling to the PIC container 320 and the quencher container 325, respectively. The dotted line indicates that a whole blood (WB) composition can be provided to the kit 300 via a blood collection container 350.

[0155] An alternative configuration of the exemplary kit of the present disclosure is Figure 3c Similar to Figure 3b , an exemplary kit 300 includes: a first container 305 including a first inlet 310 and a first outlet 315; a PIC container 320; a quenchant container 325; a conduit 330 configured to couple the PIC container 320 and the quenchant container 325 to the first container 305; a displacement device (e.g., an air displacement device) 355; a second container 307 including a first inlet 316, a first outlet 317, and an optional second outlet 319; a conduit 332; and an optional infusion line assembly 345. This configuration includes, at one end of the conduit 330 (e.g., via a conduit branch), the PIC container 320 coupled to the conduit, the quenchant container 325 coupled to the conduit, and the displacement device (e.g., an air displacement device) 355 coupled to the conduit, which is suitable for improving (e.g., increasing) the transfer of the PIC and quenchant into the container 305 (e.g., through the conduit 330 and into the container 305). The dashed lines indicate that a whole blood (WB) composition may be provided to the kit 300 via a blood collection container 350 .

[0156] Figure 4aThe exemplary kit 400 shown comprises (e.g., includes): a first container 405 suitable for mixing a whole blood composition, a pathogen inactivating compound (PIC), and an optional quencher, wherein the first container 405 comprises a first inlet 410, a second inlet 412, and a first outlet 415; a second container 407 suitable for storing the pathogen-inactivated whole blood composition, wherein the second container comprises a first inlet 416 and a first outlet 417, and an optional second outlet 419 (e.g., coupled to or configured to be coupled to a bypass transfer bag); a container 420 (PIC container) comprising a pathogen inactivating compound; 407 ); a container 425 containing a quencher (quencher container); a conduit 430 coupled at one end to the first container 405 (e.g., the second inlet 412 of the first container 405) and configured at the other end to couple the PIC container 420 and the quencher container 425 (e.g., to provide fluid communication paths between the first container 405 and the PIC container 420 and the quencher container 425, respectively); a conduit 432 coupling the first container 405 and the second container 407; and an optional infusion line assembly 445 for infusing the pathogen-inactivated whole blood composition into a subject. In some embodiments, the conduit 430 includes at one end (e.g., via a 2-way conduit branch): (a) a first adapter 435 for coupling (e.g., configured to couple) the PIC container 420 to the conduit, and (b) a second adapter 440 for coupling (e.g., configured to couple) the quencher container 425 to the conduit, wherein the conduit is coupled (e.g., at the other end) to the second inlet 412 of the first container 405. In some embodiments, the infusion line assembly 445 is configured to couple to the second container 407 (e.g., the first outlet 417 of the second container; via a nosepiece port). The dashed lines indicate that a whole blood (WB) composition can be provided to the kit 400 from or therein a blood collection container 450 and can be added to the first container 405 of the kit 400 by coupling (e.g., aseptically docking) the blood collection container (e.g., a tube 452 extending from the blood collection container) to the first inlet 410 of the first container 405 (e.g., directly coupling to the first inlet 410, coupling through a conduit 442 extending from the first inlet 410).

[0157] An alternative configuration of the exemplary kit of the present disclosure is Figure 4b Similar to Figure 4a, an exemplary kit 400 includes: a first container 405 including a first inlet 410, a second inlet 412, and a first outlet 415; a PIC container 420; a quenchant container 425; a conduit 430 coupled to the first container 405 at one end and configured to couple the PIC container 420 and the quenchant container 425 at the other end; a second container 407 including a first inlet 416, a first outlet 417, and an optional second outlet 419; a conduit 432; an optional infusion line assembly 445; and optionally including at one end of the conduit 430 (e.g., via a conduit branch): (a) a first adapter 435 for coupling the PIC container 420 to the conduit; and (b) a second adapter 440 for coupling the quenchant container 425 to the conduit. This configuration also includes a displacement device (e.g., an air displacement device) 455 adapted to improve (e.g., increase) the transfer of the PIC and quenchant into the container 405 (e.g., through the conduit 430 and into the container 405). In some embodiments, a displacement device (e.g., an air displacement device) 455 is coupled to a conduit 430 (e.g., via a 3-way conduit branch), which includes a first adapter 435 and a second adapter 440 at the ends of the conduit for coupling to the PIC container 420 and the quencher container 425, respectively. The dotted lines indicate that a whole blood (WB) composition can be provided to the kit 400 via a blood collection container 450.

[0158] An alternative configuration of the exemplary kit of the present disclosure is Figure 4c Similar to Figure 4b , an exemplary kit 400 includes: a first container 405 including a first inlet 410, a second inlet 412, and a first outlet 415; a PIC container 420; a quenchant container 425; a displacement device (e.g., an air displacement device) 455; a conduit 430 coupling the first container 405 with (e.g., and providing a fluid communication path therebetween) the PIC container 420 and the quenchant container 425; a second container 407 including a first inlet 416, a first outlet 417, and an optional second outlet 419; a conduit 432; and an optional infusion line assembly 445. This configuration includes, at one end of the conduit 430 (e.g., via a conduit branch), the PIC container 420 coupled to the conduit, the quenchant container 425 coupled to the conduit, and the displacement device (e.g., an air displacement device) 455 coupled to the conduit, and is suitable for improving (e.g., increasing) the transfer of the PIC and quenchant into the container 405 (e.g., through the conduit 430 and into the container 405). The dashed lines indicate that a whole blood (WB) composition may be provided to the kit 400 via a blood collection container 450 .

[0159] An alternative configuration of the exemplary kit of the present disclosure is Figure 4d Similar to Figure 4c, an exemplary kit 400 includes: a first container 405 including a first inlet 410, a second inlet 412, and a first outlet 415; a PIC container 420; a quencher container 425; a displacement device 455; a conduit 430 coupling the first container 405 with the PIC container 420 and the quencher container 425 (e.g., and providing a fluid communication path therebetween); a second container 407 including a first inlet 416, a first outlet 417, and optionally a second outlet 419; and a conduit 432. This configuration also includes an infusion line assembly 445, wherein the infusion line assembly is coupled to the second container 407 (e.g., coupled to the first outlet 417 of the second container). The dashed line indicates that a whole blood (WB) composition can be provided to the kit 400 via a blood collection container 450.

[0160] Figure 5aThe exemplary kit 500 shown comprises (e.g., includes): a first container 505 suitable for mixing a whole blood composition, a pathogen inactivating compound (PIC), and an optional quencher, wherein the first container 505 includes a first inlet 510, a second inlet 512, and a first outlet 515; a vascular access device 560 for withdrawing whole blood from a donor; a conduit 561 extending between the vascular access device 560 and the first inlet 510 of the first container; a second container 507 suitable for storing the pathogen inactivated whole blood composition, wherein the second container 507 includes a first inlet 516 and a first outlet 517, and optionally a second outlet 519; and a second container 507 for storing the pathogen inactivated whole blood composition in the first container 505 (e.g., the second inlet 512 of the first container).

[00145] The present invention also includes a conduit 532 extending between the first container 505 (e.g., the first outlet) and the second container 507 (e.g., the first inlet of the second container); a container 520 containing a pathogen-inactivating compound (PIC container); a container 525 containing a quencher (quencher container); a conduit 530 coupled at one end to the first container 505 (e.g., the second inlet 512 of the first container 505) and configured at the other end to couple the PIC container 520 and the quencher container 525 (e.g., to provide fluid communication paths between the first container 505 and the PIC container 520 and the quencher container 525, respectively); and an optional infusion line assembly 545 for infusing the pathogen-inactivated whole blood composition into a subject. In some embodiments, the conduit 530 includes at one end (e.g., via a two-way conduit branch): (a) a first adapter 535 for coupling (e.g., configured to couple) the PIC container 520 to the conduit; and (b) a second adapter 540 for coupling (e.g., configured to couple) the quencher container 525 to the conduit. In some embodiments, vascular access device 560 and catheter 561 together comprise a needle assembly. In some embodiments, infusion line assembly 545 is configured to couple to second container 507 (e.g., first outlet 517 of the second container; via a tip port). In some embodiments, container 505 contains an anticoagulant (e.g., an anticoagulant solution) 565.

[0161] An alternative configuration of the exemplary kit 500 of the present disclosure is Figure 5b Similar to Figure 5a, an exemplary kit 500 includes: a first container 505, which includes a first inlet 510, a second inlet 512 and a first outlet 515; a PIC container 520; a quencher container 525; a catheter 530, which is connected to the first container 505 at one end and is configured at the other end to connect the PIC container 520 and the quencher container 525; a second container 507, which includes a first inlet 516, a first outlet 517 and an optional second outlet 519; a catheter 532; a vascular access device 560; a catheter 561; an optional infusion line assembly 545; and optionally including at one end of the catheter 530 (e.g., via a catheter branch): (a) a first adapter 535 for connecting the PIC container 520 to the catheter; and (b) a second adapter 540 for connecting the quencher container 525 to the catheter. This configuration also includes a displacement device (e.g., an air displacement device) 555, which is suitable for improving (e.g., increasing) the transfer of PIC and quencher into container 505 (e.g., through conduit 530 and into container 505). In some embodiments, displacement device (e.g., an air displacement device) 555 is coupled to conduit 530 (e.g., through a 3-way conduit branch), which includes a first adapter 535 and a second adapter 540 at the ends of the conduit for coupling to PIC container 520 and quencher container 525, respectively. In some embodiments, container 505 contains anticoagulant 565.

[0162] An alternative configuration of the exemplary kit 500 of the present disclosure is Figure 5c Similar to Figure 5b , an exemplary kit 500 includes: a first container 505, which includes a first inlet 510, a second inlet 512, and a first outlet 515; a PIC container 520; a quencher container 525; a displacement device (e.g., an air displacement device) 555; a conduit 530, which connects the first container 505 to the PIC container 520 and the quencher container 525 (e.g., and provides a fluid communication path therebetween); a second container 507, which includes a first inlet 516, a first outlet 517, and an optional second outlet 519; a conduit 532; a vascular access device 560; a conduit 561; and an optional infusion line assembly 545. This configuration includes, at one end of the conduit 530 (e.g., through a conduit branch), a PIC container 520 coupled to the conduit, a quencher container 525 coupled to the conduit, and a displacement device (e.g., an air displacement device) 555 coupled to the conduit, adapted to improve (e.g., increase) the transfer of the PIC and quencher into the container 505 (e.g., through the conduit 530 and into the container 505). In some embodiments, the container 505 contains an anticoagulant 565.

[0163] An alternative configuration of the exemplary kit 500 of the present disclosure is Figure 5d Similar to Figure 5b , an exemplary kit 500 includes: a first container 505 including a first inlet 510, a second inlet 512, and a first outlet 515; a PIC container 520; a quencher container 525; a catheter 530 coupled to the first container 505 at one end and configured to couple the PIC container 520 and the quencher container 525 at the other end; a displacement device 555; a second container 507 including a first inlet 516, a first outlet 517, and optionally a second outlet 519; a catheter 532; a vascular access device 560; a catheter 561; an infusion line assembly 545; and optionally including at one end of the catheter 530 (e.g., via a catheter branch): (a) a first adapter 535 for coupling the PIC container 520 to the catheter; and (b) a second adapter 540 for coupling the quencher container 525 to the catheter. This configuration includes the infusion line assembly 545, wherein the infusion line assembly is coupled to the second container 507 (e.g., coupled to the first outlet 515 of the second container). In some embodiments, container 505 contains anticoagulant 565 .

[0164] An alternative configuration of the exemplary kit 500 of the present disclosure is Figure 5e Similar to Figure 5c , an exemplary kit 500 includes: a first container 505 including a first inlet 510, a second inlet 512, and a first outlet 515; a PIC container 520; a quencher container 525; a displacement device 555; a conduit 530 coupling the first container 505 with (e.g., and providing a fluid communication path therebetween) the PIC container 520 and the quencher container 525; a conduit 532; a second container 507 including a first inlet 516, a first outlet 517, and optionally a second outlet 519; a vascular access device 560; a conduit 561; and an infusion line 545 coupled to the container 507. In some embodiments, the container 505 contains an anticoagulant 565.

[0165] An alternative configuration of the exemplary kit 500 of the present disclosure is Figure 5f Similar to Figure 5e , an exemplary kit 500 includes: a first container 505 including a first inlet 510, a second inlet 512, and a first outlet 515; a PIC container 520; a replacement set 555; a catheter 530 coupling the first container 505 to the PIC container 520 (e.g., and providing a fluid communication path therebetween); a catheter 532; a second container 507 including a first inlet 516, a first outlet 517, and optionally a second outlet 519; a vascular access device 560; a catheter 561; and an infusion line assembly 545 coupled to the container 507. In some embodiments, the container 505 contains an anticoagulant 565.

[0166] An alternative configuration of the exemplary kit 500 of the present disclosure is Figure 5g Similar to Figure 5f , an exemplary kit 500 includes: a first container 505 including a first inlet 510, a second inlet 512, and a first outlet 515; a PIC container 520; a conduit 530 coupling the first container 505 to the PIC container 520 (e.g., and providing a fluid communication path therebetween); a conduit 532; a second container 507 including a first inlet 516, a first outlet 517, and optionally a second outlet 519; a vascular access device 560; a conduit 561; and an infusion line assembly 545 coupled to the container 507. In some embodiments, the container 505 contains an anticoagulant 565.

[0167] As described herein, when a conduit (e.g., a tube) is described as being coupled or extending (e.g., connected) between two containers (e.g., bags), such as in a kit (e.g., a processing device of the kit), it should be understood that the conduit can be joined at some point between the two containers (e.g., bags) by another component or connector at the connection between them. For example, a blood collection container coupled (e.g., connected) to a first container by a conduit includes instances where the conduit includes a filter (e.g., a leukocyte reduction filter) between the two containers, i.e., the tubes are separated by the filter such that fluid flows (e.g., aseptically transferred) from one container to the other through the conduit and the filter. Alternatively, or in addition, the conduit between the containers allows fluid to flow from one container to the other (e.g., a fluid communication path, aseptic transfer) and can be blocked to prevent flow until necessary or desired, such as, for example, preventing fluid from one container from flowing to the next container as part of processing until required or desired for the next step in the method (e.g., an openable fluid communication path). Due to such openable seal, such as clamp, stopper, valve etc. are included in or on the conduit of connection container, wherein clamp, stopper, valve etc. can be selectively opened as needed, for example, to transfer fluid from one container to the next container. In certain embodiments, the conduit between the containers comprises a breakable seal, such as a breakable valve or a frangible connector, and the rupture of the breakable seal allows the blood product solution to flow between the containers by conduit. It should be understood that the breakable seal is included in the joint between the containers so that the sterility of the system is maintained. It should also be understood that the conduit comprising a filter or a breakable seal comprises the situation where conduit can be interrupted by a filter or seal, for example, conduit extends from a container and is connected to a filter or seal (the inflow portion (incoming portion) of conduit), and conduit continues to extend to another container (the outflow portion (outgoing portion) of conduit) from another part of a filter or seal. In such a structure, fluid flows through the inflow portion of conduit, flows through a filter or seal from the first container, and flows through the outflow portion of conduit and flows into another container. In certain embodiments, the coupling or connection between the catheter and the inlet or outlet (e.g., port) of the container, or the inlet or outlet itself, may include a rupturable or pierceable seal, such as a rupturable valve or frangible connector or a pointed port, whereby rupture or piercing of the seal allows the blood product solution to flow between the catheter and the container (e.g., sterile transfer).

[0168] Different containers (e.g., bags) within the pathogen inactivation treatment device can be used for different steps of the method. For example, a system of containers to be used for pathogen inactivation of a whole blood composition can include a container containing a pathogen inactivating compound, a container containing a quencher, a container for receiving a whole blood unit (e.g., a whole blood donation) and the pathogen inactivating compound and quencher (e.g., a first container), and one or more containers (e.g., product bags, storage bags) for containing the final whole blood product, i.e., one or more pathogen-inactivated whole blood units (e.g., therapeutic dose units) with the concentration of the pathogen inactivating compound and / or its byproducts reduced to below a desired concentration, ready for use or stored for later use. Blood containers can generally be designed to hold various volumes of fluid, including but not limited to volumes in the range of 50 mL to 2 liters, for example, having a capacity of up to about 350 mL, 450 mL, 500 mL, 600 mL, 700 mL, 800 mL, 900 mL, 1 liter, up to 1.5 liters, or up to 2 liters. Each bag in the system is typically made of a plastic material. For example, the container for containing the pathogen-inactivating compound solution can be made of a suitable plastic (such as PL2411 (Baxter Healthcare)) or other plastics (such as polyvinyl chloride, polyolefins, ethylene-vinyl acetate, ethylene-vinyl acetate blended with other plastics, etc.). Suitable bag materials are discussed, for example, in PCT Publication No. WO 2003078023 and U.S. Patent No. 7025877, the disclosures of which are hereby incorporated by reference in their entirety. In all cases, the materials used to prepare the treatment device should be sterilizable by known methods, such as steam and gamma or electron beam radiation for ensuring the sterility of the treatment device. Although these are exemplary materials for making bags, the method described herein is applicable to the process of using any suitable bag material that will be readily available to those skilled in the art, and can also be used with containers other than bags. The bags used herein can be designed to allow gases such as oxygen and carbon dioxide to enter and exit the blood bag so that the blood products therein have sufficient oxygen supply and carbon dioxide levels during processing and storage.

[0169] The present disclosure also provides a method of transfusing whole blood into a subject in need thereof (eg, a human subject), comprising transfusing the aforementioned pathogen-inactivated whole blood composition or the aforementioned therapeutic dose unit of pathogen-inactivated whole blood into the subject.

[0170] Those skilled in the art will also understand that changes may be made to the form and details of the embodiments described herein without departing from the scope of the present disclosure. In addition, although various advantages, aspects, and objects have been described with reference to various embodiments, the scope of the present disclosure should not be limited by reference to such advantages, aspects, and objects. Rather, the scope of the present disclosure should be determined with reference to the appended claims.

[0171] The present invention is further illustrated by the following examples, which are not to be construed as limiting the invention in scope or spirit to the specific procedures they describe.

[0172] Example

[0173] Example 1: Preparation of pathogen-inactivated whole blood

[0174] In the kit of the present disclosure, pathogen-inactivated whole blood is prepared using the pathogen-inactivating compound amatalin and the quencher glutathione (Henschler et al., 2011, Transfus Med Hemother 38:33–42), the kit comprising a processing container and a tube having a 3-way branch for introducing amatalin / GSH (e.g., a “filter set”), which comprises two in-line filters for coupling (e.g., connecting) to containers (e.g., syringes) of PIC and quencher, and a third branch (e.g., connected to) a sample bypass transfer bag. Figure 3b ). More specifically, four ABO-matched whole blood units containing anticoagulant were pooled and divided into 8 half units (225 mL / half unit), which were added to each of the eight processing bags by connecting the pooled donor blood bags containing whole blood to the processing container with the aid of a tube and using a tube welder. After the whole blood was transferred to the processing container, the pooled donor blood bags were removed by sealing and cutting the tubes, and the filter set was aseptically connected to the same input port of the processing container through the remaining tubes using a tube welder. Amataline and GSH were added to final concentrations of 0.2 mM and 2.0 mM, respectively. GSH was added first, and then amataline was added within 5 minutes of GSH addition or after a holding time of at least 15 minutes, as shown in Table 1. The mixture was incubated at 22°C (units 1-4) or 25°C (units 5-8), and samples were taken out at the indicated times and analyzed by liquid chromatography-mass spectrometry (LCMS) to determine the level of residual unreacted amataline.

[0175] Table 1: Residual Amatalin Levels (nM) in Pathogen-Inactivated Whole Blood

[0176] unit GSH 21hr 24hr 27hr 30hr 33hr 36hr 42hr 1 >15min 3.50 2.62 2.27 2.11 2.61 1.62 1.30 2 >15min 4.09 2.66 3.07 2.95 2.29 1.38 1.35 3 <5min 3.45 3.04 3.29 2.67 2.16 1.73 1.66 4 <5min 3.85 3.53 3.7 2.84 2.22 1.41 1.57 5 >15min 2.91 2.44 2.55 2.04 1.76 1.89 1.62 6 >15min 3.21 2.58 2.71 2.49 1.47 1.87 1.22 7 <5min 3.43 2.86 2.74 2.34 1.64 2.28 1.11 8 <5min 4.15 3.39 3.30 2.59 2.83 1.97 2.96

[0177] In addition, 50 mL of the mixture was removed from each of Units 2 and 8 at 24 hr and divided into two 25 mL samples in separate containers to compare storage at 4°C versus room temperature and to assess residual amatarin levels at 2, 4, 6, and 8 hours of storage under those conditions (Table 2).

[0178] Table 2: Residual Amatalin Levels (nM) in Pathogen-Inactivated Whole Blood

[0179] unit temperature 2hr 4hr 6hr 8hr 2 Room temperature BLOQ BLOQ BLOQ BLO 8 Room temperature BLOQ BLOQ BLOQ BLOQ 2 4℃ 1.69 1.66 1.49 1.09 8 4℃ 2.69 1.77 1.89 1.64

[0180] BLOQ = below limit of quantitation (e.g., <0.75 nM)

[0181] As shown in Table 1, at 21hr, the residual amatalin level was in the range of 2.91 to 4.15nM concentration and further reduced to a range of 1.11 to 2.96nM at the final 42hr sampling time. In contrast, it was surprisingly observed that at 21hr, the treatment mixture was transferred from units 2 and 8 to different containers and then further stored at room temperature, resulting in a reduction of residual amatalin to below the limit of quantification (Table 2), which could not be achieved by continuing to incubate the initial units in the treatment containers at 22°C or 25°C (Table 1). Transferring and incubating at 4°C did not result in an equivalent decrease in residual amatalin (Table 2). These results indicate the effect of the transfer step and further incubation at room temperature on the residual amatalin level.

[0182] Example 2: Preparation of pathogen-inactivated whole blood

[0183] After pathogen inactivation of whole blood using a 3-container kit of the present disclosure that allows one or two transfer steps from the initial (e.g., first) container and a filter set similar to Example 1, another study was conducted to evaluate the effect of container transfer on residual amatarin levels. Six ABO-matched whole blood units in anticoagulant were pooled and divided into six ~450 mL units, which were transferred to the first blood bag container of the kit containing Transfufol 3226 PVC membrane (Renolit), and then the pooled donor blood bag was disconnected and sealed, and then connected by welding the filter set tubing to the same input port (e.g., attachment tube) of the first container to transfer amatarin and GSH to the whole blood. Amatarin and GSH were added to the whole blood units to final concentrations of 0.2 mM and 2.0 mM, respectively, and the mixture was then transferred to the second bag (Transfufol 3226 PVC membrane) and incubated at 22°C. At 22 hr, a sample was taken for analysis and the mixture was transferred to a third bag (Transfufol 3226 PVC membrane) and then further incubated at 22°C, with additional samples taken for analysis at 24 hr (i.e., 2 hr after the final transfer) and 26 hr (i.e., 4 hr after the final transfer).

[0184] Table 3: Residual Amatalin Levels (nM) in Pathogen-Inactivated Whole Blood

[0185]

[0186]

[0187] BLOQ = below limit of quantitation (e.g., <0.75 nM)

[0188] As previously described, samples were analyzed by LCMS to determine the level of residual, unreacted amataline. As shown in Table 3, transferring the mixture to a second bag immediately after adding amataline / GSH to whole blood and then incubating at 22°C for 22 hours showed residual amataline levels ranging from 1.35 to 1.96 nM. After incubation for 22 hours, the mixture was transferred to a third bag and then incubated at 22°C for an additional 2 hours (24 hour column) or 4 hours (26 hour column), residual amataline levels decreased to less than 1 nM for all samples and to below the limit of quantification in all but one sample.

[0189] Example 3: Preparation of pathogen-inactivated whole blood

[0190] In another study, pathogen-inactivated whole blood was prepared using amatarin and GSH in a 1-container or 3-container kit of the present disclosure, wherein the 3-container kit included a second and third bag (e.g., transfer / storage container) from three different sources (e.g., different plastic materials). The three different transfer / storage containers included a 600 mL container containing Transfufol 3226 PVC film ("PVC1"), a second 600 mL container from a different manufacturer containing Transfufol 3226 PVC film ("PVC2"), and a 750 mL container from Cerus containing EVA (ethylene vinyl acetate) 1800 film ("EVA"). Six ABO-matched whole blood units in anticoagulant were pooled and divided into seven units, each Seven whole blood units were subjected to pathogen inactivation (Table 4) with no transfer steps between containers (e.g., processing performed in a 1-container kit) or with two transfer steps between the containers of the indicated storage / transfer bag source (e.g., processing performed in a 3-container kit). As previously described, amatharine and GSH were added to the whole blood units by aseptically docking the filter set to the tubing of the input port of the first container containing the whole blood. Unit 1 was treated with amatharine and GSH at final concentrations of 0.2 mM and 2.0 mM, respectively, in the first container and incubated at 22°C, with samples taken at 18, 20, 22, and 24 hours without container transfer. For units 2-7, amatharine and GSH were added to the whole blood in the first container (final concentrations of 0.2 mM and 2.0 mM, respectively), then transferred to the second container and incubated at 22°C for 18 hours. At 18 hours, a sample was taken and the mixture was transferred to a third container and further incubated at 22°C with additional sampling at 2 hour intervals (20, 22 and 24 hr).

[0191] Table 4: Residual Amatalin Levels (nM) in Pathogen-Inactivated Whole Blood

[0192] unit transfer bag Transfer* 18hr 20hr 22hr 24hr 1 N / A none 2.16 1.99 1.46 1.54 2 PVC1 2x 1.75 0.88 BLOQ BLOQ 3 PVC1 2x 1.52 0.78 BLOQ BLOQ 4 PVC2 2x 1.48 0.87 BLOQ BLOQ 5 PVC2 2x 1.45 0.85 BLOQ BLOQ 6 EVA 2x 3.11 1.14 BLOQ BLOQ 7 EVA 2x 3.51 1.22 BLOQ BLOQ

[0193] *No transfer = 1 bag kit; 2x transfer = 3 bag kit

[0194] BLOQ = below limit of quantitation (e.g., <0.75 nM)

[0195] As previously described, all samples were analyzed by LCMS to determine the level of residual, unreacted amataline. As shown in Table 4, the lowest residual amataline levels were observed at 18 hours for Units 2-5, where the whole blood / amataline / GSH mixture had been transferred to one of two container types in the three-container kit prior to the 18-hour incubation, but not for the third container type (Units 6-7). Following a second transfer (i.e., to the third container) at 18 hours and further incubation at 22°C, residual amataline at the 20-hour time point (i.e., 2 hours after bag transfer) was less than 1 nM for the kits with two different container types and approached 1 nM for the kit with the third container type. Residual amataline levels for all samples processed in the three-container kit (with each of the three different container types) were below the limit of quantification at both 22 and 24 hours (i.e., 4 or 6 hours of incubation after bag transfer). In contrast, following treatment using the 1-vessel device and in the absence of transfer, amafil levels at 20, 22, and 24 hr remained in the range of 1.54 to 1.99 nM.

[0196] Example 4: Preparation of pathogen-inactivated whole blood

[0197] Another study was conducted to evaluate the preparation of pathogen-inactivated whole blood using amatalin and GSH in a 1-container, 2-container, or 3-container kit of the present disclosure. Six ABO-matched whole blood units in anticoagulant were pooled and divided into seven Units. Seven whole blood units were inactivated for pathogens under one of the following three conditions - no transfer step between containers, one transfer step between two containers, or two transfer steps between three containers (Table 5). As previously described, amatharine and GSH were added to the whole blood units by aseptically docking the filter group to the tube of the input port of the first container containing whole blood. In the first container, unit 1 was treated with amatharine and GSH at final concentrations of 0.2mM and 2.0mM, respectively, and incubated at 22°C. Samples were obtained at 20, 22, 24, and 26 hours without further container transfer. For units 2-4, amatharine and GSH (final concentrations of 0.2mM and 2.0mM, respectively) were added to whole blood in the first container and incubated at 22°C for 20 hours. Samples were obtained at 20 hours and transferred to the second bag and further incubated at 22°C for additional sampling at 2 hour intervals (22, 24, and 26hr). For units 5-7, amatarin and GSH (final concentrations of 0.2 mM and 2.0 mM, respectively) were added to whole blood in the first container, and the mixture was transferred to the second container and then incubated for 20 hours at 22° C. A sample was taken at 20 hours, transferred to a third bag, and further incubated at 22° C., with additional sampling at 2-hour intervals (22, 24, and 26 hr).

[0198] Table 5: Residual Amatalin Levels (nM) in Pathogen-Inactivated Whole Blood

[0199] unit Transfer* 20hr 22hr 24hr 26hr 1 none 1.47 1.54 1.39 1.32 2 1x 1.53 BLOQ BLOQ BLOQ 3 1x 1.65 BLOQ BLOQ BLOQ 4 1x 1.66 BLOQ BLOQ BLOQ 5 2x 1.53 BLOQ BLOQ BLOQ 6 2x 1.47 BLOQ BLOQ BLOQ 7 2x 1.46 BLOQ BLOQ BLOQ

[0200] *No transfer = 1 bag kit; 1x transfer = 2 bag kit; 2x transfer = 3 bag kit BLOQ = below limit of quantitation (e.g., <0.75 nM)

[0201] As previously described, samples were analyzed by LCMS to determine the levels of residual, unreacted amatalin. As shown in Table 5, amatalin levels after PI treatment of whole blood using the 2- and 3-container kits were below the limit of quantification, while amatalin levels after treatment using the 1-container kit remained within the range of 1.32 to 1.54 nM.

[0202] Example 5: Preparation of pathogen-inactivated whole blood

[0203] Another study was conducted to evaluate additional conditions for preparing pathogen-inactivated whole blood using amatalin and GSH in a 2-container kit of the present disclosure (e.g., Figure 3bAmatalin and GSH were added to the whole blood unit in a first container to final concentrations of 0.2 mM and 2.0 mM, respectively. The mixture was then incubated in the first container (e.g., a processing container) at 22°C for 18, 20, or 22 hours and then transferred to a second container, where samples were taken for analysis as indicated in Table 6 until the time point of transfer to the second container. After transfer to the second container, the mixture was further incubated at 22°C and samples were taken for analysis at 2-hour intervals, as shown in Table 6.

[0204] Table 6: Residual Amatalin Levels (nM) in Pathogen-Inactivated Whole Blood

[0205] unit Transfer time 18hr 20hr 22hr 24hr 26hr 1 18hr 1.51 2.51 2.01 2.22 0.98 2 18hr 1.21 1.06 0.91 0.75 BLOQ 3 18hr 1.07 1.04 0.81 BLOQ BLOQ 4 18hr 1.05 1.02 1.03 BLOQ BLOQ 5 18hr 34.50 8.29 1.67 1.06 BLOQ 6 18hr 3.45 1.64 1.79 0.95 BLOQ 7 18hr 2.04 2.08 2.04 1.35 BLOQ 8 20hr N / A 1.71 1.27 1.50 1.21 9 20hr N / A 1.29 0.79 BLOQ BLOQ 10 20hr N / A 1.48 0.89 0.91 BLOQ 11 20hr N / A 1.29 0.78 0.86 BLOQ 12 20hr N / A 1.92 0.91 0.86 BLOQ 13 20hr N / A 1.82 0.90 0.78 BLOQ 14 20hr N / A ** 1.13 0.95 0.77 15 22hr N / A N / A 1.52 BLOQ BLOQ 16 22hr N / A N / A 1.63 BLOQ BLOQ 17 22hr N / A N / A 1.78 BLOQ BLOQ 18 22hr N / A N / A 2.21 BLOQ BLOQ 19 22hr N / A N / A 4.29 BLOQ BLOQ 20 22hr N / A N / A 2.76 BLOQ BLOQ 21 22hr N / A N / A 3.27 BLOQ BLOQ

[0206] BLOQ = below limit of quantitation (e.g., <0.75 nM)

[0207] **Sample not available for analysis

[0208] As previously described, samples were analyzed by LCMS to determine the level of residual unreacted amatalin. As shown in Table 6, and compared to control units 1, 8, and 14, a reduction in the level of amatalin after PI treatment to less than 1 nM or lower (e.g., below the limit of quantitation) was most consistently achieved by the transfer step at 20 or 22 hr.

[0209] Example 6: Collection and preparation of pathogen-inactivated whole blood

[0210] In some embodiments, the whole blood pathogen inactivation kits of the present disclosure are designed to collect whole blood from a donor directly into a first container (e.g., a processing container) of the kit, thereby eliminating the need to collect whole blood into a donor blood bag and then connect the donor blood bag containing the whole blood to the kit. For example, a 1-container, 2-container, or 3-container kit as provided herein may further include an integral vascular access device, such as a venipuncture needle (e.g., a venipuncture needle) coupled to a processing container (e.g., containing an anticoagulant) of the kit via one or more flexible tubing segments. Figures 5a-5d In this configuration, whole blood is drawn from the donor directly into the kit's processing container, and a pathogen-inactivating compound is added, mixed with the whole blood, and incubated in the same container. A study was conducted to demonstrate the feasibility of treating whole blood with amatalin and GSH directly in the blood collection container from a commercially available collection device.

[0211] More specifically, whole blood units were collected from donors and processed in Teruflex (Terumo, BB*AGD506A2) blood collection bags (units 2-7) or Leucoflex (MacoPharma, LQE7230LD) blood collection bags (units 9-14) containing anticoagulant and integrated into the kit of the present disclosure. Six ABO-matched whole blood units were pooled and divided into a control unit and six test units, each ~450 mL. Control units 1 and 8 used the kit described in the previous example. The filter set as previously described was aseptically docked to a whole blood transfer tube that had been sealed and disconnected from the container, and then amatalin and GSH were added to final concentrations of approximately 0.2 mM and 2.0 mM, respectively. GSH was added first, followed by amatalin within 30 minutes. The mixture was then incubated at 22°C for 20 hr, samples were taken for analysis and the mixture was transferred to a second integrally connected container (for test units 2-7 and 9-14), and then further incubated and sampled at 2 hr intervals (i.e., 22 hr, 24 hr) (for all units) as shown in Table 7.

[0212] Table 7: Residual Amataline Levels (nM) in Pathogen-Inactivated Whole Blood

[0213] unit Collection Bag 20hr 22hr 24hr 1 T 2.35 1.61 1.81 2 T 1.29 BLOQ BLOQ 3 T 1.62 BLOQ BLOQ 4 T 1.53 BLOQ BLOQ 5 T 1.92 BLOQ BLOQ 6 T 2.12 BLOQ BLOQ 7 T 2.32 BLOQ BLOQ 8 M 2.62 1.93 1.85 9 M 1.36 BLOQ BLOQ 10 M 1.32 BLOQ BLOQ 11 M 1.44 BLOQ BLOQ 12 M 1.34 BLOQ BLOQ 13 M 1.49 BLOQ BLOQ 14 M 1.40 BLOQ BLOQ

[0214] BLOQ = below limit of quantitation (e.g., <0.75 nM)

[0215] The residual amatalin levels in the pathogen-inactivated whole blood preparations were analyzed by LCMS at 20 hr and were found to be below the limit of quantitation for each unit transferred to the second bag.

[0216] In another example, 450 mL of whole blood is collected from a donor through a first inlet port into a first container containing 63 mL of anticoagulant CPDA-1 (e.g., a blood:anticoagulant ratio of approximately 7:1) using a 2-container kit including a processing container and a filter set for introducing amafil / GSH (e.g., as described above). The collection line is then closed using a medical tube clamp, and the whole blood and anticoagulant solution are mixed. Next, the pathogen-inactivating compound amafil and the quencher GSH are added to the whole blood in the first container through a second port to final concentrations of approximately 0.2 mM and 2.0 mM, respectively. The mixture of whole blood, PIC, and quencher is incubated at 22° C. for 20 hours, and then the mixture is transferred to a second container, the tube between the first and second containers is clamped, and the mixture is incubated at 22° C. for an additional 2-4 hours to provide a pathogen-inactivated whole blood unit.

[0217] The residual amafilin level in the pathogen-inactivated whole blood preparation is determined by sampling from the second container and performing LCMS analysis to demonstrate a level below 1 nM (eg, below the limit of quantitation of 0.75 nM).

[0218] Example 7: Transfusion of pathogen-inactivated whole blood

[0219] Using standard transfusion methods known in the art, the pathogen-inactivated whole blood prepared as provided herein (e.g., ABO compatible, RhD compatible, O type) is transfused into a subject in need (e.g., for trauma resuscitation). More specifically, in one example, a commercially available blood transfusion line (e.g., a blood transfusion line assembly with an integrated filter) is connected to the second container of the kit via a nozzle port outlet, the second container containing the pathogen-inactivated whole blood prepared as described above. According to the generally accepted medical practice of such transfusion, the unit of pathogen-inactivated whole blood is transfused from the kit into the subject with acute bleeding caused by trauma by gravity flow. Additional units of pathogen-inactivated whole blood are transfused as needed.

[0220] Example 8: Preparation of pathogen-inactivated RBCs and cryoprecipitate

[0221] Pathogen-inactivated whole blood can be used to produce blood products, such as pathogen-inactivated red blood cells (RBCs), plasma and / or cryoprecipitate preparations. More specifically, in one example, the pathogen-inactivated whole blood preparation produced as previously described is centrifuged under conditions that separate RBCs and plasma. More specifically, the pathogen-inactivated whole blood unit is centrifuged at 4200 g for 10 minutes at 4° C. to separate plasma from RBCs, and then most of the plasma layer is removed using an expression device and transferred to a container separate from a thick RBC concentrate (packed RBC concentrate). A small amount of plasma is maintained with RBCs to produce a unit with a desired hematocrit (e.g., 60%), and the RBC unit is refrigerated at 1-6° C. The pathogen-inactivated plasma unit is then frozen at -30° C. for the preparation of cryoprecipitate.

[0222] In addition, a study was conducted to evaluate plasma and red blood cells prepared from pathogen-inactivated whole blood compositions, prior to preparing the plasma and red blood cell components, shortly after the whole blood PI process, or after storing the pathogen-inactivated whole blood at 4°C. Donor whole blood units were pooled and divided into seven units (e.g., to standardize between units) and subjected to the following treatment conditions. Unit 1 was a control and treated with saline only, while units 2-7 were treated with amatharine and GSH at concentrations of approximately 0.2 mM and 2.0 mM, respectively. After amatharine and GSH or saline were added to the first container (bag), the units were incubated at 22°C for 22 hours and then transferred to the second container (bag). Samples were taken at the time of transfer and at 24 and 26 hours (2 or 4 hours after transfer, also incubated at 22°C) to analyze the levels of residual amatharine, hemolysis and / or fibrinogen, and Factor VIII. After transfer (and further incubation) 4 hours, then unit 1-4 is stored at 4 ℃, and by being separated into plasma and red blood cell component by unit 5-7 at 22 ℃ with 3800rpm centrifugation for 6 minutes, and then after separation with plasma, in red blood cell component, add SAGM additive solution.Then to the plasma component sampling from unit 5-7, to carry out fibrinogen and factor VIII analysis (for example, using standard analytical method), and plasma component is stored at 4 ℃.To the red blood cell component sampling from unit 5-7, to carry out residual amatarin and hemolysis analysis, and red blood cell component is also stored at 4 ℃.At the 2nd day, the 5th day and the 7th day, also carry out the sampling of plasma and red blood cell component.In addition, at the 2nd day, the 5th day and the 7th day, from the whole blood of the pathogen inactivation of storage, take out sample, centrifuge to obtain plasma and red blood cell, and carry out similar analysis.Residual amatarin concentration (nM) is shown in table 8 below, and the level of fibrinogen and factor VIII is shown in table 9a and 9b. Hemolysis levels were within acceptable ranges and are not shown.Other plasma coagulation factors and / or properties analyzed may include, for example, Factor II, Factor V, Factor VII, Factor IX, Factor X, Factor XI, PT, PTT, and / or TT.

[0223] Table 8: Residual Amataline Levels (nM) in Pathogen-Inactivated Whole Blood

[0224] unit 22hr 24hr 26hr 2 1.39 BLOQ BLOQ 3 1.35 0.75 BLOQ 4 1.96 BLOQ BLOQ 5 1.84 BLOQ BLOQ 6 1.55 BLOQ BLOQ 7 1.46 BLOQ BLOQ

[0225] BLOQ = below limit of quantitation (e.g., <0.75 nM)

[0226] Table 9a: Fibrinogen and Factor VIII in Plasma from Pathogen-Inactivated Whole Blood

[0227]

[0228]

[0229] Table 9b: Fibrinogen and Factor VIII in Plasma from Pathogen-Inactivated Whole Blood

[0230]

[0231] To prepare cryoprecipitate, units of frozen plasma prepared from pathogen-inactivated whole blood are thawed in a temperature-controlled water bath at 4°C, with a total thawing time of approximately 4 hours and 30 minutes. The thawed units are then centrifuged at 4200 rcf for 12 minutes at 4°C with slow deceleration. The cryoprecipitate-reduced plasma supernatant is removed from the cryoprecipitate, while a small amount of plasma is retained for resuspending the cryoprecipitate. After resuspension, the cryoprecipitate units can be used for transfusion or frozen for storage at -30°C.

[0232] The disclosed examples and embodiments disclosed herein further describe methods, kits, and compositions for preparing pathogen-inactivated whole blood compositions and other pathogen-inactivated blood product compositions (including, for example, pathogen-inactivated whole blood and other blood product compositions suitable for transfusion into an individual). The illustrated components and steps are described to explain the exemplary embodiments shown, and it is expected that ongoing technological developments will change the manner in which specific functions are performed. These examples are provided herein for illustrative purposes rather than limiting purposes. In addition, for ease of description, the boundaries of the functional building blocks have been arbitrarily defined herein. Alternative boundaries can be defined as long as specific functions and their relationships are appropriately performed. Based on the teachings contained herein, alternatives (including equivalents, extensions, variations, deviations, etc. of those described herein) will be apparent to those skilled in the relevant art. Such alternatives fall within the scope and spirit of the disclosed embodiments.

[0233] Although the embodiments have been fully described with reference to the accompanying drawings, it should be noted that various changes and modifications will be apparent to those skilled in the art. Such changes and modifications should be understood to be included within the scope of the various embodiments as defined by the appended claims.

[0234] All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein.

[0235] Unless otherwise indicated herein or clearly contradicted by context, the use of the terms "a / an" and "the" and similar designations (especially in the context of the following claims) should be understood to encompass both the singular and the plural. Unless otherwise indicated, the terms "comprising," "having," "including," and "containing" should be understood as open-ended terms (i.e., meaning "including but not limited to") wherever open-ended terms are used to describe features or elements, it is specifically contemplated that closed terms may be used in place of the open-ended terms without departing from the spirit and scope of the present disclosure. Unless otherwise indicated herein, the recitation of ranges of values ​​herein is intended merely to serve as a shorthand method of referring individually to each individual value falling within the range, and each individual value is incorporated into the specification as if it were individually recited herein. Reference herein to "about" a value or parameter includes (and describes) variations on the value or parameter itself. For example, a reference to "about X" includes a description of "X." All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples or exemplary language (e.g., "such as") provided herein is intended merely to better illuminate the present description and does not limit the scope of the present description unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the methods, systems, and compositions disclosed herein.

[0236] Preferred embodiments are described herein. After reading the foregoing description, variations of those preferred embodiments may become apparent to those skilled in the art. It is expected that a skilled person will be able to appropriately adopt such variations, as well as the practice of the methods, systems, and compositions described herein, except as specifically described herein. Therefore, the methods, systems, and compositions described herein include all modifications and equivalents of the subject matter cited in the claims appended hereto as permitted by applicable law. In addition, this specification encompasses any combination of the above-mentioned elements in all possible variations thereof, unless otherwise indicated herein or otherwise clearly contradicted by the context. The following is a list of specific embodiments of the present disclosure. The list is exemplary and is not intended to limit the disclosure provided herein.

[0237] Embodiment 1. A method for preparing a pathogen-inactivated whole blood composition, comprising:

[0238] (a) Mix in the first container

[0239] (i) a whole blood composition;

[0240] (ii) an effective amount of a pathogen inactivating compound (PIC); and

[0241] (iii) optionally an effective amount of a quencher;

[0242] (b) incubating the mixture of the whole blood composition, the PIC, and optionally the quencher in the first container under conditions and for a duration sufficient to inactivate pathogens, if present;

[0243] (c) transferring the mixture to a second container under sterile conditions; and

[0244] (d) incubating the mixture in the second container under conditions and for a duration sufficient to reduce the concentration of the PIC to

[0245] (i) a concentration that is at least 25% less than the concentration of said PIC in said mixture at the time of transferring step (b);

[0246] (ii) at least 10 less than the concentration of said PIC in said mixture of step (a) 5 times the concentration; and / or

[0247] (iii) a concentration of about 1 nm or less,

[0248] Thereby producing the pathogen-inactivated whole blood composition.

[0249] Embodiment 2. The method of embodiment 1, wherein the concentration of the PIC in the mixture of step (a) is at least about 0.1 mM.

[0250] Embodiment 3. The method of embodiment 1 or embodiment 2, wherein the concentration of the PIC in the mixture of step (a) is about 0.1 mM to about 1.0 mM.

[0251] Embodiment 4. The method of any one of embodiments 1-3, wherein the concentration of the PIC in the mixture of step (a) is about 0.2 mM.

[0252] Embodiment 5. The method of any one of embodiments 1-4, wherein the concentration of the PIC after step (d) is less than about 2 nM.

[0253] Embodiment 6. The method of any one of embodiments 1-5, wherein the concentration of the PIC after step (d) is less than about 1 nM.

[0254] Embodiment 7. The method of any one of Embodiments 1-6, wherein the PIC comprises a functional group that is or forms a reactive electrophilic group.

[0255] Embodiment 8. The method of embodiment 7, wherein the functional group is selected from the group consisting of mustard, mustard intermediate, and mustard equivalent.

[0256] Embodiment 9. The method of embodiment 7 or embodiment 8, wherein the functional group is or is capable of forming an aziridinium ion.

[0257] Embodiment 10. The method of any one of embodiments 1-9, wherein the reactive electrophilic group is capable of reacting with nucleic acids.

[0258] Embodiment 11. The method of any one of embodiments 1-10, wherein the PIC comprises a nucleic acid binding ligand.

[0259] Embodiment 12. The method of embodiment 11, wherein the nucleic acid binding ligand is an intercalator.

[0260] Embodiment 13. The method of embodiment 12, wherein the intercalator is acridine.

[0261] Embodiment 14. The method of any one of embodiments 11-13, wherein the PIC comprises a frangible linker connecting the functional group and the nucleic acid binding ligand.

[0262] Embodiment 15. The method of any one of embodiments 1-14, wherein the PIC is β-alanine, N-(acridin-9-yl), 2-[bis(2-chloroethyl)amino]ethyl ester.

[0263] Embodiment 16. The method of any one of Embodiments 1-15, wherein step (a) comprises mixing a whole blood composition, an effective amount of PIC, and an effective amount of a quencher in the first container.

[0264] Embodiment 17. The method of embodiment 16, wherein the concentration of the quencher in the mixture of step (a) is at least about 1 mM.

[0265] Embodiment 18. The method of embodiment 16 or embodiment 17, wherein the concentration of the quencher in the mixture of step (a) is about 1 mM to about 10 mM.

[0266] Embodiment 19. The method of any one of embodiments 16-18, wherein the concentration of the quencher in the mixture of step (a) is about 2 mM.

[0267] Embodiment 20. The method of any one of embodiments 16-19, wherein the concentration of the PIC in the mixture of step (a) is about 0.2 mM and the concentration of the quencher in the mixture of step (a) is about 2 mM.

[0268] Embodiment 21. The method of any one of Embodiments 16-20, wherein the quencher comprises a sulfhydryl group, wherein the sulfhydryl group is capable of reacting with the reactive electrophilic group of the pathogen-inactivating compound.

[0269] Embodiment 22. The method of any one of Embodiments 16-21, wherein the quencher comprises cysteine ​​or a derivative of cysteine.

[0270] Embodiment 23. The method of any one of embodiments 16-22, wherein the quencher is glutathione or a pharmaceutically acceptable salt thereof.

[0271] Embodiment 24. The method of any one of embodiments 16-23, wherein the quencher is glutathione monosodium salt.

[0272] Embodiment 25. The method of any one of embodiments 1-24, wherein the mixture of step (a) further comprises an anticoagulant.

[0273] Embodiment 26. A method as described in any of Embodiments 1-25, wherein the whole blood composition further comprises an anticoagulant.

[0274] Embodiment 27. The method of any one of Embodiments 1-25, wherein the first container comprises an anticoagulant and the whole blood composition is mixed with the anticoagulant in the first container.

[0275] Embodiment 28. The method of any one of Embodiments 1-27, wherein the whole blood composition has a volume of about 250 mL to about 700 mL.

[0276] Embodiment 29. The method of any one of Embodiments 25-28, wherein the ratio of anticoagulant to whole blood in the whole blood composition is about 1:5 to about 1:9.

[0277] Embodiment 30. The method of any one of Embodiments 1-29, wherein the whole blood composition is added to the first container through a first inlet of the first container.

[0278] Embodiment 31. The method of any one of Embodiments 1-30, wherein the whole blood composition is added to the first container from a donor collection container containing the whole blood composition.

[0279] Embodiment 32. The method of any one of Embodiments 1-30, wherein the whole blood composition is added to the first container by collecting the whole blood composition from a whole blood donor into the first container.

[0280] Embodiment 33. A method as described in embodiment 32, wherein the whole blood composition is collected from a whole blood donor into the first container via a first conduit extending between a vascular access device and a first inlet of the first container and providing a fluid communication path for transferring the whole blood composition from the vascular access device to the first container.

[0281] Embodiment 34. The method of any one of Embodiments 1-33, wherein the PIC and optionally the quencher are added to the first container through a first inlet of the first container.

[0282] Embodiment 35. The method of any one of Embodiments 1-33, wherein the PIC and optionally the quencher are added to the first container through a second inlet of the first container.

[0283] Embodiment 36. The method of embodiment 34 or embodiment 35, wherein the PIC and optionally the quencher are added to the first vessel via a conduit extending between a vessel containing the PIC (PIC vessel) and an optional vessel containing the quencher (quencher vessel) and an inlet of the first vessel and providing a fluid communication path for transferring the PIC from the PIC vessel to the first vessel and optionally for transferring the quencher from the optional quencher vessel to the first vessel.

[0284] Embodiment 37. The method of embodiment 36, wherein the PIC and the quencher are added to the first container via a conduit extending between the PIC container and the quencher container and an inlet of the first container, and wherein the conduit comprises at one end: (a) a first adapter for coupling the PIC container to the conduit, (b) a second adapter for coupling the quencher container to the conduit, and optionally (c) an air displacement device for increasing transfer of the PIC and the quencher through the conduit and into the first container.

[0285] Embodiment 38. The method of any one of embodiments 1-37, wherein the incubation step (b) is from about 2 hours to about 24 hours.

[0286] Embodiment 39. The method of embodiment 38, wherein the incubation step (b) is from about 18 hours to about 22 hours.

[0287] Embodiment 40. The method of any one of embodiments 1-39, wherein the incubation step (b) is at room temperature.

[0288] Embodiment 41. The method of any one of Embodiments 1-40, wherein the transferring step (c) is performed via a conduit extending between the first container and the second container.

[0289] Embodiment 42. The method of embodiment 41, wherein the conduit connects the first outlet of the first container to the first inlet of the second container.

[0290] Embodiment 43. The method of any one of embodiments 1-42, wherein the incubation step (d) is from about 1 hour to about 8 hours.

[0291] Embodiment 44. The method of embodiment 43, wherein the incubation step (d) is from about 2 hours to about 4 hours.

[0292] Embodiment 45. The method of any one of embodiments 1-44, wherein the incubation step (d) is at room temperature.

[0293] Embodiment 46. The method of any one of Embodiments 1-45, further comprising transferring the pathogen-inactivated whole blood composition from the second container to a third container.

[0294] Embodiment 47. The method of any one of Embodiments 1-46, further comprising (e) storing the pathogen-inactivated whole blood composition at about 2-6°C.

[0295] Embodiment 48. The method of any one of Embodiments 1-47, wherein the pathogen-inactivated whole blood composition is suitable for transfusion.

[0296] Embodiment 49. The method of any one of Embodiments 1-48, further comprising separating the pathogen-inactivated whole blood composition into one or more of a red blood cell composition, a plasma composition, or a platelet composition.

[0297] Embodiment 50. The method of any one of Embodiments 1-49, further comprising separating the pathogen-inactivated whole blood composition into a plasma composition, and preparing a cryoprecipitate composition from the plasma composition.

[0298] Embodiment 51. A pathogen-inactivated whole blood composition prepared by the method of any one of embodiments 1-48.

[0299] Embodiment 52. The pathogen-inactivated whole blood composition of embodiment 51, wherein the pathogen-inactivated whole blood composition is suitable for transfusion.

[0300] Embodiment 53. A red blood cell composition prepared by the method of embodiment 49.

[0301] Embodiment 54. A cryoprecipitate composition prepared by the method of embodiment 50.

[0302] Embodiment 55. A method of transfusing a pathogen-inactivated whole blood composition into a subject in need thereof, the method comprising transfusing a pathogen-inactivated whole blood composition prepared by the method of any one of embodiments 1-48 or the pathogen-inactivated whole blood composition of embodiment 51 or embodiment 52 into the subject.

[0303] Embodiment 56. The method of embodiment 55, wherein the method comprises coupling an infusion line to the first outlet of the second container and infusing the pathogen-inactivated whole blood composition into the subject.

[0304] Embodiment 57. A kit for preparing a pathogen-inactivated whole blood composition, comprising:

[0305] (a) A vascular access device for withdrawing whole blood from a donor;

[0306] (b) a first container suitable for mixing the whole blood and a pathogen inactivating compound (PIC), wherein the first container comprises at least a first inlet and a first outlet;

[0307] (c) a first conduit extending between the vascular access device and the first inlet of the first container and providing a fluid communication path for transferring the whole blood from the vascular access device to the first container; and

[0308] (d) Containers containing pathogen-inactivating compounds (PIC containers).

[0309] Embodiment 58. The kit of embodiment 57, wherein the PIC container is configured to be coupled to the first inlet of the first container via a fluid communication path.

[0310] Embodiment 59. The kit of Embodiment 58, wherein the PIC container is configured to be coupled to the first conduit.

[0311] Embodiment 60. The kit of embodiment 57, wherein the first container comprises a second inlet and the PIC container is configured to be coupled to the second inlet of the first container via a fluid communication path.

[0312] Embodiment 61. The kit of any one of Embodiments 57-60, further comprising a container comprising a quencher (quencher container).

[0313] Embodiment 62. The kit of Embodiment 61, wherein the quencher container is configured to be coupled to the first inlet of the first container via a fluid communication path.

[0314] Embodiment 63. The kit of Embodiment 62, wherein the quencher container is configured to be coupled to the first conduit.

[0315] Embodiment 64. The kit of Embodiment 61, wherein the quencher container is configured to be coupled to the second inlet of the first container via a fluid communication path.

[0316] Embodiment 65. The kit of any one of Embodiments 60-64, wherein the PIC container and the quencher container are the same container.

[0317] Embodiment 66. The kit of any one of Embodiments 60-65, further comprising a second conduit, wherein the second conduit comprises at one end at least one adapter for coupling the PIC container and / or the quencher container to the second conduit.

[0318] Embodiment 67. The kit of Embodiment 66, wherein the second conduit comprises, at one end, a first adapter for coupling the PIC container to the second conduit and a second adapter for coupling the quencher container to the second conduit.

[0319] Embodiment 68. The kit of any one of Embodiments 60-65, further comprising a second conduit, wherein the second conduit is coupled at one end to the PIC container and / or the quencher container.

[0320] Embodiment 69. The kit of any one of Embodiments 66-68, wherein the second conduit is configured to couple to the first container, thereby providing a fluid communication path between the PIC container and / or quencher container and the first container.

[0321] Embodiment 70. The kit of Embodiment 69, wherein the second conduit is configured to couple to the first inlet of the first container.

[0322] Embodiment 71. The kit of Embodiment 69 or Embodiment 70, wherein the second conduit is coupled to the second inlet of the first container.

[0323] Embodiment 72. The kit of any one of Embodiments 66-68, wherein the second conduit is configured to couple to the first conduit to provide a fluid communication path between the PIC container and / or quencher container and the first container.

[0324] Embodiment 73. The kit of any one of Embodiments 66-72, further comprising an air displacement device for increasing transfer of the PIC and / or the quencher through the second conduit and into the first container.

[0325] Embodiment 74. The kit of any one of Embodiments 57-73, wherein the first container comprises an anticoagulant.

[0326] Embodiment 75. The kit of any one of Embodiments 57-74, further comprising a second container suitable for storing the pathogen-inactivated whole blood composition, wherein the second container comprises at least a first inlet and a first outlet.

[0327] Embodiment 76. The kit of embodiment 75, further comprising a third conduit extending between the first outlet of the first container and the first inlet of the second container and providing a fluid communication path for transferring the mixture comprising the whole blood composition and the PIC from the first container to the second container.

[0328] Embodiment 77. The kit of embodiment 75 or embodiment 76, wherein the kit comprises a third container suitable for storing a pathogen-inactivated whole blood composition, wherein the third container comprises at least a first inlet and a first outlet, and wherein the kit further comprises a fourth conduit extending between the first outlet of the second container and the first inlet of the third container and providing a fluid communication path for transferring a mixture comprising a whole blood composition and PICs from the second container to the third container.

[0329] Embodiment 78. A kit as described in any of Embodiments 1-77, further comprising an infusion line for infusing the pathogen-inactivated whole blood composition into a subject, wherein the infusion line comprises a second vascular access device and an infusion line conduit extending between the first outlet of the second container and the second vascular access device, wherein the infusion line conduit provides a fluid communication path for aseptically transferring the pathogen-inactivated whole blood composition from the second container to the vascular access device.

[0330] Embodiment 79. A kit for preparing a pathogen-inactivated whole blood composition, comprising:

[0331] (a) a first container suitable for mixing a whole blood composition, a pathogen inactivating compound (PIC), and a quencher, wherein the first container comprises at least a first inlet and a first outlet;

[0332] (b) a second container suitable for storing the pathogen-inactivated whole blood composition, wherein the second container comprises at least a first inlet and a first outlet;

[0333] (c) Containers containing pathogen-inactivating compounds (PIC containers);

[0334] (d) a container containing a quencher (quencher container);

[0335] (e) a first conduit, wherein the first conduit is coupled or configured to be coupled at one end to a PIC container and a quencher container, and wherein the first conduit further comprises an air displacement device for increasing transfer of the PIC and the quencher through the first conduit; and

[0336] (f) a second conduit extending between the first container and the second container and providing a fluid communication path for transferring the whole blood composition from the first container to the second container.

[0337] Embodiment 80. The kit of Embodiment 79, wherein the first conduit further comprises at least one adapter for coupling the second conduit to a PIC container and a quencher container.

[0338] Embodiment 81. The kit of Embodiment 80, wherein the first conduit comprises a first adapter for coupling a PIC container to the first conduit and a second adapter for coupling a quencher container to the first conduit.

[0339] Embodiment 82. The kit of any one of Embodiments 79-81, wherein the first conduit is coupled to a PIC container and a quencher container.

[0340] Embodiment 83. The kit of any one of Embodiments 79-82, wherein the PIC container and the quencher container are the same container.

[0341] Embodiment 84. The kit of any one of Embodiments 79-83, wherein the first conduit is configured to couple to the first container, thereby providing a fluid communication path between the PIC container and the quencher container and the first container.

[0342] Embodiment 85. The kit of Embodiment 83, wherein the first conduit is configured to couple to an inlet of the first container.

[0343] Embodiment 86. A kit as described in any of Embodiments 79-85, further comprising a vascular access device for withdrawing whole blood from a donor and a third conduit extending between the vascular access device and the first inlet of the first container and providing a fluid communication path for transferring the whole blood from the vascular access device to the first container.

[0344] Embodiment 87. The kit of Embodiment 86, wherein the first container further comprises a second inlet, and wherein the first conduit is configured to couple to the second inlet of the first container.

[0345] Embodiment 88. The kit of Embodiment 86, wherein the first conduit is configured to couple to the third conduit to provide a fluid communication path between the PIC container and quencher container and the first container.

[0346] Embodiment 89. The kit of any one of Embodiments 79-88, wherein the first container comprises an anticoagulant.

[0347] Embodiment 90. A kit as described in any of Embodiments 79-89, wherein the kit includes a third container suitable for storing a pathogen-inactivated whole blood composition, wherein the third container contains (e.g., includes) at least a first inlet and a first outlet, and wherein the kit further includes a fourth conduit extending between the first outlet of the second container and the first inlet of the third container and providing a fluid communication path for transferring the whole blood composition from the second container (e.g., under sterile conditions) into the third container.

[0348] Embodiment 91. A kit as described in any of Embodiments 79-90, further comprising an infusion line for infusing the pathogen-inactivated whole blood composition into a subject, wherein the infusion line comprises a second vascular access device and an infusion line conduit extending between the first outlet of the second container and the second vascular access device, wherein the infusion line conduit provides a fluid communication path for aseptically transferring the pathogen-inactivated whole blood composition from the second container to the vascular access device.

Claims

1. A kit for preparing a pathogen-inactivated whole blood composition, comprising: (a) a vascular access device for withdrawing whole blood from a donor; (b) a first container suitable for mixing the whole blood and a pathogen inactivating compound (PIC), wherein the first container comprises at least a first inlet and a first outlet; (c) a first conduit extending between the vascular access device and the first container and providing a fluid communication path for transferring the whole blood from the donor to the first container; and (d) containers containing pathogen inactivating compounds (PIC containers); Optionally, the PIC container is configured to be coupled to the first container via a fluid communication path; Optionally, the PIC container is configured to be coupled to the first conduit; Optionally, the PIC container is coupled to the first container via a fluid communication path; Optionally, the first container further comprises a second inlet and the PIC container is configured to be coupled to the second inlet of the first container via a fluid communication path; Optionally, the first container further comprises a second inlet and the PIC container is coupled to the second inlet of the first container via a fluid communication path.

2. The kit of claim 1, further comprising a container comprising a quencher (quencher container); Optionally, the quencher container is configured to be coupled to the first container via a fluid communication path; Optionally, the quencher vessel is configured to be coupled to the first conduit; Optionally, the quencher vessel is coupled to the first vessel via a fluid communication path; Optionally, the quencher vessel is configured to be coupled to the second inlet of the first vessel via a fluid communication path; Optionally, the quencher vessel is coupled to the second inlet of the first vessel by a fluid communication path.

3. The kit of claim 1 or 2, wherein the PIC container and the quencher container are the same container.

4. The kit of any one of claims 1-3, further comprising a second conduit, wherein the second conduit comprises at one end at least one adapter for coupling a PIC container to the second conduit; Optionally, the second conduit comprises at one end a first adapter for coupling a PIC container to the second conduit and a second adapter for coupling a quencher container to the second conduit; Optionally, the kit further comprises a second conduit, wherein the second conduit is coupled at one end to the PIC container and / or the quencher container; Optionally, the second conduit is configured to be coupled to the first container; Optionally, the second conduit is coupled to the first container; Optionally, the second conduit is configured to couple to the first conduit, thereby providing a fluid communication path between the PIC container and / or quencher container and the first container; Optionally, the kit further comprises a displacement device adapted to increase transfer of the PIC and / or the quencher into the first container; Optionally, the first container comprises an anticoagulant; Optionally, the kit further comprises a second container suitable for storing the pathogen-inactivated whole blood composition, wherein the second container comprises at least a first inlet and a first outlet; Optionally, the kit further comprises a third conduit extending between the first container and the second container and providing a fluid communication path for transferring a mixture comprising a whole blood composition and a PIC from the first container to the second container; Optionally, the kit comprises a third container suitable for storing a pathogen-inactivated whole blood composition, wherein the third container comprises at least a first inlet and a first outlet, and wherein the kit further comprises a fourth conduit extending between the second container and the third container and providing a fluid communication path for transferring a mixture comprising a whole blood composition and a PIC from the second container to the third container.

5. The kit of any one of claims 1-4, further comprising an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject; Optionally, the kit further comprises an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject, wherein the infusion line assembly is coupled to the second container; Optionally, the infusion line assembly comprises a second vascular access device and an infusion line catheter extending between the first outlet of the second container and the second vascular access device, wherein the infusion line catheter provides a fluid communication path for aseptically transferring the pathogen-inactivated whole blood composition from the second container to the vascular access device.

6. The kit of any one of claims 1 to 5, wherein the PIC comprises a functional group that acts as or forms a reactive electrophilic group; Optionally, the PIC comprises a nucleic acid binding ligand; Optionally, the PIC is β-alanine, N-(acridin-9-yl), 2-[bis(2-chloroethyl)amino]ethyl ester; Optionally, the quencher comprises a thiol group, wherein the thiol group is capable of reacting with a reactive electrophilic group of the PIC; Optionally, the quencher is glutathione or a pharmaceutically acceptable salt thereof.

7. A kit for preparing a pathogen-inactivated whole blood composition, comprising: (a) a first container suitable for mixing a whole blood composition, a pathogen inactivating compound (PIC), and a quencher, wherein the first container comprises at least a first inlet and a first outlet; (b) a second container suitable for storing a pathogen-inactivated whole blood composition, wherein the second container comprises at least a first inlet and a first outlet; (c) containers containing pathogen inactivating compounds (PIC containers); (d) a container containing a quencher (quencher container); (e) a first conduit, wherein the first conduit is coupled or configured to be coupled at one end to a PIC container and a quencher container, and wherein the first conduit further comprises a displacement device adapted to increase transfer of the PIC and the quencher into the first container; as well as (f) a second conduit extending between the first container and the second container and providing a fluid communication path for transferring the whole blood composition from the first container to the second container; Optionally, the first conduit further comprises at least one adapter for coupling the first conduit to a PIC container and a quencher container; Optionally, the first conduit comprises a first adapter for coupling a PIC container to the first conduit and a second adapter for coupling a quencher container to the first conduit; Optionally, the first conduit is coupled to a PIC container and a quencher container; Optionally, the PIC container and the quencher container are the same container; Optionally, the first conduit is coupled to or configured to be coupled to the first container, thereby providing a fluid communication path between the PIC container and the quencher container and the first container; Optionally, the first conduit is configured to be coupled to an inlet of the first container; Optionally, the first conduit is coupled to an inlet of the first container; Optionally, the first container further comprises a second inlet, and wherein the first conduit is coupled to or configured to be coupled to the second inlet of the first container.

8. The kit of claim 7, further comprising a vascular access device for withdrawing whole blood from a donor and a third conduit extending between the vascular access device and the first inlet of the first container and providing a fluid communication path for transferring the whole blood from the donor into the first container; Optionally, the first conduit is configured to couple to the third conduit, thereby providing a fluid communication path between the PIC container and the quencher container and the first container; Optionally, the first container comprises an anticoagulant.

9. A kit as described in claim 7 or 8, wherein the kit includes a third container suitable for storing a pathogen-inactivated whole blood composition, wherein the third container includes a first inlet and a first outlet, and wherein the kit also includes a fourth conduit extending between the second container and the third container and providing a fluid communication path for transferring the whole blood composition from the second container to the third container.

10. The kit of any one of claims 7-9, further comprising an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject; Optionally, the kit further comprises an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject, wherein the infusion line assembly is coupled to the second container; Optionally, the kit further comprises an infusion line assembly for infusing the pathogen-inactivated whole blood composition into a subject, wherein the infusion line assembly comprises a second vascular access device and an infusion line catheter extending between the first outlet of the second container and the second vascular access device, wherein the infusion line catheter provides a fluid communication path for aseptically transferring the pathogen-inactivated whole blood composition from the second container to the vascular access device.

11. The kit of any one of claims 7 to 10, wherein the PIC comprises a functional group that acts as or forms a reactive electrophilic group; Optionally, the PIC comprises a nucleic acid binding ligand; Optionally, the PIC is β-alanine, N-(acridin-9-yl), 2-[bis(2-chloroethyl)amino]ethyl ester; Optionally, the quencher comprises a thiol group, wherein the thiol group is capable of reacting with a reactive electrophilic group of the PIC; Optionally, the quencher is glutathione or a pharmaceutically acceptable salt thereof.

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