Abstract:
Method and apparatus for separating plasma from blood in a separation vessel, separating the separated plasma into desired plasma proteins in a plasma separator fluidly connected to the separation vessel to receive the separated plasma, and adding photosensitizer to the desired plasma proteins for viral inactivation treatment of such proteins.

Description:
FIELD 
       [0001]    This patent application relates to on-line pathogen reduction treatment of a separated plasma product. 
       BACKGROUND 
       [0002]    For transfusions of blood and blood components, whole blood from a single donor is typically separated into three components: plasma, red blood cells and platelets. Each component may be used to treat a multiplicity of specific conditions and disease states. For example, the red blood cell component may be used to treat anemia and replace red blood cell loss due to bleeding, the concentrated platelet component may be used to control bleeding, and the plasma component may be given to patients to increase blood volume, or may be separated off-line after collection into individual plasma proteins such as fibrinogen, von Willebrand factor, Factor VIII, Factor IX, Anti-thrombin III, Fibrin sealant, thrombin, Alpha I and IVIG. Plasma from multiple donors may also be collected and combined or pooled together, and the combined plasma pool fractionated into the desired plasma proteins. 
         [0003]    The separation of the collected plasma component into various protein or plasma components or fractions is called plasma fractionation. Such fractionation is typically done by large scale fractionators which combine plasma from many donors and concentrate plasma proteins from the collected plasma by using the known techniques of cold alcohol fractionation (also known as Cohn fractionation) and chromatography. 
         [0004]    There are traditionally two ways to obtain separated blood components from single donors. One way is to collect whole blood from donors and separate it into components some time period after the whole blood collection. Using this method, whole blood is collected into approved containers that are pyrogen-free and sterile, with sufficient anticoagulant for the quantity of blood to be collected. Whole blood which is collected in this way is separated into components in a lab by a technician, and separation typically occurs from between about 2 and 8 hours after collection in the United States, and between about 2 to 24 hours in Europe. 
         [0005]    Another way to separate whole blood into components is by using an apheresis device. Such apheresis devices separate whole blood from a single donor connected on-line to the device into components automatically, and return any uncollected and unneeded blood components back to the donor during the collection procedure. 
         [0006]    Apheresis devices may be used to separate the plasma component from the cellular components of a blood donation. Apheresis devices permit more frequent donations by a single donor due to the return of uncollected components. US Publication No.: US 2010-0042037 discloses separation of plasma proteins on-line or while connected to a donor using an apheresis system. 
         [0007]    Pathogen reduction treatment may be used to reduce pathogens in whole blood or collected and separated blood products. Such treatments may include the addition of a photosensitizer and activation of such photosensitizer with light. One such treatment using riboflavin is described in U.S. Pat. No. 6,258,577. 
       BRIEF SUMMARY 
       [0008]    The embodiments relate to a method of collecting plasma fractions from whole blood comprising rotating a separation vessel; separating plasma from other blood components of the whole blood in the rotating separation vessel; providing separated plasma from the rotating separation vessel to a plasma separator; separating the plasma into at least one fraction including desired plasma proteins using the plasma separator; mixing the at least one fraction with photosensitizer; collecting the at least one fraction with photosensitizer after the mixing step. 
         [0009]    Another aspect is an apheresis plasma separation system comprising: a rotor; a separation vessel mounted on the rotor for rotating therewith wherein blood is separated into plasma and other components in the separation vessel during rotation of the rotor; a plasma separator fluidly connected to the separation vessel to receive the separated plasma from the rotating separation vessel; the plasma separator comprising: a hollow fiber membrane wherein the hollow fiber membrane can separate at least some plasma proteins from the separated plasma; photosensitizer fluidly connected to the plasma separator; a collection container fluidly connected to the plasma separator for collecting the separated plasma proteins from the plasma separator with the photosensitizer. 
         [0010]    A further embodiment is an integrated pre-connected disposable set for an apheresis system comprising: a removal/return assembly for removing and returning blood and blood components to a donor; a separation vessel fluidly connected to the removal/return assembly and adapted for mounting on a centrifuge for rotation wherein whole blood is separated in the separation vessel during rotation of the centrifuge into at least plasma and other components; a plasma collect assembly fluidly connected to the separation vessel to receive separated plasma comprising: a membrane plasma separator for separating the separated plasma into at least a plasma protein fraction; photosensitizer in the plasma collect assembly; and a collection container fluidly connected to the plasma separator to receive the plasma protein fraction and the photosensitizer. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0011]      FIG. 1  is a schematic view of an apheresis system including a plasma separation unit. 
           [0012]      FIG. 2  is a schematic view of the blood separator/plasma separation tubing set for the system of  FIG. 1 . 
           [0013]      FIG. 3  is a schematic view of the blood separator/plasma separation tubing set of  FIG. 2  with optional dry cartridge. 
           [0014]      FIG. 4  is a schematic view of a detail of the tubing set cassette of  FIGS. 2 and 3 . 
           [0015]      FIG. 5  is a simplified schematic view of the plasma separation unit of  FIG. 2  with optional pump location. 
           [0016]      FIG. 6  is a simplified schematic view of the plasma separator unit of  FIG. 3  with optional pump location. 
           [0017]      FIG. 7  is a schematic detail of an alternative plasma protein collection assembly with prion filter for the tubing set of  FIG. 2 . 
           [0018]      FIG. 8  is a schematic detail of an alternative plasma protein collection assembly with prion filter for the tubing set of  FIG. 3 . 
           [0019]      FIG. 9  is a schematic view of the plasma protein collection assembly of  FIG. 8  with on-line illumination. 
           [0020]      FIG. 10  is a schematic detail of the tubing set of  FIG. 8  showing an alternate location for the cartridge and prion filter. 
           [0021]      FIG. 11  is a schematic view of an alternative blood separator/tubing set with additional plasma component collection. 
           [0022]      FIG. 12  is a schematic view of an alternative blood separator/tubing set with plasma protein collection from separation from the extracapilary side of the plasma separator. 
           [0023]      FIG. 13  is a schematic view of a bag of collected plasma proteins being treated for pathogen reduction. 
           [0024]      FIG. 14  is a block diagram illustrating a process using pathogen reduction treatment and collected apheresis plasma proteins for further fractionation. 
       
    
    
     DETAILED DESCRIPTION 
       [0025]    It should be noted that like elements are represented using like numerals in all the figures and the description. One embodiment is described with reference to the TRIMA Accel® automated collection system (manufactured and sold by Terumo BCT, Inc., Lakewood, Colo., USA) but it should be noted that any apheresis system, such as, but not limited to the COBE® SPECTRA system, SPECTRA OPTIA® system and the TRIMA® automatic collection system all also manufactured and sold by Terumo BCT, Inc. may be used without departing from the spirit and scope of the invention. 
         [0026]    The embodiments described herein may also may be used with the apheresis systems of other manufacturers such as the Autopheresis C system manufactured by Fenwal, Inc. Lake Zurich, Ill., U.S.A. or the PCS system as manufactured by Haemonetics Corp. of Bainbridge, Mass. 
         [0027]    The embodiments may also use an endogenous photosensitizer, though other photosensitizers could be used. A “photosensitizer” is defined as any compound which absorbs radiation of one or more defined wavelengths and subsequently utilizes the absorbed energy to carry out a chemical process. Examples of such photosensitizers include porphyrins, psoralens, dyes such as neutral red, methylene blue, acridine, toluidines, flavine (acriflavine hydrochloride) and phenothiazine derivatives, coumarins, quinolones, quinones, and anthroquinones. 
         [0028]    Also, endogenous photosensitizers may be used. The term “endogenous” means naturally found in a human or mammalian body, either as a result of synthesis by the body or because of ingestion as an essential foodstuff (e.g. vitamins) or formation of metabolites and/or byproducts in vivo. Examples of such endogenous photosensitizers are alloxazines such as 7,8-dimethyl-10-ribityl isoalloxazine (riboflavin), 7,8,10-trimethylisoalloxazine (lumiflavin), 7,8-dimethylalloxazine (lumichrome), isoalloxazine-adenine dinucleotide (flavine adenine dinucleotide [FAD]), alloxazine mononucleotide (also known as flavine mononucleotide [FMN] and riboflavine-5-phosphate), vitamin Ks, vitamin L, their metabolites and precursors, and napththoquinones, naphthalenes, naphthols and their derivatives having planar molecular conformations. The term “alloxazine” includes isoalloxazines. 
         [0029]    The fluid containing the photosensitizer is exposed to photoradiation of the appropriate wavelength to activate the photosensitizer, using an amount of photoradiation sufficient to activate the photosensitizer, but less than that which would cause non-specific damage to the biological components or substantially interfere with biological activity of other proteins present in the fluid. The wavelength used will depend on the photosensitizer selected. 
         [0030]    The activated photosensitizer inactivates any microorganisms contained in the fluid. As used herein, the term “inactivation of a microorganism” means totally or partially preventing the microorganism from replicating, either by killing the microorganism or otherwise interfering with its ability to reproduce. 
         [0031]    One embodiment further includes an optional prion filter. The P-Capt® filter manufactured by Macopharma of Mouvaux, France removes prions from at least one blood component. This filter uses ligand technology attached to resin with filter media. Prion proteins attach to the ligands for removal from a blood component product. 
         [0032]    A blood apheresis system  2  is illustrated in  FIG. 1  and allows for a continuous blood component separation process. Generally, in a continuous system or an on-line system, whole blood is withdrawn from a donor/patient  4  and provided to a blood component separation device  6  where the blood is separated into the individual blood components with at least one of these blood components being removed from the device  6  with the other components being returned to the donor. The continuous system  2  also provides for further separation or concentration of plasma into plasma proteins and the addition of photosensitizer to the plasma proteins for collection. Also, the continuous system may provide for filtration for removal of prions. 
         [0033]    In the blood apheresis system  2 , blood is withdrawn from the donor/patient  4  and directed through a pre-connected disposable set  8  (the disposable set embodiment of  FIG. 2  is shown in  FIG. 1 ) which includes an extracorporeal tubing circuit  10 , a blood processing or separation vessel  352  and a plasma separator or concentrator  205  which defines a completely closed and sterile system. The disposable set  8  is mounted on the blood component separation device  6  which includes a pump/valve/sensor assembly  1000  for interfacing with the extracorporeal tubing circuit  10 , and a channel assembly  200  for interfacing with the disposable blood processing vessel  352 . 
         [0034]    The channel assembly  200  includes a channel housing  204  which is rotatably interconnected with a rotatable centrifuge rotor assembly  568  which provides the centrifugal forces required to separate blood into its various blood component types by centrifugation. The blood processing vessel  352  is inter-fitted into the channel housing  204  to fit with a groove or channel in the channel housing. Blood thus flows from the donor/patient  4 , through the extracorporeal tubing circuit  10 , and into the rotating blood processing vessel  352 . The blood within the blood processing vessel  352  is separated into various blood component types and at least one of these blood component types (e.g., plasma) is continually removed from the blood processing vessel  352 . The plasma component may then be further concentrated or separated into plasma proteins to which photosensitizer is added as part of the continuous on-line system. Blood components which are not being retained for collection or for use in therapeutic treatments are also removed from the blood processing vessel  352  and returned to the donor/patient  4  via the extracorporeal tubing circuit  10 . The blood processing vessel  352  may optionally be used for a platelet collection although such collection will not be described. 
         [0035]    Operation of the blood component separation device  6  is controlled by one or more processors, not shown. In order to assist the operator of the apheresis system  2  with various aspects of its operation, the blood component separation device  6  includes a graphical interface  660  with a touch screen input/output device  664  connected to the processor. 
         [0036]    The apheresis system below will be described with respect to a red blood cell collection and a plasma protein collection. Although an additional red blood cell collection is described it is further understood that other blood components, such as, but not limited to, platelets could alternatively, or, in addition, be collected. It is understood, however, that a plasma collection only may also occur if desired. If plasma collection only is desired, with subsequent separation and collection of plasma proteins, and addition of photosensitizer, the system described below can be simplified. For example, the red blood cell collection assembly  950  could be deleted. Also the replacement fluid assembly  960  can be optional. Collecting plasma proteins only with no red blood cell collection can provide a simplified closed system. 
         [0037]    The apheresis system will also be disclosed with respect to pathogen reduction treatment of the desired plasma proteins to be collected and optional prion filtration. 
         [0038]    As illustrated in  FIG. 2 , and alternate embodiments  FIGS. 3 ,  11  and  12 , blood-primable pre-connected extracorporeal tubing circuit  10  comprises a cassette assembly  110  and a number of tubing assemblies  20 ,  50 ,  60 ,  950 ,  90 ,  100  and optionally  960  interconnected therewith. Generally, blood removal/return tubing assembly  20  provides a single needle interface between a donor/patient  4  and cassette assembly  110 , and blood inlet/blood component tubing subassembly  60  provides the interface between cassette assembly  110  and blood processing vessel  352 . An anticoagulant tubing assembly  50 , plasma or plasma protein collection tubing assembly  90 , red blood cell collection assembly  950  and vent bag tubing subassembly  100  are also interconnected with cassette assembly  110 . Optionally, a replacement fluid sub-assembly  960  may be included. The extracorporeal tubing circuit  10  including the assemblies or sub-assemblies above and blood processing vessel  352  are interconnected to yield a closed disposable system or pre-connected disposable for a single use. 
         [0039]    Cassette assembly  110  includes front and back molded plastic plates (not shown) that are hot-welded together to define a rectangular cassette member  115  having integral fluid passageways. The cassette assembly  110  further includes a number of outwardly extending tubing loops, described below, interconnecting various integral passageways. The integral passageways are also interconnected to the various tubing assemblies. 
         [0040]    The blood removal/return tubing assembly  20  includes a needle subassembly  30  interconnected with blood removal tubing  22 , blood return tubing  24  and anticoagulant tubing  26  via a common manifold  28 . The needle subassembly  30  includes a needle  32  having a protective needle sleeve  34  and needle cap  36 , and interconnect tubing  38  between needle  32  and manifold  28 . Needle subassembly  30  further includes a D sleeve  40  and tubing clamp  42  positioned about the interconnect tubing  38 . Blood removal tubing  22  may be provided with a Y-connector  44  interconnected with a blood sampling subassembly  46 . 
         [0041]    As shown in  FIG. 4  the blood removal/return assembly includes first integral passageway  190   a  connected to the bottom of reservoir  150 , tubing loop  192  and second integral fluid passageway  190   b  interconnected with tubing loop  192  and blood return tubing  24 . 
         [0042]    As seen in  FIG. 4 , cassette assembly  110  of  FIGS. 2 ,  3 ,  11  and  12  includes a first integral anticoagulant passageway  120   a  interconnected with the anticoagulant tubing  26  of the blood removal/return tubing assembly  20 . The cassette assembly  110  further includes a second integral anticoagulant passageway  120   b  and a pump-engaging, anticoagulant tubing loop  122  between the first and second integral anticoagulant passageways  120   a ,  120   b . The second integral anticoagulant passageway  120   b  is interconnected with anticoagulant tubing  54 . The anticoagulant tubing assembly  50  includes a spike drip chamber  52 , ( FIGS. 2 ,  3 ,  11  and  12 ) connectable to an anticoagulant source, anticoagulant feed tubing  54  and a sterile barrier filter  56 . During use, the anticoagulant tubing assembly  50  supplies anticoagulant to the blood removed from a donor/patient  4  to reduce or prevent any clotting in the extracorporeal tubing circuit  10 . 
         [0043]    Cassette assembly  110  also includes a first integral blood inlet passageway  130   a  interconnected with blood removal tubing  22  of the blood removal/return tubing assembly  20 . The cassette assembly  110  further includes a second integral blood inlet passageway  130   b  and a pump-engaging, blood inlet tubing loop  132  between the first and second integral blood inlet passageways  130   a ,  130   b . The first integral blood inlet passageway  130   a  includes a first pressure-sensing module  134  and inlet filter  136 , and the second integral blood inlet passageway  130   b  includes a second pressure-sensing module  138 . The second integral blood inlet passageway  130   b  is interconnected with blood inlet tubing  62  of the blood inlet/blood component tubing assembly  60 . 
         [0044]    Blood inlet tubing  62  is also interconnected with input port  392  of blood processing vessel  352  to provide whole blood thereto for processing. To return separated blood components to cassette assembly  110 , the blood inlet/blood component tubing assembly  60  further includes a red blood cell (RBC) outlet tubing  64  with outlet port  520  and plasma outlet tubing  68  with outlet port  456 . Alternatively the outlet tubing and outlet ports could be for other blood components such as platelets. A control port for controlling the interface is shown at  61 . 
         [0045]    The blood inlet tubing  62 , RBC outlet tubing  64 , and plasma outlet tubing  68  all pass through first and second strain relief members  72  and  74  and a braided bearing member  76  there between. This advantageously allows for a sealess interconnection, as taught in U.S. Pat. No. 4,425,112 incorporated by reference herein. As shown, multi-lumen connectors  78  can be employed in the various tubing lines. 
         [0046]    An optional replacement fluid tubing assembly  960  may be provided for delivery of replacement fluid such as sterile saline solution(s) (or replacement/exchange RBCs or plasma, e.g.) to the donor/patient  4 . As shown, the replacement fluid assembly  960  includes at least a replacement fluid inlet tubing line  962  attached to the cassette  110  in fluid communication with an internal replacement fluid passageway  140   a  which is in turn connected to a replacement fluid tubing loop  142  which is connected back to the cassette  110  and an internal replacement fluid passageway  140   b . Further internal passageways or spurs  144   a  and  144   b  and a tubing loop  146  are also shown. Internal passageway  144   b  is blocked off to disallow any fluid flow therein or therethrough. No outlet tubing line is preferably connected thereto and passageway  144   b  may also be omitted. 
         [0047]    The replacement fluid assembly  960  further preferably includes one or more spike assemblies  964   a - 964   b  with optional associated sterile barrier devices  966   a - 966   b  and tubing connection lines  968   a - 968   b  which may be connected to tubing line  962  via a Y-connector  969  as shown. One or more slide clamp(s)  970  may also be included. As the plasma proteins may be frozen before use the sterile barrier devices  966   a - 966   b  are optional. 
         [0048]    Although the replacement fluid assembly is shown as introducing such fluid through  140   a  and tubing loop  142  such is only exemplary. In other words the fluid could be introduced through other tubing loops for return such as tubing loop  162  or such fluid could even be aspirated through tubing into the system. 
         [0049]    The plasma outlet tubing  68  of blood inlet/blood component tubing assembly  60  interconnects with a first integral plasma passageway  160   a  of cassette assembly  110 . Cassette assembly  110  further includes a pump-engaging, plasma tubing loop  162  interconnecting the first integral plasma passageway  160   a  and a second integral plasma passageway  160   b . The second integral plasma passageway  160   b  includes first and second spurs  164   a  and  164   b . The first spur  164   a  is interconnected to the plasma collection tubing assembly  90 . The plasma collection tubing assembly  90  may be employed to collect plasma proteins during use and includes plasma collector tubing  92 , plasma separator or plasma separator unit  205 , plasma collector tubing  93  and one or more plasma collection bags, containers or reservoirs  94 . A slide clamp  96  may be provided on plasma collector tubing  93 . The plasma collection tubing assembly  90  may also be employed for further separation of the plasma component as will be described in more detail below. 
         [0050]    The second spur  164   b  of the second integral plasma passageway  160   b  is interconnected to a plasma return tubing loop  166  to return plasma to donor/patient  4 . For such purpose, the plasma return tubing loop  166  is interconnected to the top of the blood return reservoir  150  of the cassette assembly  110 . 
         [0051]    The plasma return assembly also returns plasma after separation or concentration. The post separation return includes tubing  963  which connects to tubing  962 , spurs  140   a  and  140   b  as well as pumps engaging plasma tubing loop  142 . Spur  144   a  is connected to plasma return loop or tubing  146  to deliver plasma to cassette reservoir  150  for ultimate delivery to the donor/patient  4 . Spur  140   b  is not used in the configuration shown and is closed off. Similarly this sub-assembly can be used to provide replacement fluid through  962 ,  140   a , tubing loop  142 ,  140   b  and  146  to return reservoir  150 . If no replacement fluid is required the portion of tubing  962  related to the replacement fluid sub-assembly above the connection with tubing  963  may be omitted. 
         [0052]    Although the plasma return assembly is shown returning plasma through tubing loop  142 , the plasma could also be returned through another pump loop arrangement such as  162 . 
         [0053]    The plasma collection tubing assembly further includes a plasma separation sub-assembly shown in FIGS.  2 , 3 , 5 , 6 ,  11  and  12  including hollow fiber membrane separator or concentrator  205 . The hollow fiber membrane separator is the plasma separator or plasma separator unit. Tubing  92  is interconnected to the inlet  203  of the separator  205 . Tubing  93  is interconnected to the outlet  206  of the membrane separator  205 . Plasma collection tubing assembly  90  also includes tubing  963 , interconnected to second outlet  208  for returning plasma or proteins that are not to be collected. The plasma return assembly including return tubing  963  connects to tubing  962  and spur  140   a  as described above. 
         [0054]    The plasma protein membrane concentrator or plasma membrane separator  205  includes inlet  203  in first end cap  214  and outlet  206  in the opposite end cap  216 . 
         [0055]    Hollow fiber membranes are arranged between the two end caps  214  and  216 . Such hollow fiber membranes include inter-capillary space (IC) within the hollow fibers and an extra-capillary space (EC) outside the hollow fibers. The pore size of the membrane forming the hollow fibers may be selected so that components such as plasma or optionally, protein of selected molecular weight may pass between the IC and EC spaces. Thus, if plasma separated from whole blood enters through tubing  92  and inlet  203  into the IC space, plasma and any proteins able to pass through the membrane pores to the EC space will pass through outlet  208  and tubing  963 . 
         [0056]    Table 1 below shows various protein factions and their molecular weight in kilodalton. The pore size of the membrane can be chosen to have a cut off value to pass through the membrane all but the desired protein fractions such as those given in the table. For example, the pore size could be such to pass all below 50 kilodaltons or the pore size could be selected to pass through the membrane those in a range in which the cut off value is selected between 50 kDa and 1300 kDa. The pore size could be selected to have a cut off value even lower than 50 kilodaltons such that only plasma liquid and sodium chloride pass through the membrane with all plasma proteins being collected. 
         [0000]    
       
         
               
               
               
             
               
               
               
             
           
               
                   
                 TABLE 1 
               
               
                   
                   
               
               
                   
                 Constituent 
                 Molecular Weight (kDa) 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Cholesterol 
                 1,300 
               
               
                   
                 IgM 
                 950 
               
               
                   
                 Fibrinogen 
                 340 
               
               
                   
                 Factor VIII 
                 100-340 
               
               
                   
                 IgE 
                 190 
               
               
                   
                 IgD 
                 175 
               
               
                   
                 IgA 
                 160 
               
               
                   
                 IgG 
                 150 
               
               
                   
                 Haptogloblin 
                 100 
               
               
                   
                 Albumin 
                 66 
               
               
                   
                 A1Antitrysin 
                 54 
               
               
                   
                 Factor VII 
                 50 
               
               
                   
                   
               
             
          
         
       
     
         [0057]    For example, a membrane having a pore size such that only constituents with a molecular weight of less than 50 kDa will pass, all proteins in Table 1 will be collected through outlet  206  with only plasma less proteins being returned through outlet  208 . 
         [0058]    For another example, a membrane having a pore size such that only constituents having a molecular weight of less than 150 kDa will be returned, only a portion of the proteins with a molecular weight greater than 150 kDa will be collected. Plasma and other plasma proteins of less than 150 kDa molecular weight will pass through the membrane to outlet  208 . 
         [0059]    In the embodiment of  FIGS. 2 and 5 , plasma proteins to be collect are mixed with photosensitizer in bag or container  94 . Outlet  206  of the membrane separator  205  is fluidly connected to the collection container or bag  94 . Photosensitizer for pathogen reduction may be included in plasma collection bags, containers or reservoirs  94  as part of the tubing assembly and the plasma collection assembly. The pathogen reduction process will be more fully described below. The photosensitizer may be in liquid form in bag  94  or it, alternatively could be in dry form for mixing with the plasma proteins. The photosensitizer may be present in plasma collection bag  94  at any desired concentration from about 1 μM up to the solubility of the photosensitizer in the plasma proteins. For 7,8-dimethyl-10-ribityl isoalloxazine a concentration range between about 1 μM and about 160 μM may be used. The amount of photosensitizer to be mixed with the plasma proteins will be an amount sufficient to adequately inactivate microorganisms therein, but less than a toxic, (to humans or other mammals), or insoluble amount. 
         [0060]    If the bag, container or reservoir  94  is the container used for illumination, it should be photo-permeable. Blood bag or photo-permeable container  94  may be prepackaged to contain the photosensitizer in the either dry or aqueous form as shown in  FIGS. 2 and 5 . The dry form may be in a dry powder form, a pill, capsule, tablet form or in various combinations therefore. The term dry solid or dry form envisions the components being in a loose powered state or in a solid state such as a pill, capsule, tablet capable of dissolving in fluid or any equivalent thereof known to one skilled in the art. 
         [0061]    In the alternative embodiment of  FIGS. 3 and 6 , the photosensitizer is in dry form in cartridge  98  which is also part of the tubing assembly and the plasma collection assembly. The plasma proteins pass through cartridge  98 , mixing with the dry photosensitizer, on the way to bag or container  94 . The cartridge  98  could be another bag, a flask, a reservoir, a small cylinder or any similar container known in the art. Also the tubing  93  itself could contain certain forms of prepackaged photosensitizer. Cartridge  98  is located in plasma collector tubing  93  as shown in  FIGS. 3 and 6 . As described above the dry form may be in a dry powder form, a pill, capsule, tablet form or in various combinations therefore. 
         [0062]    The plasma collection assembly  90  may also include an optional prion filter. Such a filter may be a media filter as shown or, alternatively could be a membrane filter. As shown in  FIG. 7  the filter  95  may be located in tubing  93  between the outlet  206  of the membrane separator  205  and the collection bag  94 . If the cartridge  98  is used to introduce dry form photosensitzer into the system and into bag  94 , the filter  95  (shown in solid lines in  FIG. 8 ), may be alternatively between the outlet  206  of the membrane separator and the dry cartridge  98  in tubing  93 . Alternatively the filter may be between the cartridge  98  and the collection bag  94  in tubing  93  as shown in dashed lines in  FIG. 8 . 
         [0063]    As shown in  FIGS. 2 ,  3 , and  4  the RBC outlet tubing  64  of the blood inlet/blood component tubing assembly  60  is interconnected with integral RBC passageway  170  of cassette assembly  110  ( FIG. 4 ). The integral RBC passageway  170  includes first and second spurs  170   a  and  170   b , respectively. The first spur  170   a  is interconnected with RBC return tubing loop  172  to return separated RBC to a donor/patient  4 . For such purpose, the RBC return tubing loop  172  is interconnected to the top of blood return reservoir  150  of the cassette assembly  110 . The second spur  170   b  may be closed off if red blood cells are not to be collected or may be connected with an RBC collection tubing assembly  950 . 
         [0064]    RBC collection tubing assembly  950  includes RBC collector tubing  952 , at least one RBC collection reservoir, container, or bag  954 , and sterile barrier filter/drip spike assembly  956 . One or a larger practical number (not shown) of RBC bag(s)  954  may be connected to the collector tubing  952 . Moreover, although not shown here one or more white blood cell (WBC) filtration devices and/or RBC storage solution connections and/or bags may also be pre-connected to and/or be included as component parts of the RBC collection tubing assembly  950 . 
         [0065]    Vent bag tubing assembly  100  is also interconnected to the top of blood return reservoir  150  of cassette assembly  110 . The vent bag tubing assembly  100  includes vent tubing  102  and a vent bag  104 . During use, sterile air present since packaging within cassette assembly  110 , and particularly within blood return reservoir  150 , cyclically passes into and back out of vent tubing  102  and vent bag  104 , as will be further described. 
         [0066]    As illustrated in  FIG. 4 , pump-engaging tubing loops  122 ,  132 ,  142 ,  162  and  192  extend from cassette member  115  to yield an asymmetric arrangement thereby facilitating proper mounting of cassette assembly  110  on blood component separation device  6  for use. 
         [0067]    In normal operation, whole blood will pass through needle assembly  30 , blood removal tubing  22 , cassette assembly  110  and blood inlet tubing  62  to processing vessel  352 . The whole blood will then be separated into blood components in vessel  352 . During product collection, plasma, and optionally RBCs will be passed out of vessel  352  through corresponding ports  520  and  456  for collection or further separation. The plasma to be further separated will pass into the plasma separator  205  with photosensitizer being added to any collected plasma proteins. 
         [0068]    In the cassette assembly the reservoir  150  having upper and lower ultrasonic sensors (not shown) is provided such that, during the blood processing mode, return blood will be removed from reservoir  150  during each blood return/replacement delivery sub-mode and accumulated during each blood removal sub-mode. When uncollected platelets and plasma (and potentially white blood cells) or red blood cells not collected and/or replacement fluid(s) have accumulated in reservoir  150  up to upper ultrasonic level sensor (not shown), operation of the pump  1090  associated with pump loop  192  will be initiated to remove the blood or replacement components from reservoir  150  through  190   a ,  192 , and  190   b  and transfer the same back to the donor/patient  4  via the return/delivery tubing  24  and needle assembly  20 . When the fluid level in the reservoir  150  drops down to the level of the lower ultrasonic level sensor, the return/delivery peristaltic pump  1090  will automatically turn off reinitiating blood removal sub-mode. The cycle between blood removal and blood return/replacement delivery sub-modes will then continue until a predetermined amount of plasma, and RBCs or other collected blood components have been harvested or collected. 
         [0069]    Pump  1040  is associated with tubing pump loop  142 , pump  1066  is associated with tubing loop  162 , pump  1030  is associated with tubing loop  132 , pump  1020  is associated with tubing loop  122 , and pump  1090  is associated with tubing loop  192  when the cassette  110  is mounted on pump/valve/sensor assembly  1000 . 
         [0070]    The channel assembly  200  includes a channel housing  204  which is disposed on the rotatable centrifuge rotor assembly  568  ( FIG. 1 ) and which receives a disposable blood processing vessel  352 . 
         [0071]    The channel housing  204  provides a mounting for the blood processing vessel  352  such that the blood may be separated into the blood component types in a desired manner. In this regard, the channel housing  204  includes a generally concave channel (not shown) in which the blood processing vessel  352  is positioned. 
         [0072]    The blood processing channel vessel  352  is disposed within the channel housing  204  such that blood can be provided to the blood processing vessel  352  during rotation of the channel housing  204 , to be separated into its various blood component types by centrifugation, and to have various blood component types removed from the blood processing vessel  352  during rotation of the channel housing  204 . In addition, the channel allows for a blood priming of the blood processing vessel  352  (i.e., using blood as the first liquid which is provided to the blood processing vessel  352  in an apheresis procedure). 
         [0073]    The blood processing vessel  352  is disposed within the channel of the channel housing  204  for directly interfacing with and receiving a flow of blood in an apheresis procedure. Further details of the blood processing vessel and parts of the apheresis system are described in U.S. Pat. No. 6,514,189B1. 
         [0074]    As shown in  FIGS. 2 ,  3 ,  11  and  12 , blood is introduced into the interior of the blood processing vessel  352  through a blood inlet port  392  from inlet tubing  62 . The blood inlet port  392  extends into an interior portion of the blood processing vessel  352 . 
         [0075]    Blood which is provided to the blood processing vessel  352  by the blood inlet port  392  is separated into at least plasma and optionally RBCs under centrifugal forces upon rotation of centrifuge rotor assembly  568  at an rpm for separation. 
         [0076]    Separated plasma exits the blood processing vessel through port  456  and tubing  68 . Separated red blood cells exit the blood through port  520  and tubing  64 . 
         [0077]    The apheresis system includes various valve assemblies shown schematically at  1120 ,  1110 , and  1100  in  FIG. 4 . These valves are part of the pump/valve/sensor assembly  1000 . 
         [0078]    The apheresis system described herein provides for continuous separation of plasma and optionally red blood cells (RBCs) and/or plasma with continuous plasma separation and photosensitizer mixing. Both the plasma separation and photosensitizer mixing occurs on-line when the blood removal/return assembly is on the rotor assembly  568  with respect to the apheresis system step. Prion filtration may also occur on-line. For example, continuous separation may be provided with contemporaneous collection of plasma proteins with photosensitizer and/or with collection of RBCs. It is anticipated that four to eight tranfusable dosage double plasma protein products may be collected in a single apheresis procedure from a single donor. 
         [0079]    The plasma proteins to be treated may be illuminated in container and/or bag  94  or transferred to a photo-permeable container. The container to be illuminated is optionally agitated and exposed to photoradiation for a time sufficient to substantially inactivate the microorganisms. The photo-permeable container is made of transparent or semitransparent plastic, and the agitating device is preferably a shaker table.  FIG. 13  illustrates a bag or container  94  under illumination. The illuminator is shown at  400 . Bag  94  rests on support platform  406  which could be a shaker table. The radiation sources are shown at  403  and  404 . The radiation emitting elements  401  and  402  may be visible or ultraviolet light or a combination thereof. The control unit  407  for the illuminator  400  controls the radiation sources as well as any shaker table. 
         [0080]    Replacement fluid(s) are also optionally administrable within the procedures of the present embodiments. Sterile saline solution(s) is one of the optional replacement fluids considered for use herein. Thus, if/when large fluid amounts of plasma and/or RBCs are taken from a donor/patient, replacement fluid(s) may be delivered in return to leave the donor/patient adequately hydrated. 
         [0081]    The initiation of blood processing provides for the collection of plasma product or plasma protein product in one or more reservoir(s)  94  containing photosensitizer optionally with collection of red blood cells in one or more reservoir(s)  954 . Alternatively, either RBC collection in reservoir(s)  954  or plasma collection in reservoir(s)  94  may also be selectively completed in separate procedures. During either collection procedure, blood component separation device  6  preferably controls the initiation and termination of successive blood removal and blood return. Additionally, blood component separation device  6  will control the plasma and RBC collection processes according to predetermined protocols, preferably including control over the valve assemblies  1100 ,  1110  and  1120  of the pump/valve/sensor assembly  1000 , and/or the appropriate pumps  1020 ,  1030 ,  1040 ,  1066  and/or  1090 . 
         [0082]    Initially, blood priming is carried out to prime the disposable system  10 . During blood priming, it may be desirable that the component separation begins even during the priming stage, and that some plasma enters plasma protein collection tubing assembly  90 . Thus plasma may flow out through the outlet port  456  to tubing  68 . 
         [0083]    Following and/or contemporaneously with the blood priming phase, blood separation control device  6  provides control signals to pump/valve/sensor assembly  1000  so that the optional replacement fluid lines may also be primed. In particular, replacement fluid valve assembly  1100  is opened and replacement fluid inlet pump  1040  is switched on to provide for the pumping of saline solution (or other replacement fluid(s)) through replacement fluid inlet tubing  962  and the replacement fluid tubing loop  142  into replacement fluid introduction tubing line  146  for initial collection in cassette reservoir  150 , though this initial priming collection will likely and preferably does constitute a small amount of replacement fluid(s). 
         [0084]    After priming is completed, yet still during the set-up phase, blood component separation device  6  may provide appropriate control signals to the pump/valve/sensor assembly  1000  such that all separated blood components flowing out of processing vessel  352  will first pass to return/delivery reservoir  150 . Optionally, one or more cycles of separation and return of all blood components back to the donor may be performed before collection. Also, blood component separation device  6  may continue operation of blood inlet pump assembly  1030  associated with pump loop  132  during one or more these initial blood component return sub-modes. 
         [0085]    To establish the desired AC ratio, blood component separation device  6  provides appropriate control signals to anticoagulant peristaltic pump  1020  so as to introduce anticoagulant into the blood inlet flow at a predetermined rate. The inlet flow rate of anti-coagulated blood to blood processing vessel  352  may be limited by a predetermined, maximum acceptable anticoagulant infusion rate (ACIR) to the donor/patient  4 . 
         [0086]    When collection begins, blood component separation device  6  may provide control signals so that plasma divert valve assembly  1110  switches to divert the flow of separated plasma pumped from vessel  352  through plasma outlet tubing  68  and plasma tubing loop  162  into plasma collector tubing  92  and into inlet  203  of membrane separator  205 . See also  FIGS. 5 and 6  which show simplified views of the apheresis system. Additionally, if plasma is to be collected alone, red blood cells will continue to flow from vessel  352  through outlet tubing  64  through return tubing loop  172  and into blood return reservoir  150 . However, if RBCs are to be collected, contemporaneously with plasma, then red blood cell valve  1120  switches to divert the flow of separated RBCs flowing from tubing  64  to and through spur  170   b  (of cassette  110 ) and into and through tubing line  952  to the one or more RBC collection reservoir(s)  954 . 
         [0087]    During any of the collection processes, one or more replacement fluid(s) may also be delivered to the donor/patient  4 . Thus, whenever the separation device  6  is in a collection rather than the return mode, the replacement fluid inlet valve assembly  1100  may also be opened and the replacement fluid pump  1040  starts to flow replacement fluids from the fluid source (not shown) through tubing line  962 , cassette passageways  140   a  and  140   b , and tubing loops  142  and  146  into the reservoir  150 . 
         [0088]    During separation and collection, channel housing  204  can be typically driven at a rotational velocity of about 3000 rpms to achieve the desired hematocrit during the both the setup and component collection phases. Correspondingly, the blood inlet flow rate to vessel  352  may be established at below about 64.7 ml/min. The desired hematocrit can be reliably stabilized by passing about two whole blood volumes of vessel  352  through vessel  352  before the RBC and/or plasma collection phases are initiated. 
         [0089]    With respect to plasma collection, which may occur separate from or continuously with red blood cell collection, the separated plasma is pumped via pump  1066  through the plasma collect line  92  through filter separator or concentrator  205  to plasma component collection bag  94  through line  93 . The pore size of the filter  205  determines whether all proteins are collected in container  94  or only those proteins of sufficiently high molecular weight. 
         [0090]    The separated plasma is pumped out of rotor  352  through port  456 , line  68 , passageway  160   a , tubing  162 , passageway  160   b , by pump  1066  around which tubing  162  extends and flows via plasma collect line  92  into filter or separator  205 . The fraction of plasma proteins that do not pass through the filter membrane from the IC to the EC side enter tubing  93  and flow into storage bag  94 . In the embodiments of  FIGS. 2 and 5 , photosensitizer contained in plasma component collection bag  94  mixes in the bag  94  with the collected plasma component or plasma proteins. This photosensitizer may be in liquid form for ease of mixing though dry form may be used. In the embodiment of  FIGS. 3 and 6 , the photosensitizer, in dry form, is dissolved and mixed with the plasma component or plasma proteins as they pass through cartridge  98  in tubing  93  on their way to bag or container  94 . Thus, the bag or container  94  will contain plasma proteins mixed with photosensitizer. 
         [0091]    If a prion filter is included in tubing  93  as shown in  FIGS. 7 ,  8  and  9  additional filtration of the desired plasma proteins will occur to remove the specific prion proteins. As the plasma proteins are pushed through the filter  95 , (or optionally  97 ) by the continuous process the filter media or filter ligands will capture and remove the prion proteins prior to collection of the desired plasma proteins in bag  94 . 
         [0092]    The remainder of the plasma and/or proteins that pass to the EC side flow out of the filter  205  through outlet  208 , tubing  963 ,  962 , passageway  140   a , tubing loop  142 , passageway  140   b , tubing  146 , to reservoir  150  and back to the donor  4 . 
         [0093]    An enriched plasma product, which may contain several times the normal amount or an increased concentration of the desired protein, could be produced by simply processing more plasma through the filter, concentrator or separator  205 . 
         [0094]    Following collection of the desired quantity of red blood cells, (if any), the separation and collection of plasma proteins, and after blood separation device  6  has provided control signals to divert assemblies  1110  and  1120  so as to divert the respective separated plasma and separated RBC flows to reservoir  150 , if further blood processing is not desired, rinse back procedures may then be completed. The plasma pump  1066  is set at the full plasma rate equal to rate of the return/delivery pump  1090  for rinse back. 
         [0095]    At the end of the procedures, the plasma bag(s)  94  and the red blood cell reservoir(s), if any,  954  may be disconnected from the extracorporeal tubing circuit  10 . 
         [0096]    After disconnection of the plasma bag(s)  94 , such bags may be placed in an illuminator  400  to activate the photosensitizer to inactivate any pathogens contained therein as shown in  FIG. 13 . Alternatively, the plasma proteins could be transferred to a photopermeable bag for illumination. Radiation sources  403 ,  404  illuminate bag  94  (or a subsequent bag containing the contents of bag  94 ) with optional agitation of such bag. 
         [0097]      FIG. 9  indicates another option for photoradiation. In this embodiment the plasma proteins with photosensitizer are exposed to the required radiation to activate the photosensitizer on-line. As shown in  FIG. 9 , the photosensitizer is added to the plasma proteins to be collected when such proteins pass through the cartridge  98 , (as described for the embodiment of  FIGS. 3 and 6 ). However tubing  93  is made of sufficiently photo-transmissive material such that the needed radiation for photosensitizer activation can occur in the tubing  93 . Radiation source  99  adds the required radiation. This embodiment illustrates a flow-through system which permits radiation while the fluid or plasma proteins flows by the source illuminator. 
         [0098]      FIG. 10  illustrates an alternative plasma collection assembly. In this embodiment the separated plasma from the blood processing vessel  352  is filtered and mixed with photosensitizer prior to further plasma separation. As shown in  FIG. 10 , prion filter  195  is in tubing  92  to filter separated plasma from the blood processing vessel  352 . After filtration, the filtered plasma mixes with photosensitizer as it passes through cartridge  198  containing dry form photosensitizer. This variation is particularly advantageous when both separated fractions of the plasma are collected as described below with respect to  FIG. 11 . Alternatively only the prion filter  195  or the cartridge  198  could be in tubing  92  with the other being in tubing  93 . Cartridge  198  could also be omitted if the photosensitizer is in the plasma collection bag  94 . 
         [0099]      FIG. 11  illustrates an alternative embodiment wherein both separated fractions of plasma are collected. In this embodiment replacement fluid would typically be provided to the donor. As shown in  FIG. 11  the cartridge  198  may provide the photosensitizer to be mixed with the plasma. The plasma, after separation in plasma separator  205 , is provided to collection bags  94  and  294 . As described previously plasma proteins that do not pass through the membrane of the membrane separator  205  may be collected in container or collection bag  94 . Plasma liquid or ultrafiltrate as well as any proteins with molecular weights sufficiently low to pass through the membrane separator will pass through tubing  293  and open slide clamp  296  to second collection bag  294 . If the cartridge  198  is not used, liquid or dry form photosensitizer may be in containers  94  and  294  for subsequent mixing with the plasma and proteins for collection. 
         [0100]    The embodiment of  FIG. 12  illustrates a further separation tubing set wherein the separated plasma enters the EC side of the plasma separator  205  through inlet port  1203 . Thus plasma proteins that do not pass through the membrane, in this variation may be collected by exiting the EC side passing through tubing  193  to plasma collection container  94 . Although this embodiment shows plasma and plasma proteins that pass through the filter being returned to the donor from the IC side through tubing  1963  and  962  such low molecular weight elements could alternatively be collected in another collection container. 
         [0101]    The simplified  FIGS. 5 and 6  also indicate another option. As shown in  FIGS. 5 and 6 , the plasma entering the plasma separator  205  is pumped on the inlet side  203  by pump  1066  and also the outlet (EC) side  208  by pump  1040 . However the locations of the pumps can be varied. For example, as shown in  FIGS. 5 and 6  there also may be a pump on the IC exit side, (illustrated in phantom lines as  1040   a ). This pump may be used with the inlet pump  1066  alone, (no pumping through  1040  on the EC side) or it may be used with pump  1040  alone, (no pumping through  1066  into the inlet or IC side. Thus two pumps are utilized but the exact locations of such pumps may be varied. 
         [0102]    Having pump  1040   a  pump on the IC side from  206  provides flow through the membrane by positive pressure on the IC side thus avoiding any degassing of the fluid as may occur using pumps  1066  and  1040  which exerts negative pressure on the EC side. If the membrane becomes blocked when pump  1040   a  is used the compression force of the rollers of pump  1040   a  could be such that they will lift sufficiently and provide less occlusion for the over volume or pressure. Thus it can function as a pressure relief valve. 
         [0103]    This continuous apheresis procedure permits desirable proteins to be collected, with on-line preparation for subsequent pathogen treatment, and removed from a donor with the remainder of the plasma proteins being returned to the donor. This enables maximum collection and concentration of the desired proteins, instead of the smaller amount of desired protein contained within a single donation. 
         [0104]    Using this procedure, plasma protein fractions with photosensitizer may be collected at the same time as other cellular components. Specifically, desired plasma proteins may be collected from a donor, while the undesired components may be returned to the donor. This would enable greater amounts of desired plasma proteins to be collected from a single donor, without increasing the risk to the donor as the amount of fluid volume removed from a donor would not be detrimental. More plasma can be processed resulting in the collection of increased amounts of plasma protein. However, from the donor perspective the increased collection of proteins can be collected with the same volume removal as a typical plasma collection. 
         [0105]    The final concentration of the protein-enriched product could be adjusted by adjusting the ratio of the plasma flow into the filter and the plasma flow out of the filter. This can be done by adjusting pump speed of pump  1066 ,  1040  or  1040   a . For example, if the membrane excluded all proteins and the flow rate through the filter was half that of the plasma flow into the filter, the resultant concentration of the proteins would be double that of normal donor plasma. 
         [0106]    If it is desired to collect high molecular weight proteins, the filter/column  205  could separate on a continuous basis the albumin and other low molecular weight proteins and return them to the donor, while collecting higher molecular weight fractions such as fibrinogen, IgG, von Willebrand factor and factor VIII. 
         [0107]    Alternatively, if lower molecular weight proteins are desired for collection, the higher molecular weight proteins could be returned to the donor, while the lower molecular weight proteins are collected by changing the tubing so that those that pass through the membrane are collected in a reservoir containing photosensitizer rather than returned. In this configuration outlet  208  would be connected to tubing  93  with outlet  206  being connected to tubing  963  for return to the donor. 
         [0108]    The separation specificity can be accomplished by selecting membranes which have pore sizes which correspond to the molecular weight of the desired protein. 
         [0109]    The high concentration protein collected could be used to enrich the plasma of a patient for therapeutic purposes. The high-concentration product may also be used for additional fractionation as described below where its yield of proteins would be much higher compared to normal plasma and thus produce an increased amount of protein concentrate products. 
         [0110]      FIG. 14  illustrates, in block diagram form, the process of taking whole blood from a donor  11  (whole blood) and using apheresis apparatus  12  as described above to collect a concentrated protein fraction. The on-line process includes the addition of photosensitizer  13 . The collected plasma proteins  14  are illuminated  15  to inactivate the photosensitizer. The collected plasma proteins from the apheresis process may optionally be provided to plasma fractionation center  16 , and optionally pooled with other collections which have been treated for pathogen inactivation, for further fractionation or concentration of such product utilizing a known plasma fractionation process such as cold alcohol fractionation  18  (also known as Cohn fractionation) or chromatography  17 . Other known fractionation processes could be used. This process could be used to provide a highly concentrated plasma protein infusion product  19  such as IVIG or clotting factor that has been treated for pathogen inactivation. 
         [0111]    It will be apparent to those skilled in the art that various modifications and variations to the methods and structure of the present invention without departing from its scope. Thus it should be understood that the invention is not be limited to the specific examples given. Rather, the invention is intended to cover modifications and variations provided they come within the scope of the following claims and their equivalents.