Method for storage of reagent red blood cells using carbon monoxide - Patent Application 20070233633

Abstract

This application relates to methods, compositions, and kits for use in blood typing and in preparing improved red blood cell-containing reagents for use in blood typing of blood prior to use in transfusion medicine.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority from U.S. Provisional Patent Application No. 62 / 769,367, filed November 19, 2018, which is incorporated by reference in its entirety.

[0002] The present invention relates to the field of blood typing and the preparation of improved red blood cell-containing reagents for use in determining the type of blood prior to use in transfusion medicine. [Background technology]

[0003] Red blood cell (RBC) reagents are manufactured from blood collected from established donors with well-characterized phenotypes. The RBCs are diluted in a diluent and packaged to be used as a standard for determining the blood type of processed RBCs in the blood center / blood bank. In most cases, automated devices are used for blood typing and characterization of other important RBC quality parameters. To preserve surface antigens, the RBCs are not fixed, making them unstable due to accumulation of storage damage. These time-dependent changes limit the shelf life of these important RBC preparations to approximately 9 weeks (63 days).

[0004] RBCs evolved to provide and transport oxygen throughout the entire body except to the surface where diffusion from the atmosphere cannot be relied upon. RBCs achieve this function by enriching the cytosol with very high concentrations of hemoglobin containing oxidizable ferrous iron. During the roughly 120 days of circulation, RBCs are maintained to transport oxygen by a dense web of metabolic / redox enzymes. However, when RBCs are removed from the circulation and stored at low temperatures, these dense evolutionary optimizations are no longer operational, resulting in storage-induced damage (storage damage) that accumulates over the useful life of the stored RBCs. One of the two main drivers of the development of storage damage is oxidative damage, which has been known but not systematically addressed until recently.

[0005] Chemical oxidation of iron in hemoglobin is the central reaction initiating oxidative stress in stored RBCs and is a major component of the development of storage damage. RBCs contain high concentrations of reactive ferrous iron in the prosthetic group of hemoglobin along with high concentrations of dissolved oxygen. The four iron moieties in hemoglobin (ferrous state) react chemically with oxygen to form methemoglobin (ferric state). As a by-product, superoxide anion is generated, which is converted by superoxide dismutase to form H2O2, the major reactive oxygen species (ROS) and substrate for hydroxyl radical (OH·) generation. In vivo, methemoglobin is reduced to hemoglobin by reductases, but the activity of these enzymes is inhibited under low-temperature storage conditions. Combined with higher dissolved oxygen concentrations due to increased solubility at low temperatures, this phenomenon enhances the generation of methemoglobin and superoxide anion. ROS molecules react with lipids and structural proteins in RBCs, compromising their integrity and reducing their in vivo circulation time. ROS also attack important enzymes or surface molecules that make the "product" RBCs valuable, reducing their effectiveness and utility.

[0006] Hemoglobin has an affinity for CO approximately 400 times higher than O2, and CO does not readily chemically react with heme. The heme-CO complex is highly stable, which significantly reduces oxidative RBC storage damage, as hemoglobin oxidation by oxygen is the primary driver of oxidative stress during low-temperature RBC storage. Unlike O2, CO does not readily react with ferrous iron, but prevents Hb oxidation by stabilizing it as Hb-CO2, significantly reducing the occurrence of oxidative storage damage in RBCs stored at low temperatures (i.e., 1-6 °C). ROS damage can be further reduced by storing RBCs under anoxic conditions, either under CO2 or an inert gas such as nitrogen.

[0007] Although the high affinity binding of CO to Hb renders RBCs containing Hb and Hb-CO useless as oxygen carriers until CO is released, stabilization of Hb by CO is beneficial during storage and can extend the shelf life of reagent RBCs not only before they are opened or reconstituted for use, but also after they are opened. Much more than RBCs for transfusion, reagent RBCs are highly characterized and carefully controlled reagents of great value. Even small improvements in shelf life can significantly reduce the costs of blood banking operations. CO treatment of reagent RBCs reduces storage damage accumulation and extends shelf life. Summary of the Invention

[0008] The present disclosure provides and includes a method for storing reagent red blood cells (RBCs) comprising obtaining packed red blood cells, flushing the packed red blood cells with a gas comprising carbon monoxide to prepare carbon monoxide saturated RBCs (CO-Hb RBCs), and storing the CO-Hb RBCs under anaerobic conditions in the presence of carbon monoxide (CO), whereby surface antigens of the CO-Hb RBCs are stabilized.

[0009] The present disclosure provides and includes a kit comprising one or more vials of carbon monoxide saturated RBCs (CO-Hb RBCs) having a plurality of CO-Hb RBCs with a common set of surface antigens in a buffer solution, and instructions for use.

[0010] The present disclosure relates to a buffer solution and blood type O cells, and D, C, c, E, e, CW, K, k, P1, Fy a ,Fy b , J.K. a , J.K. b , Le a , Le b and CO-Hb RBCs selected from the group consisting of CO-Hb RBCs that are positive for a surface antigen selected from the group consisting of D, C, c, E, e, CW, K, k, P1, Fy, Hb, Hg, Hb ... a ,Fy b, J.K. a , J.K. b , Le a , Le b ,M.,N.,S.,s.,I.,Lu a , Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs are blood group O cells that are positive for a surface antigen selected from the group consisting of D, C, c, E, e, CW, K, k, P1, Fy a ,Fy b , J.K. a , J.K. b , Le a , Le b ,M.,N.,S.,s.,I.,Lu a , Lu b , J.S. b , Kp b , and Yt a and the surface antigen Js a , Kp a , Wr a , Di a , V w ,V.,Lu a , and C w CO-Hb RBCs are blood type O cells that are negative for surface antigens D, C, c, E, e, f, CW, K, k, P1, Fy a ,Fy b , J.K. a , J.K. b , Le a , Le b ,M.,N.,S.,s.,Lu a , and Lu b CO-Hb RBCs are type O cells that are negative for Rh antigens D, C, and e, and CO-Hb RBCs are type O cells that are positive for Rh antigens D, C, and e, I, Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs are type O cells positive for Rh antigens D, C, and e, I, Lu b , J.S. b , Kp b , and Yta Positive for and surface antigen Js a , Kp a , Wr a , Di a V w ,V.,Lu a and C w CO-Hb RBCs are type O cells that are positive for surface antigen A1, CO-Hb RBCs are type A cells that are positive for surface antigen A1 and negative for surface antigens D, C, and E, CO-Hb RBCs are type A cells that are positive for surface antigen A2, CO-Hb RBCs are type A cells that are positive for surface antigen A2 and negative for surface antigens D, C, and E, CO-Hb RBCs are type B cells that are positive for surface antigen B and negative for surface antigens D, C, and E, CO-Hb RBCs are type O cells that are positive for surface antigen D, C, and e and have the Rh phenotype R1R1, and CO-Hb RBCs are type A cells that are positive for surface antigen A1 and negative for surface antigens D, C, and E. w , and e positive and Rh phenotype R1 w CO-Hb RBCs are O cells positive for surface antigens D, c, and E and have Rh phenotype R2R2; CO-Hb RBCs are O cells positive for surface antigens d, c, and e and have Rh phenotype rr; CO-Hb RBCs are O cells positive for surface antigens D, C, and e and have Rh phenotype R1R1 and have surface antigen Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs are type O cells positive for surface antigens D, C w , and e positive and Rh phenotype R1 w R1 and surface antigen Lu b , J.S. b , Kp b , and Yt aCO-Hb RBCs are positive for surface antigens D, c, and E and have the Rh phenotype R2R2 and are positive for surface antigen Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs are positive for surface antigens d, c, and e and have the Rh phenotype rr and surface antigen Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs are positive for surface antigens D, C, and e and have the Rh phenotype R1R1 and the surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w CO-Hb RBCs are type O cells that are negative for surface antigens D, C w , and e positive and Rh phenotype R1 w R1 and surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w CO-Hb RBCs are positive for surface antigens D, c, and E and have the Rh phenotype R2R2 and are negative for surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w CO-Hb RBCs are positive for surface antigens d, c, and e and have the Rh phenotype rr and are negative for surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C wand the CO-Hb RBCs are positive for surface antigens D, C, and e and have Rh haplotype R1, or are positive for surface antigens D, c, and e and have Rh haplotype R2. The CO-Hb RBCs are positive for surface antigens D, C, and e and have Rh haplotype R1, and are negative for surface antigens Lu b , J.S. b , Kp b , and Yt a or positive for surface antigens D, c, and e and have Rh haplotype R2CO-Hb RBCs and surface antigen Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs are positive for surface antigens D, C, and e and have the Rh haplotype R1 and the surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w or are positive for surface antigens D, c, and e and have the Rh haplotype R2 and surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w The CO-Hb RBCs were ficin-treated and negative for the surface antigens M, N, Fy, a ,Fy b , S, s, Xg a , Pr, Ch a , Rg a and Yk a The CO-Hb RBCs were ficin-treated and negative for the surface antigens M, N, Fy, and a ,Fy b , S, s, Xg a , Pr, Ch a , Rg a and Yk aNegative for D, C, E, c, e, f, Jk a , J.K. b , Le a , Le b , P1, I, IH, Vel, PP1P k and P antigens, and CO-Hb RBCs were ficin-treated and positive for surface antigens M, N, Fy, and P antigens. a ,Fy b , S, s, Xg a , Pr, Ch a , Rg a Negative for Fya, Fyb, S, s, M, N, Xg a , Pr, Ch a , Rg a , and Yk a CO-Hb RBCs are positive for antibody binding to D antigen, said cells being O cells sensitized with anti-D(Rh0) serum, CO-Hb RBCs are A cells positive for antibody binding to A2 antigen, CO-Hb RBCs are B cells positive for antibody binding to B antigen and negative for anti-D(Rh0) antibody binding, CO-Hb RBCs are A cells positive for antibody binding to A1 antigen and negative for anti-D(Rh0) antibody binding, CO-Hb RBCs are AB cells positive for antibody binding to A1, B antigen, and Rh antigens d, c, and e of the Rh blood group rr(dce / dec), CO-Hb RBCs are type O cells positive for antibody binding to Rh antigens D, d, C, c, and e and have the Rh phenotype R1r (DCe / dce), whereas CO-Hb RBCs are type D (RH1), C (RH2), E (RH3), c (RH4), e (RH5), M, N, S, s, P1, K, k, Fy a ,Fy b , J.K. a , J.K. b , Le a and Le bThe CO-Hb RBCs are type O cells positive for antibody binding to Hb, and the CO-Hb RBCs are type D (RH1), type C (RH2), type E (RH3), type c (RH4), type e (RH5), type M, type N, type S, type s, type P1, type K, type k, type Fy a ,Fy b , J.K. a , J.K. b , Le a and Le b Positive for antibody binding to Lu b , J.S. b , Kp b , and Yt a Type O cells positive for binding of antibodies to antigens, CO-Hb RBCs are D(RH1), C(RH2), E(RH3), c(RH4), e(RH5), M, N, S, s, P1, K, k, Fy a ,Fy b , J.K. a , J.K. b , Le a and Le b Positive for antibody binding to Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w These are type O cells that are negative for antibody binding to antigens. [Brief description of the drawings]

[0011] [Figure 1] 2 is a graph showing deoxygenation and carbon monoxide conversion of red blood cell concentrates in an embodiment according to Example 2(a). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] Blood group serology requires the determination of blood cell compatibility between a blood donor and a patient-recipient prior to a blood transfusion or organ transplant with the patient. Blood cell compatibility is determined by the absence of an immunological reaction between antibodies contained in the patient's serum and antigens present on blood cells from the donor. Blood cell typing and compatibility tests are generally of two types: (1) agglutination tests, which determine whether a specific antibody added to cells causes their agglutination, and (2) cytolytic tests, which determine whether a specific antibody added to test cells together with serum complement results in hemolysis. In commonly used blood cell typing and compatibility testing procedures, both agglutination and cytolytic tests are performed manually by a skilled technician or using automated devices. The presence of an immunological reaction is incompatible with blood transfusion and transplantation therapy.

[0013] The International Society of Blood Transfusion lists 33 blood group systems that represent more than 300 antigens. See Logdberg et al., “Human blood group genes 2004: Chromosomal locations and cloning strategies,” Transfus.Med.Rev.19:45-57 (2005) and Logdberg et al., “Human blood group genes 2010: Chromosomal locations and cloning strategies revisited,” Transfus.Med.Rev.25:36-46 (2011). Cloning and sequencing have demonstrated that the genes for these blood group systems are autosomal, with the exception of XG and XK, which are X-mediated, and MIC2, which is present on both the X and Y chromosomes. Many different blood group antigens are found on the surface of red blood cells in every individual. These antigens, which are the products of inherited genes, are present in combinations that are likely to be unique between all individuals except for identical twins.

[0014] A number of significant blood group systems are known in the art and are presented in Table 1. [Table 1]

[0015] Blood typing is generally a process of testing red blood cells to determine which antigens are present and which are not, usually utilizing antibodies against the antigen being tested. In addition, if a person does not have a particular red blood cell antigen on their red blood cells, their serum may contain antibodies against that antigen. Whether antibodies are present in the serum depends on whether their immune system has previously been triggered by and responded to that particular antigen or something very similar to it. For example, a person whose red blood cells are type A, i.e., who has the "A" antigen on their red blood cells, will have anti-B antibodies in their serum. Thus, if such a person is administered type B blood, an immunological reaction will occur with possible severe clinical consequences.

[0016] Prior to transfusion therapy, collected blood is tested for compatibility by analyzing blood types. Blood type testing is performed by testing RBCs for various antigens (A, B, etc.) and testing serum for antibodies to the antigens. In the former test, RBCs from a sample blood are incubated with antibodies that recognize each of the blood group antigens and scored for binding using agglutination or any of the other known immunological techniques. Serum testing can be performed in a variety of ways, such as, for example, by ELISA, where antigens are provided and antibody binding is indirectly detected through an enzyme or fluorescent reporter. A more traditional approach uses RBCs that have been previously characterized to express a particular antigen in an agglutination assay called a hemagglutination assay. Briefly, an agglutination assay involves mixing reagent RBCs with serum or plasma from a test sample of blood. Antibodies in the test sample, typically IgM with five antigen binding sites, crosslink the cells together, causing clumps that are visible to the naked eye. Bivalent IgG antibodies are also suitable for agglutination assays. Agglutination assays, including those directed to blood typing, are well known in the art. See Low et al., “Antiglobulin test in low-ionic strength salt solution for rapid antibody screening and cross-matching,” Vox Sang 26:53-61 (1974); Malyska et al., “The gel test,” Laboratory Medicine 25:81 (1994); and Technical manual. 14th ed. Bethesda, MD: American Association of Blood Banks, 2002.

[0017] Of the more common reagent RBCs typically used for reverse blood grouping, those with A, A, B, or no ABO antigens (A1, A2, B, O) are useful for detection of preformed antibodies that cause agglutination of the reagent RBCs. Forward testing uses monoclonal anti-A and anti-B to detect the presence of their respective antigens on the sample red cell surface. Another well-known blood group is the Rh blood group, which has 58 different antigen types, although nine types are the most common. Blood group and antigen designations are presented in Tables 2 and 3. Byrne et al., “Review: other blood group systems--Diego, Yt, Xg, Scianna, Dombrock, Colton, Landsteiner-Wiener, and Indian,” Immunohematology 20(1):50-8(2004); Daniels, “The molecular genetics of blood group polymorphism,” Transpl. Immunol. 14(3-4):143-53(2005);Daniels,“Functions of red cell surface proteins,”Vox Sang.93(4):331-40(2007);Eyler et al.,“The Lutheran glycoprotein:a multifunctional adhesion receptor.”Transfusion46(4):668-77(2006);Palacajornsuk,“Review:molecular basis of MNS blood group variants,”Immunohematology22(4):171-82(2006);Quill,“Medicine.Blood-matching goes genetic,” Science 14;319(5869):1478-9(2008); Westhoff, “The structure and function of the Rh antigen complex,” Semin Hematol 44(1):42-50(2007); Westhoff, “Review: the Kell, Duffy, and Kidd blood group systems,” Immunohematology 20(1):37-49(2004); and Yamamoto, “Review: ABO blood group system--ABH oligosaccharide antigens, anti-A and anti-B, A and B glycosyltransferases, and ABO genes,” Immunohematology 20(1):3-22(2004), each of which is incorporated herein by reference in its entirety. [Table 2] TIFF0007616992000003.tif93141 Table 3. Blood group antigens [Table 3]

[0018] The present disclosure provides and includes a method for reducing the degradation of blood group antigens during storage. More specifically, the present disclosure provides for reducing the formation of storage damage in RBCs during storage, including but not limited to ROS-induced damage. That is, RBCs are harvested and exposed to an atmosphere of carbon monoxide (CO) for a period of time sufficient to remove oxygen (O2) bound to the heme group of hemoglobin. As shown in Example 2 and FIG. 1, the exchange of oxygen with CO occurs rapidly and is essentially complete in 30 minutes and three gas exchanges. Methods for contacting CO with blood, particularly methods that increase the surface-to-volume ratio of the CO / liquid interface, would greatly reduce the length of time required to exchange the bound oxygen with carbon monoxide. As discussed above, carbon monoxide has a significantly higher affinity for hemoglobin than oxygen (e.g., 400 times), so exposure of RBCs to CO at any level and time will initiate the exchange process and sufficient CO is provided to facilitate the exchange for completion.

[0019] The present disclosure provides and includes, but is not limited to, exchanging oxygen for carbon monoxide according to the method shown in Example 2. In one embodiment, red blood cell concentrate (RCC, also known as packed red blood cells) is held in a container, CO is introduced, and the container is gently shaken or mixed for a period of time. In one embodiment, the container is a standard blood storage bag containing polyvinyl chloride. In one embodiment, CO gas is replaced and mixing is repeated one or more times until the hemoglobin is saturated with CO (e.g., Hb-CO RBCs are produced). In another embodiment, a volume of CO is provided to the container containing the RCC and left overnight with or without occasional mixing. Mixing improves the exchange rate, but is not required. In another embodiment, the blood and CO are separated by a gas-permeable membrane. This can be done using methods known in the art, for example, using a SorinD100 oxygen generator to provide CO rather than oxygen. The advantage of the membrane-based approach is that by continuously replacing the gas, the concentration gradient is maintained and the exchange rate is increased.

[0020] In another embodiment, CO gas can be bubbled into a container or bag of RBCs. Without being limited by theory, it is believed that by keeping the bubbles less than 1 μm in diameter, lysis of red blood cells can be prevented or minimized. The "bubble" method shares the same advantages as the membrane-based approach, since the CO bubbles provide the greatest concentration difference, thereby promoting the kinetics of the exchange reaction. The bubble approach also offers the added advantage of mixing of the RBCs.

[0021] Although it may be preferred to perform carbon monoxide exchange on RCC, the present specification provides and includes exchanging CO with oxygen on blood at any stage of the process. In some embodiments, CO is exchanged on whole blood, for example, before removing platelets or leukocytes. In some embodiments, CO is exchanged on leukocyte-removed blood. The method of the present specification can be applied to RBCs prepared by apheresis or collected using conventional methods. The method may also be performed after processing of RBCs into buffers suitable for reagent use and storage. See, for example, U.S. Patent Publication No. 2011 / 0045455, published February 26, 2001, and International Patent Publication No. WO1983 / 003477, published October 13, 1983. Other storage solutions compatible with blood typing methods are known in the art.

[0022] The present disclosure provides and includes a method for preparing CO-Hb RBCs, further comprising storing said CO-Hb RBCs under anaerobic conditions in the presence of CO. In one embodiment, the cells are prepared as described above and transferred to a vial under an atmosphere containing carbon monoxide. In another embodiment, the CO-Hb RBCs are transferred to a container for storage having a nitrogen atmosphere. As provided herein, any suitable non-oxygen-containing gas is suitable during storage. In certain embodiments, additional CO is provided prior to sealing the container for CO-Hb RBC storage.

[0023] The present disclosure provides and includes a method of storing reagent red blood cells (RBCs), comprising obtaining concentrated red blood cells selected from, but not limited to, the following 40 immunological types: 1. Blood type O cells and D, C, c, E, e, CW, K, k, P1, Fy a ,Fy b , J.K. a , J.K. b , Le a , Le b positive for a surface antigen selected from the group consisting of: M, N, S, and s; 2.D, C, c, E, e, CW, K, k, P1, Fy a ,Fy b , J.K. a , J.K. b , Le a , Le b ,M.,N.,S.,s.,I.,Lu a , Lu b , J.S. b , Kp b , and Yt a blood group O cells positive for a surface antigen selected from the group consisting of 3.D, C, c, E, e, CW, K, k, P1, Fy a ,Fy b , J.K. a , J.K. b , Le a , Le b ,M.,N.,S.,s.,I.,Lu a , Lu b , J.S. b , Kp b , and Yt a and the surface antigen Js a , Kp a , Wr a , Di a , V w ,V.,Lu a , and C w Blood type O cells that are negative for 4. Surface antigens D, C, c, E, e, f, CW, K, k, P1, Fy a ,Fy b , J.K. a , J.K.b , Le a , Le b ,M.,N.,S.,s.,Lu a , and Lu b Blood type-O cells that are negative for 5. Blood type-O cells that are positive for Rh antigens D, C, and e, 6. Rh antigens D, C, and e, I, Lu b , J.S. b , Kp b , and Yt a Blood type-O cells that are positive for 7. Rh antigens D, C, and e, I, Lu b , J.S. b , Kp b , and Yt a Positive for and surface antigen Js a , Kp a , Wr a , Di a V w ,V.,Lu a and C w Blood type-O cells that are negative for 8. Blood group-A cells positive for surface antigen A1, 9. Blood group-A cells that are positive for surface antigen A1 and negative for surface antigens D, C, and E; 10. Blood group A- cells positive for surface antigen A2, 11. Blood group-A cells that are positive for surface antigen A2 and negative for surface antigens D, C, and E; 12. Blood group-B cells positive for surface antigen B, 13. Blood group-B cells positive for surface antigen B and negative for surface antigens D, C, and E; 14. Blood group-O cells positive for surface antigens D, C, and e and with Rh phenotype R1R1; 15. Surface antigens D, C w , and e positive and Rh phenotype R1 w Blood type-O cells with R1, 16. Blood group-O cells positive for surface antigens D, c, and E and with Rh phenotype R2R2; 17. Blood group-O cells positive for surface antigens d, c, and e and with Rh phenotype rr; 18. Patients who are positive for surface antigens D, C, and e and have the Rh phenotype R1R1 and have surface antigen Lu b , J.S. b , Kp b , and Yt a Blood type-O cells that are positive for 19. Surface antigens D, C w , and e positive and Rh phenotype R1 w R1 and surface antigen Lu b , J.S. b , Kp b , and Yt a Blood type-O cells that are positive for 20. A patient who is positive for surface antigens D, c, and E and has the Rh phenotype R2R2 and has surface antigen Lu b , J.S. b , Kp b , and Yt a Blood type-O cells that are positive for 21. A patient who is positive for surface antigens d, c, and e and has the Rh phenotype rr and has surface antigen Lu b , J.S. b , Kp b , and Yt a Blood type-O cells that are positive for 22. A patient is positive for surface antigens D, C, and e, has the Rh phenotype R1R1, and has surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w Blood type-O cells that are negative for 23. Surface antigens D, C w , and e positive and Rh phenotype R1 w R1 and surface antigen Js a , Kp a , Wr a , Dia , Vw, V, Lu a and C w Blood type-O cells that are negative for 24. A patient is positive for surface antigens D, c, and E, has the Rh phenotype R2R2, and has surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w Blood type-O cells that are negative for 25. A patient who is positive for surface antigens d, c, and e and has the Rh phenotype rr and has surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w Blood type-O cells that are negative for 26. Blood group-O cells positive for surface antigens D, C, and e and with Rh haplotype R1, or positive for surface antigens D, c, and e and with Rh haplotype R2; 27. A patient who is positive for surface antigens D, C, and e and has the Rh haplotype R1 and has surface antigen Lu b , J.S. b , Kp b , and Yt a or positive for surface antigens D, c, and e and have Rh haplotype R2CO-Hb RBCs and surface antigen Lu b , J.S. b , Kp b , and Yt a Blood type-O cells that are positive for 28. A person who is positive for surface antigens D, C, and e and has the Rh haplotype R1 and surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C wor are positive for surface antigens D, c, and e and have the Rh haplotype R2 and surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w Blood type-O cells that are negative for 29. Ficin-treated and surface antigens M, N, and Fy a ,Fy b , S, s, Xg a , Pr, Ch a , Rg a and Yk a Blood type-O cells, which are negative for 30. Ficin-treated and surface antigens M, N, and Fy a ,Fy b , S, s, Xg a , Pr, Ch a , Rg a and Yk a Negative for D, C, E, c, e, f, Jk a , J.K. b , Le a , Le b , P1, I, IH, Vel, PP1P k and blood group-O cells positive for binding of antibodies to the P antigen, 31. Ficin-treated and surface antigens M, N, and Fy a ,Fy b , S, s, Xg a , Pr, Ch a , Rg a Negative for Fya, Fyb, S, s, M, N, Xg a , Pr, Ch a , Rg a , and Yk a having reduced or absent binding of antibodies to blood type-O cells; 32. Blood group-O cells positive for antibody binding to D antigen and said CO-Hb RBCs are sensitized with anti-D(Rh0) serum; 33. Blood group-A cells positive for binding of antibody to A2 antigen; 34. Blood group-B cells positive for antibody binding to B antigen and negative for anti-D (Rh0) antibody binding; 35. CO-Hb RBCs are type A cells that are positive for antibody binding to the A1 antigen and negative for anti-D (Rh0) antibody binding. 36. Blood group-AB cells positive for binding of antibodies to A1, B antigens, and Rh antigens d, c, and e of the Rh blood group rr(dce / dec); 37. Blood group-O cells positive for antibody binding to Rh antigens D, d, C, c, and e and having the Rh phenotype R1r (DCe / dce); 38.D(RH1), C(RH2), E(RH3), c(RH4), e(RH5), M, N, S, s, P1, K, k, Fy a ,Fy b , J.K. a , J.K. b , Le a and Le b blood type-O cells that are positive for binding of antibodies to 39.D(RH1), C(RH2), E(RH3), c(RH4), e(RH5), M, N, S, s, P1, K, k, Fy a ,Fy b , J.K. a , J.K. b , Le a and Le b Positive for antibody binding to Lu b , J.S. b , Kp b , and Yt a Blood type-O cells that are positive for binding of antibodies to antigens, and 40.D(RH1), C(RH2), E(RH3), c(RH4), e(RH5), M, N, S, s, P1, K, k, Fy a ,Fy b , J.K. a , J.K. b , Le a and Le b Positive for antibody binding to Js a , Kp a , Wr a , Dia , Vw, V, Lu a and C w Blood type-O cells that are negative for binding of antibodies to antigens.

[0024] As can be appreciated by one of skill in the art, the selection of RBCs suitable for preparation of reagent RBCs is not limited to the blood type combinations provided above. One of skill in the art will recognize that RBCs having any combination of antigens from the groups listed in Tables 2 and 3 are useful in the methods of the present application. Indeed, one of skill in the art will recognize that any RBC, once immunologically characterized, is suitable for preparation of carbon monoxide-stored reagent RBCs (Hb-CO RBCs).

[0025] The present specification provides and includes exchanging oxygen bound on hemoglobin with carbon monoxide using any method known in the art, including but not limited to the methods shown in the Examples. Thus, as used herein, the term "flushing" refers to any method of contacting carbon monoxide gas with RBCs, including bubbling or passing through a membrane.

[0026] The present specification provides and includes kits that include carbon monoxide stabilized red blood cells (Hb-CO RBCs). The kits herein may include one or more of the 40 specific types of Hb-CO RBCs described above, but are not limited to these cells. Other suitable Hb-CO RBCs can be prepared as needed. In certain embodiments, the kits provide cells suspended in a suitable buffer, and the preparation of the cells may include various washing steps, either before or after carbon monoxide exchange. Additional reagents and buffers required to perform the immunoassay may be included in the kit. In many embodiments, the kits include instructions regarding the performance of the assay, lot characterization of Hb-CO RBCs, and other materials relevant to the skilled artisan. In some embodiments, the kits herein may include various laboratory equipment, such as tubes, pipettes, buffers, etc.

[0027] The present specification provides and includes a container containing the CO-Hb RBCs described herein. In one embodiment, the container is a vial. In another embodiment, the container is a tube. In yet another embodiment, the container is an ampoule.

[0028] This specification provides and includes the following embodiments.

[0029] Embodiment 1. A method for storing reagent red blood cells (RBCs), comprising: a) obtaining concentrated red blood cells; b) flushing the packed red blood cells with a gas containing carbon monoxide to prepare carbon monoxide saturated RBCs (CO-Hb RBCs); and c) storing the CO-Hb RBCs under anaerobic conditions in the presence of carbon monoxide (CO), wherein surface antigens of the CO-Hb RBCs are stabilized.

[0030] Embodiment 2. The method of embodiment 1, wherein the gas does not contain oxygen.

[0031] Embodiment 3. The CO-Hb RBC is selected from the group consisting of D, C, c, E, e, CW, K, k, P1, Fy a ,Fy b , J.K. a , J.K. b , Le a , Le b 3. The method of embodiment 1 or 2, wherein the cells are blood type O cells that are positive for a surface antigen selected from the group consisting of: A, M, N, S, and s.

[0032] Embodiment 4. The CO-Hb RBCs are characterized by surface antigen I, Lu a , Lu b , J.S. b , Kp b , and Yt a The method according to any one of embodiments 1 to 3, wherein the blood group O cells are further positive for .

[0033] 5. The CO-Hb RBCs are characterized by surface antigen Jsa , Kp a , Wr a , Di a , V w ,V.,Lu a , and C w The method according to any one of embodiments 1 to 4, wherein the blood group O cells are negative for .

[0034] Embodiment 6. The CO-Hb RBCs are characterized by surface antigens D, C, c, E, e, f, CW, K, k, P1, Fy a ,Fy b , J.K. a , J.K. b , Le a , Le b ,M.,N.,S.,s.,Lu a , and Lu b The method according to any one of embodiments 1 to 5, wherein the cell is type O cell that is negative for .

[0035] Embodiment 7. The method of any one of embodiments 1 to 6, wherein said CO-Hb RBCs are type O cells positive for Rh antigens D, C, and e.

[0036] Embodiment 8. The CO-Hb RBCs are characterized by surface antigen I, Lu b , J.S. b , Kp b , and Yt a The method according to any one of embodiments 1 to 7, wherein the cell is type O cell positive for .

[0037] 9. The CO-Hb RBCs are characterized by surface antigen Js a , Kp a , Wr a , Di a V w ,V.,Lu a and C w The method according to any one of embodiments 1 to 8, wherein the cell is type O cell that is negative for .

[0038] Embodiment 10. The method of any one of embodiments 1 to 9, wherein said CO-Hb RBCs are type A cells that are positive for surface antigen A1.

[0039] Embodiment 11. The method of any one of embodiments 1 to 10, wherein said CO-Hb RBCs are type A cells that are negative for surface antigens D, C, and E.

[0040] Embodiment 12. The method of any one of embodiments 1 to 11, wherein said CO-Hb RBCs are type A cells that are positive for surface antigen A2.

[0041] Embodiment 13. The method of any one of embodiments 1 to 12, wherein said CO-Hb RBCs are type A cells that are negative for surface antigens D, C, and E.

[0042] Embodiment 14 The method of any one of embodiments 1 to 13, wherein said CO-Hb RBCs are B-type cells that are positive for surface antigen B.

[0043] Embodiment 15. The method of any one of embodiments 1 to 14, wherein said CO-Hb RBCs are B-type cells that are negative for surface antigens D, C, and E.

[0044] Embodiment 16. The CO-Hb RBCs are: positive for surface antigens D, C, and e and have the Rh phenotype R1R1; Surface antigen D, C w , and e positive and Rh phenotype R1 w R1 or positive for surface antigens D, c, and E and have the Rh phenotype R2R2, or The method according to any one of the preceding embodiments, wherein the cells are type O cells positive for surface antigens d, c, and e and have the Rh phenotype rr.

[0045] Embodiment 17. The CO-Hb RBCs are characterized by the surface antigen Lu b , J.S. b , Kp b , and Yt a17. The method of any one of embodiments 1 to 16, wherein the cell is type O cell positive for .

[0046] Embodiment 18. The CO-Hb RBCs are characterized by the surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w 18. The method according to any one of embodiments 1 to 17, wherein the cell is type O cell that is negative for .

[0047] Embodiment 19. The method of any one of embodiments 1 to 18, wherein the CO-Hb RBCs are type O cells positive for surface antigens D, C, and e and have Rh haplotype R1, or positive for surface antigens D, c, and e and have Rh haplotype R2.

[0048] Embodiment 20. The CO-Hb RBCs are characterized by the surface antigen Lu b , J.S. b , Kp b , and Yt a 20. The method of any one of embodiments 1-19, wherein the cell is type O cell positive for .

[0049] Embodiment 21. The CO-Hb RBCs are characterized by surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w 21. The method of any one of embodiments 1 to 20, wherein the cell is type O cell that is negative for .

[0050] Embodiment 22. The CO-Hb RBCs are ficin-treated and contain surface antigens M, N, Fy a ,Fy b , S, s, Xg a , Pr, Ch a , Rg a and Yk a22. The method of any one of embodiments 1 to 21, wherein the cell is type O cell that is negative for .

[0051] 23. The CO-Hb RBC is selected from the group consisting of D, C, E, c, e, f, Jk a , J.K. b , Le a , Le b , P1, I, IH, Vel, PP1P k and type O cells positive for binding of the antibody to the P antigen.

[0052] 24. The CO-Hb RBC is selected from the group consisting of Fya, Fyb, S, s, M, N, Xg a , Pr, Ch a , Rg a , and Yk a 24. The method of any one of embodiments 1 to 23, wherein the cell is type O cell having reduced or absent binding of the antibody to the antibody.

[0053] Embodiment 25. The method of any one of embodiments 1 to 24, wherein the CO-Hb RBCs are positive for binding of antibodies to the D antigen, and the CO-Hb RBCs are type O cells sensitized with anti-D(Rh0) serum.

[0054] Embodiment 26 The method of any one of embodiments 1 to 25, wherein said CO-Hb RBCs are type A cells positive for binding of an antibody to the A2 antigen.

[0055] Embodiment 27. The method of any one of embodiments 1 to 26, wherein the CO-Hb RBCs are type B cells that are positive for binding of antibodies to B antigens and negative for binding of anti-D (Rh0) antibodies.

[0056] Embodiment 28. The method of any one of embodiments 1 to 27, wherein the CO-Hb RBCs are type A cells that are positive for binding of antibodies to the A1 antigen and negative for binding of anti-D (Rh0) antibodies.

[0057] Embodiment 29. The method of any one of embodiments 1 to 28, wherein the CO-Hb RBCs are AB type cells positive for binding of antibodies to A1, B antigens, and Rh antigens d, c, and e of the Rh blood type rr(dce / dec).

[0058] Embodiment 30. The method of any one of embodiments 1 to 29, wherein the CO-Hb RBCs are type O cells positive for binding of antibodies to Rh antigens D, d, C, c, and e, and have the Rh phenotype R1r(DCe / dce).

[0059] 31. The CO-Hb RBC is selected from the group consisting of D(RH1), C(RH2), E(RH3), c(RH4), e(RH5), M, N, S, s, P1, K, k, Fy a ,Fy b , J.K. a , J.K. b , Le a and Le b 31. The method of any one of embodiments 1 to 30, wherein the cells are type O cells positive for binding of an antibody to

[0060] Embodiment 32. The CO-Hb RBCs are b , J.S. b , Kp b , and Yt a The method of any one of embodiments 1-31, wherein the cells are type O cells positive for binding of the antibody to the antigen.

[0061] Embodiment 33. The CO-Hb RBCs are a , Kp a , Wr a , Di a , Vw, V, Lu a and C w The method of any one of embodiments 1 to 32, wherein the cells are type O cells that are negative for binding of the antibody to the antigen.

[0062] Embodiment 34. The method of any one of embodiments 1 to 33, wherein said CO-Hb RBCs are type O cells with no profile in the Resolve® Panel A instructions.

[0063] Embodiment 35. The method of any one of embodiments 1 to 34, wherein said CO-Hb RBCs are type O cells with no profile in the Resolve® Panel B instructions.

[0064] Embodiment 36 The method of any one of embodiments 1 to 35, wherein said CO-Hb RBCs are type cells with no profile in the Resolve® Panel C manual.

[0065] Embodiment 37. A kit comprising: a) one or more vials of carbon monoxide saturated RBCs (CO-Hb RBCs), said vials comprising a buffer; (i) Blood type O cells and D, C, c, E, e, CW, K, k, P1, Fy a ,Fy b , J.K. a , J.K. b , Le a , Le b CO-Hb RBCs that are positive for a surface antigen selected from the group consisting of M, N, S, and s; (ii) CO-Hb RBC is D, C, c, E, e, CW, K, k, P1, Fy a ,Fy b , J.K. a , J.K. b , Le a , Le b ,M.,N.,S.,s.,I.,Lu a , Lu b , J.S. b , Kp b , and Yt a are blood group O cells positive for a surface antigen selected from the group consisting of (iii) CO-Hb RBC is D, C, c, E, e, CW, K, k, P1, Fy a ,Fy b , J.K. a, J.K. b , Le a , Le b ,M.,N.,S.,s.,I.,Lu a , Lu b , J.S. b , Kp b , and Yt a and the surface antigen Js a , Kp a , Wr a , Di a , V w ,V.,Lu a , and C w are blood type O cells that are negative for (iv) CO-Hb RBCs expressing surface antigens D, C, c, E, e, f, CW, K, k, P1, and Fy a ,Fy b , J.K. a , J.K. b , Le a , Le b ,M.,N.,S.,s.,Lu a , and Lu b Type O cells are negative for (v) the CO-Hb RBCs are type O cells positive for Rh antigens D, C, and e; (vi) CO-Hb RBCs expressing Rh antigens D, C, and e, I, and Lu b , J.S. b , Kp b , and Yt a are type O cells positive for (vii) CO-Hb RBCs expressing Rh antigens D, C, and e, I, and Lu b , J.S. b , Kp b , and Yt a Positive for and surface antigen Js a , Kp a , Wr a , Di a V w ,V.,Lu a and C w Type O cells are negative for (viii) the CO-Hb RBCs are type A cells positive for surface antigen A1; (ix) the CO-Hb RBCs are type A cells that are positive for surface antigen A1 and negative for surface antigens D, C, and E; (x) the CO-Hb RBC is a type A cell that is positive for surface antigen A2; (xi) The CO-Hb RBCs are type A cells that are positive for surface antigen A2 and negative for surface antigens D, C, and E; (xii) the CO-Hb RBC is a B cell positive for surface antigen B; (xiii) the CO-Hb RBCs are B-cells that are positive for surface antigen B and negative for surface antigens D, C, and E; (xiv) The CO-Hb RBCs are positive for surface antigens D, C, and e and are type O cells with Rh phenotype R1R1; (xv) CO-Hb RBCs express surface antigens D and C w , and e positive and Rh phenotype R1 w Type O cells with R1 (xvi) The CO-Hb RBCs are type O cells that are positive for surface antigens D, c, and E and have the Rh phenotype R2R2; (xvii) The CO-Hb RBCs are positive for surface antigens d, c, and e and are type O cells with Rh phenotype rr; (xviii) CO-Hb RBCs are positive for surface antigens D, C, and e and have the Rh phenotype R1R1 and surface antigen Lu b , J.S. b , Kp b , and Yt a are type O cells positive for (xix) CO-Hb RBCs express surface antigens D and C w , and e positive and Rh phenotype R1 w R1 and surface antigen Lu b , J.S. b , Kp b , and Yta are type O cells positive for (xx) CO-Hb RBCs are positive for surface antigens D, c, and E and have the Rh phenotype R2R2 and surface antigen Lu b , J.S. b , Kp b , and Yt a are type O cells positive for (xxi) CO-Hb RBCs are positive for surface antigens d, c, and e and have the Rh phenotype rr and surface antigen Lu b , J.S. b , Kp b and Yt a are type O cells positive for (xxii) CO-Hb RBCs are positive for surface antigens D, C, and e and have the Rh phenotype R1R1 and surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w Type O cells are negative for (xxiii) CO-Hb RBCs express surface antigens D and C w , and e positive and Rh phenotype R1 w R1 and surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w Type O cells are negative for (xxiv) CO-Hb RBCs are positive for surface antigens D, c, and E and have the Rh phenotype R2R2 and surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w Type O cells are negative for (xxv) CO-Hb RBCs are positive for surface antigens d, c, and e and have the Rh phenotype rr and surface antigen Jsa , Kp a , Wr a , Di a , Vw, V, Lu a and C w Type O cells are negative for (xxvi) The CO-Hb RBCs are type O cells positive for surface antigens D, C, and e and have Rh haplotype R1, or positive for surface antigens D, c, and e and have Rh haplotype R2. (xxvii) CO-Hb RBCs are positive for surface antigens D, C, and e and have the Rh haplotype R1 and surface antigen Lu b , J.S. b , Kp b , and Yt a or positive for surface antigens D, c, and e and have the Rh haplotype R2CO-HbRBC and surface antigen Lu b , J.S. b , Kp b , and Yt a are type O cells positive for (xxviii) CO-Hb RBCs are positive for surface antigens D, C, and e and have Rh haplotype R1 and surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w or are positive for surface antigens D, c, and e and have the Rh haplotype R2 and surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w Type O cells are negative for (xxix) CO-Hb RBCs are ficin-treated and have surface antigens M, N, Fy a ,Fy b , S, s, Xg a , Pr, Ch a , Rga and Yk a are type O cells that are negative for (xxx) CO-Hb RBCs are ficin-treated and have surface antigens M, N, Fy a ,Fy b , S, s, Xg a , Pr, Ch a , Rg a and Yk a Negative for D, C, E, c, e, f, Jk a , J.K. b , Le a , Le b , P1, I, IH, Vel, PP1P k and type O cells positive for antibody binding to the P antigen. (xxxi) CO-Hb RBCs are ficin-treated and have surface antigens M, N, Fy a ,Fy b , S, s, Xg a , Pr, Ch a , Rg a Negative for Fya, Fyb, S, s, M, N, Xg a , Pr, Ch a , Rg a , and Yk a are type O cells having reduced or absent binding of antibodies to (xxxii) The CO-Hb RBCs are positive for antibody binding to D antigen, and the CO-Hb RBCs are type O cells sensitized with anti-D(Rh0) serum. (xxxiii) the CO-Hb RBCs are type A cells positive for antibody binding to the A2 antigen; (xxxiv) The CO-Hb RBCs are B type cells that are positive for antibody binding to B antigens and negative for anti-D (Rh0) antibody binding. (xxxv) The CO-Hb RBCs are type A cells that are positive for antibody binding to the A1 antigen and negative for anti-D (Rh0) antibody binding. (xxxvi) The CO-Hb RBCs are AB type cells positive for A1, B antigens, and antibody binding to Rh antigens d, c, and e of the Rh blood type rr(dce / dec); (xxxvii) The CO-Hb RBCs are type O cells that are positive for antibody binding to Rh antigens D, d, C, c, and e and have the Rh phenotype R1r(DCe / dce); (xxxviii)CO-Hb RBC is D(RH1), C(RH2), E(RH3), c(RH4), e(RH5), M, N, S, s, P1, K, k, Fy a ,Fy b , J.K. a , J.K. b , Le a and Le b are type O cells positive for binding of antibodies to (xxxix) CO-Hb RBC is D(RH1), C(RH2), E(RH3), c(RH4), e(RH5), M, N, S, s, P1, K, k, Fy a ,Fy b , J.K. a , J.K. b , Le a and Le b Positive for antibody binding to Lu b , J.S. b , Kp b , and Yt a are type O cells positive for binding of antibody to antigen, (xl)CO-Hb RBC is D(RH1), C(RH2), E(RH3), c(RH4), e(RH5), M, N, S, s, P1, K, k, Fy a ,Fy b , J.K. a , J.K. b , Le a and Le b Positive for antibody binding to Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C wa plurality of CO-Hb RBCs having a common set of surface antigens selected from the group consisting of: CO-Hb RBCs that are type O cells that are negative for binding of an antibody to the antigen; b) A kit, including instructions.

[0066] 38. A buffer solution, (i) Blood type O cells and D, C, c, E, e, CW, K, k, P1, Fy a ,Fy b , J.K. a , J.K. b , Le a , Le b CO-Hb RBCs that are positive for a surface antigen selected from the group consisting of M, N, S, and s; (ii) CO-Hb RBC is D, C, c, E, e, CW, K, k, P1, Fy a ,Fy b , J.K. a , J.K. b , Le a , Le b ,M.,N.,S.,s.,I.,Lu a , Lu b , J.S. b , Kp b , and Yt a are blood group O cells positive for a surface antigen selected from the group consisting of (iii) CO-Hb RBC is D, C, c, E, e, CW, K, k, P1, Fy a ,Fy b , J.K. a , J.K. b , Le a , Le b ,M.,N.,S.,s.,I.,Lu a , Lu b , J.S. b , Kp b , and Yt a and the surface antigen Js a , Kp a , Wr a , Di a , V w ,V.,Lu a , and C ware blood type O cells that are negative for (iv) CO-Hb RBCs expressing surface antigens D, C, c, E, e, f, CW, K, k, P1, and Fy a ,Fy b , J.K. a , J.K. b , Le a , Le b ,M.,N.,S.,s.,Lu a , and Lu b Type O cells are negative for (v) the CO-Hb RBCs are type O cells positive for Rh antigens D, C, and e; (vi) CO-Hb RBCs expressing Rh antigens D, C, and e, I, and Lu b , J.S. b , Kp b , and Yt a are type O cells positive for (vii) CO-Hb RBCs expressing Rh antigens D, C, and e, I, and Lu b , J.S. b , Kp b , and Yt a Positive for and surface antigen Js a , Kp a , Wr a , Di a V w ,V.,Lu a and C w Type O cells are negative for (viii) the CO-Hb RBCs are type A cells positive for surface antigen A1; (ix) the CO-Hb RBCs are type A cells that are positive for surface antigen A1 and negative for surface antigens D, C, and E; (x) the CO-Hb RBC is a type A cell that is positive for surface antigen A2; (xi) The CO-Hb RBCs are type A cells that are positive for surface antigen A2 and negative for surface antigens D, C, and E; (xii) the CO-Hb RBC is a B cell positive for surface antigen B; (xiii) the CO-Hb RBCs are B-cells that are positive for surface antigen B and negative for surface antigens D, C, and E; (xiv) The CO-Hb RBCs are positive for surface antigens D, C, and e and are type O cells with Rh phenotype R1R1; (xv) CO-Hb RBCs express surface antigens D and C w , and e positive and Rh phenotype R1 w Type O cells with R1 (xvi) The CO-Hb RBCs are type O cells that are positive for surface antigens D, c, and E and have the Rh phenotype R2R2; (xvii) The CO-Hb RBCs are positive for surface antigens d, c, and e and are type O cells with Rh phenotype rr; (xviii) CO-Hb RBCs are positive for surface antigens D, C, and e and have the Rh phenotype R1R1 and surface antigen Lu b , J.S. b , Kp b , and Yt a are type O cells positive for (xix) CO-Hb RBCs express surface antigens D and C w , and e positive and Rh phenotype R1 w R1 and surface antigen Lu b , J.S. b , Kp b , and Yt a are type O cells positive for (xx) CO-Hb RBCs are positive for surface antigens D, c, and E and have the Rh phenotype R2R2 and surface antigen Lu b , J.S. b , Kp b , and Yt a are type O cells positive for (xxi) CO-Hb RBCs are positive for surface antigens d, c, and e and have the Rh phenotype rr and surface antigen Lu b , J.S. b , Kp b and Yt aare type O cells positive for (xxii) CO-Hb RBCs are positive for surface antigens D, C, and e and have the Rh phenotype R1R1 and surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w Type O cells are negative for (xxiii) CO-Hb RBCs express surface antigens D and C w , and e positive and Rh phenotype R1 w R1 and surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w Type O cells are negative for (xxiv) CO-Hb RBCs are positive for surface antigens D, c, and E and have the Rh phenotype R2R2 and surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w Type O cells are negative for (xxv) CO-Hb RBCs are positive for surface antigens d, c, and e and have the Rh phenotype rr and surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w Type O cells are negative for (xxvi) The CO-Hb RBCs are type O cells positive for surface antigens D, C, and e and have Rh haplotype R1, or positive for surface antigens D, c, and e and have Rh haplotype R2. (xxvii) CO-Hb RBCs are positive for surface antigens D, C, and e and have the Rh haplotype R1 and surface antigen Lub , J.S. b , Kp b , and Yt a or positive for surface antigens D, c, and e and have Rh haplotype R2CO-Hb RBCs and surface antigen Lu b , J.S. b , Kp b , and Yt a are type O cells positive for (xxviii) CO-Hb RBCs are positive for surface antigens D, C, and e and have Rh haplotype R1 and surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w or are positive for surface antigens D, c, and e and have the Rh haplotype R2 and surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w Type O cells are negative for (xxix) CO-Hb RBCs are ficin-treated and have surface antigens M, N, Fy a ,Fy b , S, s, Xg a , Pr, Ch a , Rg a and Yk a are type O cells that are negative for (xxx) CO-Hb RBCs are ficin-treated and have surface antigens M, N, Fy a ,Fy b , S, s, Xg a , Pr, Ch a , Rg a and Yk a Negative for D, C, E, c, e, f, Jk a , J.K. b , Le a , Le b , P1, I, IH, Vel, PP1P kand type O cells positive for antibody binding to the P antigen. (xxxi) CO-Hb RBCs are ficin-treated and have surface antigens M, N, Fy a ,Fy b , S, s, Xg a , Pr, Ch a , Rg a Negative for Fya, Fyb, S, s, M, N, Xg a , Pr, Ch a , Rg a , and Yk a are type O cells having reduced or absent binding of antibodies to (xxxii) The CO-Hb RBCs are positive for antibody binding to the D antigen, and the cells are type O cells sensitized with anti-D(Rh0) serum. (xxxiii) the CO-Hb RBCs are type A cells positive for antibody binding to the A2 antigen; (xxxiv) The CO-Hb RBCs are B type cells that are positive for antibody binding to B antigens and negative for anti-D (Rh0) antibody binding. (xxxv) The CO-Hb RBCs are type A cells that are positive for antibody binding to the A1 antigen and negative for anti-D (Rh0) antibody binding. (xxxvi) The CO-Hb RBCs are AB type cells positive for A1, B antigens, and antibody binding to Rh antigens d, c, and e of the Rh blood type rr(dce / dec); (xxxvii) The CO-Hb RBCs are type O cells that are positive for antibody binding to Rh antigens D, d, C, c, and e and have the Rh phenotype R1r(DCe / dce); (xxxviii)CO-Hb RBC is D(RH1), C(RH2), E(RH3), c(RH4), e(RH5), M, N, S, s, P1, K, k, Fy a ,Fy b , J.K. a , J.K. b , Le a and Le b are type O cells positive for binding of antibodies to (xxxix) CO-Hb RBC is D(RH1), C(RH2), E(RH3), c(RH4), e(RH5), M, N, S, s, P1, K, k, Fy a ,Fy b , J.K. a , J.K. b , Le a and Le b Positive for antibody binding to Lu b , J.S. b , Kp b , and Yt a are type O cells positive for binding of antibody to antigen, (xl)CO-Hb RBC is D(RH1), C(RH2), E(RH3), c(RH4), e(RH5), M, N, S, s, P1, K, k, Fy a ,Fy b , J.K. a , J.K. b , Le a and Le b Positive for antibody binding to Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w and CO-Hb RBCs selected from the group consisting of: CO-Hb RBCs that are type O cells that are negative for binding of antibody to antigen.

[0067] Having now generally described the invention, it will be more readily understood by reference to the following examples, which are provided by way of illustration and are not intended to be limitations of the invention, unless otherwise specified.

[0068] Each journal, patent, and other document or reference cited herein is incorporated herein by reference in its entirety. EXAMPLES

[0069] Example 1: Blood Collection and Red Blood Cell Concentrate (RCC) Preparation Blood for preparation of reagent RBCs is collected from established donors into an anticoagulant solution using standard methods. Various known anticoagulants suitable for use in transfusion medicine preferably include citrate phosphate dextrose (CPD) and acid citrate dextrose (ACD), although other anticoagulants, such as ethylenediaminetetraacetic acid (EDTA) and ethylene glycol-bis(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), may be suitably used if the blood is not to be used for transfusion. The collected blood is centrifuged or filtered to separate white blood cells (WBCs) and excess plasma to prepare packed red blood cells (pRBCs) or red blood cell concentrates (RCCs).

[0070] Example 2: Preparation of Hb-CO-containing RCC (Hb-CO RCC) Preferably immediately, the red blood cell concentrate (RCC) prepared in Example 1 is converted to Hb-CO by one of the following methods.

[0071] a. Rapid gas exchange: The RCCs are kept in polyvinyl chloride or other suitable bags (150 ml or more), carbon monoxide is introduced, and the bag containing the RCCs and CO is placed in a platelet shaker for approximately 10 min. After incubation, the gas containing unexchanged CO is released, and residual oxygen, and carbon dioxide are allowed to evolve and replaced with fresh CO. After a second incubation in the platelet shaker for approximately 10 min, the gas is allowed to evolve a second time and replaced with a third volume of CO. After a final incubation with shaking in the platelet shaker, excess gas is removed and the Hb-CO RCCs are transferred under anaerobic conditions to suitable vials for further characterization, quality control, and storage.

[0072] b. Gas exchange throughout the night: The RCCs are kept in polyvinyl chloride or other suitable bags (150 ml or larger), carbon monoxide is introduced, and the bag containing the RCCs and CO is placed overnight at 4 °C with constant gentle shaking or stirring at regular intervals (e.g., 3-5 x 8 h). After incubation, the CO containing excess gas is removed and the Hb-CO RCCs are transferred to suitable vials for further characterization, quality control, and storage under anaerobic conditions.

[0073] C membrane gas exchange RCCs held in polyvinyl chloride or other suitable bags (150 ml or larger) are pumped through a Sorin D100 oxygen generator with carbon monoxide as the gas source. RCCs are pumped through the D100 for 30 minutes using either a centrifugal or peristaltic pump. Oxygen and CO levels are monitored until Hb-CO levels of >95% are achieved.

[0074] d. Microbubble gas exchange: The RCCs are kept in polyvinyl chloride or other suitable ventilated bags (150 ml or larger) and carbon monoxide is bubbled into the RCCs. Care is taken to ensure that the bubble diameter is less than 1 μm to prevent lysis of red blood cells. The resulting Hb-CO RCCs are transferred to suitable vials for further characterization, quality control, and storage under anaerobic conditions.

[0075] e. Modified HEMANEXT (登録商標) Oxygen reduction bag (ORB) HEMANEXT, as described in U.S. Patent No. 10,058,091, issued August 28, 2018 (登録商標) An oxygen reduction bag (ORB) is modified to remove the sorbent pack and the headspace filled with CO gas (100-200 ml). The CO2-containing ORB bag is agitated for 30 min at room temperature on a platelet shaker. Alternatively, the CO2-containing ORB bag is placed at 4 °C with either constant shaking or agitation at regular intervals (e.g., 3-5 x over 8 h).

[0076] Example 2: Preparation and packaging of HB-CO Reagent RBC Continue further manufacturing process with CO-treated RBCs.

[0077] Package the reagent RBCs in a reagent bottle head, fill the headspace with CO under positive pressure, and store at 4 °C.

Claims

1. 1. A method for preparing reagent red blood cells (RBCs) for use in blood typing, comprising: a) obtaining packed red blood cells from donor blood of known phenotype; b) flushing the packed red blood cells with a gas containing carbon monoxide to prepare carbon monoxide saturated RBCs (CO-Hb RBCs); and c) storing the CO-Hb RBCs under anaerobic conditions in the presence of carbon monoxide (CO) to produce CO-storage reagent RBCs for blood typing, wherein surface antigens of the CO-storage reagent RBCs are stabilized and the CO-storage reagent RBCs have an extended shelf life compared to conventionally stored RBCs.

2. The method of claim 1 , wherein the gas is oxygen-free.

3. The CO-storage reagent RBCs are D, C, c, E, e, CW, K, k, P 1 , Fy a , Fy b , J.K. a , J.K. b , Le a , Le b 2. The method of claim 1, wherein the cells are blood type O cells that are positive for one or more of the surface antigens selected from the group consisting of: , M, N, S, and s.

4. The CO-storage reagent RBCs are D, C, c, E, e, f, CW, K, k, P 1 , Fy a , Fy b , J.K. a , J.K. b , Le a , Le b ,M,N,S,s,Lu a , and Lu b The method of claim 1, wherein the cells are type O cells that are negative for one or more of the surface antigens selected from the group consisting of:

5. 2. The method of claim 1, wherein the CO-storage reagent RBCs are type A cells positive for surface antigen A2 or type B cells positive for surface antigen B.

6. The CO-storage reagent RBCs are Positive for surface antigens D, C, and e and Rh phenotype R 1 R 1 Have Surface antigen D, C w , and e positive and Rh phenotype R 1 w R 1 Have Positive for surface antigens D, c, and E and Rh phenotype R 2 R 2 Have 2. The method of claim 1, wherein the cells are type O cells that are positive for surface antigens d, c, and e and have the Rh phenotype rr.

7. The CO-storage reagent RBCs contain the surface antigen Lu b , J.S. b , Kp b , and Yt a positive for or Positive for surface antigens D, C, and e and Rh haplotype R 1 or Positive for surface antigens D, c, and e and Rh haplotype R 2 The method of claim 6, wherein the cell is type O cell,

8. The CO-storage reagent RBCs contain the surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w The method of claim 6, wherein the cells are type 0 cells that are negative for

9. The CO-storage reagent RBCs are ficin-treated and contain surface antigens M, N, Fy a , Fy b , S, s, Xg a , Pr, Ch a , Rg a and Yk a The method of claim 1, wherein the cells are type O cells that are negative for

10. CO-Storage Reagent RBCs are anti-surface antigen D (Rh 0 2. The method of claim 1, wherein the cells are type O cells that are positive for antibody binding to surface antigen D when sensitized with serum.

11. The CO-storage reagent RBC contains surface antigen A 2 The method of claim 1, wherein the cells are type A cells positive for binding of an antibody to

12. The CO-storage reagent RBCs are positive for binding of antibodies to surface antigen B and are 0 2. The method of claim 1, wherein the cells are B-type cells that are negative for antibody binding.

13. The CO-storage reagent RBC contains surface antigen A 1 Positive for antibody binding to anti-D (Rh 0 2. The method of claim 1, wherein the cells are type A cells that are negative for antibody binding.

14. The CO-storage reagent RBC contains surface antigen A 1 2. The method of claim 1, wherein the cells are type AB cells positive for antibody binding to Rh surface antigen B, and to Rh surface antigens d, c, and e of Rh blood group rr(dce / dec).

15. The CO-storage reagent RBCs are positive for antibody binding to Rh surface antigens D, d, C, c, and e, and have the Rh phenotype R. 1 r(DCe / dce), or Surface antigens D (RH1), C (RH2), E (RH3), c (RH4), e (RH5), M, N, S, s, P 1 ,K,k,Fy a , Fy b , J.K. a , J.K. b , Le a and Le b The method of claim 1, wherein the cells are type O cells positive for binding of an antibody to

16. 2. The method of claim 1, wherein the CO-storage reagent RBCs are type O cells with no profile in the Resolve® Panel A instructions or the Resolve® Panel B instructions or the Resolve® Panel C instructions.

17. A kit for use in blood typing, comprising: a) one or more vials containing a plurality of carbon monoxide saturated RBCs (CO-Hb RBCs) and a buffer solution, said plurality of CO-Hb RBCs having a common set of stabilized surface antigens, said CO-Hb RBCs being selected from the group consisting of: (i) D, C, c, E, e, CW, K, k, P 1 , Fy a , Fy b , J.K. a , J.K. b , Le a , Le b CO-Hb RBCs that are type O cells positive for one or more of the surface antigens selected from the group consisting of: (ii) D, C, c, E, e, CW, K, k, P 1 , Fy a , Fy b , J.K. a , J.K. b , Le a , Le b ,M,N,S,s,I,Lu a , Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs, which are blood group O cells positive for a surface antigen selected from the group consisting of: (iii) D, C, c, E, e, CW, K, k, P 1 , Fy a , Fy b , J.K. a , J.K. b , Le a , Le b ,M,N,S,s,I,Lu a , Lu b , J.S. b , Kp b , and Yt a and the surface antigen Js a , Kp a , Wr a , Di a , V w , V., Lu a , and C w CO-Hb RBCs, which are blood type O cells that are negative for (iv) Surface antigens D, C, c, E, e, f, CW, K, k, P 1 , Fy a , Fy b , J.K. a , J.K. b , Le a , Le b ,M,N,S,s,Lu a , and Lu b CO-Hb RBCs, which are type O cells negative for (v) CO-Hb RBCs, which are type O cells positive for Rh surface antigens D, C, and e; (vi) Rh surface antigens D, C, and e, and surface antigens I, Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs, which are type O cells positive for (vii) Rh surface antigens D, C, and e, and surface antigens I, Lu b , J.S. b , Kp b , and Yt a Positive for and surface antigen Js a , Kp a , Wr a , Di a V w , V., Lu a and C w CO-Hb RBCs, which are type O cells negative for (viii) CO-Hb RBC, which is a type A cell positive for surface antigen A1; (ix) CO-Hb RBCs, which are type A cells positive for surface antigen A1 and negative for surface antigens D, C, and E; (x) CO-Hb RBC, which is a type A cell positive for surface antigen A2; (xi) CO-Hb RBCs, which are type A cells positive for surface antigen A2 and negative for surface antigens D, C, and E; (xii) CO-Hb RBC, which is a B-type cell positive for surface antigen B; (xiii) CO-Hb RBCs, which are type B cells positive for surface antigen B and negative for surface antigens D, C, and E; (xiv) positive for surface antigens D, C, and e and Rh phenotype R 1 R 1 CO-Hb RBCs, which are type O cells having (xv) Surface antigens D, C w , and e positive and Rh phenotype R 1 w R 1 CO-Hb RBCs, which are type O cells having (xvi) positive for surface antigens D, c, and E and Rh phenotype R 2 R 2 CO-Hb RBCs, which are type O cells having (xvii) CO-Hb RBCs that are positive for surface antigens d, c, and e and are type O cells with Rh phenotype rr; (xviii) positive for surface antigens D, C, and e and Rh phenotype R 1 R 1 having a surface antigen Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs, which are type O cells positive for (xix) Surface antigens D, C w , and e positive and Rh phenotype R 1 w R 1 having a surface antigen Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs, which are type O cells positive for (xx) Surface Positive for antigens D, c, and E and Rh phenotype R 2 R 2 having a surface antigen Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs, which are type O cells positive for (xxi) positive for surface antigens d, c, and e, and have the Rh phenotype rr, and positive for surface antigen Lu b , J.S. b , Kp b and Yt a CO-Hb RBCs, which are type O cells positive for (xxii) Positive for surface antigens D, C, and e and Rh phenotype R 1 R 1 having a surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w CO-Hb RBCs, which are type O cells negative for (xxiii) Surface antigens D, C w , and e positive and Rh phenotype R 1 w R 1 having a surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w are type O cells that are negative for (xxiv) positive for surface antigens D, c, and E and Rh phenotype R 2 R 2 having a surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w CO-Hb RBCs, which are type O cells negative for (xxv) positive for surface antigens d, c, and e and have the Rh phenotype rr and surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w CO-Hb RBCs, which are type O cells negative for (xxvi) Positive for surface antigens D, C, and e and Rh haplotype R 1 or positive for surface antigens D, c, and e and Rh haplotype R 2 CO-Hb RBCs, which are type O cells having (xxvii) Positive for surface antigens D, C, and e and Rh haplotype R 1 having a surface antigen Lu b , J.S. b , Kp b , and Yt a or positive for surface antigens D, c, and e and Rh haplotype R 2 CO-Hb RBC with surface antigen Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs, which are type O cells positive for (xxviii) positive for surface antigens D, C, and e and Rh haplotype R 1 having a surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w or positive for surface antigens D, c, and e and Rh haplotype R 2 having a surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w CO-Hb RBCs, which are type O cells negative for (xxix) ficin-treated and having surface antigens M, N, Fy a , Fy b , S, s, Xg a , Pr, Ch a , Rg a and Yk a CO-Hb RBCs, which are type O cells negative for (xxx) ficin-treated and surface antigens M, N, Fy a , Fy b , S, s, Xg a , Pr, Ch a , Rg a and Yk a Negative for surface antigens D, C, E, c, e, f, Jk a , J.K. b , Le a , Le b , P 1 ,I,IH,Vel,PP 1 P k and CO-Hb RBC, which is a type O cell positive for binding of antibodies to P; (xxxi) ficin-treated and having surface antigens M, N, Fy a , Fy b , S, s, Xg a , Pr, Ch a , Rg a Negative for Fya, Fyb, S, s, M, N, Xg a , Pr, Ch a , Rg a , and Y a CO-Hb RBCs, which are type O cells with reduced or no antibody binding to the (xxxii) Anti-surface antigen D (Rh 0 ) CO-Hb RBCs, which are type O cells that are positive for antibody binding to surface antigen D when sensitized with serum; (xxxiii) Surface antigen A 2 CO-Hb RBC, which is a type A cell positive for antibody binding to (xxxiv) positive for antibody binding to surface antigen B and anti-D (Rh 0 ) CO-Hb RBC, which are B-type cells negative for antibody binding; (xxxv) Surface antigen A 1 Positive for antibody binding to anti-D (Rh 0 ) CO-Hb RBCs, which are type A cells that are negative for antibody binding; (xxxvi) Surface antigen A 1 CO-Hb RBCs, which are type AB cells positive for antibody binding to the Rh surface antigen B, and to the Rh surface antigens d, c, and e of the Rh blood group rr(dce / dec); (xxxvii) positive for antibody binding to Rh surface antigens D, d, C, c, and e and Rh phenotype R 1 CO-Hb RBC, which is a type O cell with r(DCe / dce); (xxxviii) Surface antigens D (RH1), C (RH2), E (RH3), c (RH4), e (RH5), M, N, S, s, P 1 ,K,k,Fy a , Fy b , J.K. a , J.K. b , Le a and Le b CO-Hb RBCs, which are type O cells positive for antibody binding to (xxxix) Surface antigens D (RH1), C (RH2), E (RH3), c (RH4), e (RH5), M, N, S, s, P 1 ,K,k,Fy a , Fy b , J.K. a , J.K. b , Le a and Le b Positive for antibody binding to the surface antigen Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs, which are type O cells positive for antibody binding to (xl) Surface antigens D (RH1), C (RH2), E (RH3), c (RH4), e (RH5), M, N, S, s, P 1 ,K,k,Fy a , Fy b , J.K. a , J.K. b , Le a and Le b Positive for antibody binding to surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w CO-Hb RBCs, which are type O cells that are negative for binding of antibodies to antigens; and b) instructions for use of the kit; Including the kit.

18. A vial containing carbon monoxide saturated RBCs (CO-Hb RBCs) and a buffer solution, said CO-Hb RBCs having stabilized surface antigens and selected from the group consisting of: (i) D, C, c, E, e, CW, K, k, P 1 , Fy a , Fy b , J.K. a , J.K. b , Le a , Le b CO-Hb RBCs, which are blood type O cells positive for one or more of the surface antigens selected from the group consisting of: CO-Hb RBCs, CO-Hb RBCs, (ii) D, C, c, E, e, CW, K, k, P 1 , Fy a , Fy b , J.K. a , J.K. b , Le a , Le b ,M,N,S,s,I,Lu a , Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs, which are blood type O cells positive for one or more of the surface antigens selected from the group consisting of: (iii) D, C, c, E, e, CW, K, k, P 1 , Fy a , Fy b , J.K. a , J.K. b , Le a , Le b ,M,N,S,s,I,Lu a , Lu b , J.S. b , Kp b , and Yt a and the surface antigen Js a , Kp a , Wr a , Di a , V w , V., Lu a , and C w CO-Hb RBCs, which are blood type O cells that are negative for (iv) Surface antigens D, C, c, E, e, f, CW, K, k, P 1 , Fy a , Fy b , J.K. a , J.K. b , Le a , Le b ,M,N,S,s,Lu a , and Lu b CO-Hb RBCs, which are type O cells negative for (v) CO-Hb RBCs, which are type O cells positive for Rh surface antigens D, C, and e; (vi) positive for Rh surface antigens D, C, and e and positive for surface antigens I, Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs, which are type O cells positive for (vii) positive for Rh surface antigens D, C, and e and positive for surface antigens I, Lu b , J.S. b , Kp b , and Yt a Positive for and surface antigen Js a , Kp a , Wr a , Di a V w , V., Lu a and C w CO-Hb RBCs, which are type O cells negative for (viii) CO-Hb RBC, which is a type A cell positive for surface antigen A1; (ix) CO-Hb RBCs, which are type A cells positive for surface antigen A1 and negative for surface antigens D, C, and E; (x) CO-Hb RBC, which is a type A cell positive for surface antigen A2; (xi) CO-Hb RBCs, which are type A cells positive for surface antigen A2 and negative for surface antigens D, C, and E; (xii) CO-Hb RBC, which is a B-type cell positive for surface antigen B; (xiii) CO-Hb RBCs, which are type B cells positive for surface antigen B and negative for surface antigens D, C, and E; (xiv) positive for surface antigens D, C, and e and Rh phenotype R 1 R 1 CO-Hb RBCs, which are type O cells having (xv) Surface antigens D, C w , and e positive and Rh phenotype R 1 w R 1 CO-Hb RBCs, which are type O cells having (xvi) positive for surface antigens D, c, and E and Rh phenotype R 2 R 2 CO-Hb RBCs, which are type O cells having (xvii) CO-Hb RBCs that are positive for surface antigens d, c, and e and are type O cells with Rh phenotype rr; (xviii) positive for surface antigens D, C, and e and Rh phenotype R 1 R 1 having a surface antigen Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs, which are type O cells positive for (xix) Surface antigens D, C w , and e positive and Rh phenotype R 1 w R 1 having a surface antigen Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs, which are type O cells positive for (xx) Surface Positive for antigens D, c, and E and Rh phenotype R 2 R 2 having a surface antigen Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs, which are type O cells positive for (xxi) positive for surface antigens d, c, and e, and have the Rh phenotype rr, and positive for surface antigen Lu b , J.S. b , Kp b and Yt a CO-Hb RBCs, which are type O cells positive for (xxii) Positive for surface antigens D, C, and e and Rh phenotype R 1 R 1 having a surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w CO-Hb RBCs, which are type O cells negative for (xxiii) Surface antigens D, C w , and e positive and Rh phenotype R 1 w R 1 having a surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w CO-Hb RBCs, which are type O cells negative for (xxiv) positive for surface antigens D, c, and E and Rh phenotype R 2 R 2 having a surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w CO-Hb RBCs, which are type O cells negative for (xxv) positive for surface antigens d, c, and e and have the Rh phenotype rr and surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w CO-Hb RBCs, which are type O cells negative for (xxvi) Positive for surface antigens D, C, and e and Rh haplotype R 1 or positive for surface antigens D, c, and e and Rh haplotype R 2 CO-Hb RBCs, which are type O cells having (xxvii) Positive for surface antigens D, C, and e and Rh haplotype R 1 having a surface antigen Lu b , J.S. b , Kp b , and Yt a or positive for surface antigens D, c, and e and Rh haplotype R 2 having a surface antigen Lu b , J.S. b , Kp b , and Yt a CO-Hb RBCs, which are type O cells positive for (xxviii) positive for surface antigens D, C, and e and Rh haplotype R 1 having a surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w or positive for surface antigens D, c, and e and Rh haplotype R 2 having a surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w CO-Hb RBCs, which are type O cells negative for (xxix) ficin-treated and having surface antigens M, N, Fy a , Fy b , S, s, Xg a , Pr, Ch a , Rg a and Yk a CO-Hb RBCs, which are type O cells negative for (xxx) ficin-treated and surface antigens M, N, Fy a , Fy b , S, s, Xg a , Pr, Ch a , Rg a and Yk a Negative for surface antigens D, C, E, c, e, f, and Jk a , J.K. b , Le a , Le b , P 1 ,I,IH,Vel,PP 1 P k and CO-Hb RBC, which is a type O cell positive for binding of antibodies to P; (xxxi) ficin-treated and having surface antigens M, N, Fy a , Fy b , S, s, Xg a , Pr, Ch a , Rg a Negative for surface antigens Fya, Fyb, S, s, M, N, Xg a , Pr, Ch a , Rg a , and Y a CO-Hb RBCs, which are type O cells with reduced or no antibody binding to the (xxxii) Anti-surface antigen D (Rh 0 ) CO-Hb RBCs, which are type O cells that are positive for antibody binding to surface antigen D when sensitized with serum; (xxxiii) Surface antigen A 2 CO-Hb RBC, which is a type A cell positive for antibody binding to (xxxiv) positive for antibody binding to surface antigen B and anti-D (Rh 0 ) CO-Hb RBC, which are B-type cells negative for antibody binding; (xxxv) Surface antigen A 1 Positive for antibody binding to anti-D (Rh 0 ) CO-Hb RBCs, which are type A cells that are negative for antibody binding; (xxxvi) Surface antigen A 1 CO-Hb RBCs, which are type AB cells positive for antibody binding to surface antigen B, and to Rh surface antigens d, c, and e of the Rh blood group rr(dce / dec); (xxxvii) positive for antibody binding to Rh surface antigens D, d, C, c, and e and Rh phenotype R 1 CO-Hb RBC, which is a type O cell with r(DCe / dce); (xxxviii) Surface antigens D (RH1), C (RH2), E (RH3), c (RH4), e (RH5), M, N, S, s, P 1 ,K,k,Fy a , Fy b , J.K. a , J.K. b , Le a and Le b CO-Hb RBCs, which are type O cells positive for antibody binding to (xxxix) Surface antigens D (RH1), C (RH2), E (RH3), c (RH4), e (RH5), M, N, S, s, P 1 ,K,k,Fy a , Fy b , J.K. a , J.K. b , Le a and Le b Positive for antibody binding to the surface antigen Lu b , J.S. b , Kp b , and Yt a CO-Hb RBC, which is a type O cell positive for binding of antibody to antigen; (xl) Surface antigens D (RH1), C (RH2), E (RH3), c (RH4), e (RH5), M, N, S, s, P 1 ,K,k,Fy a , Fy b , J.K. a , J.K. b , Le a and Le b Positive for antibody binding to surface antigen Js a , Kp a , Wr a , Di a , Vw, V, Lu a and C w CO-Hb RBCs, which are type O cells that are negative for antibody binding to Hb.

19. 20. The vial of claim 18, wherein the vial further comprises an atmosphere of oxygen-free gas.

20. 20. The vial of claim 19, wherein the oxygen-free gas comprises carbon monoxide or nitrogen.

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