Blood screening methods

Abstract

The present invention relates to a method for identifying donor blood suitable for cancer patients receiving anti-CD38 antibodies as treatment. [Solution] In particular, the present invention addresses problems related to cross-matching tests between patient blood and donor blood when the patient blood contains anti-CD38 antibodies that interfere with cross-matching methods of the art.

Description

[Technical Field]

[0001] The present invention relates to a method for identifying donor blood suitable for cancer patients receiving anti-CD38 antibodies as treatment. In particular, the present invention addresses problems related to cross-matching tests between patient blood and donor blood when the patient blood contains anti-CD38 antibodies that interfere with cross-matching methods of the art. [Background technology]

[0002] CD38 is a type II membrane receptor glycoprotein that possesses enzymatic activity as an important ADP-ribosyl cyclase, particularly in producing cyclic adenosine diphosphate-ribose (cADPR) from nicotinamide adenine dinucleotide. CD38 is found on the surface of many cell types involved in immunological responses (abbreviated as immune cells), including not only effector cells such as T lymphocytes, B lymphocytes, and NK cells, but also immunosuppressive cells such as regulatory T cells and regulatory B cells, myeloid-derived suppressor cells (MDSCs), or tumor-associated macrophages (Non-Patent Literature 1). CD38 is also expressed on the surface of red blood cells (RBCs).

[0003] CD38 is highly expressed by cancer cells in patients with multiple myeloma at all stages and in patients with chronic lymphocytic leukemia (CLL) with a poor prognosis. Anti-CD38 monoclonal antibody therapy has been developed for targeted, direct killing of CD38-expressing tumor cells. Daratumumab and isatuximab are both anti-CD38 monoclonal antibodies approved for the treatment of multiple myeloma. However, such anti-CD38 antibodies are known to interfere with various hematological tests performed by blood banks to screen a patient's blood for compatibility before red blood cell transfusions, as they also bind to CD38 expressed on the surface of red blood cells (RBCs). Red blood cell transfusions are particularly important for multiple myeloma patients who require frequent RBC transfusions as part of supportive care.

[0004] Anti-CD38 antibodies such as daratumumab and isatuximab are known to cause interference in indirect antiglobulin (IAT), antibody detection (screening) tests, antibody identification panels, and anti-human globulin (AHG) crossmatching tests (Non-Patent Literature 2). When a patient is treated with an anti-CD38 antibody, interference caused by the anti-CD38 antibody in the patient's serum can lead to the false indication that the patient's serum contains clinically significant antibodies, namely antibodies that cause the destruction of donor red blood cells (known as hemolytic transfusion reactions) when transfused. Anti-CD38 antibodies in the patient's serum also cause interference in blood crossmatching tests, which are a process that checks for the compatibility of compatible donor RBCs with patient RBCs. If a patient is treated with daratumumab or isatuximab before a crossmatching test, compatible crossmatches may appear as incompatible crossmatches. Therefore, interference caused by anti-CD38 antibodies can delay the identification of donor red blood cells suitable for transfusion and may mask the presence of clinically significant antibodies in the patient's serum, thereby increasing the risk of hemolytic transfusion reactions. Furthermore, interference can persist for up to six months after discontinuation of anti-CD38 antibody treatment (Darzalex® prescribing information, Non-Patent Literature 3, Non-Patent Literature 4).

[0005] Strategies currently available to minimize interference caused by anti-CD38 antibodies include treating donor RBCs or reagent RBCs with antigen stripping agents that remove CD38, such as dithiothreitol (DTT), trypsin, or α-chymotrypsin. Furthermore, patient RBCs can be phenotyped or genotyped before initiating daratumumab. This allows blood banks to provide phenotypic or genotypic-matched RBCs in cases of urgent transfusion needs or when daratumumab interference cannot be immediately resolved by other means. This becomes possible. However, genotyping and phenotyping methods are time-consuming and expensive, and may not guarantee compatibility with donor RBCs. DTT and trypsin also denature some RBC antigens, particularly Kell system antigens, weakening their reactivity, which can make it difficult to perform accurate cross-matching tests. A further strategy to address interference by anti-CD38 is, for example, to neutralize anti-CD38 antibodies in the patient's plasma by treating a plasma sample or serum sample with soluble CD38 antigen. However, such an approach is expensive and not suitable for routine use. Therefore, there is a need for anti-CD38 antibodies that do not cause interference in hematological tests such as cross-matching tests and can be used in the treatment of patients with cancer, particularly multiple myeloma.

Prior Art Documents

Non-Patent Documents

[0006]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Summary of the Invention

[0007] In a first aspect, the present invention is a method for screening a blood sample obtained from a patient administered with an anti-CD38 antibody or an antigen-binding fragment thereof, comprising: a) preparing a blood sample from the patient; b) preparing a blood sample from a donor, where the donor blood sample contains donor red blood cells; c) Screening patient blood samples, including determining the presence or absence of one or more patient antibodies in a patient blood sample from a patient that specifically bind to one or more red blood cell antigens expressed on the surface of donor red blood cells, This provides a method that includes [something].

[0008] An anti-CD38 antibody or its antigen-binding fragment is generally an anti-CD38 antibody that, when present in a mixture of patient blood (e.g., patient serum or plasma) and donor red blood cells, does not cause agglutination of donor red blood cells when an agglutinating agent (such as an anti-human globulin antibody) is added to the mixture, provided that the mixture does not contain any patient-derived antibodies that bind to red blood cell antigens expressed on the donor red blood cells. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment binds to an epitope containing amino acids 65-79 of SEQ ID NO: 29 (human CD38).

[0009] Patient antibodies can be alloantibodies. For example, in some embodiments, the antibodies are alloantibodies, particularly alloantibodies that specifically bind to erythrocyte antigens. Because the antibodies are alloantibodies, they specifically bind to erythrocyte antigens other than any erythrocyte antigen expressed by the patient's erythrocytes.

[0010] In some embodiments, the method is (a) Prepare a blood sample from the patient, (b) Prepare a blood sample from the blood donor, Here, the donor blood sample contains donor red blood cells; (c) Preparing a patient blood / donor red blood cell mixture by contacting a patient blood sample with one or more donor red blood cells from a donor blood sample, (d) Optionally, incubate a mixture of patient blood / donor red blood cells to conjugate any one or more patient antibodies in the patient blood sample to one or more red blood cell antigens present on one or more donor red blood cells, if present, to form one or more patient antibody / donor red blood cell antigen complexes. To make it possible to form, (e) Optionally, if present, separate any one or more patient alloantibody / donor red blood cell antigen complexes from the patient blood / donor red blood cell mixture, Here, optionally, the separation step includes centrifugation; (f) Determining the presence or absence of patient antibodies in a patient blood sample that specifically bind to one or more erythrocyte antigens expressed on one or more donor erythrocytes, Includes.

[0011] In a second embodiment, the present invention provides a method for treating cancer in a patient, comprising preparing a blood sample from the patient and screening the blood sample according to a screening method of the present invention. In some embodiments, the patient has been administered an anti-CD38 antibody or its antigen-binding fragment (i.e., the patient has already received the anti-CD38 antibody or its antigen-binding fragment at an earlier point in time). In other embodiments, the method comprises a series of administrations of the anti-CD38 antibody or its antigen-binding fragment to the patient. In some embodiments, the method may optionally or additionally include a step of obtaining a sample from the patient.

[0012] In a third embodiment, the present invention provides an anti-CD38 antibody or its antigen-binding fragment for use in a method for treating cancer in a patient according to the present invention. [Brief explanation of the drawing]

[0013] [Figure 1] This figure shows the dose-response curves for the binding of CID103 (aCD38-b-348) to Daudi cells. Daudi cells were incubated with various concentrations of the anti-CD38 antibody CID103 or daratumumab (Darzalex), and binding was detected using an Alexa-647 conjugate F(ab')2 secondary antibody. A human IgG1 monoclonal antibody was used as a negative control for isotype control. Each data point is represented as the mean of a triple series, and error bars represent SEM. [Figure 2] This figure shows the dose-response curves for the binding of CID103 (aCD38-b-348) to Raji cells. Raji cells were incubated with various concentrations of the anti-CD38 antibody CID103 or daratumumab (Darzalex), and binding was detected using an Alexa-647 conjugate F(ab')2 secondary antibody. A human IgG1 monoclonal antibody was used as a negative control for isotype control. Each data point is represented as the mean of a triple series, and error bars represent SEM. [Figure 3] This figure shows the dose-response curves for the binding of CID103 (aCD38-b-348) to Ramos cells. Ramos cells were incubated with various concentrations of the anti-CD38 antibody CID103 or daratumumab (Darzalex), and binding was detected using an Alexa-647 conjugate F(ab')2 secondary antibody. A human IgG1 monoclonal antibody was used as a negative control for isotype control. Each data point is represented as the mean of a triple series, and error bars represent SEM. [Figure 4A] This figure shows the dose-response curve for the binding of CID103 (aCD38-b-348) to RBCs from donor 1. Blood substrates from donor 1 (5% RBC suspension in PBS) were incubated with various concentrations of the anti-CD38 antibody CID103 (aCD38-b-348) and daratumumab (Darzalex). Human IgG1 antibody as an isotype control was used as a negative control. Figure 4A shows the dose-response curve for antibody binding to RBCs. Each data point is represented as the mean of a triple series. [Figure 4B] This figure shows the dose-response curve for the binding of anti-CD47 to RBCs from donor 1. Blood substrates from donor 1 (5% RBC suspension in PBS) were incubated with various concentrations of Alexa Fluor 647 conjugated anti-CD47 antibody. The anti-CD47 antibody was used as a positive control to demonstrate specific binding to red blood cells. Figure 4B shows the dose-response curve for the binding of the control anti-CD47 antibody to RBCs. Each data point is represented as the mean of a triple series. [Figure 5A] This figure shows the dose-response curve for the binding of CID103 (aCD38-b-348) to RBCs from donor 2. Blood substrates from donor 2 (5% RBC suspension in PBS) were incubated with various concentrations of the anti-CD38 antibody CID103 (aCD38-b-348) and daratumumab (Darzalex). Human IgG1 antibody as an isotype control was used as a negative control. Figure 5A shows the dose-response curve for antibody binding to RBCs. Each data point is represented as the mean of a triple series. [Figure 5B] This figure shows the dose-response curve for the binding of anti-CD47 to RBCs from donor 2. Blood substrates from donor 2 (5% RBC suspension in PBS) were incubated with various concentrations of Alexa Fluor 647 conjugated anti-CD47 antibody. The anti-CD47 antibody was used as a positive control to demonstrate specific binding to red blood cells. Figure 5B shows the dose-response curve for the binding of the control anti-CD47 antibody to RBCs. Each data point is represented as the mean of a triple series. [Figure 6A] This figure shows the dose-response curve for the binding of CID103 (aCD38-b-348) to RBCs from donor 3. Blood substrates from donor 3 (5% RBC suspension in PBS) were incubated with various concentrations of the anti-CD38 antibody CID103 (aCD38-b-348) and daratumumab (Darzalex). Human IgG1 antibody as an isotype control was used as a negative control. Figure 6A shows the dose-response curve for antibody binding to RBCs. Each data point is represented as the mean of a triple series. [Figure 6B] This figure shows the dose-response curve for the binding of anti-CD47 to RBCs from donor 3. Blood substrates from donor 3 (5% RBC suspension in PBS) were incubated with various concentrations of Alexa Fluor 647 conjugated anti-CD47 antibody. The anti-CD47 antibody was used as a positive control to demonstrate specific binding to red blood cells. Figure 5B shows the dose-response curve for the binding of the control anti-CD47 antibody to RBCs. Each data point is represented as the mean of a triple series. [Figure 7] This figure shows IgG gel cards illustrating the interference of daratumumab in tests using Rh-positive and Rh-negative RBCs. Donor RBCs (Rh phenotypes: R1R1 (Figure 7A), R2R2 (Figure 7B), rr (Figure 7C)) were incubated with various concentrations of daratumumab (DARA) in inactive AB plasma and assayed for interference (i.e., the presence of agglutination despite the absence of clinically significant alloantibodies in the sample) using an IgG gel card assay (Ortho MTS). Figures 7A–7C show the results for each Rh phenotype at each concentration tested. Daratumumab concentrations are shown at the top of each microtube, with Φ indicating the absence of the drug. Rh phenotypes are shown at the bottom of each well. IgG gel cards were scored by an expert according to the degree of agglutination observed. Here, 4+, 3+, 2+, and 1+ all indicate the presence of agglutination. A rating of 0 or 0? indicates no agglutination or suspected agglutination. The agglutination rating is shown at the bottom of each well. [Figure 8] This figure shows the IgG gel card assay of CID103(aCD38-b-348) using Rh-positive and Rh-negative RBCs. Donor RBCs (Rh phenotypes: R1R1 (Figure 8A), R2R2 (Figure 8B), rr (Figures 8C, 8D)) were incubated with various concentrations of CID103(aCD38-b-348) in inactive AB plasma and assayed for interference (i.e., agglutination despite the absence of clinically significant alloantibodies in the sample) using an IgG gel card assay (Ortho MTS). Figures 8A-8D show the results for each Rh phenotype at each concentration tested. Daratumumab concentrations are shown at the top of each microtube, with Φ indicating the absence of the drug. Rh phenotypes are shown at the bottom of each microtube. IgG gel cards were scored by a skilled operator according to the degree of agglutination observed. Here, 4+, 3+, 2+, and 1+ all indicate the presence of agglutination. A rating of 0 or 0? indicates no agglutination or suspected agglutination. The agglutination rating is shown at the bottom of each microtube. [Figure 9]This figure shows IgG gel card assays of daratumumab and CID103 (aCD38-b-348) using untreated and pretreated RBCs. RhD-negative (rr) RBCs were incubated with various concentrations of CID103 (aCD38-b-348) or daratumumab (DARA) either untreated or after treatment with papain, trypsin, or ficin. Samples were assayed for interference (i.e., agglutination despite the absence of clinically significant alloantibodies in the sample) using an IgG gel card assay (Ortho MTS). Figures 9A to 9D show the results for each RBC treatment condition at each concentration tested. RBC treatment is indicated at the top of each microtube. IgG gel cards were scored by an expert according to the degree of agglutination observed. Here, 4+, 3+, 2+, and 1+ all indicate the presence of agglutination. A rating of 0 or + / - indicates no agglutination or suspected agglutination. Aggregation ratings are indicated at the bottom of each microtube. [Figure 10] This figure shows the screening of CID103(aCD38-b-348) using untreated RBCs on the automated platform IH-1000. Untreated RBCs were incubated with 250 μg / ml of CID103(aCD38-b-348) in inactive AB plasma and assayed for interference (i.e., agglutination despite the absence of clinically significant alloantibodies in the sample) using the automated IH-1000 platform (BioRad). Figure 10 shows the image of the results. The IH-1000 returned a result of "uninterpretable". According to expert evaluation, the result was determined to be non-aggregative. [Modes for carrying out the invention]

[0014] This invention makes it possible for the first time to test patient blood samples using standard techniques, even when the patient has been treated with anti-CD38 antibodies (also referred to herein as CD38-modulated antibody agents). Most therapeutic anti-CD38 antibodies (including, for example, daratumumab and isatuximab) interfere with tests to identify antibodies in a patient's blood (i.e., clinically significant antibodies) that can render a given donor blood unsuitable due to the possibility of hemolysis caused by the presence of antibodies in the patient's blood that bind to antigens on donor red blood cells. This interference occurs because therapeutic anti-CD38 antibodies bind to CD38 expressed on the surface of donor red blood cells, and then agglutinate when an anti-human globin reagent or analogue is added. This reagent typically causes agglutination by binding any clinically significant alloantibodies in the patient's serum that have bound to any antigen on the donor red blood cells. However, the presence of anti-CD38 antibodies in the patient's serum can lead to false positive results because agglutination occurs even if no clinically significant alloantibodies are present.

[0015] In particular, the inventors surprisingly discovered that while most therapeutic antibodies cause this interference, certain anti-CD38 antibodies do not. Without wanting to be constrained by theory, the inventors found that antibodies binding to specific anti-CD38 epitopes do not cause interference, and therefore, allow blood typing and cross-matching tests to be performed using standard techniques without the need for antigen stripping agents or equivalents.

[0016] The method of the present invention detects the presence or absence of clinically significant patient antibodies in a patient blood sample. In some embodiments, the method of the present invention detects the presence or absence of alloantibodies (e.g., clinically significant alloantibodies) in a patient. As used herein, the term "alloantibody" refers to an antibody that specifically binds to erythrocyte antigens that are not present on the subject's own erythrocytes. Therefore, an alloantibody is an anti-erythrocyte antigen alloantibody. Alloantibodies can be distinguished from "autoantibodies," which refer to antibodies that specifically bind to antigens present on the subject's own erythrocytes. Both alloantibodies and autoantibodies can be detected by the method of the present invention. For alloantibodies to develop, the individual must be exposed to non-self RBC antigens and possess an HLA-binding motif capable of presenting a portion of the non-self antigen. (Tormey & Hendrickson, 2019). Exposure to non-self antigens can occur, for example, through pregnancy, blood transfusion, or transplantation. The process of forming alloantibodies is called "alloimmunization." Alloantibodies can be clinically significant, and if a mother harbors alloantibodies against antigens on the baby's red blood cells, it can lead to either the destruction (hemolysis) of transfused RBCs or harm to the fetus or newborn. In fact, alloimmunization can be a direct cause of transfusion-related death. Alloimmunization also presents further problems in patient care, such as delayed transfusions, difficulty in identifying compatible blood for highly alloimmunized individuals, and delayed or acute hemolytic transfusion reactions. Alloimmunization is particularly clinically important for tumor patients who are at greater risk of developing alloantibodies due to frequent transfusions as part of supportive care (Hendrickson & Tormey, 2016). Therefore, it is crucial to be able to accurately and rapidly screen patient samples for the presence of alloantibodies. Patients treated with anti-CD38 antibodies such as daratumumab or isatuximab may experience delays in alloantibody screening because their serum contains anti-CD38 antibodies or their antigen-binding fragments, which bind to CD38 on RBCs and falsely indicate the presence of alloantibodies. The antibody screening and blood cross-matching tests used for alloantibody detection can be modified to incorporate steps that avoid this interference by anti-CD38 antibodies. For example, RBCs can be treated with an antigen stripping agent (such as DTT), or patient samples can be treated with an anti-CD38 neutralizing agent (such as soluble CD38). However, such additional reagents incur additional costs, are not widely available, and the extra process steps are time-consuming, leading to delays in antibody screening. The method of the present invention enables the detection of alloantibodies in patient samples without requiring additional processing of RBCs or patient blood samples. Using the method of the present invention, anti-CD38 antibodies or their antigen-binding fragments do not cause interference (e.g., agglutination in the absence of clinically significant alloantibodies) in antibody screening or cross-matching tests, thereby minimizing costs and avoiding delays in identifying transfusion-compatible blood products.

[0017] definition The following are some of the definitions of terms, technical means, and embodiments used herein, most of which are confirmations of the common understanding of those skilled in the art.

[0018] Administration: As used herein, the term “administration” refers to the administration of a composition to a subject. Administration to an animal subject (e.g., human) may be by any suitable route. For example, in some embodiments, administration may be by the bronchus (including by bronchial infusion), buccal, intestinal, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, intra-organ or tissue (e.g., intrahepatic, intratumoral, peritumoral, etc.), mucosa, nasal, oral, rectal, subcutaneous, sublingual, topical, trachea (including by intratracheal infusion), percutaneous, vaginal, and intravitreous. Administration may include intermittent dosing. Alternatively, administration may include continuous dosing (e.g., perfusion) over at least a selected period. As is known in the art, antibody therapeutic agents are generally administered parenterally, for example, by intravenous injection, subcutaneous injection, or intratumoral injection (e.g., particularly intratumoral where high doses are desired).

[0019] Substances that inhibit the binding of anti-CD38 antibodies to membrane-bound CD38: As already mentioned, treatment of patients with anti-CD38 antibodies such as daratumumab or isatuximab, or their antigen-binding fragments, can lead to inaccurate results in blood screenings such as blood antibody screening and cross-matching tests. For example, if daratumumab or isatuximab is present in a patient sample, it will agglutinate when incubated with RBCs in an indirect antiglobulin test (IAT), thus falsely indicating the presence of clinically significant antibodies in the patient sample. Therefore, measures must be taken to inhibit the binding of anti-CD38 antibodies or their antigen-binding fragments to membrane-bound CD38. This may include treating donor or panel RBCs with an active agent that inhibits the binding of anti-CD38 antibodies or their antigen-binding fragments to membrane-bound CD38. Such an active agent may be called an “anti-CD38 antagonist” or “CD38 antagonist.” An example of a CD38 antagonist is an antigen stripping agent. As used herein, the term “antigen stripping agent” may refer to any active agent used to remove an antigen from the surface of an RBC. In particular, an antigen stripping agent can be used to remove CD38 from an RBC. In some embodiments, the antigen stripping agent is a redox reagent or an enzyme. In some embodiments, the antigen stripping agent is dithiothreitol (DTT). DTT is a thiol reducing agent that denatures CD38 on the RBC surface by breaking the disulfide bond in the extracellular domain of the molecule, thereby preventing anti-CD38 from binding to the RBC. In some embodiments, the antigen stripping agent is an enzyme. In some embodiments, the antigen stripping agent is a protease. In some embodiments, the antigen stripping agent is trypsin. Trypsin is a protease, which is less efficient than DTT treatment for cleaving cell surface CD38. In some embodiments, the antigen stripping agent is α-chymotrypsin. In some embodiments, the antigen stripping agent is papain. In some embodiments, the antigen stripping agent is ficin. In some embodiments, the method of the present invention does not include the step of treating RBCs with an anti-CD38 antibody or a substance that inhibits the binding of its antigen-binding fragment to membrane-bound CD38; for example, the method of the present invention does not include the step of contacting the reaction mixture with a CD38 neutralizing agent such as an antigen stripping agent. In some embodiments, the method of the present invention does not include the step of treating RBCs with an antigen stripping agent. In some embodiments, the method of the present invention does not include the step of removing CD38 from RBCs.

[0020] Patient samples, such as serum or plasma samples, may also be treated with an "anti-CD38 neutralizer." As used herein, an anti-CD38 neutralizer is any substance used to neutralize anti-CD38 antibodies or their antigen-binding fragments in patient samples by binding or by other means. Anti-CD38 neutralizers can be used to bind anti-CD38 antibodies or their antigen-binding fragments in serum, plasma, or whole blood samples from patients before blood screening. In some embodiments, the anti-CD38 neutralizer may be a soluble CD38 antigen. In some embodiments, the anti-CD38 neutralizer may be an anti-CD38 idiotype antibody. In some embodiments, the method of the present invention does not include the step of treating a sample from a patient with an anti-CD38 neutralizer. In some embodiments, the method of the present invention does not include the step of treating a sample from a patient with a substance that inhibits the binding of anti-CD38 to membrane-bound CD38.

[0021] The methods of the present invention, which involve the use of an anti-CD38 antibody disclosed herein (not daratumumab or isatuximab, but rather aCD38-b-348 antibody or aCD38-b-329 antibody or antibodies derived therefrom (such as variants described elsewhere)), advantageously do not require the use of any agent (either an anti-CD38 antagonist or a CD38 antagonist) that inhibits the binding of the anti-CD38 antibody or its antigen-binding fragment to membrane-bound CD38.

[0022] Agglomeration: As used herein, the terms “aggregation” or “hemaglutination” refer to any process in which a large number of RBCs are bound by one or more antibodies and aggregate with each other. Aggregation is a reversible chemical reaction that is thought to occur in two stages: 1) sensitization when antibodies attach to erythrocyte antigens, and 2) agglutination when sensitized erythrocytes bridge each other to form a lattice. Yes. When a patient sample contains antibodies specific to the RBC antigen present on the donor's RBCs, and the patient sample and donor RBCs are mixed, the antibodies will bind to the RBC antigen (sensitization). Some antibodies, such as IgM antibodies, can directly cause agglutination by binding to each other. Other antibodies, such as IgG antibodies, may require the addition of a coagulant to bridge or link the bound antibodies to each other and cause agglutination. A coagulant is an antibody that binds any antibody present in the patient's blood sample to each other. In particular, a coagulant can bind any patient-derived antibody and / or any anti-CD38 antibody to each other. If the patient-derived antibody and / or anti-CD38 antibody to be agglutinated is bound to red blood cells, the coagulant will also cause agglutination of RBCs.

[0023] The method of the present invention may use certain agglutinants, such as anti-human globulin agents. Since patient-derived antibodies that specifically bind to the RBC antigen are usually IgG antibodies, anti-human globulin agents may be anti-human IgG antibodies. However, anti-human globulin agents may optionally or additionally include anti-C3 antibodies. Generally, agglutinants are substances that cause any antibody containing a human constant region (particularly the human IgG constant region) to conjugate to each other. Although the anti-CD38 antibodies described herein (i.e., aCD38-b-348 and aCD38-b-329, or antibodies derived therefrom) may contain a human constant region (for example, these may be human IgG isotypes), the presence of agglutinants surprisingly does not cause agglutination in the anti-CD38 antibodies that would interfere with the screening process.

[0024] Agglutinants (e.g., anti-human globulin reagents) cause patient antibodies bound to donor red blood cells (RBCs) to bind to each other. Antibody-bound RBCs can shrink to a visible pellet when centrifuged, for example, in an indirect antiglobulin test (IAT) performed in a tube. Agglutination is evaluated on a scale from 0 to 4+, where 0 represents no agglutination and 4+ indicates very strong agglutination. Agglutination forms the basis of most tests to determine whether a donor red blood cell sample is compatible with a patient's blood sample, and therefore whether the donor RBCs are suitable for transfusion. The occurrence of agglutination may indicate incompatibility between patient and donor red blood cells because the patient sample contains alloantibodies (clinically significant antibodies) that bind to the RBC antigen on the donor RBCs. Agglutination results of 1+, 2+, 3+, and 4+ may indicate incompatibility between patient and donor RBCs and that the donor RBCs should not be transfused to the patient. A 0 or 0¹ agglutination result may indicate compatibility between the patient's RBCs and the donor's RBCs, suggesting that the donor's RBCs can be safely transfused to the patient.

[0025] Agglutination may occur if a patient sample contains certain anti-CD38 antibodies, for example, if the patient is being treated with daratumumab or isatuximab. When a patient sample is incubated with donor RBCs, anti-CD38 antibodies or their antigen-binding fragments present in the patient sample can bind to CD38 expressed on the surface of the donor RBCs. Agglutination may occur when an agglutinating agent (e.g., an anti-human globulin reagent) is added to the patient sample / donor RBC mixture. Agglutination caused by the combination of the presence of anti-CD38 antibodies or their antigen-binding fragments in the patient sample and an agglutinating agent can occur regardless of the presence of clinically significant patient antibodies in the patient sample against the RBC antigen on the donor RBCs. Thus, the presence of anti-CD38 antibodies such as daratumumab or isatuximab or their antigen-binding fragments in the patient sample can cause interference with blood cross-matching tests, even if the patient does not have alloantibodies against the donor RBC antigen, making the donor RBCs appear unsuitable for transfusion to the patient. This problem can be avoided by using the anti-CD38 antibodies described herein (i.e., aCD38-b-348 or aCD38-b-329, or antibodies derived therefrom).

[0026] antibody: As used herein, the term “antibody” refers to a polypeptide containing standard immunoglobulin sequence elements sufficient to result in specific binding to a particular target antigen, such as CD38, particularly human CD38 and the extracellular domain of human CD38. As known in the art, naturally produced intact antibodies are approximately 150 kD tetramers composed of two identical heavy-chain polypeptides (each about 50 kD) and two identical light-chain polypeptides (each about 25 kD) that associate with each other to form a structure commonly referred to as a “Y-shaped” structure. Each heavy chain consists of at least four domains (each about 110 amino acids long): an amino-terminal variable (VH) domain (located at the tip of the Y structure) followed by three constant domains: CH1, CH2, and carboxy-terminal CH3 (located at the base of the stem of the Y). A short region known as a “switch” connects the variable and constant regions of the heavy chain. The CH2 and CH3 domains are connected to the rest of the antibody by a “hinge.” Two disulfide bonds in this hinge region connect the two heavy-chain polypeptides to each other in the intact antibody. Each light chain consists of two domains: an amino-terminal variable (VL) domain followed by a carboxy-terminal constant (CL) domain, separated from each other by another "switch." An intact antibody tetramer consists of two heavy-chain-light-chain dimers, where the heavy and light chains are linked to each other by a single disulfide bond, and the other two disulfide bonds connect the heavy chain hinge regions, causing the dimers to connect and form a tetramer. Furthermore, naturally produced antibodies are typically glycosylated at the CH2 domain, and each domain has a structure characterized by an "immunoglobulin fold" formed from two β-sheets (e.g., 3-chain, 4-chain, or 5-chain sheets) packed together in a compressed antiparallel β-barrel. Each variable domain has three hypervariable loops known as “complementary determination regions” (CDR1, CDR2, and CDR3; determined, for example, according to the Kabat numbering scheme as understood in the art) and four somewhat constant “framework” regions (FR1, FR2, FR3, and FR4).When native antibodies fold, the FR region forms a β-sheet that provides a structural framework to the domain, and the CDR loop regions of both the heavy and light chains assemble in three-dimensional space to create a single hypervariable antigen-binding site located at the tip of the Y structure. The Fc region of native antibodies binds to elements of the complement system, including receptors on effector cells, such as effector cells that mediate cytotoxicity. As is known in the art, the affinity and / or other binding attributes of the Fc region to Fc receptors can be modulated by glycosylation or other modifications that can improve the developability of the antibody (Jarasch A et al., 2015).

[0027] In some embodiments, antibodies produced and / or utilized according to the present invention include glycosylated Fc domains, which include Fc domains whose glycosylation has been modified or manipulated. For the purposes of the present invention, in certain embodiments, any polypeptide or polypeptide complex containing sufficient immunoglobulin domain sequences, as found in natural antibodies, may be referred to as and / or used as an “antibody,” regardless of whether such polypeptides are naturally produced (e.g., by organisms reacting to an antigen) or produced by recombinant operations, chemical synthesis, or other artificial systems or methodologies. In some embodiments, the antibody is a polyclonal or oligoclonal antibody produced as a panel of antibodies, each associated with a single antibody sequence, that bind to somewhat different epitopes within an antigen (e.g., different epitopes within the extracellular domain of human CD38 that associate with different reference anti-CD38 antibodies).

[0028] Polyclonal or oligoclonal antibodies can be provided in a single preparation for medical uses as described in the literature (Kearns JD et al., 2015). In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody has a constant region sequence characteristic of mouse, rabbit, primate, or human antibodies. In some embodiments, the antibody sequence element is humanized, primatized, chimeric, etc., as known in the art. Furthermore, the term “antibody” is used herein in reference to the definition of “antibody” in this specification. Where used, in appropriate embodiments (unless otherwise specified or evident from the context), the structural and functional characteristics of an antibody may refer to any of the constructs or formats known or developed in the art for use as an alternative presentation, for example, as an antigen-binding fragment as defined below. For example, antibodies used in accordance with the present invention are, but are not limited to, formats selected from intact IgG, IgE and IgM, bispecific or multispecific antibodies (e.g., Zybodies®), single-chain variable domains (scFv), polypeptide-Fc fusions, Fab, camel-like antibodies, heavy-chain shark antibodies (IgNAR), masked antibodies (e.g., Probodies®), or fusion proteins with polypeptides that enable expression and exposure on the cell surface (such as scFv in a construct for obtaining an artificial T cell receptor used to confer the specificity of a monoclonal antibody to T cells). In some embodiments, the antibody may lack covalent modifications (e.g., glycan attachments) that it may have if naturally produced. Alternatively, the antibody may have covalent modifications (e.g., attachment of glycans, payloads (e.g., detectable portion, therapeutic portion, catalytic portion, etc.), or other pendant groups (e.g., polyethylene glycol, etc.)).

[0029] Antibody screening: As used herein, the term “antibody screening” refers to any test performed to detect the presence or absence of antibodies in a patient's sample using a panel of red blood cells that specifically bind to one or more red blood cell antigens. Antibodies detected by antibody screening may be clinically significant antibodies that, for example, could cause a hemolytic transfusion reaction if the patient receives a transfusion of RBCs expressing antigens to which the antibodies can specifically bind. Antibodies detected by antibody screening may be alloantibodies, i.e., antibodies that specifically bind to antigens not present on the subject’s own RBCs, or autoantibodies, i.e., antibodies that specifically bind to antigens present on the subject’s own RBCs. Antibody screening is particularly useful for patients who are likely to have been alloimmunized as a result of prior transfusions, for example, patients being treated for hematological cancers. Antibody screening can be performed according to any of the methods described herein. Antibodies can be detected using an indirect antiglobulin test (IAT). Antibody screening can be performed using a column agglutination assay, a tube assay, or a solid-phase assay.

[0030] Antibody screening can be performed using a panel of red blood cells, where the red blood cells in the panel are known to express specific blood group antigens. A red blood cell panel may include red blood cells (RBCs) expressing any of the blood group antigens described herein. A red blood cell panel may include RBCs expressing any of the blood group antigens selected from the following groups: Ab, ABO, Cromer, Diego, Duffy, Gerbich, GLOB, Indian, Kell, Kidd, Knops, Lewis, Lutheran, LW, MNS, P1, Rh, XK, Xg, or Yt. In particular, a red blood cell panel may include RBCs expressing Kell group antigen, Duffy group antigen, Kidd group antigen, Lewis group antigen, P group antigen, MNS group antigen, Lutheran group antigen, and Xg group antigen. Screening patient samples against a red blood cell panel enables the detection of common clinically relevant patient antibodies. After performing antibody screening, patient samples can be cross-matched to donor RBCs from specific donors. Antibody screening improves the likelihood of identifying compatible donors in blood cross-matching tests because it allows for the selection of donor RBCs that do not express the RBC antigens identified in the antibody screening.

[0031] In some embodiments of the present invention, the method includes a step of antibody screening against a panel of red blood cells. In some embodiments of the present invention, the method includes cross-matching a patient blood sample with a candidate donor red blood cell sample. In this method, the above method first involves performing an antibody screening step using a panel of red blood cells, and then cross-matching a patient blood sample (from the same patient) with a candidate donor red blood cell sample, wherein the donor red blood cells do not express any red blood cell antigens identified in the antibody screening step as being found by patient-derived antibodies.

[0032] Anti-CD38 antibody: The term “anti-CD38 antibody” (also referred to herein as “CD38 modulated antibody”) is used herein to refer to an anti-CD38 antibody exhibiting the specific characteristics described herein. Unless otherwise indicated by the context, references to an anti-CD38 antibody herein include its antigen-binding fragment. In some embodiments, any antigen-binding fragment of the anti-CD38 antibody used may include an Fc portion, such as the human IgG constant region.

[0033] In many embodiments, the desirable anti-CD38 antibodies described herein are characterized by stimulating immune effector cells and / or modifying immune cell function, and being cytotoxic to or inducing phagocytosis of CD38-expressing cells (e.g., expressing high levels of CD38), such as immunosuppressive cells or tumor cells (e.g., expressing CD38 on their surface, in each case). In some embodiments, the anti-CD38 antibodies are characterized by activity (e.g., level and / or type) that is reasonably equivalent to the activity of aCD38-b-348 or aCD38-b-329 with respect to immune cells (e.g., when in contact with immune cells, particularly immune cells expressing CD38) and tumor cells. In some embodiments, the relevant activities are or include ADCP, ADCC in the absence of CDC, direct killing, depletion of certain CD38-expressing cells (e.g., high-expressing cells), activation of effector immune cells, promotion of T cell, B cell, or NK cell proliferation, regulation of immune cell activity (e.g., repolarization of suppressive macrophages into inflammatory macrophages), bias in the T cell repertoire, etc., and combinations thereof. In some embodiments, the anti-CD38 antibody is an entity or part whose presence or level correlates with the level and / or activity of CD38, and / or one or more features or consequences characteristic of CD38 activity. In some embodiments, the increase in level and / or activity is evaluated or determined by comparing it to what is observed in the absence of the entity(s) or part(s) (may be more than one) otherwise under equivalent conditions. Alternatively or additionally, in some embodiments, the increase in level and / or activity is equal to or greater than that observed under equivalent conditions when a reference anti-CD38 antibody (in many embodiments, a CD38 agonist antibody such as IB4) is present. In many embodiments, the anti-CD38 antibody used in accordance with this disclosure is an entity or portion that binds directly or indirectly to CD38, typically its extracellular domain, or includes them.In some embodiments, the anti-CD38 antibody is the anti-CD38 antibody disclosed herein, its antigen-binding fragment (e.g., including one or more CDRs, all heavy-chain CDRs, all light-chain CDRs, all CDRs, heavy-chain variable regions, light-chain variable regions, or both heavy-chain variable regions and light-chain variable regions), their affinity-mature variants (or their antigen-binding fragments), or any alternative form of any of the above (e.g., chimeric, humanized, multispecific, alternative isotype, etc.), including them, or binding to CD38 by them. Competing in combination. Alternatively or additionally, in some embodiments, the anti-CD38 antibodies disclosed herein may be characterized by one or more features that may be advantageous for screening, manufacturing, (pre)clinical trials, and / or identification of relevant epitopes within human CD38, such as sequences identified as aCD38-b-ep, and / or formulation, administration, and / or efficacy in specific situations disclosed herein (e.g., in the case of cancer therapy).

[0034] antigen: Where used herein, the term "antigen" refers to an activator that elicits an immune response and / or binds to T cell receptors (e.g., when presented by MHC molecules) and / or B cell receptors. Antigens that elicit a humoral response may be used to produce antigen-specific antibodies or to screen antibody libraries and identify candidate antibody sequences for further characterization, as shown in the examples for the CD38 extracellular domain.

[0035] Antigen-binding fragment: As used herein, the term “antigen-binding fragment” includes (include or comprise) an active substance that comprises one or more portions of an antibody described herein sufficient to impart to the antigen-binding fragment the ability of the antibody to specifically bind to the target antigen. For example, in some embodiments, this term encompasses any polypeptide or polypeptide complex containing sufficient immunoglobulin structural elements to result in specific binding. Exemplary antigen-binding fragments include small modular immunopharmaceuticals ("SMIPs"); single-chain antibodies; camel-like antibodies; and single-dominant antibodies. Examples of antibody fragments include, but are not limited to, in antibodies (e.g., shark single-domain antibodies), single-stranded or tandem diabodies (TandAb®), VHH, Anticalins®, Nanobodies®, minibodies, BiTE®, ankyrin repeat proteins, i.e., DARPINs®, Avimers®, DART, TCR-like antibodies, Adnectins®, Affilins®, Trans-bodies®, Affibodies®, TrimerX®, MicroProteins, Centyrins®, CoVX bodies, BiCyclic peptides, Kunitz domain-derived antibody constructs, or any other antibody fragments that exhibit the desired biological activity. In some embodiments, this term may refer to staple peptides, antibody-like conjugated peptide mimetic drugs, antibody-like conjugated scaffold proteins, monobodies, and / or other proteins such as other non-antibody protein scaffolds outlined in the literature (Vazquez-Lombardi R et al., 2015), for example. The structure is included. In some embodiments, the antigen-binding fragment is or includes a polypeptide having an amino acid sequence that is recognized by those skilled in the art as a complementarity-determining region (CDR) of one or more structural elements. In some embodiments, the antigen-binding fragment is or includes a polypeptide having an amino acid sequence that is substantially identical to that found in the anti-CD38 antibodies described herein (e.g., the amino acid sequence elements of aCD38-b-348 or aCD38-b-329), including at least one reference CDR (e.g., at least one heavy chain CDR and / or at least one light chain CDR), in particular at least one heavy chain CDR, e.g., HCDR3 (e.g., the HCDR3 sequence of aCD38-b-348 or aCD38-b-329). In some embodiments, the antigen-binding fragment is a polypeptide comprising at least one CDR (e.g., at least one heavy-chain CDR and / or at least one light-chain CDR) whose amino acid sequence is identical to or slightly (e.g., one, two, three, or four) more amino acid changes (e.g., substitutions, additions, or deletions; often substitutions) than such reference CDR, but which maintains the binding of the reference CDR to the target of the obtained antibody (e.g., aCD38-b-348 or aCD38-b-329). In some embodiments, the antigen-binding fragment is a polypeptide or complex thereof comprising all three CDRs (or, in some embodiments, substantially identical sequences) derived from the heavy or light chain (e.g., aCD38-b-348 or aCD38-b-329) of the reference antibody. In some embodiments, the antigen-binding fragment is or comprises a polypeptide or complex thereof containing all six CDRs (or, in some embodiments, substantially identical sequences) derived from a reference antibody (e.g., aCD38-b-348 or aCD38-b-329). In some embodiments, the antigen-binding fragment comprises the heavy chain and / or light chain variable domains (or, in some embodiments, substantially identical sequences) of the reference antibody (e.g., aCD38-b-348 or aCD38-b-329). Polypeptides or complexes thereof, or comprising them. In some embodiments, the term “antigen-binding fragment” encompasses non-peptide and non-protein structures such as nucleic acid aptamers, e.g., RNA aptamers and DNA aptamers. Aptamers are oligonucleotides (e.g., DNA, RNA, or their analogs or derivatives) that bind to specific targets such as polypeptides. Aptamers are short, synthetic single-stranded oligonucleotides that specifically bind to a variety of molecular targets, including small molecules, proteins, nucleic acids, and even cells and tissues. These small nucleic acid molecules can specifically bind to proteins or other cellular targets and can form secondary and tertiary structures that are essentially chemical equivalents of antibodies. Aptamers are highly specific, relatively small in size, and non-immunogenic. Aptamers are generally used in biopanning methods known as SELEX (Systematic Evolution of Ligands by Exponential Enrichment). Therefore, it is selected (see, for example, Ellington et al. Nature. 1990; 346(6287): 818-822, Turk et al., Science. 1990; 249(4968):505-510, Ni et al., Curr Med Che 2011; 18(27):4206-14). A method for generating an aptamer for any given target is: This is known in the art. Peptide aptamers, including affimers, are also included. Affimers are small, highly stable proteins engineered to exhibit peptide loops that provide a high-affinity binding surface to a specific target protein. Affimers are low molecular weight proteins, 12kDa–14kDa, derived from the cystatin cysteine ​​protease inhibitor family. Affimer proteins consist of a scaffold, which is a stable protein based on the cystatin protein fold. Affimer proteins exhibit two peptide loops and an N-terminal sequence that can be randomized to bind to different target proteins with high affinity and specificity similar to antibodies. Stabilization of the peptide on the protein scaffold restricts the possible conformations the peptide can adopt, thereby increasing binding affinity and specificity compared to a free peptide library.

[0036] Products and kits: In some embodiments of the present invention, the anti-CD38 antibody described herein is provided in a separate product. In some embodiments of the present invention, the product comprising the anti-CD38 antibody is provided in or with a labeled container. Suitable containers include, for example, bottles, vials, syringes, and test tubes. In some embodiments, the container may be formed from any or a variety of materials such as glass or plastic. In some embodiments, the container contains a composition effective for treating a particular disease, disorder, or condition, or its stage or type. In some embodiments, the container may have a sterile access port (for example, the container may be a vial with a stopper that can be punctured by an intravenous solution bag or a subcutaneous needle). For example, in some embodiments, the composition comprising the anti-CD38 antibody described herein is packaged in a clear glass vial with a rubber stopper and an aluminum seal. A label on or attached to the container indicates that the composition is used for the treatment of a selected condition.

[0037] In some embodiments, the product may further include separate containers containing pharmaceutically acceptable buffers such as phosphate-buffered saline, Ringer's solution, and dextrose solution, and / or other materials desirable from a commercial and user perspective, including other buffers, diluents, filters, needles, syringes, and accompanying documentation including instructions for use. For example, in some embodiments, the product may enable the provision of each active ingredient in an intravenous formulation as a sterile aqueous solution containing a total of 2 mg, 5 mg, 10 mg, 20 mg, 50 mg, or more, formulated with appropriate diluents and buffers at final concentrations of 0.1 mg / ml, 1 mg / ml, 10 mg / ml, or higher.

[0038] In some embodiments, the anti-CD38 antibody described herein is used in a kit. It may be provided in a kit of parts in a lyophilized form reconstituted with any suitable aqueous solution provided or not provided, or in other types of dosing units using any suitable pharmaceutical carrier. One or more unit dosage forms can be provided in a pack or dispenser device. Such packs or devices may include metal foil or plastic foil, such as a blister pack. To ensure the correct use of such a kit of parts, it may further include a package insert containing buffers, diluents, filters, needles, syringes, and instructions for use in the treatment of cancer.

[0039] In some embodiments, instructions accompanying the products or kits described herein may be in the form of labels, leaflets, documents, records, charts, or any other means that can be used to provide information about the correct use and / or monitoring of possible effects of the active ingredients, formulations, and other materials contained in the product and / or kit. Instructions may be provided with the product and / or within the kit.

[0040] Automated testing: As used herein, the terms “automated test,” “automated platform,” and “automated assay” may refer to any automated system for detecting antigen-antibody reactions between a patient or recipient blood sample and a donor blood sample or reagent RBC. Examples of automated platforms include Tango (BioRad) and IH-1000 (BioRad). Automated tests can be used in solid-phase assays, column agglutination assays, tube assays, and / or other assay types. The methods described herein can be appropriately carried out using automated tests.

[0041] Biological specimens: Where used herein, the terms “biological sample” or “sample” (used without distinction) typically refer to a sample obtained from or derived from a biological source of interest (e.g., tissue or organism or cell culture), as described herein. The source of interest may be an animal or a human organism. A biological sample may include biological tissue or biological fluid. The methods described herein relate to the screening of blood samples obtained from a patient. In some embodiments, the patient blood sample is a whole blood sample. In some embodiments, the patient blood sample is a red blood cell sample. In some embodiments, the patient blood sample is a plasma sample. In some embodiments, the patient blood sample is a serum sample. In some embodiments, the patient sample does not contain any red blood cells of the patient.

[0042] In some embodiments, the methods described herein include the step of preparing a blood sample from a donor. In some embodiments, the donor blood sample is a whole blood sample. In some embodiments, the donor blood sample is a red blood cell sample. In some embodiments, the donor blood sample is a plasma sample. In some embodiments, the donor blood sample is a serum sample. In some embodiments, the patient sample does not contain any of the patient's own red blood cells.

[0043] In some embodiments of the present invention, the screening method is performed on a sample obtained from a patient at an earlier point in time. In other embodiments of the present invention, the method may include the step of obtaining a sample from a patient using any suitable method.

[0044] The method of the present invention can be applied to multiple samples from the same patient. For example, in a method that includes both RBC panel screening and a cross-matching assay, RBC panel screening is performed on a first sample from the patient, and the cross-matching assay is performed on the same patient. This can be performed on a second, different sample from the patient. For this purpose (or to perform the same assay multiple times on the same patient), a single sample can be obtained from the patient and divided into multiple subsamples, or multiple different samples can be obtained from the same patient.

[0045] After processing the patient sample, it can be mixed with donor red blood cells to remove patient RBCs, for example. Further processing steps, such as diluting the patient sample with a buffer, may be used.

[0046] Generally, patients have been administered anti-CD38 antibodies beforehand, so patient samples contain anti-CD38 antibodies.

[0047] cancer: "Cancer," "malignant tumor," "neoplasm," "tumor," and "carcinoma" The term is used herein without distinction to refer to cells exhibiting a phenotype of abnormal growth characterized by a marked loss of control over cell proliferation, as they exhibit relatively abnormal, uncontrolled, and / or autonomous growth. Generally, cells targeted for detection or treatment in this application include precancerous (e.g., benign) cells, malignant cells, premetastatic cells, metastatic cells, and nonmetastatic cells. The teachings of this disclosure may be relevant to all cancers. To give some non-limiting examples, in some embodiments, the teachings of the present disclosure apply to one or more cancers such as, for example, hematopoietic cancers including leukemia, lymphoma (Hodgkin and non-Hodgkin), myeloma, and myeloproliferative disorders; urogenital cancers such as sarcoma, melanoma, adenoma, solid tissue carcinoma, squamous cell carcinoma of the oral cavity, throat, larynx, and lung; liver cancer, prostate cancer, cervical cancer, bladder cancer, uterine cancer, and endometrial cancer; and benign lesions such as renal cell carcinoma, bone cancer, pancreatic cancer, skin cancer, cutaneous melanoma or intraocular melanoma, endocrine cancer, thyroid cancer, parathyroid cancer, head and neck cancer, breast cancer, gastrointestinal cancer, and nervous system cancer, papilloma, etc.

[0048] In some embodiments, cancer is a cancer containing cells that express CD38 on their cell surface, i.e., CD38-expressing cancer. In some embodiments, cancer may be a solid tumor, such as a solid tumor, that expresses CD38 on its cell surface. In some embodiments, cancer may be a hematological malignancy, such as a CD38-expressing hematological malignancy. In some embodiments, cancer may be selected from the group consisting of T-cell or B-cell non-Hodgkin lymphoma, chronic lymphocytic leukemia, plasmacytoma, and multiple myeloma.

[0049] Column aggregation technology: Where used herein, the terms “column agglutination assay” or “column agglutination technique” refer to a technique used to identify patient-derived antibodies (alloantibodies, particularly clinically significant alloantibodies, etc.) in a patient sample that bind to antigens expressed on donor RBCs or reagent RBCs (i.e., red blood cells in an red blood cell panel). IAT can be performed using column agglutination technique. Column agglutination technique uses a microtube or column containing an agglutinant, such as an anti-human globulin (anti-IgG and / or anti-C3, etc.) gel, and allows donor cells to be incubated with patient plasma or serum using the wells on the microtube. The sample is then centrifuged through a column containing anti-human globulin, e.g., anti-human IgG. During incubation, any relevant patient-derived antibodies present in the patient's plasma or serum may bind to the antigens expressed on the donor RBCs. Donor RBCs to which antibodies have bound may be called “sensitized” RBCs. Patient-derived antibodies that bind to RBCs may be IgG antibodies. During centrifugation, patient-derived antibodies bound to donor RBCs react with anti-human globulin reagents present in the gel, hindering or delaying the passage of bound RBCs through the microtube or column. A strongly positive agglutination reaction results in a layered line of RBCs on the top of the gel. A positive reaction will be accompanied by varying degrees of visible hemagglutination suspended in the gel. Therefore, donor RBCs that do not have patient-derived antibodies bound to them will easily pass through the gel during centrifugation and form a pellet at the bottom of the microtube or column. Indirect antiglobulin testing (IAT) can be performed using column agglutination technology. The results of assays performed using column agglutination technology can be read by an automated platform such as Tango (BioRad) or IH-1000 (BioRad).

[0050] Combination therapy: As used herein, the term “combination therapy” refers to a situation in which a subject is simultaneously exposed to two or more treatment regimens (e.g., two or more therapeutic agents). In some embodiments, two or more active ingredients can be administered simultaneously. Alternatively, such active ingredients can be administered sequentially, or in overlapping drug regimens.

[0051] Equivalent: As used herein, the term “comparable” means two or more sets of active substances, entities, situations, effects, conditions, etc., that are not identical to one another but are similar enough to allow comparison (e.g., by level and / or activity) to enable reasonable conclusions to be drawn based on observed differences or similarities. Such comparable sets of conditions, effects, situations, individuals, or groups are characterized by several substantially identical features and one or a few diverse features. Those skilled in the art will understand, in context, what degree of identity is required in any given situation for two or more sets of such active substances, entities, situations, conditions, effects, or groups, etc., to be considered equivalent.

[0052] include: A composition or method described herein as "comprising" one or more specified elements or steps is open-ended, meaning that while the specified elements or steps are essential, other elements or steps may be added within the scope of the composition or method. Any composition or method described as "comprising" (or which "comprises") one or more specified elements or steps may include the same specified elements or steps. "Consisting essentially of" (or which "consists essentially of") can also be understood as describing a more restrictive composition or method, meaning that the composition or method includes the specified essential elements or steps, and may also include additional elements or steps that do not substantially affect the basic novel characteristics (which may be multiple) of the composition or method.

[0053] Cross-fitting test: As used herein, the term “cross-matching test” refers to any test of compatibility between a patient blood sample and a donor blood sample. Cross-matching can refer to testing donor red blood cells for compatibility with the serum or plasma of a recipient or patient, and is also known as primary cross-matching. Cross-matching tests can be used to identify the presence of clinically significant antibodies in samples from a patient or transfusion recipient (e.g., plasma or serum samples). Clinically significant antibodies are any antibodies that are likely to cause adverse side effects, such as hemolytic transfusion reactions, after donor blood has been transfused to a recipient. Examples of clinically significant antibodies include alloantibodies that specifically bind to antigens not expressed on the red blood cells of the transfusion recipient or the patient themselves. Examples of clinically significant antibodies include autoantibodies that specifically bind to antigens expressed on the red blood cells of the transfusion recipient or the patient themselves. In some embodiments, incompatibility between a patient blood sample and a donor blood sample is indicated by agglutination when the samples are mixed. In some embodiments, compatibility between a patient blood sample and a donor blood sample is determined by the absence of agglutination when the samples are mixed. This is indicated as follows. In some embodiments, incompatibility between a patient blood sample and a donor blood sample is indicated by hemolysis when the samples are mixed. In some embodiments, compatibility between a patient blood sample and a donor blood sample is indicated by the absence of hemolysis when the samples are mixed. Crossmatch testing can be performed according to any of the methods described herein. Crossmatch testing can be performed using a column agglutination assay, an indirect antiglobulin test (IAT) tube assay, or a solid-phase assay.

[0054] Daratumumab: As used herein, the term "daratumumab" includes antibodies that are human IgG1 monoclonal antibodies having the VH and VL sequences published in International Publication No. 2006 / 099875. For example, antibodies having a variable heavy chain sequence and a variable light chain sequence including the sequences shown below: Heavy chain: EVQLLESGGGLVQPGGSLRLSCAVSGFTFNSFAMSWVRQAPGKGLEWVSAISGSGGGTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYFCAKDKILWFGEPVFDYWGQG TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence ID 31) Light chain: EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (Sequence ID 32)

[0055] Dosage form: As used herein, the term “dosage form” refers to a physically separated unit of an active agent (e.g., a therapeutic or diagnostic agent) administered to a subject. Each unit contains a predetermined amount of the active agent. In some embodiments, such an amount is a unit dose (or the entire amount thereof) appropriate for administration according to a drug regimen (i.e., a therapeutic drug regimen) that is judged to correlate with a desired or beneficial outcome when administered to the relevant population. Those skilled in the art will understand that the total amount of a therapeutic composition or active agent administered to a particular subject may be determined by one or more attending physicians and may involve administration of multiple dosage forms.

[0056] Medication and administration: Pharmaceutical compositions comprising the anti-CD38 antibody described herein (e.g., anti-CD38 or its antigen-binding fragment, including the amino acid sequence of HCDR3 of aCD38-b-348 or aCD38-b-329, for use in accordance with the present invention) can be prepared for storage and / or delivery using any of the various techniques and / or technologies known and / or available to those skilled in the art. In some embodiments, the disclosed anti-CD38 antibody is administered in accordance with a dosing regimen approved by a regulatory authority such as the U.S. Food and Drug Administration (FDA) and / or the European Medicines Agency (EMA) for the relevant indication. In some embodiments, the disclosed anti-CD38 antibody is administered in combination with one or more other agents or therapies that can be administered by themselves in accordance with a dosing regimen approved by a regulatory authority such as the U.S. Food and Drug Administration (FDA) and / or the European Medicines Agency (EMA) for the relevant indication. However, in some embodiments, the use of the disclosed anti-CD38 antibody may enable a reduction in the dosage of the approved active ingredient or therapy used in combination with anti-CD38 antibody therapy (e.g., smaller amounts of the active ingredient in one or more doses, fewer doses, and / or reduced frequency of administration). In some embodiments, the dosage and / or administration may be adapted to other drugs administered similarly, the patient's condition, and / or the format of the anti-CD38 antibody (e.g., modified as an immune complex, nanobody, or bispecific antibody). It is possible.

[0057] Furthermore, in some embodiments, it may be desirable to tailor the drug regimen to a specific cell type, a specific tumor or its type, or a specific patient population (e.g., one possessing a genetic marker), and in particular to design a sequential drug regimen based on timing and / or threshold expression levels of CD38. In some such embodiments, the therapeutic drug regimen may be combined with, or tailored to, a detection method that evaluates the expression of one or more inducible markers or other criteria before and / or during therapy.

[0058] In some embodiments, the drug administration and administration according to the present invention utilize an active agent having a desired degree of purity in any or various forms in combination with one or more physiologically acceptable carriers, additives, or stabilizers. These forms include, for example, liquid, semi-solid, and solid dosage forms, such as liquid solutions (e.g., injection solutions and infusion solutions), dispersions or suspensions, tablets, pills, powders, liposomes, and suppositories. The preferred form may vary depending on the intended method of administration and / or therapeutic use, but may typically be in the form of an injection solution or infusion solution, such as a composition similar to those used for the treatment of human subjects with antibodies.

[0059] In some embodiments, the components(s) may be prepared using carriers that protect the active substance(s) from rapid release and / or degradation, such as controlled-release formulations including implantable injections, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as polyanhydrides, polyglycolic acid, polyorthoesters, and polylactic acid may be used. Generally, each activator is formulated, administered, and given in therapeutically effective doses using appropriate pharmaceutical compositions and dosing regimens that are compatible with the appropriate medical practice and the relevant active substance(s) (e.g., an antibody). Pharmaceutical compositions containing the active agent may be administered by any suitable method known in the art, including, but not limited to, oral administration, mucosal administration, inhalation administration, topical administration, buccal administration, nasal administration, rectal administration, or parenteral administration (e.g., intravenous, intradrip, intratumoral, intranodal, subcutaneous, intraperitoneal, intramuscular, intradermal, percutaneous, or other types of administration involving physical breaching of the subject's tissue and administration of the pharmaceutical composition through such breach).

[0060] In some embodiments, a drug regimen for a particular activator may include intermittent or continuous (e.g., by a perfusion or sustained-release system) administration to achieve a specific desired pharmacokinetic profile or other exposure pattern in, for example, one or more tissues or body fluids of the subject. In some embodiments, different active ingredients administered in combination may be administered by different delivery routes and / or according to different schedules. Alternatively or additionally, in some embodiments, one or more doses of the first activator may be administered substantially simultaneously with one or more other activators, and in some embodiments, by a common route and / or as part of a single composition.

[0061] Factors to consider when optimizing the route and / or medication schedule for a given treatment regimen may include, for example, the specific cancer being treated (e.g., type, stage, location, etc.), the subject's clinical condition (e.g., age, overall health, weight, etc.), the site of delivery of the active ingredient, the nature of the active ingredient (e.g., antibody or other protein-based compound), the method and / or route of administration of the active ingredient, the presence or absence of combination therapy, and other factors known to the physician.

[0062] Those skilled in the art will understand that, for example, a particular delivery route may affect the dose, and / or the required dose may affect the delivery route. For example, when targeting particularly high concentrations of the active ingredient at a specific site or location (e.g., within a tissue or organ), concentrated delivery (e.g., intratumoral delivery) may be desired and / or useful. In several embodiments, one or more features of a particular pharmaceutical composition and / or drug regimen used can be modified over time to optimize a desired therapeutic effect or response (e.g., a therapeutic or biological response related to the functional features of the anti-CD38 antibody described herein) (e.g., increasing or decreasing the amount of activator in any individual dose, increasing or decreasing the time interval between doses, etc.). Generally, the type, amount, and frequency of activator administration according to the present invention are subject to the safety and efficacy requirements applicable when the relevant active agents (which may be more than one) are administered to mammals, preferably humans. Generally, the features of such administration are selected to produce a specific, typically detectable therapeutic response compared to that observed in the absence of therapy. Under the context of the present invention, exemplary desirable therapeutic responses may include, but are not limited to, inhibition and / or reduction of one or more tumor growth, tumor size, metastasis, tumor-associated symptoms and side effects, as well as increased apoptosis of cancer cells, therapeutically relevant decreases or increases of one or more cellular or circulating markers, etc. Such criteria can be readily assessed by any of the various immunological, cytological, and other methods disclosed in the literature. For example, the therapeutically effective dose of an anti-CD38 antibody, either alone or in combination with other active agents, can be determined as sufficient to enhance the killing of cancer cells, as described in the examples.

[0063] The therapeutically effective dose of an anti-CD38 antibody as an activator or composition containing such active substance can be readily determined using techniques available in the art, which include considering one or more factors such as the disease or condition being treated, the stage of the disease, the age and health and physical condition of the mammal being treated, the severity of the disease, and the specific compound being administered.

[0064] In some embodiments, the therapeutically effective dose is an effective dose (and / or unit dose) of the activator that may be at least about 0.01 μg / kg (body weight), at least about 0.05 μg / kg (body weight), at least about 0.1 μg / kg (body weight), at least about 1 μg / kg (body weight), at least about 5 μg / kg (body weight), at least about 10 μg / kg (body weight), at least about 15 μg / kg (body weight), at least about 20 μg / kg (body weight), or at least about 25 μg / kg (body weight) or more (e.g., about 100 μg / kg (body weight)). In some embodiments, the therapeutically effective dose is an effective dose (and / or unit dose) of the activator, which may be at least about 0.01 mg / kg(body weight), at least about 0.05 mg / kg(body weight), at least about 0.1 mg / kg(body weight), at least about 1 mg / kg(body weight), at least about 5 mg / kg(body weight), at least about 10 mg / kg(body weight), at least about 15 mg / kg(body weight), at least about 20 mg / kg(body weight), or at least about 25 mg / kg(body weight) or more (e.g., about 100 mg / kg(body weight)). It will be understood by those skilled in the art that in some embodiments, such guidelines may be adjusted to the molecular weight of the activator. Dosage may also be modified according to a dose escalation protocol that can be used to determine the maximum tolerated dose and dose-dependent toxicity (if any) in relation to the route of administration, the treatment cycle, or the administration of an isolated antibody or its antigen-binding fragment containing the amino acid sequence of aCD38-b-348 or aCD38-b-329 in escalating doses.

[0065] Therapeutic compositions are typically sterile and stable under manufacturing and storage conditions. Compositions can be formulated as solutions, microemulsions, dispersions, liposomes, or other regular structures suitable for high drug concentrations. Sterile injectable solutions can be prepared by incorporating the required amount of antibody with one or a combination of the components listed above into a suitable solvent, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle containing a basic dispersion medium and other required components from those listed above. For powders used in the preparation of sterile injectable solutions, preferred preparation methods are vacuum drying and freeze-drying, which yield powders of the active ingredient and any additional desired components from a pre-sterilized filtered solution. Appropriate fluidity of the solution is maintained, for example, by the use of coating agents, maintaining the required particle size in the case of dispersions, and using surfactants. This is possible. Sustained absorption of an injectable composition can be achieved by adding substances that slow down absorption, such as monostearate and gelatin, to the composition.

[0066] Preparations of each active ingredient shall be sterile, preferably achieved by filtration through a sterile filtration membrane, and then packaged or sold in a form suitable for bolus or continuous administration. Injectable preparations may be prepared, packaged or sold in unit dosage forms, such as ampoules with preservatives or multi-dose containers. Preparations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and implantable sustained-release or biodegradable preparations as discussed herein. Sterile injectable preparations may be prepared using water or a non-toxic, parenterally acceptable diluent or solvent such as 1,3-butanediol. Other useful parenterally administerable preparations include those containing the active ingredient in microcrystalline form, in liposome preparations, or as a component of a biodegradable polymer system. Compositions for sustained-release or implantation may include pharmaceutically acceptable polymer materials or hydrophobic materials such as emulsions, ion exchange resins, poorly soluble polymers, or salts.

[0067] Each pharmaceutical composition used in accordance with the present invention may contain pharmaceutically acceptable dispersants, wetting agents, suspending agents, isotonic agents, coating agents, antibacterial and antifungal agents, carriers, additives, salts, or stabilizers that are nontoxic to the subject at the dosage and concentration used. Such an inexhaustible list of additional pharmaceutically acceptable compounds includes buffers such as phosphates, citrates and other organic acids; antioxidants including ascorbic acid and methionine; salts containing pharmaceutically acceptable anions (acetates, benzoates, bicarbonates, bisulfates, isethionates, lactates, lactobionates, laurates, malates, maleates, salicylates, stearates, basic acetates, succinates, tannates, tartrates, theoclates, tosylates, triethiozides and valersates, etc.); preservatives (octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; sodium chloride; phenol, butyl alcohol or benzyl alcohol; methylparaben or propyl This includes alkylparabens such as parabens; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol, etc.; low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrin; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants such as TWEEN®, PLURONICS®, or polyethylene glycol (PEG).

[0068] In some embodiments utilizing two or more activators according to the present invention, such activators may be administered simultaneously or sequentially. In some embodiments, the administration of one activator is specifically timed to coincide with the administration of another activator. In some embodiments, a desired relative dosing regimen of activators administered in combination may be evaluated or empirically determined, for example, using ex vivo, in vivo, and / or in vitro models. In some embodiments, such evaluation or empirical determination is performed in vivo in a specific patient or patient population (e.g., so that a correlation can be obtained).

[0069] In some embodiments, one or more activators used in carrying out the present invention are administered according to an intermittent dosing regimen comprising at least two cycles. When two or more active agents are combined and each is administered according to such an intermittent cycle regimen, active agents with different individual doses can be combined with each other. In some embodiments, 1 One or more doses of a second active agent are administered a certain time after the administration of the anti-CD38 antibody described herein. In some embodiments, each dose of the second active agent is administered a certain time after the administration of the anti-CD38 antibody described herein. In some embodiments, the anti-CD38 antibody described herein is administered in a regimen that includes not only subsequent administrations via the same route, but also subsequent administrations via alternative routes such as subcutaneous (or intramuscular) and intratumor administration, in addition to subsequent administrations via the same route in one or more treatment cycles lasting one, two, four weeks or more. The drug is administered in a single cycle, and such cycles can be repeated with the same regimen (or with longer intervals between doses) depending on the patient's response. In some embodiments, the exact regimen followed (e.g., number of doses, intervals between doses (e.g., other events such as the administration of another therapy or other therapies), dosage, etc.) may differ in one or more cycles compared to one or more other cycles.

[0070] By using any of the administration routes, dosages, and / or regimens described herein, the anti-CD38 antibodies described herein can be identified, characterized, and / or validated in consideration of one or more criteria measured in patients, for example, using biopsies, blood samples, and / or other clinical criteria. In some embodiments, instead of or in addition to direct assessment of tumor size and / or metastasis, the therapeutic efficacy of the anti-CD38 antibodies described herein can be determined by methods that evaluate one or more different general criteria, including: direct cytotoxicity against cancer cells (apoptosis and necrosis of cancer cells), an increase in tumor-infiltrating immune cells (such as CD4-positive and / or CD8-positive tumor-infiltrating T cells), an increase in circulating immune cells (such as lymphocytes, NK cells, monocytes, dendritic cells, macrophages, B cells, etc., either the whole population or specific subpopulations), and / or the presentation of some differential expression before versus after treatment in either responsive or non-responsive patients only (determined by RNA sequencing, mass flow cytometry, and / or other mass sequencing approaches). Alternatively or additionally, in some embodiments, such identification, characterization, and / or verification may include tracking at the molecular level by screening the protein expression of mRNA and / or one or more specific proteins or sets of proteins. In some embodiments, one or more such techniques may enable the identification of relevant information for evaluating the response to the anti-CD38 antibodies described herein, which may be related to tissue distribution and / or markers for specific cell populations in (or near) a tumor and / or circulating in the blood.

[0071] Such approaches and immunobiological data can not only enable the determination of one or more efficacy and / or safety parameters or characteristics, but in some embodiments, they can provide a rationale for selecting specific doses, routes or drug regimens that can be used in one or more clinical trials for a given indication, for example, in combination with other drugs, standard treatment protocols or immunotherapies that may produce further therapeutic effects, either alone or in combination with other drugs. Therefore, in a series of further embodiments of the present invention, the anti-CD38 antibody described herein is used in a method to treat a patient with a disease (such as cancer) or prevent a disease (such as cancer) after determining the combination of the presence (and / or absence) of the expression of one or more genes in the patient's cells or tissues (such as tumors, blood samples or blood fractions) at the RNA and / or protein level, either before or after treatment with such formulation. Accordingly, such a method may enable the determination of one or more biomarkers or more complex gene expression signatures (or cell population distributions) associated with a desired therapeutically effective dose of anti-CD38 antibody, therapeutically relevant biomarkers (may be more than one) that predict whether a subject may have an antitumor or anti-infective response after treatment with an anti-CD38 antibody described herein, or therapeutically relevant biomarkers (may be more than one) that predict whether a subject may respond to treatment with a compound after treatment with an anti-CD38 antibody.

[0072] Alternatively or additionally, in some embodiments, specific information disclosed herein may be used. The administration and administration of anti-CD38 antibodies can be pre-established and / or later evaluated by collecting data on CD38 distribution in various cancer, tissue and / or patient immune subsets, for example, stromal subsets, and / or in immune subsets of different cancers, tissues and / or patients, taking into account CD38 expression in human cancers and / or other human tissues. Such data can be generated using common techniques (flow cytometry, mass cytometry, immunohistochemistry or mRNA expression libraries, etc.) across common oncologies and / or tissues (central nervous system, esophagus, stomach, liver, colon, rectum, lung, bladder, heart, kidney, thyroid, pancreas, uterus, skin, breast, ovary, prostate and testis) to identify the relationship of CD38 expression in various immune and non-immune subpopulations and / or the relationship of a measure of cell infiltration associated with subsets of cancer cells or immune cells and / or cancer-related markers (Foxp3 and PD-1 / PD-L1, etc.). CD38 expression can be restricted (or not restricted) to an immune subset of tumor tissue (e.g., NK cells and other effector or regulatory immune cells), and a correlation between CD38 expression and immune checkpoint inhibitors can be determined if positive, suggesting appropriate applications for anti-CD38 antibodies in combination with compounds targeting such immune checkpoint inhibitors.

[0073] Medication regimen: As used herein, the term “medication regimen” typically refers to a set of unit doses (typically two or more) administered individually to a subject at intervals. In some embodiments, a given therapeutic agent has a recommended drug regimen, which may comprise one or more doses. In some embodiments, the drug regimen comprises multiple doses, each spaced equally apart. Alternatively, the drug regimen comprises multiple doses, each separated by at least two different time intervals. In some embodiments, all doses within the drug regimen are of the same unit dose. Alternatively, different doses within the drug regimen are of different amounts. In some embodiments, the drug regimen comprises one or more additional doses of a second dose different from the first dose, following a first dose of a first dose. The drug regimen may comprise one or more additional doses of a second dose, the same as the first dose, following a first dose of a first dose. In some embodiments, the medication regimen correlates with a desired or beneficial outcome when implemented across the relevant population (i.e., it is a therapeutic medication regimen).

[0074] Epitope: As used herein, the term “epitope” refers to a portion of an antigen to which an antibody or antigen-binding fragment binds. In some embodiments, when the antigen is a polypeptide, the epitope is a conformational epitope, which consists of portions of the antigen that are not covalently contiguous in the antigen but are in close proximity to each other in three-dimensional space when the antigen is in a relevant conformation. For example, for CD38, the conformational epitope is an epitope consisting of non-contiguous amino acid residues in the extracellular domain of CD38, and the linear epitope is an epitope consisting of contiguous amino acid residues in the extracellular domain of CD38. In some embodiments, the epitopes utilized in accordance with the present invention are provided by referring to the epitopes to which the anti-CD38 antibodies disclosed herein (defined, for example, as aCD38-b-348 or aCD38-b-329 and aCD38-b-ep) bind. Means for determining the precise sequence and / or specific amino acid residues of the aCD38-b-348 or aCD38-b-329 epitope are known from the literature and examples and include competition with peptides derived from antigen sequences, binding to, cleavage, and / or mutagenesis of CD38 sequences from different species (e.g., by alanine scanning or other site-directed mutagenesis), phage display-based screening, or (co)crystal structure analysis.

[0075] Indirect antiglobulin test (IAT): As used herein, the terms “indirect antiglobulin test” or “IAT” may refer to a method for testing for any patient-derived antibodies that specifically bind to erythrocyte antigens expressed by donor erythrocytes in a donor blood sample. The patient blood sample may include whole blood, plasma, or serum. The donor blood sample may include whole blood or erythrocytes. The indirect antiglobulin test can be performed using a column agglutination assay, a tube assay, or a solid-phase assay. The IAT may include the following steps: A erythrocyte suspension can be incubated with a plasma or serum sample from the patient, or with a blood typing reagent or control. The erythrocyte suspension can be obtained from the donor blood sample. Incubation can be performed at room temperature (approximately 15°C to approximately 25°C). Alternatively, incubation can be performed at approximately 37°C. Incubation can be performed for a period determined according to the manufacturer's instructions. During the incubation process, if patient-derived anti-erythrocyte antigen antibodies are present in the patient sample and their specific antigen is also present on the RBCs, binding of patient-derived antibodies to the erythrocyte antigen may occur. The process by which patient-derived antibodies bind to the erythrocyte antigen is sometimes called sensitization. Following sensitization, a washing step can be performed to separate unbound antibodies from the antibody-bound RBCs in the solution. A coagulant, such as an anti-human globulin reagent, is added to the solution containing the RBCs and any bound antibodies (if present). The anti-human globulin reagent contains anti-human IgG antibodies and may further contain anti-C3. Any anti-human globulin reagent can be used in this invention. When patient-derived antibodies bind to RBCs, the anti-human globulin reagent will bind to the patient-derived antibodies on the RBCs, causing agglutination of the RBCs. A separation step may be included to separate the agglutinated RBCs from the remaining solution. The separation step may include centrifugation. Centrifugation may be performed at a rate and time determined according to the specific equipment used or any conditions sufficient to separate the agglutinated RBCs from the remaining solution. Under normal testing conditions, an agglutination reaction in the IAT test indicates the presence of patient-derived antibodies (clinically significant antibodies) against RBC antigens in the patient's sample.Under normal testing conditions, agglutination in the IAT test indicates incompatibility between donor RBCs and the patient's serum and / or plasma. Hemolysis (destruction of red blood cells) may also be observed in the IAT test, which is also an indicator of incompatibility between donor RBCs and the patient's serum and / or plasma. If the patient is being treated with daratumumab or isatuximab, agglutination of RBCs may occur in the IAT test regardless of whether the patient's sample contains alloantibodies against any RBC antigen. This is because anti-human globulins agglutinate the anti-CD38 antibodies of daratumumab or isatuximab, which in turn causes agglutination of red blood cells to which the anti-CD38 antibodies bind, and this can be detected visually (with the naked eye). The method of the present invention is characterized in that, when the patient sample contains an anti-CD38 antibody or its antigen-binding fragment (i.e., not daratumumab or isatuximab) as described herein, the presence of the anti-CD38 antibody surprisingly does not cause RBC agglutination in the IAT test. This is also true when the anti-CD38 antibody is, for example, an IgG antibody and the anti-human globulin contains anti-human IgG.

[0076] interference: As used herein, the term “interference” may refer to any type of false-positive or false-negative result obtained in blood antibody screening or blood cross-matching tests. For example, interference may refer to the occurrence of an agglutination reaction in an IAT test when no clinically significant alloantibodies are present in the patient sample. Interference may falsely indicate incompatibility between the patient’s serum or plasma and the donor’s RBCs. Interference may falsely indicate the presence of alloantibodies in the patient’s serum or plasma sample that specifically bind to the donor RBC antigen. For example, most anti-CD38 antibodies (e.g., daratumumab and isatuximab) are known to cause interference in blood antibody screening and blood cross-matching tests. When present in the patient’s plasma or serum sample, daratumumab and / or isatuximab may bind to CD38 on donor RBCs when the patient sample and donor sample are mixed. Agglutination occurs when an anti-human globulin reagent is added to a mixture (derived from patient serum) containing daratumumab or isatuximab conjugated to donor RBCs. Agglutination usually indicates incompatibility between the patient and the donor, but agglutination may occur even if the patient blood sample does not contain clinically significant antibodies, such as alloantibodies that bind to any erythrocyte antigen present on the donor RBCs, when the patient is being treated with daratumumab or isatuximab. The anti-CD38 antibodies described herein (i.e., aCD38-b-348 and aCD38-b-329, and antibodies derived therefrom) do not cause interference to blood antibody screening or blood cross-matching tests. In some embodiments, the presence of the anti-CD38 antibodies described herein or their antigen-binding fragments in the patient's serum or plasma sample causes less interference to blood antibody screening or blood cross-matching tests compared to daratumumab and / or isatuximab. In some embodiments, the presence of the anti-CD38 antibody or its antigen-binding fragment described herein in a patient's serum or plasma sample causes less agglutination in blood antibody screening or blood cross-matching tests compared to daratumumab and / or isatuximab. In some embodiments, treatment of a patient with the anti-CD38 antibody described herein does not result in agglutination in blood antibody screening or blood cross-matching tests.

[0077] Isatsukimab: Isatuximab is a human monoclonal IgG1 anti-CD38 antibody. Isatuximab is known to interfere with blood antibody screening and blood cross-matching tests (i.e., agglutination when clinically significant patient antibodies that specifically bind to erythrocyte antigens are absent). Isatuximab may contain the following variable heavy chain and variable light chain sequences: Heavy chain: QVQLVQSGAEVAKPGTSVKLSCKASGYTFTYWMQWVKQRPGQGLEWIGTIYPGDGDTGYAQKFQGKATLTADKSSKTVYMHLSSLASEDSAVYYCARGDYYGSNSLDYWGQGTS VTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(Sequence ID 33) Light chain: DIVMTQSHLSMSTSLGDPVSITCKASQDVSTVVAWYQQKPGQSPRRLIYSASYRYIGVPDRFTGSGAGTDFTFTISSVQAEDLAVYYCQQHYSPPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (Sequence ID 34)

[0078] patient: Where used herein, the terms “patient” or “subject” refer to any organism to which the provided composition is administered or can be administered for, for example, experimental, diagnostic, preventive, cosmetic and / or therapeutic purposes. Typical patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates and / or humans). In some embodiments, the patient is human. In some embodiments, the patient has or is susceptible to one or more disorders or conditions. The patient may exhibit one or more symptoms of a disorder or condition, or may have been diagnosed with one or more disorders or conditions (e.g., cancer or the presence of one or more tumors). In some embodiments, the patient is receiving or has received certain therapies for the diagnosis and / or treatment of such disease, disorder or condition. In preferred embodiments, the patient is a human cancer patient, for example, a multiple myeloma patient.

[0079] Percentage (%) sequence identity: The percentage (%) of "sequence identity" between two sequences is known in the art. Sequence identity for peptide, polypeptide, or antibody sequences can be determined using the following methods. Sequence identity for peptide, polypeptide, or antibody sequences can be defined as the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues in a particular peptide or polypeptide sequence, after aligning the sequences and introducing gaps to achieve the maximum possible percentage of sequence identity, without considering any conservative substitutions as part of the sequence identity. Alignment aimed at determining a percentage of amino acid sequence identity can be achieved using default parameters in various methods within the scope of the skills in the art, for example, using publicly available computer software such as BLAST-2, including BLAST, GapBLAST and BLASTp (for proteins) (Altschul SF et al (1997)), or FASTA.

[0080] Patient antibodies: Where used herein, the term “patient antibody” or “patient antibodies” refers to antibodies produced in vivo by the patient themselves. Therefore, patient antibodies are patient-derived antibodies. Thus, patient antibodies are distinguished from anti-CD38 antibodies, for these are therapeutic antibodies (exogenous antibodies administered to a patient as part of a treatment regimen, e.g., treatment for CD38-expressing cancer). Patient antibodies may be alloantibodies or autoantibodies. Generally, the method of the present invention determines the presence or absence of clinically significant patient antibodies in a patient blood sample. “Clinically significant” means the ability of patient antibodies to react with donor red blood cells to cause an adverse reaction when donor red blood cells are administered to the patient. Adverse reactions may include the destruction (hemolysis) of donor red blood cells. Adverse reactions may include acute or delayed hemolytic transfusion reactions, or hemolytic diseases of the fetus and neonatal disease (HDFN).

[0081] Typically, patient-derived antibodies that specifically bind to one or more red blood cell antigens are human IgG antibodies.

[0082] An "alloantibody" refers to an antibody that specifically binds to red blood cell antigens that are not present on the subject's own red blood cells. Therefore, an alloantibody is an anti-red blood cell antigen alloantibody. Alloantibodies can be distinguished from "autoantibodies," which refer to antibodies that specifically bind to antigens present on the subject's own red blood cells. Both alloantibodies and autoantibodies can be detected by the method of the present invention. For alloantibodies to develop, the individual must be exposed to non-self RBC antigens and possess an HLA-binding motif that can present a portion of the non-self antigen (Tormey & Hendrickson, 2019). Exposure to non-self antigens can occur through pregnancy, blood transfusion, or transplantation. The process of forming alloantibodies is called "alloimmunization." Alloantibodies can be clinically significant antibodies, and if a mother possesses alloantibodies against antigens on her baby's red blood cells, it can lead to either the destruction (hemolysis) of transfused RBCs or harm to the fetus or newborn. In fact, alloimmunization can be a direct cause of transfusion-related death. Alloimmunization also presents further challenges in patient care, such as delayed transfusions, difficulty in identifying compatible blood in highly alloimmunized individuals, and delayed or acute hemolytic transfusion reactions. Alloimmunization is particularly clinically important for tumor patients at greater risk of developing alloantibodies, who frequently receive transfusions as part of supportive care (Hendrickson & Tormey, 2016). Therefore, it is crucial to be able to accurately and rapidly screen patient samples for the presence of alloantibodies. Screening for alloantibodies may be time-consuming in patients treated with anti-CD38 antibodies such as daratumumab or isatuximab, because the presence of anti-CD38 antibodies in their serum, which bind to CD38 on RBCs, can falsely indicate the presence of alloantibodies. Modifications to antibody screening and blood cross-matching tests used for alloantibody detection can incorporate steps to avoid this interference by anti-CD38 antibodies. For example, RBCs can be treated with an antigen stripping agent (such as DTT), or patient samples can be treated with an anti-CD38 neutralizing agent (such as soluble CD38). However, However, such additional reagents incur additional costs, are not widely available, and the extra process steps are time-consuming, leading to delays in antibody screening. The method of the present invention enables the detection of alloantibodies in patient samples without requiring additional processing of RBCs or patient blood samples. Using the method of the present invention, anti-CD38 antibodies or their antigen-binding fragments do not cause interference (e.g., agglutination in the absence of clinically significant alloantibodies) in antibody screening or cross-matching tests, thereby minimizing costs and avoiding delays in identifying transfusion-compatible blood products.

[0083] Patient antibodies (e.g., patient alloantibodies) do not specifically bind to CD38.

[0084] Pharmaceutically acceptable: When used herein, the term “pharmaceutically acceptable” as applied to carriers, diluents, or additives used in the formulation of compositions disclosed herein means that the carrier, diluent, or additive must be compatible with the other components of the composition and not harmful to its recipient.

[0085] Pharmaceutical composition: As used herein, the term “pharmaceutical composition” refers to a composition comprising an active agent with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in a unit dose appropriate for administration in a therapeutic regime that demonstrates a statistically significant potential to achieve a predetermined therapeutic effect when administered to the relevant population. Pharmaceutical compositions may be formulated for administration in solid or liquid forms, including: oral administration, e.g., drenches (aqueous or nonaqueous solutions or suspensions), tablets, e.g., tablets intended for oral, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, e.g., sterile solutions or suspensions, or as sustained-release formulations by subcutaneous, intramuscular, intravenous, intratumoral, or epidural injection; topical application, e.g., as creams, ointments, or controlled-release patches or sprays applied to the skin, lungs, or oral cavity; and application to the vagina, rectum, sublingual, intraocular, transdermal, transnasal, transpulmonary, and other mucosal surfaces.

[0086] plasma: As used herein, the term “plasma” refers to the liquid component of blood that contains few red blood cells, white blood cells, and platelets. Plasma may also contain albumin, as well as fibrinogen and other coagulation factors. The plasma used in the present invention may be prepared from whole blood using any appropriate or standard preparation protocol. In the present invention, plasma may be provided by or derived from a patient who is to receive a transfusion. To prepare the plasma to be used, whole blood may be collected in a tube treated with an anticoagulant. Red blood cells and platelets are removed or separated by centrifugation, and the resulting supernatant is called plasma. The plasma sample used in the present invention may contain, for example, a volume of about 10 μl to about 3 ml. For example, about 100 μl, 150 μl, 160 μl, 200 μl, 250 μl, or 300 μl of plasma may be used. The plasma and / or serum used in the method of the present invention may be diluted with an appropriate buffer or diluent before use. Plasma and / or serum can be prepared and used at dilutions of 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10. Suitable diluents include, for example, phosphate-buffered saline (PBS) and / or low ionic strength solution (LISS).

[0087] Red blood cells: In this specification, the terms "red blood cells," "RBC," or "red" are used. The term "erythrocytes" refers to blood cells that can transport oxygen. The red blood cells used in this invention, i.e., donor red blood cells, are prepared in any appropriate or standard preparation. The protocol allows for obtaining whole blood from any suitable source. In this invention, red blood cells can be obtained from donor blood sources intended for use, for example, in transfusions to patients. Therefore, donor red blood cells are generally derived from human donors. Donor blood can be collected and stored in flexible plastic bags. The bags may contain compounds and chemicals (e.g., sodium citrate, phosphates, dextrose, and sometimes adenine) to prevent blood clotting and facilitate storage. The tubes that carry the blood through the storage bag can be divided after collection to provide "pigtail" compartments containing smaller volumes of blood. Guter volume donor blood is suitable for use in cross-matching assays, including the assay of the present invention. Small volumes of whole blood can be provided as a source of red blood cells used in the assay of the present invention. For example, about 1 μl to about 500 μl of donor red blood cells can be used. The method of the present invention can use about 10 μl, about 20 μl, about 30 μl, about 40 μl, about 50 μl, about 60 μl, about 70 μl, about 80 μl, about 90 μl, about 100 μl, about 150 μl, or about 200 μl (e.g., about 10 μl to about 200 μl) of donor whole blood. Before use, the red blood cells can be diluted with any suitable diluent or buffer. The method of the present invention can use donor red blood cells prepared in about 10 μl, about 20 μl, about 30 μl, about 40 μl, about 50 μl, about 60 μl, about 70 μl, about 80 μl, about 90 μl, about 100 μl, about 150 μl, or about 200 μl (for example, about 10 μl to about 200 μl) of a suitable diluent or buffer.

[0088] serum: As used herein, the term “serum” refers to the liquid component of blood that contains few clotting factors, platelets, and blood cells. The serum used in the present invention may be prepared from whole blood using any suitable or standard preparation protocol. In the present invention, the serum may be provided by or derived from a patient who is to receive a blood transfusion. To prepare the serum to be used, whole blood is collected and allowed to coagulate for a certain period of time. Red blood cells and platelets can be removed by centrifugation, and the resulting supernatant is called serum. The plasma and / or serum used in the method of the present invention may be diluted with a suitable buffer or diluent before use. Plasma and / or serum may be prepared and used as 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10 dilutions. Suitable diluents include, for example, phosphate-buffered saline (PBS) and / or low ionic strength solution (LISS).

[0089] Solid-phase assay: As used herein, the terms “solid-phase test,” “solid-phase assay,” or “solid-phase method” refer to methods suitable for detecting patient-derived antibodies (alloantibodies, especially clinically significant alloantibodies, etc.) that specifically bind to erythrocyte antigens in patient samples, particularly in blood cross-matching tests or RBC panel antibody screening. Solid-phase systems such as ect can be used in antibody screening to detect the presence of patient-derived antibodies. Solid-phase assays may involve binding RBCs to a solid surface, such as a microplate well. RBCs are selected based on the known expression of erythrocyte antigens on which patient-derived antibodies are commonly formed. The RBCs attached to the solid surface are then incubated with the patient's plasma or serum sample, followed by a washing step, and indicator cells (e.g., cells coated with anti-IgG) are added. If patient-derived antibodies from the patient sample bind to the immobilized RBCs on the solid surface, the anti-IgG on the indicator cells will also bind to the patient antibodies bound to the immobilized RBCs. Indicator cells may be erythrocytes coated with an agglutinant such as anti-human globulin (e.g., anti-IgG and / or anti-C3). The presence of relevant patient-derived antibodies in the patient sample is indicated by a faint red color covering the solid surface to which the indicator RBCs are bound. A negative solid-phase assay in which relevant patient-derived antibodies are not detected in the patient sample is indicated by the indicator cell pellet at the bottom of the well. It is indicated by the mark.

[0090] Solid-phase testing can also be used for blood cross-matching (compatibility testing). In solid-phase cross-matching, a coagulant such as anti-human globulin (e.g., anti-IgG and / or anti-C3) is directly attached to or adhered to a solid surface. Serum or plasma from the patient and the donor RBCs to be tested are incubated together and brought into contact with the solid surface to which the coagulant is attached. All antibodies from the patient sample bound to the donor RBCs will adhere to the coagulant on the solid surface. Therefore, the faint red color covering the solid surface of the well indicates incompatibility between the patient and the donor due to the presence of patient-derived antibodies in the patient sample that specifically bind to the antigen on the donor RBCs.

[0091] Solid tumors: As used herein, the term “solid tumor” usually refers to an abnormal mass of tissue that does not have a cyst or fluid area. Solid tumors can be benign or malignant. The various types of solid tumors are named after the type of cells that form them. Examples of solid tumors include sarcomas (including cancers arising from mesenchymal-derived transformed cells in tissues such as cancellous bone tissue, cartilage tissue, adipose tissue, muscle tissue, vascular tissue, hematopoietic tissue, or fibrous connective tissue), carcinomas (including tumors arising from epithelial cells), melanoma, lymphoma, mesothelioma, neuroblastoma, retinoblastoma, etc. Cancers associated with solid tumors include, but are not limited to, brain cancer, lung cancer, gastric cancer, duodenal cancer, esophageal cancer, breast cancer, colon and rectal cancer, kidney cancer, bladder cancer, renal cancer, pancreatic cancer, prostate cancer, ovarian cancer, melanoma, oral cancer, sarcoma, eye cancer, thyroid cancer, urethral cancer, vaginal cancer, cervical cancer, lymphoma, etc.

[0092] Therapeutic effective dose: As used herein, the term “therapeutic dose” means an amount (e.g., an active substance or pharmaceutical composition) sufficient to treat a disease and / or condition when administered in accordance with a therapeutic drug regimen to a population suffering from or susceptible to such disease and / or condition. A therapeutic dose is an amount that reduces, stabilizes, and / or delays the onset of one or more symptoms of a disease, disorder and / or condition. Those skilled in the art will understand that a “therapeutic dose” does not require that actual therapeutic success be achieved in a particular subject.

[0093] Treatment: In the foregoing, the term “treatment” (as used herein) "Treatment" also means to partially or completely reduce, improve, or alleviate one or more symptoms. : Any administration of a substance (e.g., an anti-CD38 antibody disclosed, exemplified by aCD38-b-348 or aCD38-b-329, or any other active agent) that causes, inhibits, delays the onset of, reduces the severity of, and / or reduces the incidence of, the disease. In some embodiments, the treatment is either the direct administration of an anti-CD38 antibody such as aCD38-b-348 or aCD38-b-329 (e.g., as an aqueous composition for injection optionally containing a pharmaceutically acceptable carrier, additives and / or adjuvant used for intravenous, subcutaneous, intratumoral or peritumoral injection), or obtaining cells from a subject (e.g., from blood, tissue or tumor, with or without selection based on the presence or absence of marker expression), and extinguishing the cells with an anti-CD38 antibody such as aCD38-b-348 or aCD38-b-329. This may include administration using a regimen that involves in vivo contact and administration of such cells to a subject (with or without selection based on the presence or absence of marker expression).

[0094] Tube assay: As used herein, the terms "tube assay" or "tube method" are used. The term refers to a method of performing an indirect antiglobulin test (IAT) in a tube, such as a test tube. Using a tube assay, patient-derived antibodies (alloantibodies, especially clinically significant alloantibodies, etc.) that specifically bind to erythrocyte antigens in patient samples, such as serum or plasma samples, can be detected as part of antibody screening against an RBC panel or as part of a cross-matching test with donor RBCs. A tube assay generally includes the following steps: RBCs can be suspended in a suitable solution, such as isotonic saline (NISS), phosphate-buffered saline (PBS), polyethylene glycol (PEG) solution, or low ionic strength saline (LISS). Patient samples, such as serum or plasma samples, can be added to a tube, followed by the RBC suspension. The patient samples and RBC suspension can be incubated under conditions sufficient to allow binding of any patient antibody to any RBC antigen expressed on donor RBCs. For example, incubation can be performed for at least about 5 minutes. Incubation can be performed for at least approximately 5 minutes, at least approximately 10 minutes, at least approximately 15 minutes, at least approximately 20 minutes, at least approximately 30 minutes, at least approximately 40 minutes, at least approximately 50 minutes, at least approximately 1 hour, at least approximately 90 minutes, or at least approximately 2 hours. The incubation period may vary depending on the reagents used; for example, using LISS may result in a shorter reaction time and thus a shorter required incubation time. Incubation can be performed at approximately 37°C. Incubation can be performed at near room temperature (approximately 15°C to approximately 25°C). After incubation, a washing step can be performed to remove unbound patient antibodies. The tube assay may further include a step of adding an active agent that specifically binds any patient-derived antibodies present in the patient blood sample to each other. The active agent that specifically binds any antibodies present in the patient blood sample to each other may be an antiglobulin, such as anti-human IgG (and / or anti-C3). When patient-derived antibodies bind to RBC antigens on donor RBCs, antiglobulin reagents cause the patient-derived antibodies to bind to each other, leading to agglutination of RBCs bound to the patient antibodies.RBC agglutination in a tube assay indicates the presence of patient-derived antibodies that specifically bind to the RBC antigen on donor RBCs. RBC agglutination in a tube assay may indicate that donor RBCs are incompatible with those of the patient.

[0095] Screening method In a first embodiment of the present invention, a method for screening blood samples obtained from patients who have been administered an anti-CD38 antibody or its antigen-binding fragment, a) Prepare a blood sample from the patient, b) Prepare a blood sample from the blood donor, Here, the donor blood sample contains donor red blood cells; c) Screening a patient blood sample, including determining the presence or absence of one or more patient antibodies in the patient blood sample that specifically bind to one or more red blood cell antigens expressed on the surface of donor red blood cells, A method is provided that includes this.

[0096] Advantageously, the methods of the present invention generally do not involve contacting a patient blood sample or donor blood sample with an active agent that inhibits the binding of an anti-CD38 antibody or its antigen-binding fragment to membrane-bound CD38 that may be present on the surface of donor red blood cells (i.e., that may be expressed by donor red blood cells). In prior art methods, particularly in methods for screening blood from patients administered with anti-CD38 antibodies (other than those disclosed herein), additional method steps are required to prevent other therapeutic anti-CD38 antibodies from interfering with the screening of patient samples for patient antibodies that can bind to red blood cell antigens expressed on the surface of donor red blood cells. In particular, the step of adding an active agent that inhibits the binding of an anti-CD38 antibody or its antigen-binding fragment to membrane-bound CD38 present on the surface of donor red blood cells is necessary to prevent false positives in the screening assay. The active agent that inhibits the binding of an anti-CD38 antibody or its antigen-binding fragment to membrane-bound CD38 is, for example, soluble It may be a CD38 antigen, an anti-CD38 idiotype antibody, or an antigen stripping agent.

[0097] Patients are generally those who, prior to obtaining a blood sample from the patient, have not been administered any anti-CD38 antibodies (and optionally anti-erythrocyte antigen antibodies) for at least 6 months, preferably at least 1 year, other than anti-CD38 antibodies that do not interfere with screening assays such as the anti-CD38 antibodies disclosed herein (particularly aCD38-b-348 antibody or aCD38-b-329 antibody, or antibodies or antigen-binding fragments derived therefrom). Antibodies derived from aCD38-b-348 or aCD38-b-329 include antibodies comprising any of aCD38-b-348 or aCD38-b-329, or their variants aCD38-b-348-m1, aCD38-b-348-m2, aCD38-b-348-m3, aCD38-b-348-m5, aCD38-b-329-m6, or aCD38-b-329-m7, or at least one, but preferably all six, CDRs of any anti-CD38 antibody that binds to the same epitope as aCD38-b-348 or aCD38-b-329. In some embodiments, the patient is a patient who has not received daratumumab or isatuximab for 6 months, preferably at least 1 year, prior to obtaining a blood sample from the patient. Naturally, especially if the anti-CD38 antibody disclosed herein is the first anti-CD38 antibody treatment (or anti-erythrocyte antigen antibody treatment) a patient has received in their lifetime, it is possible that the patient has never received any other anti-CD38 antibody (and optionally never received any anti-erythrocyte antigen antibody) other than the anti-CD38 antibody disclosed herein that does not interfere with the screening assays.

[0098] In some embodiments, the screening in step (b) is performed using an assay selected from the group consisting of column agglutination assays, indirect antiglobulin (IAT) tube assays, and solid-phase assays. These are described in more detail elsewhere.

[0099] In some embodiments, the method includes preparing a patient blood / donor red blood cell mixture by contacting a patient blood sample with donor red blood cells from a donor blood sample prior to screening in step (b). The method may then further include incubating the patient blood / donor red blood cell mixture under conditions sufficient to allow, for example, one or more patient antibodies in the patient blood sample to bind to one or more red blood cell antigens present on the donor red blood cells, if present, to form one or more patient antibody / donor red blood cell antigen complexes. Optionally, the method may further include separating any one or more patient antibody / donor red blood cell antigen complexes, if present, from the patient blood / donor red blood cell mixture, for example, by centrifugation of the patient blood / donor red blood cell mixture. Even if a patient antibody / donor red blood cell antigen complex does not form (for example, because the patient blood sample did not contain any patient antibodies that specifically bind to any RBC antigen on the donor RBCs), the "separation step" (e.g., centrifugation) can still be performed after mixing the patient sample and donor red blood cells. However, this may not become clear until after the separation step (e.g., centrifugation) has been performed.

[0100] Anti-CD38 antibody or its antigen-binding fragment The anti-CD38 antibody or its antigen-binding fragment does not cause interference when cross-matching patient blood samples with donor red blood cells or when performing any antibody-RBC panel assay. The anti-CD38 antibody or its antigen-binding fragment is not daratumumab or isatuximab.

[0101] In some embodiments, the anti-CD38 antibody or its antigen-binding fragment may be aCD38-b-348 or aCD38-b-329, or may be derived from the above antibody. For example, in some embodiments, the anti-CD38 antibody or its antigen-binding fragment The fragment contains the amino acid sequence of SEQ ID NO: 3 or SEQ ID NO: 19 as a variable heavy chain complementarity determining region 3 (HCDR3). In some embodiments, the anti-CD38 antibody or antigen-binding fragment contains amino acid sequence elements, for example, a) The amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 17 as variable heavy chain complementarity determination region 1 (HCDR1), and / or b) The amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 18 as variable heavy chain complementarity determining region 2 (HCDR2), It may further include the following.

[0102] In some embodiments, the anti-CD38 antibody or its antigen-binding fragment is a) The amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 20 as variable light chain complementarity determination region 1 (LCDR1), b) The amino acid sequence of SEQ ID NO: 5 or SEQ ID NO: 21 as variable light chain complementarity determination region 2 (LCDR2), and, c) An amino acid sequence selected from the group consisting of SEQ ID NOs: 6, SEQ ID NOs: 22, SEQ ID NOs: 9, SEQ ID NOs: 10, SEQ ID NOs: 11, SEQ ID NOs: 12, SEQ ID NOs: 25, and SEQ ID NOs: 26 as a variable light chain complementarity determining region 3 (LCDR3), It may further include the following.

[0103] In some embodiments, the anti-CD38 antibody or its antigen-binding fragment comprises a variable heavy chain containing the amino acid sequence of SEQ ID NO: 7 or SEQ ID NO: 23. Preferably, the anti-CD38 antibody or its antigen-binding fragment further comprises a variable light chain containing an amino acid sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 24, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 27, and SEQ ID NO: 28.

[0104] As will be discussed in more detail below, the antigen-binding fragments of mutant antibodies, such as mutants having a certain percentage of identity and / or one or more amino acid substitutions, can also be used.

[0105] Certain characteristics of anti-CD38 antibodies In some embodiments, the anti-CD38 antibody or antigen-binding fragment modulates one or more characteristics of CD38. That is, in some embodiments, the level and / or activity of CD38, and / or one or more downstream effects thereof, are detected to change when the provided antibody is present compared to when it is not present. Alternatively or additionally, in some embodiments, the level and / or activity of CD38, and / or one or more downstream effects thereof, when the provided antibody is present, are equivalent to or greater than those observed under equivalent conditions in the case of a reference anti-CD38 antibody (such as IB-4, which has a known ability to agonize one or more characteristics of CD38, known to be a known desired characteristic).

[0106] In many embodiments, one or more characteristics of CD38 are enhanced in the presence of an anti-CD38 antibody or its antigen-binding fragment used in the present invention. For example, in some embodiments, the presence of an anti-CD38 antibody or its antigen-binding fragment correlates with increased activation and / or proliferation of immune cells. Thus, anti-CD38 antibodies are often referred to herein as “agonists.” However, those skilled in the art will understand that the teachings of this disclosure are not limited by the specific mechanism of action of the antibodies or their antigen-binding fragments provided. The relevant structural and / or functional characteristics of the antibodies provided are described herein and will be self-evident.

[0107] In some embodiments, an anti-CD38 antibody or its antigen-binding fragment is used to exert an effect, for example, on certain immune effector cells (e.g., NK cells and / or T cells). It can be characterized by the results. Alternatively or additionally, in some embodiments, the anti-CD38 antibody or antigen-binding fragment thereof can be characterized, for example, by the effect on immunosuppressive cells. For example, in some embodiments, the anti-CD38 antibody or antigen-binding fragment thereof exhibits activation properties with respect to immune effector cells such as NK cells and T cells, and exhibits cytotoxic properties against CD38-high expressing cells such as immunosuppressive cells. Alternatively or additionally, in some embodiments, the anti-CD38 antibody or antigen-binding fragment thereof is characterized by binding to specific epitopes in the human CD38 extracellular domain and / or one or more features that make them particularly suitable for pharmaceutical use and / or pharmaceutical manufacturing. In particular, the anti-CD38 antibody is particularly useful in methods of blood screening including cross-matching tests between patients and RBC donors, including patients undergoing therapy for certain diseases that may require RBC transfusion (including cancer patients, particularly multiple myeloma patients).

[0108] In some embodiments, the provided antibody or antigen-binding fragment thereof is 10 -8 M or less (range of 10 -9 M) binds to human CD38 with a Kd, and preferably, the antibody or antigen-binding fragment thereof binds to human CD38 with a Kd in the range of 10 -8 M to 10 -11 M. In some embodiments, the Kd is about 10 -8 to about 10 -11 . In some embodiments, the Kd is 10 -8 to 10 -9 . In some embodiments, the provided antibody or antigen-binding fragment thereof can also bind to human and cynomolgus monkey CD38 (e.g., extracellular epitopes on human and cynomolgus monkey CD38) with a Kd value in the range of 10 -8 M to 10 -11 M. The Kd for evaluating the binding affinity of the antibody or antigen-binding fragment thereof is determined by Biacore analysis or Forte Bio's Octet This can be obtained by standard methodologies, including surface plasmon resonance (SPR), such as analysis using Systems.

[0109] The antibodies (and / or their antigen-binding fragments) used herein may be particularly useful for use in medical applications (e.g., therapy and / or prevention, e.g., in the treatment of cancer) and / or in relation to methods that require or involve targeting an epitope, such as that identified as aCD38-b-ep within the extracellular domain of human CD38. The antibodies or their antigen-binding fragments provided may be prepared to represent the most appropriate isotype, in particular a human isotype from the group consisting of isotype antibodies of IgG1, IgG2, IgG3, and IgG4, more specifically human IgG1.

[0110] Antibodies can be provided in various forms. For example, in some embodiments, suitable forms may be, or include, monoclonal antibodies, domain antibodies, single-chain antibodies, Fab fragments, F(ab')2 fragments, single-chain variable fragments (scFv), scFv-Fc fragments, single-chain antibodies (scAb), aptamers, or nanobodies. In some embodiments, the antibody or its antigen-binding fragment (and monoclonal antibodies in particular) may be a rabbit antibody, mouse antibody, chimeric antibody, humanized antibody, or fully human antibody, or its antigen-binding fragment. In some embodiments, the antibody or its antigen-binding fragment provided may be an isotype of IgG, IgA, IgE, or IgM (preferably human), as this may be most appropriate for a given use. In some embodiments, the antibody or its antigen-binding fragment provided is an IgG isotype, more specifically an isotype of IgG1, IgG2, IgG3, or IgG4 (preferably human IgG1). In some embodiments, the provided antibody or its antigen-binding fragment is provided as part of a multispecific binding agent, such as an isolated antibody or antigen-binding agent, which may be included in a bispecific antibody, a multispecific antibody, or other multispecific form that may be available in the art, for example, when it is desirable to associate further binding and / or functional parts.

[0111] In some embodiments, antibodies or their antigen-binding fragments (or their variants) may be defucosylated. It is well known that glycosylation of antibodies (e.g., monoclonal antibodies, recombinant antibodies, and / or other manipulated or isolated antibodies) can affect antibody activity, pharmacokinetics, and pharmacodynamics, and by utilizing Fc fusion proteins and certain techniques, antibodies with a desired glycosylation profile can be obtained (Liu L, 2015). Effector devices supporting the cytotoxicity of antibodies used according to the present invention. The ability can be enhanced by methods that reduce the fucosylation level of the antibody. Antibodies containing specific aCD38-b-348 or aCD38-b-329 sequence elements exhibiting such properties can be produced, for example, by expressing the aCD38-b-348 or aCD38-b-329 sequence using techniques for genetically modified cell lines capable of producing antibodies with no or reduced fucosylation ability (some of which are commercially available, such as Potelligent (Lonza) and GlyMAXX (ProBiogen)), or by manipulating the manufacturing process, for example by controlling osmotic pressure, and / or by using enzyme inhibitors (see, for example, the method described in European Patent No. 2480671).

[0112] The anti-CD38 antibody or its antigen-binding fragment used in the present invention may be provided in the form of a composition (e.g., a pharmaceutical composition) comprising the provided antibody or its antigen-binding fragment having the desirable properties described herein (e.g., aCD38-b-348 antibody or aCD38-b-329 antibody, or their antigen-binding fragments, and their variants or other antibodies derived therefrom). In some embodiments, such compositions are intended for and / or used in medical applications such as therapeutic, diagnostic, or preventive applications. In some embodiments, such compositions may further comprise a pharmaceutically acceptable carrier or excipient and / or be used in the treatment of cancer. In some embodiments, the pharmaceutical composition may be formulated using one or more carriers, excipients, salts, buffers, etc., as known in the art. Those skilled in the art will recognize and readily utilize a variety of formulation techniques, including those particularly desired and / or useful for a given method and / or site of administration, such as parenteral administration (e.g., subcutaneous, intramuscular, or intravenous injection), mucosal administration, intratumoral administration, peritumoral administration, oral administration, or topical administration. In many embodiments, the pharmaceutical composition provided, comprising an anti-CD38 antibody or its antigen-binding moiety, is formulated for parenteral delivery (e.g., by injection and / or infusion). In some embodiments, such pharmaceutical compositions may be provided, for example, in the form of a pre-loaded syringe or vial. In some embodiments, such pharmaceutical compositions may be provided and / or used, for example, in a dry (e.g., lyophilized) form. Alternatively, in some embodiments, such pharmaceutical compositions may be provided and / or used in a liquid form (e.g., solution, suspension, dispersion, emulsion, etc.), a gel form, etc.

[0113] Functional characteristics of anti-CD38 antibodies In some embodiments of the present invention, antibodies (and their variants as described herein, such as mutants obtained by mutation and removal of the DG motif) may have an advantageous activity profile. For example, in one embodiment, an antibody or its antigen-binding fragment (and their variants) CD38 + It can exhibit antibody-dependent cell-mediated cytotoxicity (ADCC) activity against target cells, It may be possible to demonstrate complement-dependent cell injury (CDC), It may exhibit antibody-dependent cell phagocytosis (ADCP), and / or This may induce the activation of immune effector cells.

[0114] Preferably aCD38-b-348 or aCD38-b-329, or the same Antigen-binding fragments (or their variants) compared to daratumumab and / or isatuximab under the same or substantially the same conditions for CD38 + It shows a decrease in CDC activity against target cells.

[0115] The antibody-dependent cell-mediated cytotoxicity (ADCC) activity of an anti-CD38 antibody or its antigen-binding fragment is determined in vitro, for example, by testing CD38 cells as target cells. + This can be determined using Daudi cells and human PBMC cells as effector cells, where the target cell to effector cell ratio is approximately 50:1 to approximately 25:1.

[0116] CD38 + Complement-dependent cytotoxicity (CDC) activity against target cells is measured in vitro, for example, in the presence of 10% complement in CD38 + Daudi cells and / or Raji cells can be used to determine CDC activity. CDC activity can be determined by treating target cells with progressively increasing concentrations of antibody up to 10 μg / ml in the presence of human complement. In some embodiments, CDC activity is determined by CD38 in the presence of 10% complement. + Cells, i.e., CD38 +This can be determined by measuring the maximum percentage of cell lysis in Daudi cells. The maximum lysis for a given antibody may vary between experiments. Therefore, for example, EC 50 Maximum % lysis and / or EC compared to the value and / or reference antibody (daratumumab, etc.) 50 It is helpful to consider other metrics for measuring CDC activity, including multiplier differences in EC. Therefore, determining lower CDC activity compared to daratumumab and / or isatuximab is based on maximum % lysis, EC. 50 This may relate to the multiplier change of either value compared to daratumumab, and / or any of the values.

[0117] In one preferred embodiment of the present invention, the anti-CD38 antibody is a) EC that is at least 0.5 times (or more preferably at least 1 time) higher than daratumumab and / or isatuximab 50 And, or, b) Maximum lysates measured in Raji cells and / or Daudi cells in the presence of 10% complement, which is less than half of that shown by daratumumab. For example, CDC may be observed, particularly when the anti-CD38 antibody is aCD38-b-348, or an antibody derived therefrom, or a variant thereof.

[0118] In one preferred embodiment of the present invention, the CD38-modulated antibody agent is a) EC that is at least 0.5 times (or more preferably at least 1 time) higher than daratumumab and / or isatuximab 50 And, or, b) Maximum lysates measured in Raji cells and / or Daudi cells in the presence of 10% complement, which is less than half of that shown by daratumumab. For example, CDC may be observed, particularly when the anti-CD38 antibody is aCD38-b-329, or an antibody derived therefrom, or a variant thereof.

[0119] Naturally, the CDC of daratumumab and / or isatuximab is determined under the same or substantially the same conditions for comparison. CDC activity can be determined using antibody concentrations up to approximately 10 μg / mL. As will be understood by those skilled in the art, a concentration of 10 μg / mL is not necessarily required when determining maximum cell lysis, since maximum cell lysis can occur at lower antibody concentrations, although 10 μg / mL may be used if necessary.

[0120] In some embodiments, the decrease in CDC activity compared to daratumumab and / or isatuximab is due to the EC of the antibody or its antibody-binding fragment. 50 However, under the same or substantially the same conditions, the EC of daratumumab 50 It is at least about 0.5 times larger (i.e., at least about 1.5 times larger), or preferably at least about 1 time larger (i.e., at least about 2 times larger). For example, an antibody or its antibody-conjugated fragment. EC 50 This refers to the EC of daratumumab against Daudi cells and / or Raji cells in the presence of 10% complement. 50 It is at least about 0.5 times larger than, or preferably about 1 time larger.

[0121] In some embodiments, the antibody or its antigen-binding fragment (or variant thereof) is CD38 + Daudi cells and / or Raji cells should receive at least approximately 0.05 μg / mL of EC1. 50 Induce the CDC (optionally, by the CDC such as CD38 + (Causes lysis of less than 60% of expressing cells). In some embodiments, the antibody or its fragment is CD38 + For Daudi cells and / or Raji cells, at least about 0.05 μg / mL, at least about 0.10 μg / mL, or at least about 0.15 μg / mL of EC2. 50 This induces CDC (optionally, with an antibody concentration of up to approximately 10 μg / ml, such as CD38 by CDC). +(Causes lysis of less than 60% of expressing cells).

[0122] In some embodiments, anti-CD38 antibodies or their antigen-binding fragments (or their variants) may exhibit antibody-dependent phagocytosis (ADCP) against CD38-expressing cells. ADCP activity can be determined by a reporter cell assay that measures FcgRIIa binding in Jurkat cells, which are effector cells expressing FcgRIIa. The effector cells also express NFAT-inducible luciferase. The target cells in the assay may be CD38-expressing Raji cells. Activity can be determined by measuring NFAT signaling.

[0123] In some embodiments, anti-CD38 antibodies or their antigen-binding fragments (or their variants) can induce ADCP in Treg cells generated in vitro.

[0124] In some embodiments, anti-CD38 antibodies or their antigen-binding fragments (or their variants) can induce T cell activation in greater quantities compared to daratumumab under the same or substantially the same conditions. In some embodiments, T cell activation can be determined by measuring NFAT signaling in luc reporter Jurkat cells. In some embodiments, NFAT signaling induced by anti-CD38 antibodies or their antigen-binding fragments, as measured in luc reporter Jurkat cells, is at least about 10% higher than NFAT signaling for daratumumab, as measured under the same or substantially the same conditions. In some embodiments, NFAT signaling is at least about 15%, at least about 20%, or at least about 30% higher than NFAT signaling for daratumumab, as measured under the same or substantially the same conditions.

[0125] In a luc reporter assay for NFAT in Jurkat cells, NFAT signaling can be measured in relative luminescence units (RLUs) in the presence of a soluble CD3 monoclonal antibody. The CD3 monoclonal antibody may be at a concentration of 1 μg / ml, and Jurkat cells can be stimulated with anti-CD38 antibody at concentrations of approximately 5 μg / ml to approximately 40 μg / ml (e.g., 10 μg / ml). Using such an assay, NFAT signaling can be at least approximately 30% higher than daratumumab NFAT signaling measured under the same or substantially the same conditions, when the RLU of CD3-only stimulation is used as the baseline.

[0126] T cell activation can be further characterized by increased T cell proliferation and / or increased cytokine secretion, where cytokines may be selected from the group consisting of IL-2, TNF-α, IFN-γ, IL-10, and GM-CSF.

[0127] T cell proliferation can be determined and measured, for example, after 72 hours of incubation, at an antibody concentration of 10 μg / ml in the presence of 0.1 μg / ml or 0.5 μg / ml anti-CD3 antibody, as in the examples. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment is CD4 + Cells and / or CD8 + The treatment increases T cell proliferation in cells by at least about 20% compared to untreated cells. In some embodiments, T cell proliferation increases by at least about 25%, at least about 30%, at least about 35%, or at least about 40% compared to untreated cells.

[0128] Preferably, the anti-CD38 antibody or its antigen-binding fragment (or variant thereof) is CD4 + Cells and / or CD8 +To increase T cell proliferation in cells by at least approximately 0.5 times (i.e., at least 1.5 times), at least 1 time (i.e., at least 2 times), at least 2 times (i.e., at least 3 times), or at least 3 times (i.e., at least 4 times) compared to cells treated with human IgG1 under the same or substantially the same conditions (e.g., 72-hour incubation at the same antibody concentration).

[0129] In some embodiments, the anti-CD38 antibody or its antigen-binding fragment (or variant thereof) is CD4 + Cells and / or CD8 + In cells, the secretion of cytokines selected from the group consisting of IL-2, TNF-α, IFN-γ, IL-10, and / or GM-CSF is induced in greater quantities than that induced by daratumumab under the same or substantially the same conditions. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment increases GM-CSF secretion compared to daratumumab. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment increases IL-2 secretion compared to daratumumab. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment increases the secretion of IL-2, TNF-α, IFN-γ, IL-10, and GM-CSF compared to daratumumab. Cytokine secretion can be determined and measured, for example, at an antibody concentration of 10 μg / ml after 72 hours of incubation, as shown in the examples.

[0130] In some embodiments, anti-CD38 antibodies or their antigen-binding fragments (or their variants) may induce NK cell activation. NK cell activation may be characterized by increased NK cell proliferation. NK cell activation may also be determined, alternatively or additionally, by increased intracellular IFNg production and / or increased expression of the degranulation marker CD107a.

[0131] In some embodiments, anti-CD38 antibodies or their antigen-binding fragments (or variants thereof) may affect cyclase activity and / or NADase activity. The effect of CD38 on NADase activity can be measured, for example, by measuring the conversion of E-NAD+ to 5'-eAMP in Jurkat cells. The effect of CD38 on cyclase activity can be measured, for example, by measuring the conversion of NGD+ to cGDPR in Jurkat cells.

[0132] In some embodiments, an anti-CD38 antibody or its antigen-binding fragment (or variant thereof) has an inhibitory effect on CD38 cyclase activity. The inhibitory effect on CD38 cyclase activity can be measured, for example, by measuring the conversion of NGD+ to cGDPR in Jurkat cells. The inhibitory effect on CD38 cyclase activity, measured by the conversion of NGD+ to cGDPR in Jurkat cells, can result in CD38 activity at least 10% lower compared to CD38 cyclase activity in the presence of an IgG-unbound control antibody. In some embodiments, an anti-CD38 antibody or its antigen-binding fragment is used under the same or substantially the same conditions. The inhibitory effect on CD38 cyclase is smaller than that of daratumumab. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment reduces CD38 cyclase activity to about 25% or more of the CD38 cyclase activity in the presence of an IgG-unbound control antibody, as measured by the conversion of NGD+ to cGDPR in Jurkat cells. Preferably, the antibody reduces CD38 cyclase activity to about 30% or more, about 40% or more, or about 50% or more of the CD38 cyclase activity in the presence of an IgG-unbound control antibody. Preferably, the antibody reduces CD38 cyclase activity to between 25% and 95%, between about 30% and 90%, or between about 50% and 90% of the CD38 cyclase activity in the presence of an IgG-unbound control antibody. This means that, in the presence of anti-CD38 antibodies or their antigen-binding fragments, CD38 cyclase activity is still present in Jurkat cells, but at a reduced level compared to the presence of IgG-unbound control antibodies.

[0133] Daratumumab is known to inhibit cyclase activity and stimulate NADase activity. In contrast, the antibody of the present invention may have a smaller inhibitory effect on CD38 cyclase activity than that of daratumumab under the same or substantially the same conditions.

[0134] Therefore, anti-CD38 antibodies or their antigen-binding fragments (or their variants) are CD38 + It exhibits antibody-dependent cell-mediated cytotoxicity (ADCC) activity against target cells and, under the same or substantially the same conditions, compared to daratumumab and / or isatuximab, CD38 + It shows a decrease in CDC activity against target cells (for example, EC measured as described herein). 50 The value is the EC of daratumumab. 50These antibodies induce activation of immune effector cells (which may be at least twice the normal value), induce T cell proliferation, increase cytokine secretion including IL-2, IFNγ, TNFα, GM-CSF, and IL-10, and induce NK cell activation. Such antibodies may also show a slight inhibitory effect on CD38 cyclase activity.

[0135] Epitope Given that the selected anti-CD38 antibody does not interfere with cross-matching tests, an anti-CD38 antibody that binds to a specific epitope of anti-CD38 may be particularly useful in the present invention.

[0136] In some embodiments, an anti-CD38 antibody or its antigen-binding fragment binds to an epitope on human CD38 that is conjugated by aCD38-b-348 or aCD38-b-329. In some embodiments, such an anti-CD38 antibody or its antigen-binding fragment binds to the extracellular domain of human CD38. In some embodiments, the anti-CD38 antibody may bind to an epitope identified as aCD38-b-ep (amino acids 65-79 in the protein sequence ARCVKYTEIHPEMRH (SEQ ID NO: 30); Uniprot sequence P28907 (SEQ ID NO: 29)).

[0137] In some embodiments, the anti-CD38 antibody specifically binds to an epitope of human CD38 that includes one or more amino acid residues contained in amino acids 65-79 of SEQ ID NO: 29. In some embodiments, the anti-CD38 antibody specifically binds to an epitope of human CD38 that includes or consists of amino acids 65-79 of SEQ ID NO: 29. In some embodiments, the anti-CD38 antibody binds within the epitope of amino acids 65-79 of SEQ ID NO: 29. Any reference to “within” in this specification includes the edges of the range. For example, in this case, “within the epitope of amino acids 65-79 of SEQ ID NO: 29” includes residues 65 and 79, and therefore the epitope may include those residues.

[0138] Preferably, the epitope comprises at least 5 amino acids, at least 6 amino acids, at least 7 amino acids, at least 8 amino acids, at least 9 amino acids, at least 10 amino acids, at least 11 amino acids, at least 12 amino acids, at least 13 amino acids, or at least 14 or more amino acids, where the epitope comprises one or more amino acids included in amino acids 65 to 79 of SEQ ID NO: 29. The epitope may be linear or structural, i.e., discontinuous. In some embodiments, the anti-CD38 antibody or antigen-binding fragment specifically binds to a human CD38 epitope comprising at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or at least 14 or more amino acid residues included in amino acids 65 to 79 of SEQ ID NO: 29. In some embodiments, the anti-CD38 antibody or antigen-binding fragment binds to an epitope containing amino acids 65-79 of SEQ ID NO: 29.

[0139] In some embodiments, the provided antibody or its antigen-binding fragment binds to mutant human CD38 (compared to non-mutant human CD38 (SEQ ID NO: 29)), where in mutant human CD38, the serine residue at position 274 is substituted with phenylalanine.

[0140] In some embodiments, the provided antibody or its antigen-binding fragment binds to mutant human CD38 (compared to non-mutant human CD38 (SEQ ID NO: 29)), where in mutant human CD38, the aspartic acid residue at position 202 is replaced with a glycine residue.

[0141] In some embodiments, the provided antibody or antigen-binding fragment binds to mutant human CD38 (compared to non-mutant human CD38 (SEQ ID NO: 29)), where in mutant human CD38, the serine residue at position 274 is substituted with phenylalanine, and the aspartic acid residue at position 202 is substituted with a glycine residue.

[0142] aCD38-b-348 and antibodies, and antigen-binding fragments derived therefrom. The anti-CD38 antibody may be the anti-CD38 antibody aCD38-b-348, for example, the aCD38-b-348 antibody disclosed in International Publication No. 2018 / 224683. The aCD38-b-348 antibody is also known as CID-103. The anti-CD38 antibody may be any antibody or its antigen-binding fragment based on or derived from such an antibody. The complementarity-determining regions CDR1, CDR2, and CDR3 of the anti-CD38 antibody are determined according to the Kabat numbering scheme.

[0143] For example, in some embodiments, the anti-CD38 antibody or its antigen-binding fragment is a) Amino acid sequence of Sequence ID No. 1 as HCDR1, b) Amino acid sequence of Sequence ID No. 2 as HCDR2, c) Amino acid sequence of Sequence ID No. 3 as HCDR3, d) Amino acid sequence of sequence number 4 as LCDR1, e) The amino acid sequence of sequence number 5 as LCDR2, and f) Amino acid sequence of sequence number 6 as LCDR3 (same as in aCD38-b-348), It may include.

[0144] In some embodiments, the anti-CD38 antibody or its antigen-binding fragment is sequence It contains a variable heavy chain containing the amino acid sequence of number 7 and / or a variable light chain containing the amino acid sequence of SEQ ID NO: 8 (same as in aCD38-b-348).

[0145] In some embodiments, the variable heavy chain sequence of the anti-CD38 antibody is the variable heavy chain sequence of aCD38-b-348, i.e., QLQLQESGPGLVKPSETLSLTCTVSG GSISSSDYYWG WIRQPPGKGLEWIG SIYYSGSTYYNPSLKS RVTISVDTSKNQFSLKLSSVTAADTAVYYC ARGQYSSGWYAYPFDM The variable light chain sequence of the anti-CD38 antibody containing WGQGTMVTVSS (SEQ ID NO: 7) is the variable light chain sequence of aCD38-b-348, i.e., EIVLTQSPGTLSLSPGERATLSC RASQSVRSSYLA WYQQKPGQAPRLLIY GASSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC QQDGNVYT Includes FGGGTKVEIK (sequence number 8).

[0146] In some embodiments, the anti-CD38 antibody may be a variant of aCD38-b-348. Such variants may be aCD38-b-348-m1, aCD38-b-348-m2, aCD38-b-348-m3, or aCD38-b-348-m4 of the antibody.

[0147] For example, in some embodiments, the anti-CD38 antibody or its antigen-binding fragment is a) Amino acid sequence of Sequence ID No. 1 as HCDR1, b) Amino acid sequence of Sequence ID No. 2 as HCDR2, c) Amino acid sequence of Sequence ID No. 3 as HCDR3, d) Amino acid sequence of sequence number 4 as LCDR1, e) The amino acid sequence of sequence number 5 as LCDR2, and f) Amino acid sequence of sequence number 9 as LCDR3 (same as in aCD38-b-348-m1), It may include.

[0148] In some embodiments, the anti-CD38 antibody or its antigen-binding fragment is a) Amino acid sequence of Sequence ID No. 1 as HCDR1, b) Amino acid sequence of Sequence ID No. 2 as HCDR2, c) Amino acid sequence of Sequence ID No. 3 as HCDR3, d) Amino acid sequence of sequence number 4 as LCDR1, e) The amino acid sequence of sequence number 5 as LCDR2, and f) Amino acid sequence of sequence number 10 as LCDR3 (same as in aCD38-b-348-m2), It may include.

[0149] In some embodiments, the anti-CD38 antibody or its antigen-binding fragment is a) Amino acid sequence of Sequence ID No. 1 as HCDR1, b) Amino acid sequence of Sequence ID No. 2 as HCDR2, c) Amino acid sequence of Sequence ID No. 3 as HCDR3, d) Amino acid sequence of sequence number 4 as LCDR1, e) The amino acid sequence of sequence number 5 as LCDR2, and f) Amino acid sequence of sequence number 11 as LCDR3 (same as in aCD38-b-348-m3), It may include.

[0150] In some embodiments, the anti-CD38 antibody or its antigen-binding fragment is a) Amino acid sequence of Sequence ID No. 1 as HCDR1, b) Amino acid sequence of Sequence ID No. 2 as HCDR2, c) Amino acid sequence of Sequence ID No. 3 as HCDR3, d) Amino acid sequence of sequence number 4 as LCDR1, e) The amino acid sequence of sequence number 5 as LCDR2, and f) Amino acid sequence of sequence number 12 as LCDR3 (same as in aCD38-b-348-m4), It may include.

[0151] In some embodiments, the anti-CD38 antibody or its antigen-binding fragment is a) A variable heavy chain containing the amino acid sequence of SEQ ID NO: 7 and / or a variable light chain containing the amino acid sequence of SEQ ID NO: 13 (same as in aCD38-b-348-m1), b) A variable heavy chain containing the amino acid sequence of SEQ ID NO: 7 and / or a variable light chain containing the amino acid sequence of SEQ ID NO: 14 (same as in aCD38-b-348-m2), c) A variable heavy chain containing the amino acid sequence of SEQ ID NO: 7 and / or a variable light chain containing the amino acid sequence of SEQ ID NO: 15 (same as in aCD38-b-348-m3), or d) A variable heavy chain containing the amino acid sequence of SEQ ID NO: 7 and / or a variable light chain containing the amino acid sequence of SEQ ID NO: 16 (same as in aCD38-b-348-m4), Includes.

[0152] Therefore, the mutant antibody aCD38-b-348-m1 is QLQLQESGPGLVKPSETLSLTCTVSG GSISSSDYYWG WIRQPPGKGLEWIG SIYYSGSTYYNPSLKS RVTISVDTSKNQFSLKLSSVTAADTAVYYC ARGQYSSGWYAYPFDM A heavy chain variable region containing the sequence WGQGTMVTVSS (sequence number 7), Array: EIVLTQSPGTLSLSPGERATLSC RASQSVRSSYLA WYQQKPGQAPRLLIY GASSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC QQEANVYT A variable light chain containing FGGGTKVEIK (SEQ ID NO: 13), It can be characterized as containing

[0153] The mutant antibody aCD38-b-348-m2 is, QLQLQESGPGLVKPSETLSLTCTVSG GSISSSDYYWG WIRQPPGKGLEWIG SIYYSGSTYYNPSLKS RVTISVDTSKNQFSLKLSSVTAADTAVYYC ARGQYSSGWYAYPFDM A heavy chain variable region containing the sequence WGQGTMVTVSS (sequence number 7), Array: EIVLTQSPGTLSLSPGERATLSC RASQSVRSSYLA WYQQKPGQAPRLLIY GASSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC QQDSNVYT A variable light chain containing FGGGTKVEIK (SEQ ID NO: 14), It can be characterized as containing

[0154] The mutant antibody aCD38-b-348-m3 is, QLQLQESGPGLVKPSETLSLTCTVSG GSISSSDYYWG WIRQPPGKGLEWIG SIYYSGSTYYNPSLKS RVTISVDTSKNQFSLKLSSVTAADTAVYYC ARGQYSSGWYAYPFDM A heavy chain variable region containing the sequence WGQGTMVTVSS (sequence number 7), Array: EIVLTQSPGTLSLSPGERATLSC RASQSVRSSYLA WYQQKPGQAPRLLIY GASSRAT GIPDRFSGSGSGTDFTLTIS RLEPEDFAVYYC QQDANVYT A variable light chain containing FGGGTKVEIK (SEQ ID NO: 15), It can be characterized as containing

[0155] The mutant antibody aCD38-b-348-m4 is, QLQLQESGPGLVKPSETLSLTCTVSG GSISSSDYYWG WIRQPPGKGLEWIG SIYYSGSTYYNPSLKS RVTISVDTSKNQFSLKLSSVTAADTAVYYC ARGQYSSGWYAYPFDM A heavy chain variable region containing the sequence WGQGTMVTVSS (sequence number 7), Array: EIVLTQSPGTLSLSPGERATLSC RASQSVRSSYLA WYQQKPGQAPRLLIY GASSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC QQEGNVYT A variable light chain containing FGGGTKVEIK (SEQ ID NO: 16), It can be characterized as containing

[0156] The present invention may also utilize mutant antibodies and their antigen-binding fragments having a certain percentage of identity with respect to a reference sequence such as the CDR sequence or heavy chain variable sequence and / or light chain variable sequence of aCD38-b-348.

[0157] For example, in some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 7. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 95% sequence identity with SEQ ID NO: 7. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 98% sequence identity with SEQ ID NO: 7. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 99% sequence identity with SEQ ID NO: 7. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing the amino acid sequence of SEQ ID NO: 7.

[0158] In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable light chain sequence containing an amino acid sequence having at least 90% sequence identity with a sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable light chain sequence containing an amino acid sequence having at least 95% sequence identity with a sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable light chain sequence containing an amino acid sequence having at least 98% sequence identity with a sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 99% sequence identity with a sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable light chain that has sequence identity with respect to a sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16.

[0159] In some embodiments, the anti-CD38 antibody or its antigen-binding fragment comprises a variable heavy chain sequence having at least 90% sequence identity with respect to SEQ ID NO: 7, and SEQ ID NO: 8, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16. The anti-CD38 antibody or its antigen-binding fragment includes a variable light chain sequence containing an amino acid sequence having at least 90% sequence identity with a sequence selected from the group. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 95% sequence identity with SEQ ID NO: 7, and a variable light chain sequence containing an amino acid sequence having at least 95% sequence identity with a sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 98% sequence identity with SEQ ID NO: 7, and a variable light chain sequence containing an amino acid sequence having at least 98% sequence identity with a sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment comprises a variable heavy chain sequence containing an amino acid sequence having at least 99% sequence identity with SEQ ID NO: 7, and a variable light chain sequence containing an amino acid sequence having at least 99% sequence identity with a sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16.

[0160] In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 7, and a variable light chain sequence containing an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 8. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 95% sequence identity with SEQ ID NO: 7, and a variable light chain sequence containing an amino acid sequence having at least 95% sequence identity with SEQ ID NO: 8. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 98% sequence identity with SEQ ID NO: 7, and a variable light chain sequence containing an amino acid sequence having at least 98% sequence identity with SEQ ID NO: 8. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 99% sequence identity with SEQ ID NO: 7, and a variable light chain sequence containing an amino acid sequence having at least 99% sequence identity with SEQ ID NO: 8. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment comprises a variable heavy chain sequence containing the amino acid sequence of SEQ ID NO: 7 and a variable light chain sequence containing the amino acid sequence of SEQ ID NO: 8.

[0161] Such mutant antibodies and their antigen-binding fragments (i.e., those having a certain percentage of identity) may retain or exhibit the same (or substantially the same) functional and pharmacological properties as described herein for antibodies and their antigen-binding fragments having heavy-chain variable sequences and light-chain variable sequences disclosed herein for aCD38-b-348, for example, that bind to the same epitope as aCD38-b-348.

[0162] In some embodiments, particularly in any embodiment referring to a sequence having a specific sequence identity with respect to a reference sequence, the % sequence identity may be calculated without including the sequences of all six CDRs of the identified heavy-chain variable region or light-chain variable region. In such embodiments, sequence changes (if any) occur only in the framework region.

[0163] In some embodiments, the anti-CD38 antibody may be an anti-CD38 antibody defined by several substitutions to the amino acid sequence elements of aCD38-b-348 (or those of mutants m1 to m4) as defined above, either substituted or additionally.

[0164] For example, such an antibody may contain a variable heavy chain complementarity determination region 3 (HCDR3) containing a sequence with up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions within aCD38-b-348-HCDR3 (SEQ ID NO: 3). In further embodiments, the anti-CD38 antibody or its antigen-binding fragment may include, as variable heavy chain complementarity determination region 1, variable heavy chain complementarity determination region 2, and variable heavy chain complementarity determination region 3 (HCDR1, HCDR2, and HCDR3), sequences containing up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in sequence numbers 1, 2, and 3, respectively, and / or as variable light chain complementarity determination region 1, variable light chain complementarity determination region 2, and variable light chain complementarity determination region 3 (LCDR1, LCDR2, and LCDR3), sequences containing up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in sequences selected from the group consisting of sequence numbers 4, 5, and sequence numbers 6, 9, 10, 11, and 12, respectively.

[0165] In further embodiments, the anti-CD38 antibody or its antigen-binding fragment may, as a variable heavy chain sequence, contain up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in SEQ ID NO: 7, and / or as a variable light chain sequence, contain up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in a sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16. In some embodiments, the anti-CD38 antibody may include, as a variable heavy chain sequence, a sequence containing up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions within the framework region of the variable heavy chain sequence of SEQ ID NO: 7, and / or as a variable light chain sequence, a sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16, containing up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions within the framework region of the variable light chain sequence.

[0166] In further embodiments, the anti-CD38 antibody or its antigen-binding fragment may, as a variable heavy chain sequence, contain up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in SEQ ID NO: 7, and / or as a variable light chain sequence, contain up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in SEQ ID NO: 8. In some embodiments, the anti-CD38 antibody may, as a variable heavy chain sequence, contain a sequence containing up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in the framework region of the variable heavy chain sequence of SEQ ID NO: 7, and / or as a variable light chain sequence, contain up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in the framework region of SEQ ID NO: 8.

[0167] In some embodiments, the anti-CD38 antibody (i.e., the antibody or its antigen-binding fragment described herein and its variants, for example, a mutant in which the DG motif has been mutated and removed) is used. a) The variable heavy chain region sequence of SEQ ID NO: 7 (or its affinity-matured variants, etc.), or the variable heavy chain region sequence having up to 5 amino acid substitutions compared to the variable heavy chain region sequence of SEQ ID NO: 7 (or its affinity-matured variants, etc.), and / or b) A variable light chain region sequence (or its affinity-matured variant, etc.) selected from the group consisting of SEQ ID NOs: 8, SEQ ID NOs: 13, SEQ ID NOs: 14, SEQ ID NOs: 15, and SEQ ID NOs: 16, or a variable light chain region sequence (or its affinity-matured variant, etc.) having up to 5 amino acid substitutions compared to a variable light chain region sequence (or its affinity-matured variant, etc.) selected from the group consisting of SEQ ID NOs: 8, SEQ ID NOs: 13, SEQ ID NOs: 14, SEQ ID NOs: 15, and SEQ ID NOs: 16, It may include.

[0168] In some embodiments, the anti-CD38 antibody (i.e., the antibody or its antigen-binding fragment described herein and its variants, for example, a mutant in which the DG motif has been mutated and removed) is used. a) The variable heavy chain region sequence of SEQ ID NO: 7 (or its affinity-matured variants, etc.), or the variable heavy chain region sequence having up to 5 amino acid substitutions compared to the variable heavy chain region sequence of SEQ ID NO: 7 (or its affinity-matured variants, etc.), and / or b) The variable light chain region sequence of SEQ ID NO: 8 (or its affinity-matured variants, etc.), or the variable light chain region sequence having up to 5 amino acid substitutions compared to the variable light chain region sequence of SEQ ID NO: 8 (or its affinity-matured variants, etc.), It may include.

[0169] In some embodiments, the anti-CD38 antibody (i.e., the antibody or its antigen-binding fragment described herein and its variants, for example, a mutant in which the DG motif has been mutated and removed) is used. a) The variable heavy chain region sequence of SEQ ID NO: 7 (or its affinity-matured variants, etc.), or a variable heavy chain region sequence having up to two amino acid substitutions compared to the variable heavy chain region sequence of SEQ ID NO: 7 (or its affinity-matured variants, etc.), and / or b) A variable light chain region sequence (or its affinity-matured variant, etc.) selected from the group consisting of SEQ ID NOs: 8, 9, 10, 11, and 12, or a variable light chain region sequence (or its affinity-matured variant, etc.) having up to two amino acid substitutions compared to a variable light chain region sequence (or its affinity-matured variant, etc.) selected from the group consisting of SEQ ID NOs: 8, 13, 14, 15, and 16, It may include.

[0170] In some embodiments, the anti-CD38 antibody (i.e., the antibody or its antigen-binding fragment described herein and its variants, for example, a mutant in which the DG motif has been mutated and removed) is used. a) The variable heavy chain region sequence of SEQ ID NO: 7 (or its affinity-matured variants, etc.), or a variable heavy chain region sequence having up to two amino acid substitutions compared to the variable heavy chain region sequence of SEQ ID NO: 7 (or its affinity-matured variants, etc.), and / or b) The variable light chain region sequence of SEQ ID NO: 8 (or its affinity-matured variants, etc.), or the variable light chain region sequence having up to two amino acid substitutions compared to the variable light chain region sequence of SEQ ID NO: 8 (or its affinity-matured variants, etc.), It may include.

[0171] Amino acid substitutions preferably do not adversely affect, or substantially adversely affect, the functional properties of the antibody. Therefore, the substitutions can be considered conservative amino acid substitutions. Preferably, when amino acid substitutions are present, they exist in a 1:1 ratio so as not to change the total length of the heavy chain variable region and / or light chain variable region.

[0172] In some embodiments, arbitrary amino acid substitutions (such as conserved amino acid substitutions) may exist only within the framework region. In such embodiments, the CDR sequence remains unchanged.

[0173] Antibodies exhibiting such amino acid sequences and any substitutions may still exhibit the binding and / or functional characteristics of aCD38-b-348 and generally anti-CD38 antibodies (such as binding to the same epitope or any of the functional characteristics described herein for disclosed anti-CD38 antibodies).

[0174] The present invention also relates to an antibody or an antigen-binding fragment thereof, wherein the light chain or heavy chain of the antibody The present invention provides an antibody or its antigen-binding fragment in which the DG motif may be modified to, for example, reduce sensitivity to aspartate isomerization, and / or any methionine in the light or heavy chain of the antibody may be modified to, for example, reduce methionine oxidation. For example, the DG motif may be modified to replace one or both of the amino acids in the motif with different amino acids. For example, such a motif may be mutated to EG, DQ, or DA. The methionine residue may be modified to replace it with a different amino acid, such as leucine or phenylalanine.

[0175] Accordingly, in some embodiments, antibodies or fragments thereof provided herein may be mutated to remove or modify DG motifs, particularly DG motifs appearing in the CDR region, as is standard practice in the art for reducing sensitivity to aspartate isomerization. Such modified antibodies may require further modification (e.g., affinity maturation) before reaching their final sequence.

[0176] In one embodiment of the present invention, a variant antibody is provided that has the CDR1, CDR2, and CDR3 sequences of the antibody disclosed herein (e.g., the CDR1, CDR2, and CDR3 sequences of aCD38-b-348), or the variable heavy chain and variable light chain of any antibody disclosed herein (e.g., the variable heavy chain and variable light chain of aCD38-b-348), but in which at least one DG motif in the CDR (if present) is changed to a different motif, which is different from the identified sequence. The disclosed variant can be used and formulated as described for aCD38-b-348.

[0177] For example, aCD38-b-348 contains a DG motif in its LCDR3 sequence. In some embodiments, the aspartic acid in the DG motif can be changed to a different amino acid, and / or the glycine in the DG motif can be changed to a different amino acid. In such embodiments, the anti-CD38 antibody or its antigen-binding fragment may be, for example, aCD38-b-348, or may be derived from aCD38-b-348. In some embodiments, the mutant antibody or its antigen-binding fragment has one of the VL CDR3 sequences of SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, or SEQ ID NO: 12. For example, a variant LCDR3 sequence (e.g., the aCD38-b-348-m1 variant LCDR3 sequence similar to the one in SEQ ID NO: 9, the aCD38-b-348-m2 variant LCDR3 sequence similar to the one in SEQ ID NO: 10, the aCD38-b-348-m3 variant LCDR3 sequence similar to the one in SEQ ID NO: 11, or the aCD38-b-348-m4 variant LCDR3 sequence similar to the one in SEQ ID NO: 12) can be incorporated into an antibody containing the LCDR1 and / or LCDR2 sequences of aCD38-b-348. In one embodiment, a mutant LCDR3 sequence (e.g., the aCD38-b-348-m1 mutant LCDR3 sequence similar to that in SEQ ID NO: 9, the aCD38-b-348-m2 mutant LCDR3 sequence similar to that in SEQ ID NO: 10, the aCD38-b-348-m3 mutant LCDR3 sequence similar to that in SEQ ID NO: 11, or the aCD38-b-348-m4 mutant LCDR3 sequence similar to that in SEQ ID NO: 12) can be incorporated into an antibody containing the LCDR1, LCDR2, HCDR1, HCDR2, and HCDR3 sequences of aCD38-b-348. In some embodiments, the mutant antibody or its antibody-binding fragment may contain, along with the variable heavy chain and variable light chain sequences of aCD38-b-348, but with a mutated LCDR3 sequence from which the DG motif has been removed (e.g., SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, or SEQ ID NO: 12 may instead be present as LCDR3).The mutant anti-CD38 antibody provides a further antibody possessing all, perhaps all, of the binding and functional properties of the parent aCD38-b-348 (e.g., binding to the same epitope or any of the functional properties described herein for the disclosed anti-CD38 antibody). It can be used and formulated as described for b-348.

[0178] The present invention may also use other or further affinity-matured antibodies, for example, affinity-matured variants derived from any of the antibodies disclosed herein. In one embodiment, the affinity-matured antibody is an affinity-matured antibody having a modified DG motif and / or NG motif, and / or modified to remove or mutate any methionine residue. The disclosed affinity-matured variants can be used and formulated as described for aCD38-b-348.

[0179] In some embodiments, the present invention provides a method for preparing an anti-CD38 antibody, comprising preparing an antibody described herein (e.g., aCD38-b-348 or its antigen-binding fragment or variant) and subjecting the antibody to affinity maturation, wherein the resulting antibody binds to CD38 with a higher affinity than the parent antibody. Preferably, the resulting antibody binds to CD38 with an affinity at least 20%, at least 30%, at least 40%, more preferably at least 50%, higher than that of the parent antibody, as measured by, for example, Kd. Methods for measuring affinity are known in the art and are described in the following examples. The affinity-matured antibody produced by such a method can be formulated and used for other anti-CD38 antibody agents as described herein.

[0180] aCD38-b-329 and antibodies, and antigen-binding fragments derived therefrom. The anti-CD38 antibody can be the aCD38-b-329 of the anti-CD38 antibody, for example, the aCD38-b-329 antibody disclosed in International Publication No. 2018 / 224685. The anti-CD38 antibody can be any antibody based on or derived from such an antibody or an antigen-binding fragment thereof.

[0181] In some embodiments, the anti-CD38 antibody or an antigen-binding fragment thereof a) has the amino acid sequence of SEQ ID NO: 17 as HCDR1, b) has the amino acid sequence of SEQ ID NO: 18 as HCDR2, c) has the amino acid sequence of SEQ ID NO: 19 as HCDR3, d) has the amino acid sequence of SEQ ID NO: 20 as LCDR1, e) has the amino acid sequence of SEQ ID NO: 21 as LCDR2, and f) has the amino acid sequence of SEQ ID NO: 22 as LCDR3 (similar to that in aCD38-b-329). It may include.

[0182] In some embodiments, the anti-CD38 antibody or an antigen-binding fragment thereof includes a variable heavy chain containing the amino acid sequence of SEQ ID NO: 23 and / or a variable light chain containing the amino acid sequence of SEQ ID NO: 24 (similar to that in aCD38-b-329).

[0183] In some embodiments, the variable heavy chain sequence of aCD38-b-329 has the sequence: QLQLQESGPGLVKPSETLSLTCTVSG GSISSSDYYWG WIRQPPGKGLEWIG SIYYSGSTYYNPSLKS RVTISVDTSKNQFSLKLSSVTAADTAVYYC ARGQYSSGWYAYPFDM WGQGTMVTVSS (SEQ ID NO: 23), and the variable light chain sequence of aCD38-b-329 has the sequence: EIVLTQSPGTLSLSPGERATLSC RASQSVRSSYLA WYQQKPGQAPRLLIY GASSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC QQDGAVFTIncludes FGGGTKVEIK (sequence number 24).

[0184] In some embodiments, the anti-CD38 antibody may be a variant of aCD38-b-329. Such variants may be aCD38-b-329-m6 or aCD38-b-329-m7 of the antibody.

[0185] For example, in some embodiments, the anti-CD38 antibody or its antigen-binding fragment is a) Amino acid sequence of Sequence ID No. 17 as HCDR1, b) Amino acid sequence of sequence number 18 as HCDR2, c) Amino acid sequence of Sequence ID No. 19 as HCDR3, d) Amino acid sequence of sequence number 20 as LCDR1, e) The amino acid sequence of sequence number 21 as LCDR2, and f) Amino acid sequence of sequence number 25 as LCDR3 (same as in aCD38-b-329-m6), It may include.

[0186] In some embodiments, the anti-CD38 antibody or its antigen-binding fragment is a) Amino acid sequence of Sequence ID No. 17 as HCDR1, b) Amino acid sequence of sequence number 18 as HCDR2, c) Amino acid sequence of Sequence ID No. 19 as HCDR3, d) Amino acid sequence of sequence number 20 as LCDR1, e) The amino acid sequence of sequence number 21 as LCDR2, and f) Amino acid sequence of sequence number 26 as LCDR3 (same as in aCD38-b-329-m7), It may include.

[0187] In some embodiments, the anti-CD38 antibody or its antigen-binding fragment is a) A variable heavy chain containing the amino acid sequence of SEQ ID NO: 23 and / or a variable light chain containing the amino acid sequence of SEQ ID NO: 27 (same as in aCD38-b-329-m6), or b) A variable heavy chain containing the amino acid sequence of SEQ ID NO: 23 and / or a variable light chain containing the amino acid sequence of SEQ ID NO: 28 (same as in aCD38-b-329-m7), Includes.

[0188] Therefore, the mutant antibody aCD38-b-329-m6 is QLQLQESGPGLVKPSETLSLTCTVSG GSISSSDYYWG WIRQPPGKGLEWIG SIYYSGSTYYNPSLKS RVTISVDTSKNQFSLKLSSVTAADTAVYYC ARGQYSSGWYAYPFDM A heavy chain variable region containing the sequence WGQGTMVTVSS (sequence number 23), Array: EIVLTQSPGTLSLSPGERATLSC RASQSVRSSYLA WYQQKPGQAPRLLIY GASSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC QQDEAVFT A variable light chain containing FGGGTKVEIK (SEQ ID NO: 27), It can be characterized as containing

[0189] The mutant antibody aCD38-b-329-m7 is, QLQLQESGPGLVKPSETLSLTCTVSG GSISSSDYYWG WIRQPPGKGLEWIG SIYYSGSTYYNPSLKS RVTISVDTSKNQFSLKLSSVTAADTAVYYC ARGQYSSGWYAYPFDM A heavy chain variable region containing the sequence WGQGTMVTVSS (sequence number 23), Array: EIVLTQSPGTLSLSPGERATLSC RASQSVRSSYLA WYQQKPGQAPRLLIY GASSRAT GIPDRFSGSGSGTDFTLTIS RLEPEDFAVYYC QQDSAVFT A variable light chain containing FGGGTKVEIK (SEQ ID NO: 28), It can be characterized as containing

[0190] The present invention can also use mutant antibodies and antigen-binding fragments thereof having a certain percentage of identity to reference sequences such as the CDR sequences or heavy chain variable sequences and / or light chain variable sequences of aCD38-b-329.

[0191] For example, in some embodiments, the anti-CD38 antibody or antigen-binding fragment thereof comprises a variable heavy chain sequence comprising an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 23. In some embodiments, the anti-CD38 antibody or antigen-binding fragment thereof comprises a variable heavy chain sequence comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 23. In some embodiments, the anti-CD38 antibody or antigen-binding fragment thereof comprises a variable heavy chain sequence comprising an amino acid sequence having at least 98% sequence identity to SEQ ID NO: 23. In some embodiments, the anti-CD38 antibody or antigen-binding fragment thereof comprises a variable heavy chain sequence comprising an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 23. In some embodiments, the anti-CD38 antibody or antigen-binding fragment thereof comprises a variable heavy chain sequence comprising the amino acid sequence of SEQ ID NO: 23.

[0192] In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable light chain sequence containing an amino acid sequence having at least 90% sequence identity with a sequence selected from the group consisting of SEQ ID NO: 24, SEQ ID NO: 27, and SEQ ID NO: 28. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable light chain sequence containing an amino acid sequence having at least 95% sequence identity with a sequence selected from the group consisting of SEQ ID NO: 24, SEQ ID NO: 27, and SEQ ID NO: 28. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable light chain sequence containing an amino acid sequence having at least 98% sequence identity with a sequence selected from the group consisting of SEQ ID NO: 24, SEQ ID NO: 27, and SEQ ID NO: 28. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 99% sequence identity with a sequence selected from the group consisting of SEQ ID NO: 24, SEQ ID NO: 27, and SEQ ID NO: 28. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable light chain that has sequence identity with respect to a sequence selected from the group consisting of SEQ ID NO: 24, SEQ ID NO: 27, and SEQ ID NO: 28.

[0193] In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 23, and a variable light chain sequence containing an amino acid sequence having at least 90% sequence identity with a sequence selected from the group consisting of SEQ ID NO: 24, SEQ ID NO: 27, and SEQ ID NO: 28. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 95% sequence identity with SEQ ID NO: 23, and a variable light chain sequence containing an amino acid sequence having at least 95% sequence identity with a sequence selected from the group consisting of SEQ ID NO: 24, SEQ ID NO: 27, and SEQ ID NO: 28. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 98% sequence identity with SEQ ID NO: 23, and a variable light chain sequence containing an amino acid sequence having at least 98% sequence identity with a sequence selected from the group consisting of SEQ ID NO: 24, SEQ ID NO: 27, and SEQ ID NO: 28. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment comprises a variable heavy chain sequence containing an amino acid sequence having at least 99% sequence identity with SEQ ID NO: 23, and a sequence selected from the group consisting of SEQ ID NO: 24, SEQ ID NO: 27, and SEQ ID NO: 28. In contrast, it includes a variable light chain sequence containing an amino acid sequence having at least 99% sequence identity. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing the amino acid sequence of SEQ ID NO: 23 and a variable light chain sequence containing an amino acid sequence selected from the group consisting of SEQ ID NO: 24, SEQ ID NO: 27, and SEQ ID NO: 28.

[0194] In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 23, and a variable light chain sequence containing an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 24. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 95% sequence identity with SEQ ID NO: 23, and a variable light chain sequence containing an amino acid sequence having at least 95% sequence identity with SEQ ID NO: 24. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 98% sequence identity with SEQ ID NO: 23, and a variable light chain sequence containing an amino acid sequence having at least 98% sequence identity with SEQ ID NO: 24. In some embodiments, the anti-CD38 antibody or its antigen-binding fragment includes a variable heavy chain sequence containing an amino acid sequence having at least 99% sequence identity with SEQ ID NO: 23, and a variable light chain sequence containing an amino acid sequence having at least 99% sequence identity with SEQ ID NO: 24.

[0195] Such mutant antibodies and their antigen-binding fragments (i.e., those having a certain percentage of identity) may retain or exhibit the same (or substantially the same) functional and pharmacological properties as described herein for antibodies and their antigen-binding fragments having heavy-chain variable sequences and light-chain variable sequences disclosed herein for aCD38-b-329 that bind to the same epitope as aCD38-b-329, or any of the functional features described herein for disclosed anti-CD38 antibodies.

[0196] In some embodiments, particularly in any embodiment referring to a sequence having a specific sequence identity with respect to a reference sequence, the % sequence identity may be calculated without including the sequences of all six CDRs of the identified heavy-chain variable region or light-chain variable region. In such embodiments, sequence changes (if any) occur only in the framework region.

[0197] In some embodiments, the anti-CD38 antibody may be an anti-CD38 antibody defined by several substitutions to the amino acid sequence element of aCD38-b-329 as defined above (or to mutant m6 or mutant m7).

[0198] For example, such an antibody may contain a sequence as a variable heavy chain complementarity determination region 3 (HCDR3) containing up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions within aCD38-b-329-HCDR3 (SEQ ID NO: 19). In further embodiments, the anti-CD38 antibody or its antigen-binding fragment may include, as variable heavy chain complementarity determination region 1, variable heavy chain complementarity determination region 2, and variable heavy chain complementarity determination region 3 (HCDR1, HCDR2, and HCDR3), sequences containing up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in sequence numbers 17, 18, and 19, respectively, and / or as variable light chain complementarity determination region 1, variable light chain complementarity determination region 2, and variable light chain complementarity determination region 3 (LCDR1, LCDR2, and LCDR3), sequences selected from the group consisting of sequence numbers 20, 21, 22, 25, and 26, respectively, containing up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in sequence numbers 20, 21, 22, 25, and 26, respectively.

[0199] In further embodiments, the anti-CD38 antibody or its antigen-binding fragment may, as a variable heavy chain sequence, contain up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in SEQ ID NO: 23, and / or as a variable light chain sequence, contain up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in a sequence selected from the group consisting of SEQ ID NO: 24, SEQ ID NO: 27, and SEQ ID NO: 28. In some embodiments, the anti-CD38 antibody may include, as a variable heavy chain sequence, a sequence containing up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions within the framework region of the variable heavy chain sequence of SEQ ID NO: 23, and / or as a variable light chain sequence, a sequence selected from the group consisting of SEQ ID NO: 24, SEQ ID NO: 27, and SEQ ID NO: 28, containing up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions within the framework region of the variable light chain sequence.

[0200] In further embodiments, the anti-CD38 antibody or its antigen-binding fragment may, as a variable heavy chain sequence, contain up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in SEQ ID NO: 23, and / or as a variable light chain sequence, contain up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in SEQ ID NO: 24. In some embodiments, the anti-CD38 antibody may, as a variable heavy chain sequence, contain a sequence containing up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in the framework region of the variable heavy chain sequence of SEQ ID NO: 23, and / or as a variable light chain sequence, contain up to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions in the framework region of SEQ ID NO: 24.

[0201] In some embodiments, the anti-CD38 antibody (i.e., the antibody or its antigen-binding fragment described herein and its variants, for example, a mutant in which the DG motif has been mutated and removed) is used. a) The variable heavy chain region sequence of SEQ ID NO: 23 (or its affinity-matured variants, etc.), or a variable heavy chain region sequence having up to 5 amino acid substitutions compared to the variable heavy chain region sequence of SEQ ID NO: 23 (or its affinity-matured variants, etc.), and / or b) A variable light chain region sequence (or its affinity-matured variant, etc.) selected from the group consisting of SEQ ID NOs: 24, SEQ ID NOs: 27, and SEQ ID NOs: 28, or a variable light chain region sequence (or its affinity-matured variant, etc.) having up to 5 amino acid substitutions compared to a variable light chain region sequence (or its affinity-matured variant, etc.) selected from the group consisting of SEQ ID NOs: 24, SEQ ID NOs: 27, and SEQ ID NOs: 28, It may include.

[0202] In some embodiments, the anti-CD38 antibody (i.e., the antibody or its antigen-binding fragment described herein and its variants, for example, a mutant in which the DG motif has been mutated and removed) is used. a) The variable heavy chain region sequence of SEQ ID NO: 23 (or its affinity-matured variants, etc.), or a variable heavy chain region sequence having up to 5 amino acid substitutions compared to the variable heavy chain region sequence of SEQ ID NO: 23 (or its affinity-matured variants, etc.), and / or b) A variable light chain region sequence of SEQ ID NO: 24 (or its affinity-matured variant or other variant thereof), or a variable light chain region sequence having up to 5 amino acid substitutions compared to the variable light chain region sequence of SEQ ID NO: 24 (or its affinity-matured variant or other variant thereof), It may include.

[0203] In some embodiments, anti-CD38 antibodies (i.e., antibodies or their antigen-binding fragments as described herein and their variants, for example, mutated DG) are used. The mutant with the motif removed is, a) The variable heavy chain region sequence of SEQ ID NO: 23 (or its affinity-matured variants, etc.), or a variable heavy chain region sequence having up to two amino acid substitutions compared to the variable heavy chain region sequence of SEQ ID NO: 23 (or its affinity-matured variants, etc.), and / or b) A variable light chain region sequence (or its affinity-matured variant, etc.) selected from the group consisting of SEQ ID NOs: 24, SEQ ID NOs: 27, and SEQ ID NOs: 28, or a variable light chain region sequence (or its affinity-matured variant, etc.) having up to two amino acid substitutions compared to a variable light chain region sequence (or its affinity-matured variant, etc.) selected from the group consisting of SEQ ID NOs: 24, SEQ ID NOs: 27, and SEQ ID NOs: 28, It may include.

[0204] In some embodiments, the anti-CD38 antibody (i.e., the antibody or its antigen-binding fragment described herein and its variants, for example, a mutant in which the DG motif has been mutated and removed) is used. a) The variable heavy chain region sequence of SEQ ID NO: 23 (or its affinity-matured variants, etc.), or a variable heavy chain region sequence having up to two amino acid substitutions compared to the variable heavy chain region sequence of SEQ ID NO: 23 (or its affinity-matured variants, etc.), and / or b) The variable light chain region sequence of SEQ ID NO: 24 (or its affinity-matured variants, etc.), or the variable light chain region sequence having up to two amino acid substitutions compared to the variable light chain region sequence of SEQ ID NO: 24 (or its affinity-matured variants, etc.), It may include.

[0205] Amino acid substitutions preferably do not adversely affect, or substantially adversely affect, the functional properties of the antibody. Therefore, the substitutions can be considered conservative amino acid substitutions. Preferably, when amino acid substitutions are present, they exist in a 1:1 ratio so as not to change the total length of the heavy chain variable region and / or light chain variable region.

[0206] In some embodiments, arbitrary amino acid substitutions (such as conserved amino acid substitutions) may exist only within the framework region. In such embodiments, the CDR sequence remains unchanged.

[0207] Antibodies exhibiting such amino acid sequences and any substitutions may still exhibit the binding and / or functional characteristics of aCD38-b-329 and generally anti-CD38 antibodies (such as binding to the same epitope or any of the functional characteristics described herein for disclosed anti-CD38 antibodies).

[0208] The present invention also provides an antibody or an antigen-binding fragment thereof, wherein the DG motif in the light or heavy chain of the antibody may be modified to reduce sensitivity to, for example, aspartate isomerization, and / or any methionine in the light or heavy chain of the antibody may be modified to reduce methionine oxidation, for example. For example, the DG motif may be modified to replace one or both of the amino acids in the motif with different amino acids. For example, such a motif may be mutated to EG, DQ, or DA. The methionine residue may be modified to replace it with a different amino acid, for example, leucine or phenylalanine.

[0209] Therefore, in some embodiments, the antibodies or fragments thereof provided herein may be mutated to remove or modify the DG motif, particularly the DG motif appearing in the CDR region, as is standard practice in the art for reducing sensitivity to aspartate isomerization. The organism may require further modifications (e.g., affinity maturation) before reaching its final sequence.

[0210] In one embodiment of the present invention, a variant antibody is provided that has the CDR1, CDR2, and CDR3 sequences of the antibody disclosed herein (e.g., the CDR1, CDR2, and CDR3 sequences of aCD38-b-329), or the variable heavy chain and variable light chain of any antibody disclosed herein (e.g., the variable heavy chain and variable light chain of aCD38-b-329), but in which at least one DG motif in the CDR (if present) is changed to a different motif, which is different from the identified sequence. The disclosed variant can be used and formulated as described for aCD38-b-329.

[0211] For example, aCD38-b-329 contains a DG motif in its LCDR3 sequence. In some embodiments, the aspartic acid in the DG motif can be changed to a different amino acid, and / or the glycine in the DG motif can be changed to a different amino acid. In such embodiments, the anti-CD38 antibody or its antigen-binding fragment may be, for example, aCD38-b-329, or may be derived from aCD38-b-329. In some embodiments, the mutant antibody or its antigen-binding fragment has either the VL CDR3 sequence of SEQ ID NO: 25 or SEQ ID NO: 26. For example, a mutant LCDR3 sequence (e.g., the aCD38-b-329-m6 mutant LCDR3 sequence similar to that in SEQ ID NO: 25, or the aCD38-b-329-m7 mutant LCDR3 sequence similar to that in SEQ ID NO: 26) can be incorporated into an antibody containing the LCDR1 and / or LCDR2 sequences of aCD38-b-329. In one embodiment, a mutant LCDR3 sequence (e.g., the aCD38-b-329-m6 mutant LCDR3 sequence similar to that in SEQ ID NO: 25, or the aCD38-b-329-m7 mutant LCDR3 sequence similar to that in SEQ ID NO: 26) can be incorporated into an antibody containing the LCDR1, LCDR2, HCDR1, HCDR2, and HCDR3 sequences of aCD38-b-329. In some embodiments, the mutant antibody or its antibody-binding fragment may contain, along with the variable heavy chain and variable light chain sequences of aCD38-b-329, but with a mutated LCDR3 sequence from which the DG motif has been removed (e.g., SEQ ID NO: 25 or SEQ ID NO: 26 may instead be present as LCDR3). The mutant anti-CD38 antibody provides further antibodies having all, perhaps all, of the binding and functional properties of the parent aCD38-b-329 (e.g., binding to the same epitope or any of the functional properties described herein for the disclosed anti-CD38 antibody). The disclosed variants can be used and formulated in the same manner as described for aCD38-b-329.

[0212] Further antibodies that may be used in the present invention The present invention may also use affinity-mature antibodies, for example, affinity-mature mutants derived from any of the antibodies disclosed herein. In one embodiment, the affinity-mature antibody is an affinity-mature antibody having a modified DG motif and / or NG motif, and / or modified to remove or mutate any methionine residue. The disclosed affinity-mature mutants can be used and formulated as described for aCD38-b-348 or aCD38-b-329.

[0213] In some embodiments, the present invention provides a method for preparing an anti-CD38 antibody, comprising preparing an antibody as described herein (e.g., aCD38-b-329 or its antigen-binding fragment or variant) and subjecting the antibody to affinity maturation, wherein the generated antibody binds to CD38 with a higher affinity than the parent antibody. Preferably, the generated antibody binds to CD38 with an affinity at least 20%, at least 30%, at least 40%, more preferably at least 50%, than the parent antibody binds to CD38, as measured by, for example, Kd. The method for measuring affinity is as described herein. This method is known in the art and is described in the following examples. Affinity-matured antibodies produced by such a method can be formulated and used as described herein for other anti-CD38 antibody agents.

[0214] Affinity maturation can be carried out according to any suitable method known to those skilled in the art. For example, in vitro antibody display systems are widely used to produce specific antibodies with high affinity. In these systems, the phenotype (i.e., antibody fragment) is linked to the genotype (i.e., antibody gene), allowing for direct determination of the antibody sequence. Several systems have been developed to achieve display of the antibody repertoire and enable subsequent selection of binders, thereby increasing the stringency of selection and enabling the selection of increasingly high-affinity variants. Antibody fragments can be expressed in yeast, ribosomes, phage display particles, or by direct coupling to DNA.

[0215] This antibody affinity maturation method belongs to two categories of mutagenesis: stochastic and non-stochastic. Error-prone polymerase chain reaction (PCR), mutator bacterial strains, and saturated mutagenesis are typical examples of stochastic mutagenesis methods. Non-stochastic techniques often use alanine scanning or site-directed mutagenesis to generate a limited collection of specific variants. Furthermore, antibody affinity can be further improved using a shuffling approach to obtain shuffled variants of the parent antibody.

[0216] Therefore, in one embodiment of the present invention, the affinity maturation method is selected from the group consisting of stochastic mutagenesis (e.g., error-prone polymerase chain reaction (PCR), mutator bacterial strain, or saturated mutagenesis), non-stochastic mutagenesis (e.g., alanine scanning or site-directed mutagenesis), shuffling (e.g., DNA shuffling, strand shuffling, or CDR shuffling), and the use of a CRISPR-Cas9 system to introduce modifications.

[0217] The affinity maturation method is described not only in Rajpal et al., Proc Natl Acad Sci USA, 2005, 102(24):8466-71 and Steinwand et al., MAbs, 2014, 6(1):204-18, but also in Handbook of Therapeutic Antibodies, Wiley, 2014, Chapter 6, Antibody Affinity (pp. 115-140).

[0218] The present invention may also use an anti-CD38 antibody or its antigen-binding fragment (e.g., a fragment containing or not containing part of a specified CDR or variable chain sequence, or a variant having a certain percentage of identity and / or amino acid substitutions) that competes with any of the disclosed antibodies, such as aCD38-b-348 or aCD38-b-329 or any induced variant thereof, for binding to CD38.

[0219] Administration of anti-CD38 antibody In some embodiments, the method described herein includes obtaining a sample from a patient who has been previously administered an anti-CD38 antibody or its antigen-binding fragment (in other words, a sample provided or obtained from a patient who has been administered an anti-CD38 antibody or its antigen-binding fragment at a prior time). In other embodiments, the method described herein further includes the step of obtaining a sample from a patient after administering an anti-CD38 antibody to the patient. In all cases, since a blood sample is obtained from a patient after administering an anti-CD38 antibody, the blood sample contains the administered anti-CD38 antibody (for anti-CD38 antibodies are generally administered intravenously or subcutaneously and circulate in the patient's bloodstream).

[0220] Generally, the blood sample obtained from the patient contains a portion of the therapeutic anti-CD38 antibody molecule. The anti-CD38 antibody should be administered within the time frame intended for the patient to obtain the sample. In some embodiments, the patient is administered the anti-CD38 antibody less than one year, six months, three months, two months, or one month before obtaining the sample from the patient. In some embodiments, the patient is administered the anti-CD38 antibody less than eight weeks, seven weeks, six weeks, five weeks, four weeks, three weeks, two weeks, or one week before obtaining the sample from the patient. In some embodiments, the patient is administered the anti-CD38 antibody less than ten days, nine days, eight days, seven days, six days, five days, four days, three days, two days, or one day before obtaining the sample from the patient. In some embodiments, patients are administered anti-CD38 antibodies less than 48 hours, less than 24 hours, less than 12 hours, less than 6 hours, less than 5 hours, less than 4 hours, less than 3 hours, less than 2 hours, or less than 1 hour before obtaining a sample from the patient.

[0221] In a preferred embodiment, the patient is administered an anti-CD38 antibody less than two months prior to obtaining a sample from the patient.

[0222] In embodiments including the step of administering an anti-CD38 antibody or its antigen-binding fragment, the administration step is performed before any step of obtaining a sample from the patient. Depending on the patient's exact treatment regimen, there may be a delay between the administration of the anti-CD38 antibody or its antigen-binding fragment and the acquisition of a sample from the patient. Generally, the sample will be obtained less than two months after the administration of the anti-CD38 antibody or its antigen-binding fragment to the patient.

[0223] The anti-CD38 antibody administered to the patient before sample collection is an anti-CD38 antibody that does not cause interference when cross-matching the patient's blood sample with donor red blood cells or when performing any antibody-RBC panel assay. Therefore, the anti-CD38 antibody or its antigen-binding fragment is not (e.g.) daratumumab or isatuximab. Anti-CD38 antibodies compatible with this invention are discussed elsewhere under the heading "Anti-CD38 antibody or its antigen-binding fragment".

[0224] Assay type The methods described herein include the step of screening a blood sample from a patient. The screening may include determining the presence or absence of one or more patient antibodies in the patient blood sample. In some embodiments, the screening is performed using an assay selected from the group consisting of column agglutination assays, indirect antiglobulin (IAT) tube assays, and solid-phase assays. These are further described in the definitions given above.

[0225] Red blood cell antigen: The term "red blood cell antigen" refers to any antigen found on the surface of red blood cells or expressed by red blood cells. RBC antigens can be classified according to the region in which they are found. For example, the "K" and "k" RBC antigens are found on the "Kell" glycoprotein and belong to the "Kell group" of RBC antigens. A group of antigens may be called a "blood group" or "blood system," for example, a "Kell blood group." Each blood group may contain several different antigens, sometimes as many as 50 or more. RBCs from a single individual may be positive or negative for different antigens within the same blood group.

[0226] RBC antigens can be carbohydrates, such as ABO group antigens, or proteins, such as rhesus monkey (Rh) group antigens. When a patient is exposed to RBC antigens, the patient develops resistance to them. Antibodies against the progenitor, such as alloantibodies of the IgG or IgM subtype, may be produced. When patient antibodies bind to the RBC antigen in vivo, it may lead to the destruction (hemolysis) of RBCs. Due to this potentially harmful effect, patients receiving transfusions should be screened to identify any antibodies in the patient's plasma or serum that specifically bind to the RBC antigen. RBC antigens that can produce clinically significant patient antibodies include those of the Ab, ABO, Cromer, Diego, Duffy, Gerbich, GLOB, Indian, Kell, Kidd, Knops, Lewis, Lutheran, LW, MNS, P1, Rh, XK, Xg, and Yt blood groups. Exemplary blood group antigens are described in more detail below.

[0227] ABO blood group antigens are the most immunogenic of all blood groups and form the basis of routine blood typing in blood banks. ABO blood group antigens are RBC membrane-bound and attached to oligosaccharide chains protruding from the RBC surface. This group consists of four antigens: A, B, AB, and A1. Naturally occurring antibodies against ABO blood group antigens are frequently found in serum; for example, patients with blood group A have anti-B antibodies in their serum. Patient antibodies that specifically bind to ABO blood group antigens are naturally occurring (i.e., these antibodies arise, for example, without exposure to non-self antigens through transfusion) and are very common; therefore, all patients are screened to determine their ABO status. Antibodies against ABO blood group antigens can be IgG or IgM. Antibodies against ABO blood group antigens can cause serious acute hemolytic transfusion reactions if incompatible RBCs are transfused. ABO typing is not affected by the presence of anti-CD38 antibodies, such as daratumumab or isatuximab, in the patient's blood.

[0228] The Rh blood group is one of the most complex blood groups, containing at least 50 known antigens, of which D, C, E, c, and e are the most clinically significant. Rh group antigens are highly immunogenic. Rh group antigens are found on the proteins RhD and RhCE, both of which are transmembrane RBC proteins. Rh phenotyping is routinely performed before a patient receives a blood transfusion and typically uses monoclonal or polyclonal anti-D, anti-C, anti-E, anti-c, and anti-e reagents that bind to any Rh group antigen present on the RBC. Therefore, routine Rh phenotyping is typically not affected by the presence of anti-CD38 antibodies such as daratumumab or isatuximab in the patient's blood. Patient antibodies against Rh blood group antigens, particularly RhD, are a major cause of hemolytic disease of the fetus and neonatal fetus (HDFN).

[0229] The Kell blood group system arises from Kell glycoproteins, which are transmembrane RBC proteins containing Kell antigens. The Kell blood group system is complex, consisting of over 30 antigens, many of which are highly immunogenic. The two main Kell group antigens are K and k, with K being the most immunogenic. Antibodies against Kell group antigens are typically IgG antibody class antibodies, with IgM antibodies being less common. Antibodies known to cause adverse reactions (such as transfusion reactions or HDFN) include anti-K, anti-k, and anti-Kp. a , and anti-Js b One example is the Kell group antigen, which is included in the antigens that are denatured by treating RBCs with antigen stripping agents such as DTT or analogues. This makes it particularly difficult to accurately identify patient antibodies against the Kell group antigen in patients treated with anti-CD38 antibodies such as daratumumab or isatuximab, where CD38 is removed from RBCs using DTT or analogues to prevent agglutination caused by the presence of anti-CD38 antibodies.

[0230] The Kidd blood system arises from the Kidd (JK) glycoprotein, a transmembrane glycoprotein expressed on red blood cell cells (RBCs) that transport urea across the red blood cell membrane. There are three known Kidd antigens (Jk1(Jk a ), Jk2 (Jk b ), and Jk3) are also present. Anti-Kidd patients Antibodies, especially anti-Jk a It is known to cause delayed hemolytic transfusion reactions. Fetal Kidd antigen may also cause maternal alloimmunization.

[0231] The Duffy blood group includes six known antigens present on the Duffy transmembrane glycoprotein expressed on RBCs, also known as DARC (Duffy antigen / chemokine receptor). The six known Duffy antigens are Fy a Fy b Examples include Fy3, Fy4, Fy5, and Fy6. Patient antibodies against the Duffy antigen are mainly IgG subclass antibodies, while IgM subclass patient antibodies are rare. Patient antibodies against the Duffy antigen, especially Fy a and Fy b Patient antibodies against [the substance] are known to cause both hemolytic transfusion reactions and hemolytic blood follicle necrosis (HDFN).

[0232] The Diego blood group originates from the Diego protein, a transmembrane protein expressed on RBCs. The Diego blood group contains 21 antigens, the most important of which is Di a , Di b , and Wr a Patient antibodies against Diego blood group antigens may be IgG antibodies and / or IgM antibodies. Patient antibodies against Diego blood group antigens are known to cause both hemolytic transfusion reactions and HDFN.

[0233] The Lutheran blood group contains 24 known antigens. These are formed from two Lutheran glycoprotein isoforms expressed on the RBC membrane. Patient antibodies against Lutheran group antigens may be IgG and / or IgM antibodies. Patient antibodies against Lutheran group antigens can cause hemolytic transfusion reactions or HDFN, although these are rare and typically mild. Lutheran group antigens are included in antigens that are denatured by treating RBCs with antigen stripping agents such as DTT or analogues. This makes it particularly difficult to accurately identify patient antibodies against Lutheran group antigens in patients treated with anti-CD38 antibodies such as daratumumab or isatuximab, where CD38 has been removed from RBCs using DTT or analogues to prevent agglutination caused by the presence of anti-CD38 antibodies.

[0234] The MNS blood group system is derived from glycophorin A and glycophorin B, which are antigen-containing proteins and are both glycophorin proteins expressed on the RBC membrane. The MNS blood group system is known to contain at least 43 antigens, many of which are related to HDFN. Patient antibodies against MNS blood group antigens are also known to cause a variety of hemolytic transfusion reactions, from mild to severe. Patient antibodies against MNS blood group antigens may be IgG antibodies and / or IgM antibodies.

[0235] Other blood group antigens, such as those of the Cromer, Gerbich, GLOB, Indian, Knops, Lewis, LW, P1, XX, Xg, and Yt groups, may also be expressed on RBCs. If RBCs carrying these antigens are transfused to a patient who has antibodies that specifically bind to any of the antigens within that group, it may cause a hemolytic transfusion reaction or hemolytic hemolytic filamentous necrosis (HDFN).

[0236] The presence of patient antibodies against any of the above blood group antigens in patient samples is routinely evaluated in blood banks and is known to be influenced by the presence of anti-CD38 antibodies such as daratumumab or isatuximab in the patient's blood. In contrast, the method of the present invention enables accurate antibody screening and blood cross-matching of patient samples when the patient is being treated with anti-CD38 antibodies, and does not require an additional step to inhibit the binding of anti-CD38 antibodies to CD38 (membrane-bound CD38) expressed on RBCs.

[0237] Therefore, in some embodiments, the RBC antigen is the Ab group antigen, ABO group antigen, Cromer group antigen, Diego group antigen, Duffy group antigen, Gerbich group antigen, The antigens may be selected from the following groups: GLOB group antigen, Indian group antigen, Kell group antigen, Kidd group antigen, Knops group antigen, Lewis group antigen, Lutheran group antigen, LW group antigen, MNS group antigen, P1 group antigen, Rh group antigen, XK group antigen, Xg group antigen, and Yt group antigen.

[0238] RBC antigens are expressed by RBCs. This means that RBC antigens are present on the surface of RBCs. Because RBC antigens are incorporated into the RBC membrane with at least a portion exposed on the RBC surface, they are sometimes called membrane-bound antigens.

[0239] A more specific method of the present invention The present invention includes at least the following methods.

[0240] The present invention is a method for screening blood samples obtained from patients who have been administered an anti-CD38 antibody or its antigen-binding fragment, a) Prepare a blood sample from the patient, b) Prepare a blood sample from the blood donor, Here, the donor blood sample contains donor red blood cells; c) Preparing a patient blood / donor red blood cell mixture by contacting the patient's blood sample with donor red blood cells from the donor's blood sample, d) Incubating a mixture of patient blood / donor red blood cells to allow one or more patient antibodies in the patient blood sample to bind to one or more red blood cell antigens present on the donor red blood cells, if present, to form one or more patient antibody / donor red blood cell antigen complexes, e) Adding to a mixture of patient blood / donor red blood cells an agglutinant, such as anti-human globulin, which is a substance that specifically binds any complex of patient antibodies / donor red blood cell antigens present in the mixture of patient blood / donor red blood cells to each other. f) Optionally, if present, separate any one or more patient antibody / donor red blood cell antigen complexes from the patient blood / donor red blood cell mixture, Here, optionally, the separation step may include centrifugation; g) Determining the presence or absence of one or more patient antibodies in the patient blood sample that specifically bind to one or more red blood cell antigens expressed on the surface of donor red blood cells, This provides a method that includes [something].

[0241] The present invention also provides a method for screening blood samples obtained from patients who have been administered an anti-CD38 antibody or its antigen-binding fragment, a) Prepare a blood sample from the patient, Here, the patient blood sample contains one or more patient antibodies that specifically bind to one or more erythrocyte antigens, and these patient antibodies do not specifically bind to any erythrocyte antigen expressed by the patient's erythrocytes; b) Prepare a blood sample from the blood donor, Here, the donor blood sample contains donor red blood cells; c) Preparing a patient blood / donor red blood cell mixture by contacting the patient's blood sample with donor red blood cells from the donor's blood sample, d) Incubating a mixture of patient blood / donor red blood cells to enable the binding of one or more patient antibodies in the patient blood sample to one or more red blood cell antigens present on the donor red blood cells, thereby forming one or more patient antibody / donor red blood cell antigen complexes, e) Adding a coagulant, such as anti-human globulin, to a mixture of patient blood and donor red blood cells, which is a substance that specifically binds the complexes of patient antibodies and donor red blood cell antigens present in the mixture of patient blood and donor red blood cells to each other. f) Optionally, a complex of any one or more patient antibodies / donor red blood cell antigens is added to patient blood / donor blood. Separation from a mixture of red blood cells, Here, optionally, the separation step may include centrifugation; g) Determining the presence of one or more patient antibodies in the patient blood sample that specifically bind to one or more red blood cell antigens expressed on the surface of donor red blood cells, The method includes, wherein the presence of patient antibodies that specifically bind to one or more erythrocyte antigens present on one or more erythrocytes from a donor blood sample indicates that the donor blood sample is incompatible with the patient.

[0242] The present invention also provides a method for screening blood samples obtained from patients who have been administered an anti-CD38 antibody or its antigen-binding fragment, a) Prepare a blood sample from the patient, Here, the patient blood sample does not contain any patient antibodies that specifically bind to any red blood cell antigen; b) Prepare a blood sample from the blood donor, Here, the donor blood sample contains donor red blood cells; c) Preparing a patient blood / donor red blood cell mixture by contacting the patient's blood sample with donor red blood cells from the donor's blood sample, d) Adding an agglutinating agent, such as anti-human globulin, to a mixture of patient blood and donor red blood cells, e) Optionally, isolate donor red blood cells from a mixture of patient blood / donor red blood cells, Here, optionally, the isolation step includes centrifugation; f) Determining the absence of one or more patient antibodies in the patient blood sample that specifically bind to one or more red blood cell antigens expressed on the surface of donor red blood cells, The method includes, wherein the absence of patient antibodies that specifically bind to one or more erythrocyte antigens present on one or more erythrocytes from a donor blood sample indicates that the donor blood sample is compatible with the patient.

[0243] The present invention also provides a method for screening blood samples obtained from patients who have been administered an anti-CD38 antibody or its antigen-binding fragment, a) Prepare a blood sample from the patient, b) Screening the patient's blood sample against a red blood cell panel to determine the presence or absence of any patient antibody in the patient's blood sample that specifically binds to any red blood cell antigen present on the surface of any red blood cell in the red blood cell panel, c) Prepare a blood sample from the blood donor, Here, the donor blood sample contains donor red blood cells, and the donor red blood cells in the donor blood sample do not express any red blood cell antigens that can be specifically bound by any patient antibody identified in step (b) to any red blood cell antigen expressed on the surface of any red blood cell in the red blood cell panel; d) Screening a patient blood sample, including determining the presence or absence of one or more patient antibodies in the patient blood sample that specifically bind to one or more red blood cell antigens expressed on the surface of donor red blood cells, Step (c) may be carried out according to any suitable screening method disclosed herein.

[0244] The present invention also provides a method for screening blood samples obtained from patients who have been administered an anti-CD38 antibody or its antigen-binding fragment, a) Prepare one or more blood samples from the patient, b) Screening the patient blood sample prepared in step (a) against a red blood cell panel to determine the presence or absence of any patient antibody in the patient blood sample that specifically binds to any red blood cell antigen present on the surface of any red blood cell in the red blood cell panel, c) Prepare a blood sample from the blood donor, Here, the donor blood sample contains donor red blood cells, and the donor red blood cells in the donor blood sample do not express any red blood cell antigens that can be specifically bound by any patient antibody identified in step (b) to any red blood cell antigen expressed on the surface of any red blood cell in the red blood cell panel; d) Preparing a patient blood / donor red blood cell mixture by contacting the patient blood sample prepared in step (a) with donor red blood cells from the donor blood sample, e) Incubating a mixture of patient blood / donor red blood cells to allow one or more patient antibodies in the patient blood sample to bind to one or more red blood cell antigens present on the donor red blood cells, if present, to form one or more patient antibody / donor red blood cell antigen complexes, f) Adding an agglutinating agent, such as anti-human globulin, to a mixture of patient blood and donor red blood cells, which specifically binds any complex of patient antibodies and donor red blood cell antigens present in the mixture. g) Optionally, if present, separate any one or more patient antibody / donor red blood cell antigen complexes from the patient blood / donor red blood cell mixture, Here, optionally, the separation step may include centrifugation; h) Determining the presence or absence of one or more patient antibodies in the patient blood sample that specifically bind to one or more red blood cell antigens expressed on the surface of donor red blood cells, Includes methods.

[0245] Any of the above methods can be combined with any of the more detailed or preferred embodiments of the present disclosure described herein.

[0246] The incubation step of any of the methods described herein may be carried out under conditions sufficient to allow any patient antibody (if present) to bind to any erythrocyte antigen present on the erythrocytes of the donor blood sample, or to any erythrocyte antigen present on the erythrocytes of the erythrocyte panel. The incubation may be carried out for about 5 minutes to about 2 hours. The incubation may be carried out for at least about 5 minutes, at least about 10 minutes, at least about 15 minutes, at least about 20 minutes, at least about 30 minutes, at least about 40 minutes, at least about 50 minutes, at least about 1 hour, at least about 90 minutes, or at least about 2 hours. The incubation may be carried out at about 37°C. The incubation may be carried out at near room temperature (e.g., about 15°C to about 25°C). In some embodiments, the incubation step includes incubating at a temperature of about 15°C to about 40°C for at least about 5 minutes.

[0247] Treatment methods and medical use The present invention provides a method for treating cancer in a patient, comprising preparing a blood sample from the patient and screening the blood sample according to the method of the present invention. In some embodiments, the patient has been administered an anti-CD38 antibody or its antigen-binding fragment (i.e., the patient has already received the anti-CD38 antibody or its antigen-binding fragment at an earlier point in time). In other embodiments, the method comprises a series of administrations of the anti-CD38 antibody or its antigen-binding fragment to the patient. In some embodiments, the method may optionally or additionally include the step of obtaining a sample from the patient. The sample is a blood sample and can be obtained according to any suitable method known to those skilled in the art (e.g., simple blood collection). The sample may be screened after processing, for example, by diluting the sample or by performing other processing steps as discussed elsewhere.

[0248] In some embodiments, the above method is a) Administering the patient an anti-CD38 antibody or its antigen-binding fragment, b) Obtaining a blood sample from the patient after administration of an anti-CD38 antibody or its antigen-binding fragment, c) Screening a patient blood sample according to the method described in any one of claims 1 to X, Includes.

[0249] The method of the present invention may include preparing a report indicating the presence or absence of patient antibodies in a patient's blood sample and / or the suitability of a blood donor who provides blood or red blood cells to the patient.

[0250] The treatment method may include the step of administering blood or red blood cells from a blood donor, for example, by transfusion, if the blood donor is found to be compatible with the patient.

[0251] Therefore, the method of the present invention extends to a method for transfusing blood to a patient, wherein compatible donor blood or donor red blood cells are administered to the patient, and the donor blood or donor red blood cells are determined to be compatible with the patient according to one of the screening methods of the present invention. Therefore, the donor blood is cross-matched with the patient and is administered to the patient only if it is found to be compatible. For example, in some embodiments, the above method is a method for transfusing blood to a patient, a) Prepare donor blood or donor red blood cells, Here, donor blood or donor red blood cells are determined to be compatible with the patient according to any screening method of the present invention; b) Administering the patient compatible donor blood or compatible donor red blood cells, This method includes [something].

[0252] The patients are those who have been administered anti-CD38 antibodies or their antigen-binding fragments at an early stage, particularly before a blood sample was obtained from the patient and before the compatibility of the donor blood or donor red blood cells with the patient was determined.

[0253] The method of administering blood transfusions to a patient may include a step of screening the donor blood or donor red blood cells for compatibility with the patient (i.e., a cross-matching test).

[0254] In some embodiments, the present invention includes the treatment of cancers such as B-cell malignancies, lymphomas (Hodgkin lymphoma, non-Hodgkin lymphoma, chronic lymphocytic leukemia, acute lymphoblastic leukemia, myeloma), myeloproliferative disorders, and solid tumors (breast cancer, squamous cell carcinoma, colon cancer, head and neck cancer, lung cancer, genitourinary cancer, rectal cancer, gastric cancer, sarcoma, melanoma, esophageal cancer, liver cancer, testicular cancer, cervical cancer, mast cell tumor, hemangioma, eye cancer, laryngeal cancer, oral cancer, mesothelioma, skin cancer, rectal cancer, pharyngeal cancer, bladder cancer, breast cancer, uterine cancer, prostate cancer, lung cancer, pancreatic cancer, kidney cancer, gastric cancer, non-small cell lung cancer, and ovarian cancer, etc.). Cancer may be defined based on the presence of specific tumor-associated markers and antigens such as CD20, HER2, PD-1, PD-L1, SLAM7F, CD47, CD137, CD134, TIM3, CD25, GITR, CD38, and EGFR, or it may be cancer identified as having a biomarker called microsatellite instability-high (MSI-H) or mismatch repair deficiency (dMMR). Furthermore, such conditions may be considered to define precancerous, non-invasive states of the above cancers, such as carcinoma in situ, smoldering myeloma, monoclonal immunoglobulinemia of unknown significance, cervical intraepithelial neoplasm, MALTomas / GALTomes, and various lymphoproliferative disorders. Preferably, in some embodiments, the subject to be treated has a solid tumor. In one embodiment, the subject has a hematological malignancy. In some embodiments, the subject has a CD38-positive tumor.

[0255] The method and use of the present invention are particularly useful when a patient may require one or more blood transfusions. For example, in some embodiments, the disease being treated is cancer, such as multiple myeloma. Patients receiving anti-CD38 antibodies as treatment for multiple myeloma may require one or more blood transfusions that require blood screening (particularly cross-matching with donors) to identify a donor who is compatible with the patient.

[0256] Accordingly, in some embodiments, the present invention provides a method for treating cancer in a subject, comprising administering to the subject an effective amount of a composition comprising an anti-CD38 antibody or its antigen-binding fragment (e.g., aCD38-b-348 or aCD38-b-329 or antibodies derived therefrom) as described herein. In some embodiments, the provided method may further include administering to the subject at least one additional active agent or therapy simultaneously or sequentially in any order (i.e., so that the subject receives combination therapy). In some embodiments, such at least one additional active agent or therapy may be an anticancer drug (e.g., a chemotherapeutic agent), radiotherapy (by irradiating the body from an external source), or a radio-conjugated compound. By administering a compound, antitumor antigen or marker antibody (the antigen or marker is, for example, CD4, CD25, CA125, PSMA, c-MET, VEGF, CD137, VEGFR2, CD20, HER2, HER3, SLAMF7, CD326, CAIX, CD40, CD47, or EGF receptor), checkpoint inhibitor or immunomodulatory antibody (for example, antibodies targeting PD-1, PD-L1, TIM3, CD25, GITR, CD134, CD134L, CD137L, CD80, CD86, B7-H3, B7-H4, B7RP1, LAG3, ICOS, TIM3, GAL9, CD28, AP2M1, SHP-2, OX-40, etc.), vaccine, adjuvant, usage protocol, one or more other compounds that target cancer cells or stimulate an immune response against cancer cells, or any combination thereof, or may include them. In certain embodiments, where at least one additional active agent or therapy is or includes an antibody, the form of such antibody and / or the antigen targeted by such antibody may be selected from those listed in the literature and possibly adapted to a given cancer (Sliwkowski M & Mellman I, 2013, Redman JM et al., 2015, Kijanka M et al., 2015).

[0257] Among other methods, the present invention is a method for screening donor blood samples for compatibility with patients who have been administered an anti-CD38 antibody or its antigen-binding fragment, a) Prepare blood samples obtained from the patient, b) Prepare a blood sample from the blood donor, Here, the donor blood sample contains donor red blood cells; c) Preparing a patient blood / donor red blood cell mixture by contacting the patient's blood sample with donor red blood cells from the donor's blood sample, d) Incubate the patient blood / donor red blood cell mixture at a temperature of approximately 15°C to approximately 40°C for at least 5 minutes to allow one or more patient IgG alloantibodies in the patient blood sample to bind to one or more red blood cell antigens present on the donor red blood cells, if present, to form one or more patient IgG alloantibodies / donor red blood cell antigen complexes, e) Adding anti-human IgG antibodies to a mixture of patient blood / donor red blood cells to agglutinate any patient IgG alloantibody / donor red blood cell antigen complexes present in the mixture of patient blood / donor red blood cells, f) If present, separate any one or more patient IgG alloantibody / donor red blood cell antigen complexes from the patient blood / donor red blood cell mixture, Here, optionally, the separation step includes centrifugation; g) Determining the presence or absence of one or more patient IgG alloantibodies in the patient blood sample that specifically bind to one or more erythrocyte antigens expressed on the surface of donor erythrocytes, Here, the aggregation of the patient antibody / donor red blood cell antigen complex in step (e) is performed by the donor blood The absence of agglutination of the patient antibody / donor red blood cell antigen complex in step (e) indicates that the donor blood is compatible with the patient; Including, and further, The patient's blood sample is either a plasma sample or a serum sample. Anti-CD38 antibodies are aCD38-b-348 or antibodies derived therefrom, or their variants. Anti-CD38 antibodies are IgG isotype antibodies, When anti-human IgG antibody is added in step (e), the anti-CD38 antibody does not cause red blood cell agglutination, and The method provides a procedure in which the patient has been administered an anti-CD38 antibody at least two months prior to obtaining a blood sample from the patient.

[0258] The aspects and embodiments described herein using the term "comprising" may include other features or processes within that scope. It should also be understood that the aspects and embodiments described using "comprising" may also include aspects and embodiments in which the term "comprising" is replaced with the terms "consisting essentially of" or "consisting of."

[0259] The phrase "selected from the group comprising" is used in this specification, where it is present, to mean "selected from the group consisting of". It can be replaced with the phrase "from the group consisting of)" and vice versa. It is the same.

[0260] Furthermore, unless otherwise required by context, this application discloses all combinations of any of the above embodiments and models. Similarly, unless otherwise required by context, this application discloses all combinations of preferred features and / or any features, either individually or in any of the other embodiments.

[0261] The present invention will now be further described with reference to the drawings by the following embodiments, which are intended to help those skilled in the art to carry out the invention and are not intended to limit the scope of the invention in any way. [Examples]

[0262] Example 1: Binding of CID103 (aCD38-b-348) to CD-38-expressing malignant cell lines Materials and methods The cell lineage / primary cell type used was investigated by flow cytometry using multiple human cell lineages that endogenously express CD38 (Daudi (ATCC CCL-213), Raji (ATCC CCL-86), and Ramos (ATCC CRL-1596)). Prior to the experiment, Daudi, Raji, Ramos, and cells were cultured in RPMI-1640 medium (ATCC 30-2001) supplemented with 10% FCS, 2 mM L-glutamine, 100 IU / ml penicillin, 100 μg / ml streptomycin, and 1 mM sodium pyruvate (all from ThermoFisher) during the logarithmic phase, with a viability of over 85% as measured by an automated cell counter (Countess II FL automated cell counter; ThermoFisher; catalog number AMQAF1000).

[0263] Cell staining and flow cytometry. All experiments were performed in triplicate using 96-well plates. 50,000 cells were seeded in each well of a 96-well round-bottom tissue culture plate. FACS cell staining buffer (0.2% BSA, 0.02% NaN3) was used. Following the washing process using BioLegend (catalog number 420201), the cells are then treated with CID103 Cells were incubated on ice for 30 minutes in seven 3-fold dilutions of (aCD38-b-348), daratumumab, or human IgG1 isotype control (0.03 μg / ml, 0.08 μg / ml, 0.24 μg / ml, 0.74 μg / ml, 2.2 μg / ml, 6.6 μg / ml, and 20 μg / ml). Following a wash with FACS cell staining buffer, cells were incubated with a secondary antibody (Alexa Fluor 647-labeled rabbit anti-human Fcγ F(ab')2; Jackson ImmunoResearch catalog no. 309-606-008) at 5 μg / ml in the dark on ice for 30 minutes. Cells were washed again with FACS cell staining buffer and analyzed by flow cytometry using a BD Biosciences FACSCalibur flow cytometer (San Jose, California). Target cells were identified using FSC vs. SSC gating. A total of 3000 living single cells were recorded for each sample, and the corresponding fluorescence signal intensity histograms were analyzed. The mean fluorescence intensity (MFI) was calculated for each antibody concentration using FlowJo analysis software (Ashland, Oregon).

[0264] EC 50 Calculation of EC 50 To calculate the values, MFI was plotted against antibody concentration (semi-logarithmic graph), and the data was fitted to a nonlinear regression curve using GraphPad Prism 8 software (La Jolla, California).

[0265] result EC study on the binding of CID103 (aCD38-b-348) and daratumumab to Daudi, Raji, and Ramos cell lineages. 50 The maximum MFI values ​​are shown in Table 1 below. EC for IgG1 isotype control (negative control) 50 The maximum MFI value is also included.

[0266] [Table 1]

[0267] The dose-response curves for binding to the Daudi cell lineage, the Raji cell lineage, and the Ramos cell lineage are shown in Figures 1, 2, and 3, respectively.

[0268] Daratumumab showed typical dose-response binding curves for Daudi, Raji, and Ramos cells. CID103 (aCD38-b-348) showed similar saturable, concentration-dependent binding to all three cell lines. EC of CID103 50 The values ​​were 382 ng / ml (Daudi cells), 373 ng / ml (Raji cells), and 412 ng / ml (Ramos cells), indicating the EC of daratumumab. 50 Value (166 ng / ml ~ 199 The binding of CID103 (aCD38-b-348) to test cells was slightly higher than that of daratumumab (in the case of Raji cells) or slightly higher than that of daratumumab (in the case of Daudi and Ramos cells). The IgG1 isotype control did not show binding to any of the test cell lines.

[0269] Example 2: Binding of CID-103 to donor RBCs Materials and methods Primary cell types used. Binding of CID103 (aCD38-b-348) to human erythrocytes was examined by flow cytometry using fresh human whole blood from three (3) normal, healthy donors. Fresh heparinized whole blood was approved by the Institutional Review Board for Human Participants (IRB) for human subjects. The tissue samples were obtained from the facility for in vitro research purposes. Whole blood was visually examined for hemolysis before the experiment.

[0270] Preparation of donor RBCs. Whole blood was centrifuged and washed three times with 1×PBS before use. The washed whole blood was diluted 1:20 to prepare a 5% erythrocyte suspension in PBS (hereinafter referred to as the blood substrate). The blood substrate was used in the assay within 3 hours of preparation.

[0271] Cell staining and flow cytometry. 100 μl of blood substrate mixed with 100 μl of FACS buffer was seeded per well on a 96-well round-bottom tissue culture plate. Seven 3-fold dilutions (0.03 μg / ml, 0.08 μg / ml, 0.24 μg / ml, 0.74 μg / ml, 2.2 μg / ml, 6.6 μg / ml, and 20 μg / ml) of CID103 (aCD38-b-348), human IgG1 isotype control antibody (BioXcell; catalog number BE0297), daratumumab (Darzalex), or Alexa Fluor 647-labeled CD47 antibody (BioLegend; catalog number 32) were used. The samples were incubated on ice for 30 minutes with one of the following: 3118). Each condition was performed in three consecutive cycles. Following a washing step with FACS buffer, the samples were incubated with Alexa Fluor 647-labeled goat anti-human Fcγ F(ab')2 secondary antibody (Jackson ImmunoResearch; catalog no. 109-606-170) at 5 μg / ml in the dark on ice for 30 minutes. The samples were washed again with FACS buffer to remove unbound secondary antibody and analyzed by flow cytometry using an Intellicyt flow cytometer. Samples incubated with Alexa Fluor 647-labeled CD47 antibody were processed for flow cytometry analysis without incubation with the secondary antibody. Erythrocytes were identified using FSC vs. SSC gating. At least 3000 live erythrocytes were collected for each sample, and the corresponding fluorescence signal intensity histograms were analyzed. Mean fluorescence intensity (MFI) was determined for each antibody concentration using ForeCyt analysis software.

[0272] EC 50 Calculation of EC50 To calculate the values, MFI was plotted against antibody concentration (semi-logarithmic graph), and the data was fitted to a nonlinear regression curve using GraphPad Prism 8 software (La Jolla, California).

[0273] result EC study on the binding of CID103 (aCD38-b-348) and daratumumab to RBCs from each of the three donors. 50 The maximum MFI values ​​are shown in Table 2 below. EC for IgG1 isotype control (negative control) and anti-human CD47 (positive control) 50 The maximum MFI value is also included.

[0274] [Table 2]

[0275] The dose-response curves for binding to RBCs from each of the three donors are shown in Figures 4, 5, and 6.

[0276] Summary: Daratumumab showed a dose-dependent increase in binding to RBCs from all three donors. CID103 (aCD38-b-348) also showed a dose-dependent increase in binding to RBCs from all three donors, but its overall MFI value was lower compared to daratumumab. The IgG1 isotype control did not show binding to RBCs from any of the three donors, except at the highest concentration tested. The positive control, AF647 conjugate anti-human CD47, showed a dose-response binding curve to RBCs from all three donors.

[0277] Example 3: Pre-transfusion testing of CID103 (aCD38-b-348) and daratumumab using the IAT tube method. Materials and methods Sample preparation. CID103 (aCD38-b-348) and daratumumab were prepared in inactive AB plasma at concentrations of 250 μg / ml and 1000 μg / ml, respectively. Samples of AB plasma alone were used as a control.

[0278] RBCs were used. RBCs with the following Rh phenotypes were employed: RhD-positive (R1R1, R2R2) and RhD-negative (rr). RBCs were prepared from whole blood samples according to a standard protocol.

[0279] IAT tube method protocol. RBCs are treated with anti-human globulin (Ortho Diagnostics). The samples were incubated in polyethylene glycol (PEG)-enhanced medium containing CID103 (aCD38-b-348), daratumumab, or control plasma, according to a standard protocol.

[0280] Data analysis. After incubation, skilled personnel at the New York Blood Center will perform the tubing. The observed reactivity was visually evaluated and scored on a scale from 4+ to 0. Here, 4+, 3+, 2+, and 1+ all indicate reactivity, while 0 or 0? indicates no reactivity or questionable reactivity. Any reactivity observed in any of the experiments indicates interference with the anti-CD38 antibody present in the sample.

[0281] result The results are shown in Table 3.

[0282] [Table 3]

[0283] In summary, daratumumab showed a 1+ reading for all RBC phenotypes tested, while CID103(aCD38-b-348) showed no detectable interference even at high concentrations of 1000 μg / ml. In a blood bank context, a +1 reading indicates a mismatch between donor and recipient. Therefore, treatment of patients with daratumumab may falsely indicate a mismatch between recipient and donor RBCs, while treatment of patients with CID103(aCD38-b-348) is not expected to cause such interference.

[0284] Example 4: Pre-transfusion study of CID103 (aCD38-b-348) and daratumumab in untreated RBCs using column agglutination technology Materials and methods Sample preparation. CID103(aCD38-b-348) and daratumumab were prepared in inactive AB plasma at concentrations of 1 μg / ml, 10 μg / ml, 100 μg / ml, and 250 μg / ml. Additional samples of CID103(aCD38-b-348) were prepared in inactive AB plasma at concentrations of 625 μg / ml and 1000 μg / ml. Samples in inactive AB plasma alone were used as controls.

[0285] RBCs were used. RBCs with the following Rh phenotypes were employed: RhD-positive (R1R1, R2R2) and RhD-negative (rr). RBCs were prepared from whole blood samples according to a standard protocol.

[0286] Column aggregation technology. IgG gel cards were obtained from Ortho MTS. Following the manufacturer's instructions... RBCs were incubated with CID103 (aCD38-b-348), daratumumab, or control plasma. After incubation, the samples were centrifuged at 900 rpm for 10 minutes according to the manufacturer's instructions.

[0287] Data analysis. After centrifugation, the tubes are visually evaluated by experts at the New York Blood Center. The observed reactivity level was valued and scored on a scale from 4+ to 0. Here, 4+, 3+, 2+, and 1+ all indicate reactivity, while 0 or 0? indicates no reactivity or questionable reactivity. Φ indicates the absence of the drug in the sample (plasma control). Any reactivity observed in any of the experiments indicates interference with the anti-CD38 antibody present in the sample.

[0288] result The gel card results for CID103 (aCD38-b-348) and daratumumab at each tested concentration are shown in Figures 7A-7C and 8A-8D.

[0289] As can be seen from Figures 7A to 7C, daratumumab is 1+ even at a low concentration of 1 μg / l. Interference was definitely observed between 2+ and 3+. In the case of daratumumab, interference was seen for all RBC Rh phenotypes tested. In contrast, as can be seen from Figures 8A to 8D, CID103 (aCD38-b-348) caused minimal interference, and only questionable interference was detected at higher concentrations of 100 μg / ml and 250 μg / ml in the Rh-positive RBCs tested. Even at very high concentrations of 625 μg / ml and 1000 μg / ml, no interference was observed at all in Rh-negative RBCs (Figure 8D).

[0290] Example 5: Pre-transfusion study of CID103 (aCD38-b-348) and daratumumab in enzymatically treated or DTT-treated RBCs using column agglutination technology. Materials and methods Sample preparation. CID103 (aCD38-b-348) and daratumumab were prepared in inactive AB plasma at concentrations of 10 μg / ml and 250 μg / ml. Samples of inactive AB plasma alone were used as a control.

[0291] RBCs. RBCs with an RhD-negative (rr) phenotype were used. RBCs were prepared from whole blood samples according to a standard protocol. Both untreated and pretreated RBCs were used. For pretreated RBCs, the RBCs were treated with commercially prepared ficin according to the manufacturer's instructions, or as described in Judd, Johnson & Storry (2008) Judd's Methods in Immunohematology 3. rd Papain prepared in-house as described by Ed, The samples were treated with trypsin or 0.2M DTT.

[0292] Column aggregation technology. IgG gel cards were obtained from Ortho MTS. Following the manufacturer's instructions... RBCs were incubated with CID103 (aCD38-b-348), daratumumab, or control plasma. After incubation, the samples were centrifuged at 900 rpm for 10 minutes according to the manufacturer's instructions.

[0293] Data analysis. After centrifugation, the tubes are visually evaluated by experts at the New York Blood Center. The observed reactivity level was valued and scored on a scale from 4+ to 0. Here, 4+, 3+, 2+, and 1+ all indicate reactivity, while 0 or ± indicates no reactivity or questionable reactivity. Φ indicates the absence of the drug in the sample (plasma control). Any reactivity observed in any of the experiments indicates interference with the anti-CD38 antibody present in the sample.

[0294] result Figure 9 shows the gel card results for untreated RBCs or RBCs treated with papain, trypsin, or ficin in plasma containing daratumumab or CID103 (aCD38-b-348).

[0295] In untreated RBCs, as previously demonstrated, daratumumab caused a definite 2+ interference at both 10 μg / ml and 250 μg / ml concentrations, while CID103(aCD38-b-348) caused only minimal interference at both concentrations tested. Treatment of RBCs with papain and ficin did not resolve the observed daratumumab interference at either concentration. Treatment of RBCs with papain and ficin resulted in the observation of interference in CID103(aCD38-b-348) samples. Treatment of RBCs with trypsin resolved the interference observed with both 10 μg / ml and 250 μg / ml daratumumab. No interference with CID103(aCD38-b-348) was observed in trypsin-treated RBCs.

[0296] RBC treatment in DTT also involves both 10 μg / ml and 250 μg / ml daratumumab. The interference observed in the previous case was resolved. No interference with CID103 (aCD38-b-348) was observed in the DTT-treated RBC.

[0297] Example 6: Pre-transfusion testing of CID103 (aCD38-b-348) using an automated platform Materials and methods CID103 (aCD38-b-348) was prepared in inactive AB plasma at a concentration of 250 μg / ml. RBCs were prepared from whole blood samples according to a standard protocol. Samples were run through two different automated analyzers, IH-1000 (BioRad) and Tango (BioRad), according to the manufacturer's instructions.

[0298] result No reactivity was detected when using 250 μg / ml CID103 (aCD38-b-348) on the Tango (BioRad) automated platform. As shown in Figure 10, the IH-1000 platform (BioRad) returned an indeterminate result. This result was evaluated by an expert according to a standard protocol and reported as non-reactive.

[0299] Example 7: Amino acid sequence This disclosure refers to the following many different amino acid sequences:

[0300] [Table 4] TIFF2026136194000005.tif236170

[0301] Equivalents and range Those skilled in the art will see that the present invention is described in other descriptions of the examples or certain specific embodiments included herein. It is understood that this is not defined by the attached claims.

[0302] Similarly, unless otherwise clearly indicated by the context, the singular forms "a," "an," and "the" generally refer to multiple objects.

[0303] Unless otherwise defined above, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which the present invention pertains. Any methods and materials similar to or equivalent to those described herein may also be used in the implementation or testing of the present invention. In general, the nomenclature and techniques used herein in relation to cell and tissue culture, molecular biology, immunology, genetics, and protein and nucleic acid chemistry are known, commonly used, or in accordance with the manufacturer's specifications in the art.

[0304] All publications referenced herein constitute part of this specification by reference to disclose and explain the above methods and / or materials in connection with the publications cited herein.

[0305] References Chevrier S et al. 2017. Cell. 169:736-749 Darzalex package insert. Horsham PA: Janssen Biotech, 2015 Ellington et al. Nature. 1990; 346(6287): 818-822 Handbook of Therapeutic Antibodies, Wiley, 2014, Chapter 6, Antibody Affinity (pages 115-140) Hendrickson & Tormey, 2016, Hematol Oncol Clin N Am, 30:635-651 Jarasch A et al., 2015. J Pharm Sci. 104:1885-1898 Judd, Johnson & Storry (2008) Judd’s Methods in Immunohematology 3rd Ed. Kearns JD et al., 2015. Mol Cancer Ther. 14:1625-36 Kijanka M et al., 2015. Nanomedicine. 10:161-174. Liu L, 2015. J Pharm Sci. 104:1866-84. Ni et al., Curr Med Che 2011; 18(27):4206-14 Oostendorp et al. 2015, Transfusion, 55:1555-62 Rajpal et al., Proc Natl Acad Sci USA, 2005, 102(24):8466-71 Redman JM et al., 2015. Mol Immunol. 67: 28-45. Regan & Markowitz, 2016, American Association of Blood Banks, Bulletin #16-02 Sliwkowski M & Mellman I, 2013. Science. 341:1192-8. Steinwand et al., MAbs, 2014, 6(1):204-18 Tormey & Hendrickson, 2019, Blood, 133:1821-1830 Tuerk et al., Science. 1990; 249(4968):505-510 Vazquez-Lombardi R et al., 2015. Drug Discov Today. 20:1271-83

[0306] item This invention includes at least the following numbered items:

[0307] 1. A method for screening blood samples obtained from patients who have been administered an anti-CD38 antibody or its antigen-binding fragment, a) Prepare a blood sample from the patient, b) Prepare a blood sample from the blood donor, Here, the donor blood sample contains donor red blood cells; c) Screening a patient blood sample, including determining the presence or absence of one or more patient antibodies in the patient blood sample that specifically bind to one or more red blood cell antigens expressed on the surface of donor red blood cells, Methods that include...

[0308] 2. The method of item 1, which does not include the step of contacting a patient blood sample or a donor blood sample with an anti-CD38 antibody or an agent that inhibits the binding of its antigen-binding fragment to membrane-bound CD38 present on the surface of one or more donor red blood cells.

[0309] 3. The method of item 2, wherein the substance that inhibits the binding of anti-CD38 antibodies or their antigen-binding fragments to membrane-bound CD38 present on the surface of one or more donor red blood cells is a soluble CD38 antigen, an anti-CD38 idiotype antibody, or an antigen stripping agent.

[0310] 4. The antigen stripping agent is a redox reagent or enzyme, as described in item 3.

[0311] 5. The redox reagent is DTT, according to the method in item 4.

[0312] 6. The enzyme is a protease, as described in item 4.

[0313] 7. The protease is selected from the group consisting of trypsin, α-chymotrypsin, papain, and ficin, by the method of item 6.

[0314] 8. The anti-CD38 antibody or its antigen-binding fragment specifically binds to the epitope of human CD38, where the epitope comprises one or more amino acid residues included in amino acids 65-79 of SEQ ID NO: 29 (human CD38), in any of the above manner.

[0315] 9. The anti-CD38 antibody or its antigen-binding fragment specifically binds to the epitope of human CD38, where the epitope comprises amino acids 65 to 79 of SEQ ID NO: 29 (human CD38), as described in any of the above items.

[0316] 10. The anti-CD38 antibody or its antigen-binding fragment comprises HCDR3 containing the amino acid sequence of SEQ ID NO: 3 or SEQ ID NO: 19, by any of the methods described above.

[0317] 11. Anti-CD38 antibodies or their antigen-binding fragments are a) HCDR1 containing the amino acid sequence of SEQ ID NO: 1, HCDR2 containing the amino acid sequence of SEQ ID NO: 2, HCDR3 containing the amino acid sequence of SEQ ID NO: 3, LCDR1 containing the amino acid sequence of SEQ ID NO: 4, LCDR2 containing the amino acid sequence of SEQ ID NO: 5, and, LCDR3 containing an amino acid sequence selected from the group consisting of SEQ ID NOs. 6, SEQ ID NOs. 9, SEQ ID NOs. 10, SEQ ID NOs. 11, and SEQ ID NOs. 12, or b) HCDR1 containing the amino acid sequence of SEQ ID NO: 17, HCDR2 containing the amino acid sequence of SEQ ID NO: 18, HCDR3 containing the amino acid sequence of SEQ ID NO: 19, LCDR1 containing the amino acid sequence of SEQ ID NO: 20, LCDR2 containing the amino acid sequence of SEQ ID NO: 21, and, LCDR3 containing an amino acid sequence selected from the group consisting of SEQ ID NOs. 22, SEQ ID NOs. 25, and SEQ ID NOs. 26, Any of the methods listed above, including the above.

[0318] 12. Anti-CD38 antibodies or their antigen-binding fragments are a) HCDR1 containing the amino acid sequence of SEQ ID NO: 1, HCDR2 containing the amino acid sequence of SEQ ID NO: 2, HCDR3 containing the amino acid sequence of SEQ ID NO: 3, LCDR1 containing the amino acid sequence of SEQ ID NO: 4, LCDR2 containing the amino acid sequence of SEQ ID NO: 5, and, LCDR3 containing the amino acid sequence of SEQ ID NO: 6, or b) HCDR1 containing the amino acid sequence of SEQ ID NO: 17, HCDR2 containing the amino acid sequence of SEQ ID NO: 18, HCDR3 containing the amino acid sequence of SEQ ID NO: 19, LCDR1 containing the amino acid sequence of SEQ ID NO: 20, LCDR2 containing the amino acid sequence of SEQ ID NO: 21, and, LCDR3 containing the amino acid sequence of SEQ ID NO: 22, Any of the methods listed above, including the above items.

[0319] 13. Anti-CD38 antibodies or their antigen-binding fragments are a) A variable light chain containing an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% identity with SEQ ID NO: 7, and / or a variable heavy chain containing an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% identity with SEQ ID NO: 8, b) A variable light chain containing an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% identity with SEQ ID NO: 23, and / or a variable heavy chain containing an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% identity with SEQ ID NO: 24, c) A variable light chain containing an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% identity with SEQ ID NO: 7, and / or a variable heavy chain containing an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% identity with SEQ ID NO: 13. d) A variable light chain containing an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% identity with SEQ ID NO: 7, and / or a variable heavy chain containing an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% identity with SEQ ID NO: 14, e) A variable light chain containing an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% identity with SEQ ID NO: 7, and / or a variable heavy chain containing an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% identity with SEQ ID NO: 15. f) A variable light chain containing an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% identity with SEQ ID NO: 7, and / or a variable heavy chain containing an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% identity with SEQ ID NO: 16. g) A variable light chain containing an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% identity with SEQ ID NO: 23, and / or a variable heavy chain containing an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% identity with SEQ ID NO: 27, or h) A variable light chain containing an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% identity with SEQ ID NO: 23, and / or a variable heavy chain containing an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% identity with SEQ ID NO: 28. A method of any of the above claims, including:

[0320] 14. Anti-CD38 antibodies or their antigen-binding fragments are a) A variable light chain containing the amino acid sequence of SEQ ID NO: 7 or a variable light chain region sequence having up to two amino acid substitutions compared to SEQ ID NO: 7, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 8. Alternatively, a variable heavy chain region sequence having up to two amino acid substitutions compared to Sequence ID No. 8, b) A variable light chain containing the amino acid sequence of SEQ ID NO: 23 or a variable light chain region sequence having up to two amino acid substitutions compared to SEQ ID NO: 23, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 24 or a variable heavy chain region sequence having up to two amino acid substitutions compared to SEQ ID NO: 24, c) A variable light chain containing the amino acid sequence of SEQ ID NO: 7 or a variable light chain region sequence having up to two amino acid substitutions compared to SEQ ID NO: 7, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 13 or a variable heavy chain region sequence having up to two amino acid substitutions compared to SEQ ID NO: 13, d) A variable light chain containing the amino acid sequence of SEQ ID NO: 7 or a variable light chain region sequence having up to two amino acid substitutions compared to SEQ ID NO: 7, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 14 or a variable heavy chain region sequence having up to two amino acid substitutions compared to SEQ ID NO: 14, e) A variable light chain containing the amino acid sequence of SEQ ID NO: 7 or a variable light chain region sequence having up to two amino acid substitutions compared to SEQ ID NO: 7, and f) A variable heavy chain containing the amino acid sequence of sequence number 15 or a variable heavy chain region sequence having up to two amino acid substitutions compared to sequence number 15, g) A variable light chain containing the amino acid sequence of sequence number 7 or a variable light chain region sequence having up to two amino acid substitutions compared to sequence number 7, and a variable heavy chain containing the amino acid sequence of sequence number 16 or a variable heavy chain region sequence having up to two amino acid substitutions compared to sequence number 16, g) A variable light chain containing the amino acid sequence of sequence number 23 or a variable light chain region sequence having up to two amino acid substitutions compared to sequence number 23, and a variable heavy chain containing the amino acid sequence of sequence number 27 or a variable heavy chain region sequence having up to two amino acid substitutions compared to sequence number 27, or h) A variable light chain containing the amino acid sequence of SEQ ID NO: 23 or a variable light chain region sequence having up to two amino acid substitutions compared to SEQ ID NO: 23, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 28 or a variable heavy chain region sequence having up to two amino acid substitutions compared to SEQ ID NO: 8. Any of the methods listed above, including the above.

[0321] 15. Anti-CD38 antibodies or their antigen-binding fragments are a) A variable light chain containing the amino acid sequence of SEQ ID NO: 7, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 8, b) A variable light chain containing the amino acid sequence of SEQ ID NO: 23, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 24. c) A variable light chain containing the amino acid sequence of SEQ ID NO: 7, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 13. d) A variable light chain containing the amino acid sequence of SEQ ID NO: 7, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 14. e) A variable light chain containing the amino acid sequence of SEQ ID NO: 7, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 15. f) A variable light chain containing the amino acid sequence of SEQ ID NO: 7, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 16. g) A variable light chain containing the amino acid sequence of SEQ ID NO: 23, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 27, or h) A variable light chain containing the amino acid sequence of SEQ ID NO: 23, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 28. Any of the methods listed above, including the above.

[0322] 16. The anti-CD38 antibody or its antigen-binding fragment is a monoclonal antibody, domain antibody, single-chain antibody, Fab fragment, F(ab')2 fragment, single-chain variable fragment (scFv), scFv-Fc fragment, single-chain antibody (scAb), aptamer, or nanobody, as described in any of the above methods.

[0323] 17. The anti-CD38 antibody or its antigen-binding fragment is a rabbit antibody, mouse antibody, chimeric antibody, humanized antibody, or fully human antigen-binding antibody, using any of the methods listed above.

[0324] 18. The anti-CD38 antibody or its antigen-binding fragment is an IgG antibody, as described in any of the above items.

[0325] 19. The anti-CD38 antibody or its antigen-binding fragment is selected from the group consisting of IgG1 isotype antibody, IgG2 isotype antibody, IgG3 isotype antibody, and IgG4 isotype antibody, by any of the methods described above.

[0326] 20. The anti-CD38 antibody or its antigen-binding fragment is an IgG1 antibody, as per any of the above methods.

[0327] 21. Any of the above methods, wherein an anti-CD38 antibody or its antigen-binding fragment is included in an immunoconjugate further comprising a bispecific antibody, a multispecific antibody, or a therapeutic or diagnostic agent.

[0328] 22. The anti-CD38 antibody or its antigen-binding fragment binds to the extracellular domain of human CD38 in any of the above manner.

[0329] 23. The patient's blood sample shall be selected from the group consisting of whole blood samples, plasma samples, and serum samples, using one of the methods described above.

[0330] 24. The patient blood sample is a whole blood sample, and the method is one of the methods described above, comprising the step of removing patient red blood cells from the patient blood sample.

[0331] 25. The antibody is an alloantibody, using one of the methods listed above.

[0332] 26. The patient antibody specifically binds to any erythrocyte antigen other than any erythrocyte antigen expressed by the patient's erythrocytes, by any of the methods described above.

[0333] 27. Patient antibodies are clinically significant patient antibodies, as determined by any of the methods listed above.

[0334] 28. The presence of patient antibodies that specifically bind to one or more red blood cell antigens present on one or more red blood cells from a donor blood sample is indicated by agglutination or hemolysis, using any of the methods described above.

[0335] 29. The presence of patient antibodies that specifically bind to one or more red blood cell antigens present on one or more red blood cells from a donor blood sample indicates that the donor blood sample is incompatible with the patient, by any of the methods described above.

[0336] 30. The absence of patient antibodies that specifically bind to one or more red blood cell antigens present on one or more red blood cells from a donor blood sample indicates that the donor blood sample is compatible with the patient, by any of the methods described above.

[0337] 31. The patient's antibody is an IgG antibody and / or an IgM antibody, as determined by any of the methods listed above.

[0338] 32. The patient is human, by any of the methods listed above.

[0339] 33. The patient has cancer, is suffering from cancer, or is receiving treatment for cancer, in any of the above ways.

[0340] 34. Cancer is a solid tumor that expresses CD38 on its cell surface, according to item 33.

[0341] 35. Cancer is a hematological malignancy that expresses CD38 on the cell surface, as described in item 33.

[0342] 36. The cancer is a T-cell or B-cell non-Hodgkin lymphoma, chronic lymphocytic leukemia, plasmacytoma, or multiple myeloma, by any one of the methods described in items 33-35.

[0343] 37. One or more erythrocyte antigens expressed on the surface of donor erythrocytes are selected from the group consisting of Ab, ABO, Cromer, Diego, Duffy, Gerbich, GLOB, Indian, Kell, Kidd, Knops, Lewis, Lutheran, LW, MNS, P1, Rh, XK, Xg, and Yt, by any of the methods described above.

[0344] 38. One or more erythrocyte antigens expressed on the surface of donor erythrocytes are selected from the group consisting of Ab, Rh, MNS, P1, Lewis, Kell, Duffy, KIDD, Lutheran, and Xg, by any of the methods described above.

[0345] 39. Any of the above methods, further comprising the step of administering an anti-CD38 antibody or its antigen-binding fragment to a patient prior to step (a).

[0346] 40. The patient has been administered an anti-CD38 antibody or its antigen-binding fragment less than one year, six months, three months, two months, one month, four weeks, three weeks, two weeks, or one week prior to obtaining a sample from the patient, using any of the methods listed above.

[0347] 41. The screening in step (b) is performed using any of the above methods, selected from the group consisting of column agglutination assays, indirect antiglobulin (IAT) tube assays, and solid-phase assays.

[0348] 42. Any method of the above item, further comprising preparing a patient blood / donor red blood cell mixture by contacting a patient blood sample with donor red blood cells from a donor blood sample prior to screening in step (c).

[0349] 43. The method of item 42, further comprising the step of incubating a mixture of patient blood / donor red blood cells to enable the binding of any one or more patient antibodies in the patient blood sample to one or more red blood cell antigens present, if present, on the donor red blood cells, thereby forming one or more patient antibody / donor red blood cell antigen complexes.

[0350] 44. The method of item 42 or 43, further comprising the step of separating any one or more patient antibody / donor red blood cell antigen complexes, if present, from a mixture of patient blood / donor red blood cells, wherein optionally the separation step includes centrifugation.

[0351] 45. The method of any one of items 42-44, further comprising centrifugation of a mixture of patient blood and donor red blood cells.

[0352] 46. a) Prepare a blood sample from the patient, b) Prepare a blood sample from the blood donor, Here, the donor blood sample contains donor red blood cells; c) Preparing a patient blood / donor red blood cell mixture by contacting a patient blood sample with one or more donor red blood cells from a donor blood sample, d) Incubating a mixture of patient blood / donor red blood cells to allow one or more patient antibodies in the patient blood sample to bind to one or more red blood cell antigens present on one or more donor red blood cells, if present, to form one or more patient antibody / donor red blood cell antigen complexes, e) Optionally, if present, separate any one or more patient alloantibody / donor red blood cell antigen complexes from the patient blood / donor red blood cell mixture, Here, optionally, the separation step includes centrifugation; f) Determining the presence or absence of patient antibodies in a patient blood sample that specifically bind to one or more red blood cell antigens expressed on one or more donor red blood cells, Any of the methods listed above, including the above.

[0353] 47. The method of item 46, further comprising centrifugation of a mixture of patient blood / donor red blood cells.

[0354] 48. Any method of the above item, further comprising the step of adding a coagulant that specifically binds any antibodies present in a patient's blood sample to each other.

[0355] 49. A coagulant that specifically binds one or more patient antibodies in a patient's blood sample to each other is an anti-human globulin reagent, according to the method of item 48.

[0356] 50. Any method of the above item, further comprising preparing a report indicating the presence or absence of patient antibodies in a patient blood sample and / or the suitability of the donor who provides blood or red blood cells to the patient.

[0357] 51. Any method of the above item, further comprising the step of screening the patient blood sample against a red blood cell panel prior to step (b) to determine the presence or absence of any patient antibody in the patient blood sample that specifically binds to any red blood cell antigen present on the surface of any red blood cell in the red blood cell panel.

[0358] 52. The method of item 51, wherein the donor red blood cells in the donor blood sample do not express any red blood cell antigens that can be specifically bound by any patient antibody identified to specifically bind to any red blood cell antigen expressed on the surface of any red blood cell in the red blood cell panel.

[0359] 53. A method for treating cancer in a patient, comprising preparing a blood sample from the patient and screening the blood sample according to any of the methods described in items 1 to 52.

[0360] 54. The patient has been administered an anti-CD38 antibody or its antigen-binding fragment, according to the method of item 53.

[0361] 55. The method of item 53 or 54, which includes the step of obtaining a sample from a patient.

[0362] 56. The method is, a) Administering the patient an anti-CD38 antibody or its antigen-binding fragment, b) Obtaining a blood sample from the patient after administration of an anti-CD38 antibody or its antigen-binding fragment, c) Screen the patient's blood sample according to one of the methods described in item 1 to 52, Method 53, including item 53.

[0363] 57. Cancer is a solid tumor that expresses CD38 on its cell surface, by any one of items 53-56.

[0364] 58. Cancer is a hematological malignancy that expresses CD38 on the cell surface, by any one of items 53-56.

[0365] 59. The cancer is T-cell or B-cell non-Hodgkin lymphoma, chronic lymphocytic leukemia, plasmacytoma, or multiple myeloma, by any one of items 53-58.

[0366] 60. The method of any one of items 53 to 59, further comprising the step of administering blood or red blood cells from the donor if the donor is found to be compatible with the patient.

[0367] 61. An anti-CD38 antibody or its antigen-binding fragment used in a method of treating cancer in a patient, comprising any one of the methods described in items 53 to 60.

Claims

1. A method for screening blood samples obtained from patients who have been administered an anti-CD38 antibody or its antigen-binding fragment, a) Prepare a blood sample from the patient, b) Prepare a blood sample from the blood donor, Here, the donor blood sample includes donor red blood cells; c) Screening a patient blood sample, which includes determining the presence or absence of one or more patient antibodies in the patient blood sample that specifically bind to one or more red blood cell antigens expressed on the surface of the donor red blood cells, Methods that include...

2. The method according to claim 1, wherein the method does not include the step of contacting the patient blood sample or the donor blood sample with an active substance that inhibits the binding of the anti-CD38 antibody or its antigen-binding fragment to membrane-bound CD38 present on the surface of one or more donor red blood cells.

3. The method according to claim 2, wherein the substance that inhibits the binding of the anti-CD38 antibody or its antigen-binding fragment to membrane-bound CD38 present on the surface of one or more donor red blood cells is a soluble CD38 antigen, an anti-CD38 idiotype antibody, or an antigen stripping agent.

4. The method according to claim 3, wherein the antigen stripping agent is a redox reagent, optionally DTT, or an enzyme, or optionally a protease.

5. The method according to claim 4, wherein the protease is selected from the group consisting of trypsin, α-chymotrypsin, papain, and ficin.

6. The method according to any one of claims 1 to 5, wherein the anti-CD38 antibody or its antigen-binding fragment specifically binds to an epitope of human CD38, where the epitope comprises one or more amino acid residues included in amino acids 65 to 79 of SEQ ID NO: 29 (human CD38).

7. The anti-CD38 antibody or its antigen-binding fragment is a) HCDR1 containing the amino acid sequence of SEQ ID NO: 1, HCDR2 containing the amino acid sequence of SEQ ID NO: 2, HCDR3 containing the amino acid sequence of SEQ ID NO: 3, LCDR1 containing the amino acid sequence of SEQ ID NO: 4, LCDR2 containing the amino acid sequence of SEQ ID NO: 5, and, LCDR3 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 9, 10, 11, and 12, or b) HCDR1 containing the amino acid sequence of SEQ ID NO: 17, HCDR2 containing the amino acid sequence of SEQ ID NO: 18, HCDR3 containing the amino acid sequence of SEQ ID NO: 19, LCDR1 containing the amino acid sequence of SEQ ID NO: 20, LCDR2 containing the amino acid sequence of SEQ ID NO: 21, and, LCDR3 containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 22, 25, and 26, The method according to any one of claims 1 to 6, including the method described in any one of claims 1 to 6.

8. The anti-CD38 antibody or its antigen-binding fragment is a) A variable light chain containing the amino acid sequence of SEQ ID NO: 7, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 8, or b) A variable light chain containing the amino acid sequence of SEQ ID NO: 23, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 24, or c) A variable light chain containing the amino acid sequence of SEQ ID NO: 7, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 13, or d) A variable light chain containing the amino acid sequence of SEQ ID NO: 7, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 14, or e) A variable light chain containing the amino acid sequence of SEQ ID NO: 7, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 15, or f) A variable light chain containing the amino acid sequence of SEQ ID NO: 7, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 16, or g) A variable light chain containing the amino acid sequence of SEQ ID NO: 23, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 27, or h) A variable light chain containing the amino acid sequence of SEQ ID NO: 23, and a variable heavy chain containing the amino acid sequence of SEQ ID NO: 28, The method according to any one of claims 1 to 7, including the method described in any one of claims 1 to 7.

9. The method according to any one of claims 1 to 8, wherein the anti-CD38 antibody or its antigen-binding fragment is a monoclonal antibody, a domain antibody, a single-chain antibody, a Fab fragment, an F(ab')2 fragment, a single-chain variable fragment (scFv), an scFv-Fc fragment, a single-chain antibody (scAb), an aptamer, or a nanobody.

10. The method according to any one of claims 1 to 9, wherein the anti-CD38 antibody or its antigen-binding fragment is an IgG antibody, optionally an IgG1 antibody.

11. The method according to any one of claims 1 to 10, wherein the patient antibody is an alloantibody, and / or the patient antibody specifically binds to any erythrocyte antigen other than any erythrocyte antigen expressed by the patient's erythrocytes, and / or the patient antibody is a clinically significant patient antibody, and / or the patient antibody is an IgG antibody and / or an IgM antibody.

12. The method according to any one of claims 1 to 11, wherein the patient has cancer, is suffering from cancer, or is receiving treatment for cancer.

13. The method according to claim 12, wherein the cancer is a solid tumor expressing CD38 on its cell surface, or the cancer is a hematological malignancy expressing CD38 on its cell surface.

14. The method according to claim 12 or 13, wherein the cancer is a T-cell or B-cell non-Hodgkin lymphoma, chronic lymphocytic leukemia, plasmacytoma, or multiple myeloma.

15. The method according to any one of claims 1 to 14, wherein the one or more erythrocyte antigens expressed on the surface of the donor erythrocytes are selected from the group consisting of Ab, ABO, Cromer, Diego, Duffy, Gerbich, GLOBAL, Indian, Kell, Kidd, Knops, Lewis, Lutheran, LW, MNS, P1, Rh, XK, Xg, and Yt.

16. The method according to any one of claims 1 to 15, wherein the one or more red blood cell antigens expressed on the surface of the donor red blood cells are selected from the group consisting of Ab, Rh, MNS, P1, Lewis, Kell, Duffy, KIDD, Lutheran, and Xg.

17. The method according to any one of claims 1 to 16, wherein the patient has been administered the anti-CD38 antibody or its antigen-binding fragment less than one year prior to obtaining a sample from the patient.

18. a) Prepare blood samples obtained from the patient, b) Prepare a blood sample obtained from the blood donor, Here, the donor blood sample includes donor red blood cells; c) Preparing a patient blood / donor red blood cell mixture by contacting the patient blood sample with one or more donor red blood cells from the donor blood sample, d) Incubating the patient blood / donor red blood cell mixture to enable the binding of any one or more patient antibodies in the patient blood sample to one or more red blood cell antigens present on the one or more donor red blood cells, if present, to form one or more patient antibody / donor red blood cell antigen complexes, e) Optionally, if present, separate any one or more patient alloantibody / donor red blood cell antigen complexes from the patient blood / donor red blood cell mixture, Here, optionally, the separation step includes centrifugal separation; f) Determining the presence or absence of patient antibodies in the patient blood sample that specifically bind to one or more red blood cell antigens expressed on one or more donor red blood cells, Includes, The method according to any one of claims 1 to 17, further comprising optionally centrifuging the mixture of patient blood and donor red blood cells.

19. The method according to any one of claims 1 to 18, further comprising the step of adding a coagulant that specifically binds any antibodies present in the patient's blood sample to each other.

20. The method according to claim 19, wherein the agglutinant that specifically binds one or more patient antibodies in the patient blood sample to each other is an anti-human globulin reagent.

21. The method according to any one of claims 1 to 20, further comprising the step of screening the patient blood sample against a red blood cell panel prior to step (b) to determine the presence or absence of any patient antibody in the patient blood sample that specifically binds to any red blood cell antigen present on the surface of any red blood cell in the red blood cell panel, wherein optionally, the donor red blood cells in the donor blood sample do not express any red blood cell antigens that can be specifically bound by any patient antibody identified to specifically bind to any red blood cell antigen expressed on the surface of any red blood cell in the red blood cell panel.

22. A method for treating cancer in a patient, comprising preparing a blood sample from the patient and screening the blood sample according to the method described in any one of claims 1 to 21.

23. The method according to claim 22, wherein the patient is administered an anti-CD38 antibody or an antigen-binding fragment thereof.

24. The aforementioned method, a) Administering the patient an anti-CD38 antibody or its antigen-binding fragment, b) Obtaining a blood sample from the patient after administration of the anti-CD38 antibody or its antigen-binding fragment, c) Screening the patient blood sample according to the method described in any one of claims 1 to 21, The method according to claim 23, including the method described in claim 23.

25. The method according to any one of claims 22 to 24, wherein the cancer is a solid tumor expressing CD38 on its cell surface.

26. The method according to any one of claims 22 to 25, wherein the cancer is a hematological malignancy that expresses CD38 on the cell surface.

27. The method according to any one of claims 22 to 26, wherein the cancer is a T-cell or B-cell non-Hodgkin lymphoma, chronic lymphocytic leukemia, plasmacytoma, or multiple myeloma.

28. The method according to any one of claims 22 to 27, further comprising the step of administering blood or red blood cells from the blood donor if the blood donor is found to be compatible with the patient.

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