Method and system for implementing patient-specific immunotherapy procedures using continuous monitoring and identification of biological samples.

Abstract

This provides a method for implementing patient-specific immunotherapy procedures. [Solution] A method comprising: receiving a cell order request for producing patient T cells by a computing device; generating a patient-specific identifier associated with the cell order request; initiating a process for producing transfected T cells for injection into the patient's bloodstream, the process comprising: collecting T cells by performing a leukocyte apheresis procedure on a patient's blood sample; labeling the container holding the T cells with the patient-specific identifier; transporting the T cells to a manufacturing facility; producing transfected T cells from the T cells using cell modification techniques; receiving the transfected T cells from the manufacturing facility; and injecting the T cells into the patient's bloodstream, wherein the computing device records tracking events for each step of the process, each tracking event including the patient-specific identifier, and the tracking events include continuous management of the patient's T cells during the process.

Description

[Technical Field]

[0001] [Cross-reference of related applications] This application claims the interests of U.S. Provisional Patent Application No. 62 / 559,330, filed September 15, 2017, and U.S. Provisional Patent Application No. 62 / 556,912, filed October 2, 2017, and the entire disclosure thereof constitutes part of this specification by reference.

[0002] This application generally relates to chain-of-custody and chain-of-identity The present invention relates to a method and apparatus, including a computer program product, for implementing patient-specific immunotherapy procedures using biological sample tracking. [Background technology]

[0003] In recent years, advances in medical technology have led to the rise of immunotherapy for treating various types of diseases and disorders, including various forms of cancer. Generally, immunotherapy is a treatment of disease that involves activating or suppressing the immune response. In many cases, modified versions of the patient's own biomaterials, such as immune cells, are reintroduced into the patient to initiate and / or supplement the immune response.

[0004] For example, modified immune cells have been shown to possess desirable properties, particularly in therapeutic applications in oncology. Two main types of modified immune cells include chimeric antigen receptors (referred to as "CAR" or "CAR-T") and T cell receptors ("TCR"). These modified cells are designed to confer antigen specificity while retaining or enhancing their ability to recognize and kill target cells. A chimeric antigen receptor may include, for example, (i) an antigen-specific component ("antigen-binding molecule"), (ii) an extracellular domain, (iii) one or more costimulatory domains, and (iv) one or more activating domains. Each domain may be heterogeneous, i.e., composed of sequences derived from (or corresponding to) different protein chains.

[0005] Since many patients receiving immunotherapy are seriously ill, an important factor in the effectiveness of such immunotherapy procedures is the ability to provide the modified biomaterial to the patient as soon as possible so that the therapeutic benefit can be maximized. Also, since many types of immunotherapy are tailored to specific patients (i.e., using the patients' own cells), it is important to ensure accurate tracking of the patients' biomaterials throughout the immunotherapy process, from extraction to modification and then back to injection into the patient, in order to avoid manufacturing delays, mislabeling of materials, and patient misidentification. However, existing immunotherapy procedures generally lack a technical mechanism to automatically track the patients' biomaterials and ensure that the biomaterials are linked to the identity of a specific patient throughout the manufacturing process.

Summary of the Invention

Problems to be Solved by the Invention

[0006] Therefore, what is needed is a method and system for implementing a patient-specific immunotherapy procedure using continuous management and continuous identification of biomaterial tracking. The techniques described herein provide certain technical advantages over existing systems in providing continuous and automatic continuous management and continuous identification of patient-specific biomaterials during immunotherapy procedures, creating a computerized information portal that can be used by parties (such as patients, physicians, manufacturers, and other healthcare stakeholders) to quickly understand and track the current stage of the immunotherapy procedure and the status of the patient's biomaterials during the procedure. Such advanced tracking is an improvement over existing systems that can experience delays during the manufacturing process, which can be extremely serious for patients dealing with life-threatening illnesses, as they lack technical solutions for maintaining continuous management and continuous identification.

Means for Solving the Problems

[0007] In one aspect, the present invention features a method for performing a patient-specific immunotherapy procedure. A computing device receives a cell order request to produce transfected T cells for a patient. The computing device generates a patient-specific identifier associated with the cell order request, the patient-specific identifier including a patient identification element, a sales order identifier, and a cell order lot number. The computing device initiates a process to produce transfected T cells for injection into the patient's bloodstream. The process includes performing a leukocyte apheresis procedure on a patient's blood sample to collect T cells from the sample, transferring the collected T cells to a container, labeling the container with the patient-specific identifier, sending the collected T cells to a manufacturing facility, producing transfected T cells from the collected T cells using cell modification techniques, receiving the transfected T cells from the manufacturing facility, and injecting the transfected T cells into the patient's bloodstream. The computing device records tracking events for each step of the process, each tracking event including the patient-specific identifier. The tracking events include continuous management of the patient's T cells throughout the process.

[0008] In another aspect, the present invention features a method for tracking cell orders during an immunotherapy procedure. A computing device receives a cell order request to produce transfected T cells for a patient. The computing device generates a patient-specific identifier associated with the cell order request, the patient-specific identifier including a patient identification element, a sales order identifier, and a cell order lot number. The computing device monitors the process of producing transfected T cells for injection into the patient's bloodstream, the process including receiving an indicia indicating that a leukocyte apheresis procedure has been performed on a patient's blood sample to collect T cells from the sample; receiving an indicia indicating that the collected T cells have been transferred to a container; receiving an indicia indicating that the container has been labeled with the patient-specific identifier; receiving an indicia indicating that the collected T cells have been sent to a manufacturing facility; receiving an indicia indicating that transfected T cells have been produced from the collected T cells using cell modification techniques; receiving an indicia indicating that the transfected T cells have been received from the manufacturing facility; and receiving an indicia indicating that the transfected T cells have been injected into the patient's bloodstream. The computing device records tracking events upon receiving indicia, with each tracking event containing a patient-specific identifier. The computing device maintains continuous management of the patient's T cells by saving tracking events during the process.

[0009] In another aspect, the present invention features a method for carrying out a patient-specific immunotherapy procedure. A cell order request is received for the production of transfected T cells for the patient. An event tracking module running on a processor generates a patient-specific identifier associated with the cell order request. The process for producing transfected T cells to be injected into the patient's bloodstream is initiated. The process includes performing a leukocyte apheresis procedure on a sample of the patient's blood to collect T cells from the sample, transferring the collected T cells to a container, labeling the container with the patient-specific identifier, sending the collected T cells to a manufacturing facility, producing transfected T cells from the collected T cells using cell modification techniques, receiving the transfected T cells from the manufacturing facility, and injecting the transfected T cells into the patient's bloodstream. The event tracking module receives a first tracking event from a first client device located at the site of the leukocyte apheresis procedure, confirming the leukocyte apheresis procedure and including the patient-specific identifier. The event tracking module adds the first tracking event to a data structure relating to the patient-specific identifier. The event tracking module integrates the data structure, which is stored in a database, and the integration process records a first timestamp along with the first tracking event. The event tracking module receives a second tracking event from a second client device located in the manufacturing facility, confirming receipt of the collected T cells at the manufacturing facility and containing a patient-specific identifier. The event tracking module integrates the second tracking event into the data structure relating to the patient-specific identifier, and the integration process records a second timestamp along with the second tracking event.

[0010] In another aspect, the present invention features a method for carrying out a patient-specific immunotherapy procedure. A tracking module running on a processor receives a cell order request to produce transfected T cells for a patient. The tracking module generates a patient-specific identifier and a cell order lot associated with the cell order request, identifying the patient. A database generates data records that track the cell orders, and these data records are identified in the database according to the patient-specific identifier. The tracking module receives a first tracking event indicating that the collected T cells are ready for shipment to the manufacturing facility. The data record corresponding to the patient-specific identifier is updated according to the first tracking event. The tracking module receives a second tracking event indicating that the collected T cells have been received at the manufacturing facility, based on the container received at the manufacturing facility. The data record corresponding to the patient-specific identifier is updated according to the second tracking event. The tracking module receives a third tracking event indicating that the transfected T cells have been produced, based on the manufacturing facility that produced the transfected T cells from the collected T cells using cell modification techniques. The data record corresponding to the patient-specific identifier is updated according to the third tracking event. The tracking module receives a fourth tracking event indicating that the transfected T cells have been received, based on the transfected T cells received from the manufacturing facility. The data record corresponding to the patient-specific identifier is updated according to the fourth tracking event. The tracking module receives a fifth tracking event indicating that the transfected T cells have been injected into the patient's bloodstream, based on the transfected T cells injected into the patient's bloodstream. The data record corresponding to the patient-specific identifier is updated according to the fifth tracking event, where the first, second, third, fourth, and fifth tracking events each include a patient-specific identifier, a timestamp, and an event identifier, and the data record corresponding to the patient-specific identifier stores the first, second, third, fourth, and fifth tracking events in a regular order as the data record is updated according to each event.

[0011] Any of the above embodiments may include one or more of the following features:

[0012] In some embodiments, the transfected T cells are produced by transfecting the collected T cells with a polynucleotide encoding a chimeric antigen receptor (CAR), the CAR containing an antigen-binding molecule that specifically binds to a target molecule. Preferably, the antigen-binding molecule is a single-stranded variable fragment (scFv).

[0013] In some embodiments, the target molecule is assumed to be a blood-derived cancer-associated antigen. The blood-derived cancer-associated antigen of the present invention is associated with one or more cancers selected from the group consisting of acute myeloid leukemia (AML), chronic myeloid leukemia (CML), chronic myelomonocytic leukemia (CMML), juvenile myelomonocytic leukemia, atypical chronic myelomonocytic leukemia, acute promyelocytic leukemia (APL), acute monoblastic leukemia, acute erythroid leukemia, acute megakaryoblastic leukemia, lymphoblastic leukemia, B-lineage acute lymphoblastic leukemia, B-cell chronic lymphocytic leukemia, B-cell non-Hodgkin lymphoma, myelodysplastic syndrome (MDS), myeloproliferative disorders, myeloneoplasms, myelosarcoma, and blastic plasmacytoid dendritic cell neoplasms (BPDCN).

[0014] In some embodiments, the target molecule is a viral infection-associated antigen. The virus of the present invention Infection can be caused by any virus, including, for example, HIV.

[0015] The chimeric antigen receptor (CAR) of the present invention may further comprise at least one costimulatory domain. The co-stimulatory domains of the present invention include CD28, OX-40, 4-1BB / CD137, CD2, CD7, CD27, CD30, CD40, programmed cell death-1 (PD-1), inducible T cell costimulatory molecule (ICOS), lymphocyte function-associated antigen-1 (LFA-1, CD11a / CD18), CD3 gamma, CD3 delta, CD3 epsilon, CD247, CD276 (B7-H3), LIGHT (TNFSF14), NKG2C, Ig alpha (CD79a), DAP-10, Fc gamma receptor, MHC class I molecule, TNF receptor protein, immunoglobulin protein, cytokine receptor, integrin, signal transduction lymphocyte activating molecule (SLAM protein), activated NK cell receptor, BTLA, Toll ligand receptor, ICAM-1, B7-H3, CDS, ICAM-1, GITR, B AFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8 Alpha, CD8 Beta, IL-2R Beta, IL-2R Gamma, IL-7R Alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, C D11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA-1, IT GB7, NKG2D, TNFR2, TRANCE / RANKL, DNAM1(CD226), SLAMF4(CD244, 2B4), CD84, CD96(Tactile), CEACAM1, CRT Examples include, but are not limited to, ligands that specifically bind to AM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, ​​LAT, GADS, SLP-76, PAG / Cbp, CD19a, CD83, or signaling domains of any combination thereof, or other preferred parts.

[0016] The co-stimulatory domain preferably includes CD28. The CD28 co-stimulatory domain of the present invention includes, for example, a sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, and SEQ ID NO: 8.

[0017] In other embodiments, the co-stimulatory domain includes CD8. The CD8 co-stimulatory domain of the present invention includes, for example, SEQ ID NO: 14.

[0018] The chimeric antigen receptor (CAR) of the present invention may further comprise at least one activating domain. The activating domain of the present invention comprises, for example, CD3. The CD3 activating domain preferably comprises CD3 zeta. The CD3 zeta activating domain of the present invention comprises, for example, SEQ ID NO: 10.

[0019] In a preferred embodiment, the chimeric antigen receptor (CAR) comprises all or part of anti-CD19 scFv, CD28, and CD3 zeta.

[0020] In a further preferred embodiment, the chimeric antigen receptor (CAR) is Anti-CD19 scFv, CD8 and 4-1BB, all or part of it All or part of anti-BCMA scFv and CD8, Anti-CD19 scFv, CD28 and 4-1BB, all or part of it Anti-CD22 scFv and CD8, all or part of it All or part of anti-CD19 scFv, CD28 and EGFRt / 19-28z / 4-1BBL Anti-MUC16 scFv and CD28, all or part of it All or part of the anti-CD171 All or part of the anti-CD123 and CD28 All or part of anti-BCMA, CD8, and 4-1BB, All or part of anti-CD19 and CD28 All or part of anti-CD19 and CD8, All or part of CD28 Includes.

[0021] In a further preferred embodiment, the CAR comprises a leader sequence (CSF2RA), an anti-CD19 scFv, a Whitlow linker, a CD28 spacer, a CD28 costimulatory domain, and a CD3 zeta. An example of such a CAR is encoded by the nucleotide sequence of SEQ ID NO: 146 and the amino acid sequence of SEQ ID NO: 147.

[0022] In a further preferred embodiment, the CAR comprises a leader sequence (CD8), an anti-CD19 scFv, a Whitlow linker, a CD28T spacer, a CD28 costimulatory domain, and a CD3 zeta. An example of such a CAR is encoded by the nucleotide sequence of SEQ ID NO: 148 and the amino acid sequence of SEQ ID NO: 149.

[0023] In another preferred embodiment, the CAR comprises a leader sequence (CD8a), an anti-CD19 scFv, a Whitlow linker, a CD8a spacer and transmembrane domain, a CD28 costimulatory domain, and a CD3 zeta. An example of such a CAR is encoded by the nucleotide sequence of SEQ ID NO: 150 and the amino acid sequence of SEQ ID NO: 151.

[0024] In a further preferred embodiment, the CAR comprises a leader sequence (CD8), anti-CLL-1 scFv, a G4S linker, a minispacer, CD28T (the extracellular / transmembrane region of CD28), CD28 (the intracellular costimulatory region of CD28), and CD3 zeta. An example of such a CAR is encoded by the nucleotide sequence of SEQ ID NO: 154 and the amino acid sequence of SEQ ID NO: 155.

[0025] In a further preferred embodiment, the CAR is a leader sequence (CD8a), anti-BCMA It includes scFv, Whitlow linker, CD28T spacer, CD28 costimulatory domain, and CD3 zeta. An example of such a CAR is encoded by the nucleotide sequence of SEQ ID NO: 156 and the amino acid sequence of SEQ ID NO: 157.

[0026] In some embodiments, transfected T cells are produced by transfecting collected T cells with a polynucleotide encoding a T cell receptor (TCR). The TCR of the present invention can bind to tumor-associated antigens. Tumor-associated antigens include adrenocortical carcinoma, anal carcinoma, bladder carcinoma, bone carcinoma, brain carcinoma, breast carcinoma, carcinoma, cervical carcinoma, colon carcinoma, endometrial carcinoma, esophageal carcinoma, extrahepatic bile duct carcinoma, extracranial germ cell carcinoma, eye carcinoma, gallbladder carcinoma, and gastric carcinoma. Cancer, germ cell tumors, gestational trophoblastic tumors, head and neck cancer, hypopharyngeal cancer, islet cell carcinoma, kidney cancer, colorectal cancer, laryngeal cancer, leukemia, lip and oral cancer, liver cancer, lung cancer, lymphoma, malignant mesothelioma, Merkel cell carcinoma, mycosis fungoides, myelodysplastic syndrome, myeloproliferative disorders, nasopharyngeal cancer, neuroblastoma, oral cancer, oropharyngeal cancer, osteosarcoma, ovarian epithelial carcinoma, ovarian germ cell carcinoma, pancreatic cancer, sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pituitary cancer, plasma cell neoplasms, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, transitional cell carcinoma of the renal pelvis and ureter, salivary gland cancer, Sézary syndrome, skin cancer, small intestine cancer, soft tissue sarcoma, stomach cancer It is associated with one or more cancers selected from the group consisting of testicular cancer, thymoma, thyroid cancer, urethral cancer, uterine cancer, vaginal cancer, vulvar cancer, and Wilms' tumor.

[0027] In certain embodiments, this application may be suitable for target molecules against hematological cancers. In some embodiments, cancer is a cancer of white blood cells. In other embodiments, cancer is a cancer of plasma cells. In some embodiments, cancer is leukemia, lymphoma, or myeloma. In certain embodiments, cancer is acute lymphoblastic leukemia (ALL) (including non-T cell ALL), acute lymphoblastic leukemia (ALL), and hemophagocytic lymphohistiocytosis (HLH), B-cell prelymphoblastic leukemia, B-cell acute lymphoblastic leukemia ("BALL"), blastic plasmacytoid dendritic cell neoplasm, Burkitt lymphoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic or acute granuloma. Sexual diseases, chronic or acute leukemia, diffuse large B-cell lymphoma, diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, follicular lymphoma (FL), hairy cell leukemia, hemophagocytic syndrome (macrophage activation syndrome (MAS)), Hodgkin's disease, large cell granuloma, leukocyte adhesion disorder, malignant lymphoproliferative disorder, MALT lymphoma, mantle cell lymphoma, marginal zone lymphoma, monoclonal gammaglobulinemia of unknown significance (M GUS), multiple myeloma, myelodysplasia and myelodysplastic syndromes (MDS), bone marrow diseases including acute myeloid leukemia (AML), non-Hodgkin lymphoma (NHL), plasma cell proliferation disorders (e.g., asymptomatic myeloma (smoldering multiple myeloma or painless myeloma), plasmablastic lymphoma, plasmacytoid dendritic cell neoplasms, plasmacytomas (e.g., plasma cell hyperplasia; solitary myeloma; solitary plasmacytoma; extramedullary plasmacytoma; and multi These include plasmacytoma, POEMS syndrome (Crow-Fukase syndrome; Takatsuki disease; PEP syndrome), primary mediastinal large B-cell lymphoma (PMBCL), small cell or large cell follicular lymphoma, splenic marginal zone lymphoma (SMZL), systemic amyloid light chain amyloidosis, T-cell acute lymphoblastic leukemia (TALL), T-cell lymphoma, transformed follicular lymphoma, Waldenström macroglobulinemia, or combinations thereof.

[0028] In a further preferred embodiment, the TCR comprises the AV38-2 variable alpha chain, the BV7-2 variable beta chain, a mouse constant alpha domain, a mouse constant beta domain, TRAJ40, furin-SG SG-P2A, and TRBJ1-3. An example of such a TCR is encoded by the nucleotide sequence of SEQ ID NO: 152 and the amino acid sequence of SEQ ID NO: 153.

[0029] In a further preferred embodiment, the TCR comprises a TCR beta-chain variable region, a TCR beta-chain constant region, a P2A peptide (including a furin cleavage site and a linker), a TCR alpha-chain variable region, and a TCR alpha-chain constant region. An example of such a TCR is encoded by the nucleotide sequence of SEQ ID NO: 158 and the amino acid sequence of SEQ ID NO: 159. Another example of such a TCR is encoded by the nucleotide sequence of SEQ ID NO: 160 and the amino acid sequence of SEQ ID NO: 161.

[0030] In other embodiments, the TCR of the present invention binds to a viral infection-associated antigen. The viral infection of the present invention may be caused by any virus, including, for example, HIV.

[0031] The polynucleotides of the present invention may be components of a vector. Suitable vectors include, but are not limited to, retroviral vectors, DNA vectors, plasmids, RNA vectors, adenovirus vectors, adenovirus-related vectors, lentiviral vectors, or any combination thereof. The vector is preferably a lentiviral vector. Examples of lentiviral vectors of the present invention include, but are not limited to, pGARs and their derivatives.

[0032] In some embodiments, initiating the process of producing transfected T cells involves requesting a time slot from a remote computing device to perform a leukocyte apheresis procedure, and receiving indicia in response to the request for a time slot that is available to perform the leukocyte apheresis procedure. This further includes trusting and confirming acceptance of available time slots on the remote computing device.

[0033] In some embodiments, initiating a process to produce transfected T cells further includes receiving an indicia indicating that the transfected T cells were shipped from a first site after they were produced, and receiving an indicia indicating that the transfected T cells were received at a second site before they were injected, wherein the computing device records a tracking event when it receives an indicia indicating that the transfected T cells were shipped, and records a tracking event when it receives an indicia indicating that the transfected T cells were received at a second site.

[0034] In some embodiments, the patient identification element includes a first patient ID associated with the immunotherapy procedure and a second patient ID associated with the facility performing one or more of the leukocyte apheresis procedure or the transfected T cell infusion. In some embodiments, the computing device stores the tracking events in a regular order. In some embodiments, the regular order is arranged chronologically.

[0035] Other aspects and advantages of the present invention will become apparent from the following detailed description, together with the accompanying drawings illustrating the principles of the invention for illustrative purposes only.

[0036] The advantages of the present invention described above, along with further advantages, can be better understood by referring to the following description in conjunction with the accompanying drawings. The drawings are not necessarily to scale and instead focus on illustrating the general principles of the present invention. [Brief explanation of the drawing]

[0037] [Figure 1A]This is a block diagram of a system for implementing patient-specific immunotherapy procedures using continuous monitoring and identification of biological samples. [Figure 1B] This is a detailed block diagram of a system for implementing patient-specific immunotherapy procedures using continuous monitoring and identification of biological samples. [Figure 2] This is a flowchart of a computerized method for implementing patient-specific immunotherapy procedures using continuous monitoring and identification of biological samples. [Figure 3A] This is an exemplary screenshot generated by the user interface module to receive patient-specific information during a patient-specific immunotherapy procedure. [Figure 3B] This is an exemplary screenshot generated by the user interface module to receive patient-specific information during a patient-specific immunotherapy procedure. [Figure 4A] This is an exemplary screenshot generated by the user interface module to receive confirmation of extraction and injection sites and schedule appointments during a patient-specific immunotherapy procedure. [Figure 4B] This is an exemplary screenshot generated by the user interface module to receive confirmation of extraction and injection sites and schedule appointments during a patient-specific immunotherapy procedure. [Figure 4C] This is an exemplary screenshot generated by the user interface module to receive confirmation of extraction and injection sites and schedule appointments during a patient-specific immunotherapy procedure. [Figure 4D] This is an exemplary screenshot generated by the user interface module to receive confirmation of extraction and injection sites and schedule appointments during a patient-specific immunotherapy procedure. [Figure 5A]This is an exemplary screenshot generated by the user interface module to display the ongoing management of biomaterials during a patient-specific immunotherapy procedure. [Figure 5B] This is an exemplary screenshot generated by the user interface module to display the ongoing management of biomaterials during a patient-specific immunotherapy procedure. [Modes for carrying out the invention]

[0038] Figure 1A is a block diagram of a system 100 for implementing patient-specific immunotherapy procedures using continuous management and continuous identification of biological samples. The system in Figure 1A includes multiple client computing devices 102a-102d, a communication network 104, a server computing device 106 equipped with a user interface module 108a, an event tracking module 108b, and a continuous management module 108c, and a database 110.

[0039] The client computing devices 102a-102d communicate with the server computing device 106 by connecting to the communication network 104, providing inputs and receiving outputs related to the process of implementing patient-specific immunotherapy procedures using continuous management and continuous identification of biological samples as described herein. In some embodiments, each client computing device 102a-102d may be coupled to its respective display device to provide a detailed graphical user interface (GUI) that receives inputs from, for example, the methods and systems described herein and presents outputs. For example, the client computing devices 102a-102d may be connected to a user interface module 108a of the server computing device 106, which provides users of the client computing devices 102a-102c with a web-based portal for accessing the methods and related functions described herein.

[0040] Examples of client devices 102a-102d include, but are not limited to, desktop computers, laptop computers, tablets, mobile devices, smartphones, and internet-connected home appliances. It will be understood that other types of computing devices that can connect to the components of the system in Figure 1A may be used without departing from the scope of the present invention. It should be understood that each of the client computing devices 102a-102d may be associated with a different user type. For example, client computing device 102a may be associated with a patient who accesses the system in Figure 1A to generate a user profile and receive updates regarding patient-specific immunotherapy procedures; client computing device 102b may be associated with a physician who is treating the patient and accessing the system in Figure 1A to initiate the patient's immunotherapy; client computing device 102c may be associated with a hospital or other facility administering the immunotherapy procedure to the patient; and client computing device 102d may be associated with a manufacturing facility that is producing patient-specific immunotherapy products (as described herein) for use in the immunotherapy procedure.

[0041] The communication network 104 enables other components of system 100 to communicate with each other in order to carry out the process of implementing patient-specific immunotherapy procedures using continuous management and continuous identification of biological samples as described herein. Network 104 may be a local network such as a LAN, or a wide-area network such as the Internet and / or a cellular network. In some embodiments, network 104 consists of several separate networks and / or subnetworks (e.g., from cellular to the Internet) that enable the components of the system in Figure 1A to communicate with each other.

[0042] The server computing device 106 is a combination of hardware and software modules, including dedicated hardware and / or software modules that run on a processor and interact with the memory modules of the server computing device 106 to perform functions for carrying out patient-specific immunotherapy procedures using continuous management and continuous identification of biological samples as described herein. The server computing device 106 includes a user interface module 108a, an event tracking module 108b, and a continuous management module 108c (as described above) that run on and / or interact with the processor of the server computing device 106.

[0043] In some embodiments, the user interface module 108a, the event tracking module 108b, and the continuity management module 108c are special sets of computer software instructions programmed into one or more dedicated processors within the server computing device 106, and may include specifically designated memory locations and / or registers for executing these special computer software instructions. While modules 108a-108c are shown in Figure 1A as running within the same server computing device 106, in some embodiments, the functionality of modules 108a-108c may be distributed across multiple server computing devices. As shown in Figure 1A, the server computing device 106 enables modules 108a-108c to communicate with each other to exchange data for the purpose of performing the described functionality. It should be understood that, without departing from the scope of the invention, any number of computing devices may be used, arranged in various architectures, resources, and configurations (e.g., cluster computing, virtual computing, cloud computing). Exemplary functionality of modules 108a-108c is described in detail below.

[0044] Database 110 is a computing device (or, in some embodiments, a set of computing devices) coupled to a server computing device 106 and is configured to receive, generate, and store specific segments of data relating to the process of implementing patient-specific immunotherapy procedures using continuous management and continuous identification of biological samples as described herein. In some embodiments, all or part of Database 110 may be integrated with the server computing device 106 or may reside on one or more separate computing devices. Database 110 may comprise one or more databases configured to store portions of data used by other components of the system in Figure 1A, as will be described in more detail below. In some embodiments, Database 110 includes an enterprise business suite such as Oracle's E-Business Suite (EBS), which includes a variety of modules that enable diverse functions to support the methods and systems described herein, including logistics management, supply chain, transportation, CRM, and other types of modules.

[0045] Figure 1B is a detailed block diagram of the system in Figure 1A for implementing patient-specific immunotherapy procedures using continuous management and continuous identification of biological samples. As shown in Figure 1B, the server computing device 106 is the central component of the overall hardware architecture, interfaced with the client computing devices 102a-102e and the database 110, and also interfaced with the scheduling module 114 and the physician's master data feed 116. In some embodiments, the server computing device 106 and corresponding modules 108a-108c leverage the Salesforce platform, available from Salesforce.com, Inc. in San Francisco, California, as described herein. Specific functions are integrated. Client computing devices 102a to 102e communicate with server computing device 106 to perform patient registration in immunotherapy procedures and monitor the tracking of ongoing management and ongoing identification as described herein (e.g., via a browser-based user interface).

[0046] For example, client computing device 102a may be associated with a patient undergoing an immunotherapy procedure and may include browser and email software to allow the patient to monitor tracking and electronically sign documents necessary to participate in the immunotherapy procedure (e.g., via DocuSign or other similar technology). Similarly, client computing devices 102b-102d may be located in different hospitals where a treating physician can use browser software to enroll a patient in an immunotherapy procedure, order cells in the system, and monitor tracking by continuous management and continuous identification. Client computing devices 102b-102d may also include a single sign-on (SSO) module that allows the device to authenticate to server computing device 106 (e.g., using SSO supported by SAML2.0 or a specific username / password for the server). Client computing device 102e may be located at an administration or manufacturing site so that an administrator of server computing device 106 can use browser software to communicate with the server, receive communications such as email from other participants in the system, and monitor tracking by continuous management and continuous identification.

[0047] As described above, the database 110 may include an enterprise business suite for managing data from the server computing device 106, and may include modules that enable tracking and logistics management through continuous management and identification of biological samples. For example, the database 110 may, upon request, send approved customer sites to the server 106, receive cell order entry data from the server 106, and provide the server 106 with cell order reservations and apheresis slot update information.

[0048] The scheduling module 114 may be integrated into the server computing device 106 or reside on a separate computing device. The scheduling module 114 authenticates and communicates with the server computing device 106, receives certain information regarding cell orders and immunotherapy procedures (e.g., patient ID, apheresis site, injection site, and product code), and can provide the server 106 with calendaring and scheduling functions (e.g., treatment This allows the physician to select the date / time of apheresis and to provide a scheduled delivery date / time for the biological sample after it has undergone the manufacturing process. Additionally, the server computing device 106 may communicate with the physician's master data feed 116 (provided, for example, using a Veeva® CRM platform integrated with a Heroku® application) to receive certain information about the treating physician.

[0049] Figure 2 is a flowchart of a computerized method 200 for implementing a patient-specific immunotherapy procedure using the system of Figure 1A and / or the system of Figure 1B, with continuous management and continuous identification of the biological sample tracking. To be understood, the methods and systems described herein are presented in relation to implementing a patient-specific immunotherapy procedure. As used herein, “patient-specific immunotherapy procedure” means any procedure that uses molecular or cellular components of the immune system to target and / or destroy cells that cause cancer, pathogenicity, or other disease. An immunotherapy procedure is “patient-specific” if it utilizes components of the patient’s immune system to treat that patient’s own cancer, pathology, or other disease.

[0050] As used herein, the terms “patient” and “subject” are used interchangeably and include, alongside humans and non-human animals, those with formally diagnosed disabilities, those without formally recognized disabilities, those receiving medical treatment, those at risk of developing disabilities, etc. In addition to humans, categories of animals within the scope of this invention include, for example, farm animals, domestic animals, and laboratory animals. Some examples of farm animals include cattle, pigs, horses, and goats. Some examples of domestic animals include dogs and cats. Some examples of laboratory animals include rats, mice, rabbits, and guinea pigs.

[0051] The term "leukocyte apheresis" refers to a specific form of apheresis involving the selective separation and removal of leukocytes from a blood sample. During leukocyte apheresis, the extracted blood is passed through a cell separation device that separates nucleated leukocytes, including T cells, from red blood cells and plasma. The separated T cells can then be collected and used in the cell modification techniques of the present invention. In certain embodiments, the red blood cells and plasma are returned to the organism as part of the separation process. In further embodiments, the red blood cells and plasma are discarded or stored for further analysis.

[0052] The terms “T cell” and “T lymphocyte” as used herein are not distinguished. T cells are a subset of lymphocytes defined by their development in the thymus and by heterodimeric receptors associated with the CD3 complex protein. The T cells of the present invention include, but are not limited to, naive T cells, cytotoxic T cells, helper T cells, suppressor T cells, regulatory T cells, memory T cells, NKT cells, γδ cells, CD8αα cells, lymphokine-activated cells, TCR-expressing cells, their subtypes, and any other cell types capable of expressing chimeric receptor chains.

[0053] T cells can be modified to be specific to one or more desired targets. For example, T cells can be transduced with DNA or other genetic material encoding antigen-binding molecules, such as one or more single-strand variable fragments ("scFv") of an antibody, in conjunction with one or more signaling molecules and / or one or more activation domains, such as CD3 zeta. In addition to the ability of CAR-T cells to recognize and destroy target cells, the success of T cell therapy benefits from the ability of CAR-T cells to sustain and maintain their ability to proliferate in response to antigens.

[0054] As used herein, the term “cell modification techniques” includes, but is not limited to, transfection and transduction. The term “transfection” and its grammatical variations refer to the introduction of foreign or exogenous DNA into cells. Many transfection techniques are well known in the art and are disclosed herein. See, for example, Graham et al., 1973, Virology 52:456; Sambrook et al., 2001, Molecular Cloning: A Laboratory Manual; Davis et al., 1986, Basic Methods in Molecular Biology, Elsevier; and Chu et al., 1981, Gene 13:197. Transfection techniques include, but are not limited to, calcium phosphate-DNA coprecipitation, DEAE-dextran-mediated transfection, cationic lipid-mediated delivery, polybrene-mediated transfection, electroporation, sonoporation, microinjection, liposome fusion, lipofection (lipid transfection), polymer transfection, nanoparticles, polyplexes, receptor-mediated gene delivery, delivery mediated by polylysine, histones, chitosan, and peptides, protoplast fusion, retroviral infection, and bioristics (e.g., gene guns). The term "transduction" and its grammatical variations refer to the process by which foreign DNA is introduced into cells via a viral vector. Jones et al., (1998). Genetics: principles and analysis. See Boston: Jones & Bartlett Publ.

[0055] As used herein, the term “inject” and its grammatical variations refer to the act of injecting through the body via blood vessels. This means introducing a solution into a body. The injection of the present invention includes, but is not limited to, the therapeutic introduction of fluids other than whole blood into a blood vessel. For example, the transfected T cells of the present invention can be injected into the patient's bloodstream, for example, intramuscularly, intravenously, intraarterially, intraperitoneally, or subcutaneously.

[0056] It will be understood that chimeric antigen receptors (CARs or CAR-Ts) are genetically modified receptors, and T cell receptors (TCRs) can be genetically modified receptors. These modified receptors can be readily inserted into immune cells, including T cells, and expressed by immune cells according to techniques known in the art. Using CARs, a single receptor can be programmed to both recognize specific antigens and, upon binding to those antigens, activate immune cells to attack and destroy cells possessing those antigens. If these antigens are present in tumor cells, immune cells expressing CARs can target and kill the tumor cells.

[0057] CARs can be modified to bind to antigens (such as cell surface antigens) by incorporating antigen-binding molecules that interact with their target antigens. As used herein, “antigen-binding molecule” means any protein that binds to a specific target molecule. Antigen-binding molecules include, but are not limited to, antibodies and their binding portions, such as immunologically functional fragments. Peptibodies (i.e., Fc fusion molecules containing peptide-binding domains) are another example of suitable antigen-binding molecules.

[0058] The term “immunologically functional fragment” (or “fragment”) of an antigen-binding molecule refers to a type of antigen-binding molecule that contains a portion of an antibody (regardless of how that portion is obtained or synthesized) that lacks at least some of the amino acids present in the full-length chain but is still capable of specifically binding to an antigen. Such fragments are biologically active in that they bind to a target antigen and may compete for binding to a given epitope with other antigen-binding molecules containing intact antibodies. In some embodiments, the fragment is a neutralizing fragment. In one embodiment, such a fragment retains at least one CDR present in the full-length light or heavy chain, and in some embodiments, it comprises a single heavy chain and / or a light chain or a portion thereof. These fragments may be produced by recombinant DNA technology or by enzymatic or chemical cleavage of an antigen-binding molecule containing intact antibodies.

[0059] Immunologically functional immunoglobulin fragments include, but are not limited to, scFv fragments, Fab fragments (Fab', F(ab')2, etc.), one or more CDRs, diabodies (heavy chain variable domains on the same polypeptide as a light chain variable domain, connected via short peptide linkers too short to pair between two domains on the same chain), domain antibodies, and single-chain antibodies. These fragments may originate from any mammalian source, including but not limited to humans, mice, rats, camelids, or rabbits. As will be understood by those skilled in the art, antigen-binding molecules may contain non-protein components.

[0060] In some embodiments, the antigen-binding molecule binds to an antigen on a tumor cell. In some embodiments, the antigen-binding molecule binds to an antigen on a cell involved in hyperproliferative disease, or to a viral or bacterial antigen. In further embodiments, the antigen-binding molecule is an antibody or a fragment thereof containing one or more of its complementarity-determining regions (CDRs). In further embodiments, the antigen-binding molecule is a single-stranded variable fragment (scFv).

[0061] The antigen-binding molecule is preferably an antibody fragment. More preferably, the antigen-binding molecule contains one or more single-chain variable fragments ("scFv"). The scFv is a single-chain antibody fragment having variable regions of the heavy and light chains of the antibody linked together. See U.S. Patent Nos. 7,741,465, 6,319,494, International Publication No. 88 / 01649, U.S. Patent Nos. 4,946,778, and 5,260,203, and Eshhar et al., Cancer Immunol Immunotherapy (1997) 45: 131-136, the contents of which are further incorporated herein by reference. scFv retains the ability of the parental antibody to specifically interact with the target antigen. scFv can be modified to be expressed together with other CAR components as part of a single chain, making it preferable for use in chimeric antigen receptors (ibid.). See also Krause et al., J. Exp. Med., Volume 188, No. 4, 1998 (619-626), and Finney et al., Journal of Immunology, 1998, 161: 2791-2797.

[0062] It will be understood that antigen-binding molecules are typically contained in the extracellular portion of the CAR so that they can recognize and bind to the target antigen. Bispecific and multiplespecific CARs having specificity for two or more target organisms are intended within the scope of the present invention.

[0063] In certain embodiments, the polypeptide structure of the antigen-binding molecule is based on an antibody, including but not limited to monoclonal antibodies, bispecific antibodies, minibodies, domain antibodies, synthetic antibodies (sometimes referred to herein as “antibody mimes”), chimeric antibodies, humanized antibodies, human antibodies, antibody fusions (sometimes referred to herein as “antibody conjugates”), and fragments thereof. In some embodiments, the antigen-binding molecule includes or consists of an avimer.

[0064] An antigen-binding molecule is said to "specifically bind" to its target antigen when the dissociation constant (K d ) is about 1×10 -7 M. The antigen-binding molecule specifically binds to the antigen with "high affinity" when K d is 1×10 -9 M to 5×10 -9 M, and with "very high affinity" when K d is 1×10 -10 M to 5×10 -10 M. In one embodiment, the antigen-binding molecule has a K -9 of 10 d M. In one embodiment, the dissociation rate (off-rate) is less than 1×10 -5 .

[0065] One of ordinary skill in the art will understand that the target molecule of the present invention can be any molecule to which it is desirable to transfer specificity to transfected T cells. The terms "target molecule" or "antigen" refer to a molecule or a portion of a molecule to which an antigen-binding molecule can bind. In certain embodiments, the target may have one or more epitopes.

[0066] The target molecules of the present invention include, but are preferably not limited to, blood-derived cancer-associated antigens. Non-limiting examples of blood-derived cancer-associated antigens include antigens associated with one or more cancers selected from the group consisting of acute myeloid leukemia (AML), chronic myelogenous leukemia (CML), chronic myelomonocytic leukemia (CMML), juvenile myelomonocytic leukemia, atypical chronic myeloid leukemia, acute promyelocytic leukemia (APL), acute monoblastic leukemia, acute erythroid leukemia, acute megakaryocytic leukemia, lymphoblastic leukemia, B-lineage acute lymphoblastic leukemia, B-cell chronic lymphocytic leukemia, B-cell non-Hodgkin lymphoma, myelodysplastic syndrome (MDS), myeloproliferative disorder, myeloneoplasm, myelosarcoma), and blastic plasmacytoid dendritic cell neoplasm (BPDCN).

[0067] In some embodiments, the antigen is a tumor-associated antigen, such as 5T4, alpha-fetoprotein (AFP), B7-1 (CD80), B7-2 (CD86), BCMA, β-human chorionic gonadotropin, CA-125, carcinoembryonic antigen (CEA), CD123, CD133, CD138, CD19, CD20, CD22, CD23, CD24, CD25, CD30, CD33, CD34, CD4, CD40, CD44, CD56, CD8, CLL-1, c-Met, CMV-specific antigen, CSPG4, CTLA-4, disialoganglioside GD2, ductal epithelial mucin, EBV-specific antigen, or EGFR variant II. I (EGFRvIII), ELF2M, endoglin, ephrin B2, epidermal growth factor receptor (EGFR), epithelial cell adhesion molecule (EpCAM), epithelial tumor antigen, ErbB2 (HER2 / neu), fibroblast-related protein (fap), FLT3, folate-binding protein, GD2, GD3, glioma-related antigen, sphingoglycolipid, gp36, HBV-specific antigen, HCV-specific antigen, HER1-HER2, HER2-HER3 combination, HERV-K, high molecular weight melanoma-related antigen (HMW-MAA), HIV-1 envelope glycoprotein gp41, HPV-specific antigen, human telomerase Reverse transcriptase, IGF-II receptor, IGF-II, IL-11R-alpha, IL-13R-a2, influenza virus-specific antigen; CD38, insulin growth factor (IGFI)-1, intestinal carboxylesterase, kappa chain, LAGA-1a, lambda chain, Lassa virus-specific antigen, lectin-reactive AFP, lineage-specific or tissue-specific antigens, e.g., CD3, MAGE, MAGE-A1, major histocompatibility complex (MHC) molecules, major histocompatibility complex (MHC) molecules presenting tumor-specific peptide epitopes, M-CSF, melanoma-associated antigen, mesothelin, mesothelin, MN-CA The following are selected from IX, MUC-1, mut hsp72, mutant p53, mutant p53, mutant ras, neutrophil elastase, NKG2D, Nkp30, NY-ESO-1, p53, PAP, prostase, prostase-specific antigen (PSA), prostate cancer tumor antigen-1 (PCTA-1), prostate-specific antigen, prostain, PSMA, RAGE-1, ROR1, RU1, RU2 (AS), surface adhesion molecules, survivorin and telomerase, TAG-72, extradomain A (EDA) and extradomain B (EDB) of fibronectin, and A1 domain of tenascin C (TnC A1), thyroglobulin, tumor stromal antigen, vascular endothelial growth factor receptor-2 (VEGFR2), virus-specific surface antigens, such as HIV-specific antigen (e.g., HIV gp120), and any derivatives or variants of these surface markers.

[0068] In some embodiments, the target molecule of the present invention includes a viral infection-associated antigen. The viral infection of the present invention may be caused by any virus, including, for example, HIV.

[0069] This list of possible target molecules is not intended to be exclusive, and those skilled in the art will know of many additional molecules that may be useful in targeting the chimeric antigen receptor of the present invention.

[0070] Chimeric antigen receptors can incorporate co-stimulatory (signal transduction) domains to enhance their efficacy. (U.S. Patents 7,741,465 and 6,319,494, and Krause et al. and Finney et al. (see above), Song et al., Blood 119:696-706) See (2012), Kaloset al., Sci Transl. Med. 3:95 (2011), Porter et al., N. Engl. J. Med. 365:725-33 (2011), and Gross et al., Annu. Rev. Pharmacol. Toxicol. 56:59-83 (2016). For example, CD28 is a naturally occurring co-stimulatory stenosis on T cells. It is a protein. Various co-stimulatory molecules are defined herein, but it will be understood that additional co-stimulatory molecules are also included within the scope of the present invention.

[0071] The complete native amino acid sequence of CD28 is described in the NCBI reference sequence: NP_006130.1. The complete native CD28 nucleic acid sequence is described in the NCBI reference sequence: NM_006139.1.

[0072] A specific CD28 domain was used as a chimeric antigen receptor. In accordance with this invention, a novel CD28 extracellular (hinge) construct, referred to as "CD28T," was unexpectedly found to provide certain benefits when used in CAR constructs. This construct exhibits the ability to retain (sometimes exceed) the properties of CD28-containing CARs despite the cleavage (removal) of multiple amino acids from the extracellular CD28 sequence. These benefits include equivalent or superior cytokine production, equivalent or superior cytolytic activity, and / or equivalent Examples include having a high or excellent CAR expression level.

[0073] The nucleotide sequence of the CD28T molecule, including the extracellular domain and the CD28 transmembrane and intracellular domains, is defined in Sequence ID No. 1 below: CTTGATAATGAAAAGTCAAACGGAACAATCATTCACGTGAAGGGCAAGCACCTCTGTCCGTCACCCTTGTTCCCTGGTCCATCCAAGCCATTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTGCTCGTCACC GTGGCTTTTATAATCTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGC

[0074] The corresponding amino acid sequence is defined in Sequence ID No. 2 below: LDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSK RSRLLHSDYM NMTPRRPGPT RKHYQPYAPP RDFAAYRS

[0075] The nucleotide sequence of the extracellular portion of CD28T is defined in Sequence ID No. 3 below: CTTGATAATGAAAAGTCAAACGGAACAATCATTCACGTGAAGGGCAAGCACCTCTGTCCGTCACCCTTGTTCCCTGGTCCATCCAAGCCA

[0076] The corresponding amino acid sequence of the CD28T extracellular domain is defined in Sequence ID No. 4 below: LDNEKSNGTI IHVKGKHLCP SPLFPGPSKP

[0077] The nucleotide sequence of the CD28 transmembrane domain is defined in Sequence ID No. 5 below: TTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTGCTCGTCACCGTGGCTTTTATAATCTTCTGGGTT

[0078] The amino acid sequence of the CD28 transmembrane domain is defined in Sequence ID No. 6 below: FWVLVVVGGV LACYSLLVTV AFIIFWV

[0079] The nucleotide sequence of the CD28 intracellular signaling domain is defined in Sequence ID No. 7 below: AGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGC

[0080] The amino acid sequence of the CD28 intracellular signaling domain is defined in Sequence ID No. 8 below: RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS

[0081] Additional CD28 sequences suitable for use in the present invention include the CD28 nucleotide sequence defined in Sequence ID No. 11 below: ATTGAGGTGATGTATCCACCGCCTTACCTGGATAACGAAAAGAGTAACGGTACCATCATTCACGTGAAAGGTAAACACCTGTGTCCTTCTCCCCTCTTCCCGGGCCATCAAAGCCC

[0082] The corresponding amino acid sequence is defined in Sequence ID No. 12 below: IEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP

[0083] It will be understood that the present invention relates to antigen-binding molecules, CARs, TCRs, etc., comprising at least one isolated nucleic acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3. It will be further understood that the present invention relates to antigen-binding molecules, CARs, TCRs, etc., whose extracellular portion consists of at least one isolated nucleic acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3. In addition, it will be understood that the present invention relates to antigen-binding molecules, CARs, TCRs, etc., whose extracellular portion essentially consists of at least one isolated nucleic acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3.

[0084] It will be understood that the present invention relates to antigen-binding molecules, CARs, TCRs, etc., comprising at least one amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 4. It will be further understood that the present invention relates to antigen-binding molecules, CARs, TCRs, etc., whose extracellular portion consists of at least one amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 4. It will also be understood that the present invention relates to antigen-binding molecules, CARs, TCRs, etc., whose extracellular portion essentially consists of at least one amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 4.

[0085] Another suitable source of extracellular and / or transmembrane domains may originate from (or correspond to) some or all of CD8.

[0086] The appropriate nucleotide sequences for the extracellular and transmembrane domains of CD8 are specified in Sequence ID No. 13 below: GCTGCAGCATTGAGCAACTCAATAATGTATTTTAGTCACTTTGTACCAGTGTTCTTGCCGGCTAAGCCTACTACCACACCCGCTCCACGGCCACCTACCCCAGCTCCTACCATCGCTTCACAGCCTCTGTCCCTGCGCCCAGAG GCTTGCCGACCGGCCGCAGGGGGCGCTGTTCATACCAGAGGACTGGATTTCGCCTGCGATATCTATATCTGGGCACCCCTGGCCGGAACCTGCGGCGTACTCCTGCTGTCCCTGGTCATCACGCTCTATTGTAATCACAGGAAC

[0087] The corresponding amino acid sequence is defined in Sequence ID No. 14 below: AAALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRN

[0088] Structurally, it will be understood that the domains described herein correspond to positions on immune cells or other cells. Therefore, these domains may be (i) a "hinge" or extracellular (EC) domain, (ii) a transmembrane (TM) domain, and / or (iii) part of an intracellular / cytoplasmic (IC) domain. The intracellular component often partially contains an activating domain, such as a CD3 family member, preferably part of CD3 zeta. This domain is capable of activating T cells when an antigen-binding molecule binds to its target. It will be understood that the intracellular domain typically further contains one or more co-stimulatory molecules described herein.

[0089] As used herein, “costimulatory molecules” refer to molecules that generate signals that mediate T cell responses, including but not limited to proliferation, activation, and differentiation. Costimulatory molecules can generate signals in addition to the primary signals generated by the activating molecules described herein.

[0090] Suitable co-stimulatory domains within the scope of the present invention include, for example, CD28, CD28T, OX40, 4-1BB / CD137, CD2, CD3 (alpha, beta, delta, epsilon, gamma, zeta), CD4, CD5, CD7, CD9, CD16, CD22, CD27, CD30, CD33, CD37, CD40, CD45, CD64, CD80, CD86, CD134, CD137, CD154, PD-1, ICOS, and RI. Lymphocyte function-associated antigen-1 (LFA-1, CD11a / CD18), CD247, CD276 (B7-H3), LIGHT (tumor necrosis factor superfamily member 14; TNFSF14), NKG2C, Ig alpha (CD79a), DAP-10, Fc gamma receptor, MHC class I molecule, TNF, TNFr, integrin, signal transduction lymphocyte activating molecule, BTLA, Toll ligand receptor, ICAM-1, B7 -H3, CDS, ICAM-1, GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8 Alpha, CD8 Beta, IL-2R Beta, IL-2R Gamma, IL-7R Alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD 49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA -1, ITGB7, NKG2D, TNFR2, TRANCE / RANKL, DNAM1(CD226), SLAMF4(CD244, 2B4), CD84, CD96(Tactile), CEACAM1, CRT It may originate from (or correspond to) a co-stimulatory domain such as AM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, ​​LAT, GADS, SLP-76, PAG / Cbp, CD19a, CD83 ligand or fragment, or a combination thereof.It will be understood that additional co-stimulatory molecules or their fragments not listed above are also included within the scope of the present invention.

[0091] In some embodiments, the co-stimulatory domain may include all or part of the 4-1BB nucleic acid sequence defined in SEQ ID NO: 140 and the corresponding amino acid sequence defined in SEQ ID NO: 141. In other embodiments, the co-stimulatory domain may include all or part of the amino acid sequence of OX40 defined in SEQ ID NO: 142. See also Hombach et al., Oncoimmunology. 2012 Jul.1; 1(4):458-466. In yet another embodiment, the co-stimulatory domain may include all or part of the ICOS molecule described in Guedan et al., August 14, 2014; Blood:124 (7) and Shen et al., Journal of Hematology & Oncology (2013) 6:33. In yet another embodiment, the co-stimulatory domain may include all or part of CD27 described in Song et al., Oncoimmunology. 2012 Jul. 1;1(4):547-549.

[0092] The modified T cells of the present invention include an antigen-binding molecule (such as scFv), an extracellular domain (which may include a "hinge" domain), a transmembrane domain, and an intracellular domain. The intracellular domain includes at least a partially activated domain and may preferably be composed of CD3 family members such as CD3 zeta, CD3 epsilon, CD3 gamma, or a portion thereof.

[0093] It will become clearer that antigen-binding molecules (e.g., one or more scFvs) are modified to recognize and bind to their target(s) by being located in the extracellular portion of the molecule / construct.

[0094] It will be understood that the hinge region may contain some or all members of the immunoglobulin family, such as IgG1, IgG2, IgG3, IgG4, IgA, IgD, IgE, IgM, or fragments thereof.

[0095] In some embodiments, the extracellular domain is located between the antigen-binding molecule and the transmembrane domain.

[0096] The extracellular domains of specific use in this invention include CD28, OX-40, 4-1BB / CD137, CD2, CD7, CD27, CD30, CD40, programmed cell death-1 (PD-1), inducible T cell costimulatory molecule (ICOS), lymphocyte function-associated antigen-1 (LFA-1, CD11a / CD18), CD3 gamma, CD3 delta, CD3 epsilon, CD247, CD276 (B7-H3), LIGHT (TNFSF14), NKG2C, Ig alpha (CD79a), DAP-10, Fc gamma receptor, MHC class I molecule, TNF receptor protein, immunoglobulin protein, cytokine receptor, integrin, signaling lymphocyte activator molecule (SLAM protein), activated NK cell receptor, BTLA, Toll ligand receptor, ICAM-1, B7-H3, CDS, ICAM-1, and GIT. R, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8 Alpha, CD8 Beta, IL-2R Beta, IL-2R Gamma, IL-7R Alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD , CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA-1, I TGB7, NKG2D, TNFR2, TRANCE / RANKL, DNAM1(CD226), SLAMF4(CD244, 2B4), CD84, CD96(Tactile), CEACAM1, CRT The extracellular domain may be derived from (i.e., include) all or part of ligands that specifically bind to AM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, ​​LAT, GADS, SLP-76, PAG / Cbp, CD19a, CD83, or any combination thereof. The extracellular domain may be derived from either a natural or synthetic source.

[0097] The extracellular domain often includes a hinge region, sometimes referred to as a "spacer" region. Various hinges, including some of the molecules or derivatives described herein, can be used in accordance with the present invention.

[0098] CARs can be designed using a transmembrane domain fused to the extracellular domain of the CAR. This may also be fused to the intracellular domain of the CAR. In some cases, the transmembrane domain may be selected or modified by amino acid substitutions to avoid binding of such domain to the transmembrane domain of the same or different surface membrane proteins and to minimize interaction with other members of the receptor complex. The transmembrane domain may be derived from either a natural or synthetic source. If the source is natural, the domain may be derived from any membrane-binding or transmembrane protein. The transmembrane regions of specific use in this invention include CD28, CD28T, OX-40, 4-1BB / CD137, CD2, CD7, CD27, CD30, CD40, programmed cell death-1 (PD-1), inducible T cell costimulatory molecule (ICOS), lymphocyte function-associated antigen-1 (LFA-1, CD11a / CD18), CD3 gamma, CD3 delta, CD3 epsilon, CD247, CD276 (B7-H3), LIGHT (TNFSF14), NKG2C, Ig alpha (CD79a), DAP-10, Fc gamma receptor, MHC class I molecule, TNF receptor protein, immunoglobulin protein, cytokine receptor, integrin, signaling lymphocyte activator molecule (SLAM protein), activated NK cell receptor, BTLA, Toll ligand receptor, ICAM-1, B7-H3, CDS, ICAM-1, GITR, BAFFR, LI GHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8 Alpha, CD8 Beta, IL-2R Beta, IL-2R Gamma, IL-7R Alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11 d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB 7, NKG2D, TNFR2, TRANCE / RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRT Ligands that specifically bind to AM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, ​​LAT, GADS, SLP-76, PAG / Cbp, CD19a, CD83, or any combination thereof may be derived from (including or correspond to) these ligands.

[0099] Optionally, short linkers may form links between one or more of the extracellular, transmembrane, and intracellular domains of the CAR.

[0100] In other embodiments, the transmembrane domain in the CAR of the present invention is a CD8 transmembrane domain. In one embodiment, the CD8 transmembrane domain includes the transmembrane portion of the nucleic acid sequence of SEQ ID NO: 13. In another embodiment, the CD8 transmembrane domain includes a nucleic acid sequence encoding the transmembrane amino acid sequence contained in SEQ ID NO: 14.

[0101] In certain embodiments, the transmembrane domain in the CAR of the present invention is a CD28 transmembrane domain. In one embodiment, the CD28 transmembrane domain contains the nucleic acid sequence of SEQ ID NO: 5. In one embodiment, the CD28 transmembrane domain contains a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 6. In another embodiment, the CD28 transmembrane domain contains the amino acid sequence of SEQ ID NO: 6.

[0102] The modified intracellular (cytoplasmic) domain of T cells according to the present invention can result in the activation of at least one normal effector function of immune cells. The effector function of T cells may refer to, for example, cytolytic activity or helper activity, including cytokine secretion.

[0103] Appropriate intracellular molecules include CD28, CD28T, OX-40, 4-1BB / CD137, CD2, CD7, CD27, CD30, CD40, programmed cell death-1 (PD-1), inducible T cell costimulatory molecule (ICOS), lymphocyte function-associated antigen-1 (LFA-1, CD11a / CD18), CD3 gamma, CD3 delta, CD3 epsilon, CD247, CD276 (B7-H3), LIGHT (TNFSF14), NKG2C, Ig alpha (CD79a), DAP-10, Fc gamma receptor, MHC class I molecule, TNF receptor protein, immunoglobulin protein, cytokine receptor, integrin, signal transduction lymphocyte activating molecule (SLAM protein), activated NK cell receptor, BTLA, Tol l-ligand receptor, ICAM-1, B7-H3, CDS, ICAM-1, GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8 alpha, CD8 beta, IL-2R beta, IL-2R gamma, IL-7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, NKG2D, TNF It should be understood that this includes, but is not limited to, signaling domains derived from (or corresponding to) ligands that specifically bind to R2, TRANCE / RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRT AM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, ​​LAT, GADS, SLP-76, PAG / Cbp, CD19a, CD83, or any combination thereof.

[0104] In preferred embodiments, the intracellular / cytoplasmic domain of the CAR may be designed to include the CD3 zeta domain itself or to be combined with any other desired intracellular domain(s) useful in relation to the CAR of the present invention. For example, the intracellular domain of the CAR may include a CD3 zeta chain portion and a portion of a co-stimulatory signaling molecule. The intracellular signaling sequences in the intracellular signaling portion of the CAR of the present invention may be linked together randomly or in a specified order.

[0105] In another preferred embodiment, the intracellular domain is designed to include the activation domain of CD3 zeta and the signaling domain of CD28. In another embodiment, the intracellular domain is designed to include the activation domain of CD3 zeta and the signaling domain of 4-1BB. In yet another embodiment, the intracellular domain in CAR is designed to include CD28 and a portion of CD3 zeta, where the intracellular CD28 includes the nucleic acid sequence defined in SEQ ID NO: 7 and the amino acid sequence defined in SEQ ID NO: 8. The CD3 zeta nucleic acid sequence is defined in SEQ ID NO: 9, and the amino acid sequence is defined in SEQ ID NO: 8.

[0106] As used herein, "activation" or "stimulation" refers to a primary response induced by the binding of an activating molecule to a homologous ligand, which mediates a signaling event.

[0107] "Activating molecule" or "stimulating molecule" refers to a molecule on a T cell, such as the TCR / CD3 complex that specifically binds to a homologous stimulating ligand present on an antigen-presenting cell. Appropriate activating molecules are described herein.

[0108] It will be understood that suitable activation domains within the scope of the present invention may be derived from (or correspond to) activation / stimulation molecules, such as CD3 or CD3 zeta. CD3 is an element of the T cell receptor on native T cells and has been shown to be an important intracellular activation element in CAR.

[0109] In a preferred embodiment, CD3 is CD3 zeta, and its nucleotide sequence is defined in Sequence ID No. 9 below: AGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAG GGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG

[0110] The corresponding amino acid for intracellular CD3 zeta is defined in Sequence ID No. 10 below: RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPR RKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR

[0111] It will be understood that one preferred orientation of the CAR according to the present invention is to include an antigen-binding molecule (such as scFv) together with the extracellular and / or hinge domain, costimulatory domain, and activating domain. It will be further understood that multiple domains can be utilized together.

[0112] Table 1 shows an exemplary CAR structure according to the present invention.

[0113] [Table 1]

[0114] The term "vector" refers to any molecule or entity (e.g., nucleic acid, plasmid, bacteriophage, or virus) used to transmit protein-coding information to a host cell. The terms "expression vector" or "expression construct" refer to an expression vector suitable for the transformation of a host cell and operatively linked to the expression of one or more heterogeneous coding regions. This refers to a vector containing nucleic acid sequences that are directed towards and / or regulated (in conjunction with host cells). Expression constructs may, but are not limited to, sequences that affect or regulate transcription and translation, and, where introns are present, sequences that affect RNA splicing of coding regions manipulably ligated to them.

[0115] The term "host cell" refers to a cell that has been transformed by a nucleic acid sequence, or can be transformed by a nucleic acid sequence, thereby expressing the target gene. This term includes offspring of a parent cell, regardless of whether the offspring's morphology or genetic structure is identical to that of the original parent cell, as long as the target gene is present.

[0116] The term "transformation" refers to a change in the genetic characteristics of a cell, and a cell is transformed when it is modified to contain new DNA or RNA. For example, a cell is transformed when its genes are modified from its native state by introducing new genetic material through transfection, transduction, or other techniques. After transfection or transduction, the transformed DNA can be recombined with the cell's DNA by physically incorporating it into the cell's chromosomes, or it can be temporarily maintained as an episomal element without replication, or it can be replicated independently as a plasmid. A cell is considered "stablely transformed" if the transformed DNA is replicated by cell division.

[0117] The present invention further relates to isolated polynucleotides encoding the chimeric antigen receptor (CAR) and T cell receptor (TCR) of the present invention, and to vectors containing said polynucleotides. Any vector known in the art is suitable for the present invention. In some embodiments, the vector is a viral vector. In some embodiments, the vector is a retroviral vector (e.g., pMSVG1), a DNA vector, a mouse leukemia virus vector, an SFG vector, a plasmid, an RNA vector, an adenovirus vector, a baculovirus vector, an Epstein-Barr virus vector, a papovavirus vector, a vaccinia virus vector, a herpes simplex virus vector, an adeno-associated virus vector (AAV), a lentiviral vector (e.g., pGAR), or any derivative or combination thereof.

[0118] The pGAR sequence is as follows:

[0119] Appropriate additional exemplary vectors include, for example, pBABE-puro, pBABE-neo largeTcDNA, pBABE-hygro-hTERT, and pMKO.1. Examples include GFP, MSCV-IRES-GFP, pMSCV PIG (Puro IRES GFP empty plasmid), pMSCV-loxp-dsRed-loxp-eGFP-Puro-WPRE, MSCV IRES Luciferase, pMIG, MDH1-PGK-GFP_2.0, TtRMPVIR, pMSCV-IRES-mCherry FP, pRetroX GFP T2A Cre, pRXTN, pLncEXP, and pLXIN-Luc.

[0120] In further embodiments, a mixture of various expression vectors may be used to genetically modify a donor population of immunoeffector cells, each vector encoding one of the various CARs disclosed herein. The resulting transduced immunoeffector cells form a mixed population of modified cells, with a certain percentage of the modified cells expressing two or more CARs.

[0121] In a preferred embodiment of the present invention, the CAR comprises all or part of the anti-CD19 scFv, CD28, and CD3 zeta.

[0122] The transfected T cells of the present invention can also be produced by transfecting collected T cells with a polynucleotide encoding the T cell receptor (TCR). The T cell receptor (TCR) is a molecule found on the surface of T cells that recognizes antigen fragments as a peptide bound to the major histocompatibility complex (MHC) molecule. The TCR consists of two distinct protein chains; in approximately 95% of human TCRs, the TCR consists of an alpha (α) chain and a beta (β) chain. In approximately 5% of human T cells, the TCR consists of gamma and delta (γ / δ) chains. Each chain consists of two extracellular domains: a variable (V) region and a constant (C) region, both belonging to the immunoglobulin superfamily. Like other immunoglobulins, the variable domains of the α and β chains (or gamma and delta (γ / δ) chains) of the TCR each contain three hypervariable regions or complementarity-determining regions (CDRs). When the TCR binds to an antigen peptide and MHC (peptide / MHC), the T cell is activated, allowing it to attack and destroy the target cell.

[0123] The TCR of the present invention can, for example, bind to tumor-associated antigens. As used herein, “tumor-associated antigens” include adrenocortical carcinoma, anal cancer, bladder cancer, bone cancer, brain cancer, breast cancer, carcinoid cancer, carcinoma, cervical cancer, colon cancer, endometrial cancer, esophageal cancer, extrahepatic bile duct cancer, extracranial germ cell carcinoma, eye cancer, gallbladder cancer, gastric cancer, germ cell tumor, gestational trophoblastic tumor, head and neck cancer, hypopharyngeal cancer, islet cell carcinoma, kidney cancer, colorectal cancer, laryngeal cancer, leukemia, lip and oral cancer, liver cancer, lung cancer, lymphoma, malignant mesothelioma, Merkel cell carcinoma, mycosis fungoides, myelodysplastic syndrome, myeloproliferative disorder, and nasal cancer. This refers to any antigen associated with one or more cancers selected from the group consisting of pharyngeal cancer, neuroblastoma, oral cancer, oropharyngeal cancer, osteosarcoma, ovarian epithelial carcinoma, ovarian germ cell carcinoma, pancreatic cancer, sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pituitary cancer, plasma cell neoplasm, prostate cancer, rhabdomyosarcoma, rectal cancer, renal cell carcinoma, transitional cell carcinoma of the renal pelvis and ureter, salivary gland cancer, Sézary syndrome, skin cancer, small intestine cancer, soft tissue sarcoma, gastric cancer, testicular cancer, thymoma, thyroid cancer, urethral cancer, uterine cancer, vaginal cancer, vulvar cancer, and Wilms' tumor.

[0124] In certain embodiments, this application may be suitable for target molecules of hematological cancers. In some embodiments, cancer is a cancer of leukocytes. In other embodiments, cancer is a cancer of plasma cells. In some embodiments, cancer is leukemia, lymphoma, or myeloma. In certain embodiments, cancer is acute lymphoblastic leukemia (ALL) (including non-T cell ALL), acute lymphoblastic leukemia (ALL), and hemophagocytic lymphohistiocytosis (HLH), B-cell prelymphoblastic leukemia, B-cell acute lymphoblastic leukemia ("BALL"), blastic plasmacytoid dendritic cell neoplasm, Burkitt lymphoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic or acute granulomatous Diseases, chronic or acute leukemia, diffuse large B-cell lymphoma, diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, follicular lymphoma (FL), hairy cell leukemia, hemophagocytic syndrome (macrophage activation syndrome (MAS)), Hodgkin's disease, large cell granuloma, leukocyte adhesion disorder, malignant lymphoproliferative disorder, MALT lymphoma, mantle cell lymphoma, marginal zone lymphoma, monoclonal gammaglobulinemia of unknown significance (MG (US), multiple myeloma, myelodysplasia and myelodysplastic syndromes (MDS), bone marrow diseases including acute myeloid leukemia (AML), non-Hodgkin lymphoma (NHL), plasma cell proliferation disorders (e.g., asymptomatic myeloma (smoldering multiple myeloma or painless myeloma), plasmablastic lymphoma, plasmacytoid dendritic cell neoplasms, plasmacytomas (e.g., plasma cell proliferation disorders; solitary myeloma; solitary plasmacytoma; extramedullary plasmacytoma; and multiple These include plasmacytoma, POEMS syndrome (Crow-Fukase syndrome; Takatsuki disease; PEP syndrome), primary mediastinal large B-cell lymphoma (PMBCL), small cell or large cell follicular lymphoma, splenic marginal zone lymphoma (SMZL), systemic amyloid light chain amyloidosis, T-cell acute lymphoblastic leukemia ("TALL"), T-cell lymphoma, transformed follicular lymphoma, Waldenström macroglobulinemia, or combinations thereof.

[0125] In some embodiments, the antigens are 5T4, alpha-fetoprotein (AFP), B7-1 (CD80), B7-2 (CD86), BCMA, β-human chorionic gonadotropin, CA-125, carcinoembryonic antigen (CEA), carcinoembryonic antigen (CEA), CD123 CD133, CD138, CD19, CD20, CD22, CD23, CD24, CD25, CD30, CD33, CD34, CD4, CD40, CD44, CD56, CD8, CLL-1, c-Met, CMV-specific antigen, CSPG4, CTLA-4, disialoganglioside GD2, ductal epithelial mucin, EBV-specific antigen, EGFR variant III (EGFRvIII), ELF2M, endoglin, ephrin B2, epidermal growth factor receptor (EGFR), epithelial cell adhesion molecule (EpCAM), epithelial tumor antigen, ErbB2 (HER2 / neu), fibroblast-related protein (fap), FLT3, folate-binding protein, GD2, GD3, glioma-related antigen, sphingoglycolipid, gp36, HBV-specific antigen, HCV-specific antigen, HER1-HER2, HER 2-HER3 combination, HERV-K, high molecular weight melanoma-associated antigen (HMW-MAA), HIV-1 envelope glycoprotein gp41, HPV-specific antigen, human telomerase reverse transcriptase, IGFI receptor, IGF-II, IL-11R-alpha, IL-13R-a2, influenza virus-specific antigen, CD38, insulin growth factor (IGFI)-1, intestinal carboxylesterase, kappa chain, LAGA-1a, lambda chain, Lassa virus-specific antigen, lectin-reactive AFP, lineage-specific or tissue-specific antigens, e.g., CD3, MAGE, MAGE-A1, major histocompatibility complex (MHC) molecules, major histocompatibility complex (MHC) molecules presenting tumor-specific peptide epitopes, M-CSF, melanoma-associated antigen, mesothelin, mesothelin, MN-CA IX, MUC-1, mut hsp72, mutant p53, mutant p53, mutant ras, neutrophil elastase, NKG2D, Nkp30, NY-ESO-1, p53, PAP, prostase, prostase-specific antigen (PSA), prostate cancer tumor antigen-1 (PCTA-1), prostate-specific antigen, prostain, PSMA, RAGE-1, ROR1, RU1, RU2 (AS), surface adhesion molecule, survivor and telomerase, TAG-72, extradomain A (EDA) and extradomain B (EDB) of fibronectin, and A1 domain (TnC) of tenascin C A1) The target is tumor antigens such as thyroglobulin, tumor stromal antigens, vascular endothelial growth factor receptor-2 (VEGFR2), virus-specific surface antigens, such as HIV-specific antigen (e.g., HIV gp120), and any derivatives or variants of these surface markers.

[0126] The TCR of the present invention can also bind to viral infection-associated antigens. Examples of viral infection-associated antigens include antigens associated with any viral infection, including, for example, viral infection caused by HIV.

[0127] To initiate a patient-specific immunotherapy procedure as described herein, a physician or other healthcare professional on a client computing device 102a accesses a user interface module 108a on a server computing device 106 (e.g., via a web portal, website, or other similar platform). The user interface module 108a generates user interface screens and / or elements for presentation to the physician on the client computing device 102a to register a patient and initiate a patient-specific immunotherapy procedure. The user interface module 108a may generate UI screens that allow the physician to enter patient identification information (e.g., name, date of birth), demographic attributes (e.g., sex), and healthcare provider information (e.g., physician name, hospital name). The user interface module 108a may also provide UI elements for entering a healthcare provider-specific or hospital-specific user identifier (e.g., medical record number, hospital patient ID). Figures 3A and 3B are exemplary screenshots generated by the user interface module 108a that enable the registration of a new patient to the system. Figure 3A shows the patient registration data input screen, and Figure 3B shows the patient information review and confirmation screen.

[0128] Returning to Figure 2, the client computing device 102a generates a request to produce transfected T cells for the patient, and the server computing device 106 receives the request (202). As described above, the client computing device A physician on the client computing device 102a interacts with the user interface module 108a to register a patient by providing the necessary patient information. Once the client interface module 108a receives confirmation from the client computing device 102a that the patient information has been fully entered and is accurate, the user interface module 108a stores the data in the database 110. The user interface module 108a also generates a patient-specific identifier used as part of the sample tracking and continuous management / continuous identification process described below (204). In one embodiment, the patient-specific identifier includes a patient identification element (e.g., patient ID number), a sales order identifier, and a cell order lot number. For example, the user interface module 108a may generate the patient-specific identifier by mapping the patient identification element, sales order number, and cell order lot number to a database table indexed with an identifier (e.g., a 9-digit numeric code) that uniquely identifies the combination of patient, sales order, and cell lot.

[0129] Next, the physician on the client computing device 102a interacts with the user interface module 108a to schedule an appointment to obtain biomaterial from the patient and to confirm that the manufacturing facility has the capability to process the biomaterial immediately after it is obtained, for the time sensitivity of quickly returning the modified biomaterial to the patient. The user interface module 108a requests confirmation of the material extraction site (e.g., site name, address, contact information) to which the extraction kit (e.g., leukocyte apheresis kit) will be dropped off, as well as confirmation of the modified material delivery and treatment site (e.g., site name, address, contact information) to which the material (e.g., transfected T cells) will be delivered from the manufacturing facility. Figures 4A to 4D are exemplary screenshots generated by the user interface module 108a that enable the confirmation of these sites and scheduling of appointments. Figure 4A shows the confirmation screen for the delivery site, Figure 4B shows the confirmation screen for the material delivery site, Figure 4C shows the screen to open the appointment scheduler, and Figure 4D shows the appointment scheduler. In some embodiments, the user interface module 108a communicates with a remote computing device at the manufacturing facility in conjunction with the database 110 to coordinate the scheduling of biomaterial modifications and ensure the most efficient processing schedule so that the modified materials are quickly returned to the patient.

[0130] Returning to Figure 2, once the cell ordering process is completed as described above, process 206 is initiated to perform a biomaterial extraction procedure at the extraction site, ship the extracted material for modification to the manufacturing facility, and return the modified material to the delivery site for injection into the patient's bloodstream. First, the patient arrives at the material extraction site, and a procedure (e.g., a leukocyte apheresis procedure) is performed on the patient's blood sample (206a) to collect T cells from the sample. Once the procedure is performed, the client computing device at the extraction site (e.g., device 102b) communicates with the event tracking module 108b of the server computing device 106 to send a tracking event to module 108b corresponding to the execution of the procedure. For example, a clinician at client computing device 102b may send a tracking event by entering information into the user interface. In another example, client computing device 102b may automatically send a tracking event to module 108b (e.g., via API) when information about the procedure is captured by client computing device 102b (e.g., by scanning a barcode).

[0131] Tracking events may include a patient-specific identifier, a timestamp, an event ID (e.g., indicating that a material extraction procedure was performed), and other process-related information (e.g., cell order lot number, sales order number, site location, etc.). The event tracking module 108b tracks data based on the information received from the client computing device 102b. The tracking event is stored in the database 110. Since this is the first step in the biomaterial extraction and modification process, the event tracking module 108b notifies the continuation management module 108c of the receipt of the tracking event. The continuation management module 108c generates a continuation management data structure (for example, in the database 110) that incorporates the tracking event (and each subsequent tracking event as described herein) in a regular order so that patients, physicians, manufacturers, and other stakeholders can understand the exact state of the biomaterial and ensure that this is always taken into consideration to avoid loss or mishandling of the biomaterial. In one example, the continuation management data structure may be a linked list that connects each of the tracking events sequentially together according to the timestamp of the tracking event.

[0132] Next, the collected T cells are transferred to a container (e.g., a tube, vial, or other type of biomaterial transport device) (206b), another tracking event is captured, and it is sent to the event tracking module 108b for integration into the continuous management data structure described above. Then, the container is labeled with a patient-specific identifier (206c), another tracking event is captured, and it is sent to the event tracking module 108b for communication with the continuous management module 108c for integration into the continuous management data structure. For example, after a barcode containing a patient-specific identifier is labeled on the container containing the collected T cells, it is scanned at the extraction site, indicating that the collected T cells are ready for shipment to the manufacturing facility. Upon scanning the barcode, the client computing device 102b generates a tracking event and sends the event to the event tracking module 108b.

[0133] Next, the extraction site sends the collected T cells to the manufacturing facility (206d), where the manufacturing facility carries out procedures to generate transfected T cells. Both when the collected T cells are shipped to the manufacturing facility and when the collected T cells are received at the manufacturing facility, one or more devices used to record the shipment and receipt of the T cells communicate with the event tracking module 108b to send tracking events associated with specific activities for communication with the continuity management module 108c, thereby integrating them into continuity management. In this way, the continuity management module 108c automatically and continuously updates the continuity management data structure with the latest information, which is reflected in one or more screens generated by the user interface module 108a.

[0134] Next, the manufacturing facility produces transfected T cells from the collected T cells using cell modification technology (206e), and a client computing device (e.g., device 102c) generates one or more tracking events based on the specific cell modification technology being used. For example, the cell modification technology may include (i) quality assurance of the collected T cells before modification, (ii) modification of the T cells, (ii) release testing of the transfected T cells, and (iv) the transfected T cells to be returned to the injection site. This may include several phases, such as the final processing of cells. For each of these phases, the client computing device 102c captures tracking events and sends them to the event tracking module 108b for integration into the continuous management data structure by the continuous management module 108c.

[0135] When transfected T cells are shipped, the injection site receives the transfected T cells (206f), and a client computing device (e.g., device 102d) generates a tracking event for transmission to the event tracking module 108b for integration into the continuous management data structure by the continuous management module 108c. For example, the client computing device 102d can scan a barcode associated with the shipped and / or transfected T cells to automatically generate a tracking event and transmit the event to the server computing device 106.

[0136] After receiving the transfected T cells, the process is completed by injecting them (206g) into the patient's bloodstream. Simultaneously, the client computing device 102d generates a tracking event and sends it to the event tracking module 108b for integration into the continuous management data structure by the continuous management module 108c.

[0137] Figures 5A and 5B are exemplary screenshots generated by the user interface module 108a to allow client computing devices 102a-102d to view ongoing management related to specific patients, biomaterials, and cell modification processes. As shown in Figure 5A, ongoing management of biomaterials during the leukocyte apheresis process (including the steps of scheduling procedures, completing procedures, and preparing extracted T cells for shipment) is captured on the timeline at the top of the screen, where each step of the leukocyte apheresis process is associated with a point in time on the timeline, and ongoing management of biomaterials during the delivery process (e.g., T cells shipped from the extraction site, T cells delivered to the manufacturing facility) is captured on the timeline at the bottom of the screen. As described above, once the event tracking module 108b and the ongoing management module 108c record tracking events, the user interface module 108a scans the ongoing management data structure to display the current status of ongoing management graphically on the screen.

[0138] As shown in Figure 5B, the continuous management of the biomaterial during the manufacturing process (including QA, manufacturing, release testing, and completion of shipment) is shown on the timeline at the top of the screen, and the continuous management of the biomaterial is shown in the center of the screen during the final product delivery process (including shipment and delivery to the injection site). In addition, treatment details, including the treatment date, are displayed at the bottom of the screen. Furthermore, since the continuous management is always associated with a specific patient, complete continuous identification between the patient and the biomaterial is ensured at all phases of manufacturing.

[0139] The technologies described above may be implemented in digital and / or analog electronic circuits, or in computer hardware, firmware, software, or a combination thereof. The implementation may be a computer program product, i.e., a computer program tangibly embodied in a data processing device, such as a programmable processor, a computer, and / or a machine-readable memory device for execution by or control of the operation of multiple computers. The computer program may be written in any form of a computer language or programming language, including source code, compiled code, interpreted code, and / or machine code, and the computer program may be deployed as a standalone program or in any form including subroutines, elements, or other units suitable for use in a computing environment. The computer program may be deployed to run on one computer or on multiple computers at one or more sites. The computer program may be deployed in a cloud computing environment (such as Amazon® AWS, Microsoft® Azure, IBM®, etc.).

[0140] The steps of the method may be performed by one or more processors executing a computer program to manipulate input data and / or generate output data, thereby enabling the functions of the present invention. The steps of the method may also be performed by dedicated logic circuits, such as FPGAs (Field-Programmable Gate Arrays), FPAAs (Field-Programmable Analog Arrays), CPLDs (Composite Programmable Logic Devices), PSoCs (Programmable System-on-Chips), ASIPs (Application-Specific Instruction Set Processors), or ASICs (Application-Specific Integrated Circuits), and the device may be implemented as such. A subroutine may refer to a portion of a stored computer program and / or a processor and / or a special circuit that implements one or more functions.

[0141] A processor suitable for executing computer programs is, for example, a dedicated microprocessor specifically programmed with instructions executable to carry out the methods described herein. Generally, a processor receives instructions and data from read-only memory, random-access memory, or both. Essential elements of a computer are a processor for executing instructions, and one or more memory devices for storing instructions and / or data. Data can be temporarily stored using memory devices such as caches. Memory devices may also be used for long-term data storage. Generally, a computer also includes or is operably coupled to one or more mass storage devices (e.g., magnetic disks, magneto-optical disks, or optical disks) for storing data, or for transferring data to or from such mass storage devices. A computer may also be operably coupled to a communication network for receiving instructions and / or data from and / or transferring instructions and / or data to a network. Computer-readable storage media suitable for embodying computer program instructions and data include, for example, semiconductor memory devices such as DRAM, SRAM, EPROM, EEPROM, and flash memory devices; magnetic disks such as internal hard disks or removable disks; magneto-optical disks; and all forms of volatile and non-volatile memory, including optical disks such as CDs, DVDs, HD-DVDs, and Blu-ray discs. Processors and memory may be complemented and / or incorporated into special-purpose logic circuits.

[0142] To provide user interaction, the above technologies may be implemented on display devices, such as CRT (cathode ray tube), plasma, or LCD (liquid crystal display) monitors for displaying information to the user, displays or screens of mobile devices, holographic devices and / or projectors, and on computing devices that communicate with keyboards and pointing devices, such as mice, trackballs, touchpads, or motion sensors, which allow the user to provide input to the computer (e.g., interact with user interface elements). Other types of devices may also be used to provide user interaction; for example, the feedback provided to the user may be any form of sensory feedback, such as visual feedback, auditory feedback, or haptic feedback, and user input may be received in any form, including acoustic, voice, and / or haptic input.

[0143] The above-described technology may be implemented in a distributed computing system including backend components. The backend components may be, for example, data servers, middleware components, and / or application servers. The above-described technology may also be implemented in a distributed computing system including frontend components. The frontend components may be, for example, a client computer having a graphical user interface, a web browser that allows users to interact with the implementation, and / or other graphical user interfaces for the transmitting device. The above technology may be implemented in a distributed computing system including any combination of such backend, middleware, or frontend components.

[0144] Components of a computing system may be interconnected by a transmission medium that may include any form or medium of digital or analog data communication (e.g., communication networks). The transmission medium may include, in any configuration, one or more packet-based networks and / or one or more circuit-based networks. Packet-based networks include, for example, the Internet, carrier Internet Protocol (IP) networks (e.g., local area networks (LANs), wide area networks) Networks may include, for example, a WAN (Wide Area Network), a Campus Area Network (CAN), a Metropolitan Area Network (MAN), a Home Area Network (HAN), a Private IP Network, an IP Private Branch Exchange (IPBX), a wireless network (e.g., a Radio Access Network (RAN), Bluetooth, Near Field Communication (NFC) networks, Wi-Fi, WiMAX, General Purpose Packet Radio Services (GPRS) networks, HiperLAN), and / or other packet-based networks. Circuit-based networks may include, for example, a Public Switched Telephone Network (PSTN), a legacy Private Branch Exchange (PBX), a wireless network (e.g., a RAN, a Code Division Multiple Access (CDMA) network, a Time Division Multiple Access (TDMA) network, a Global System for Mobile Communications (GSM) network), and / or other circuit-based networks.

[0145] Information transfer via a transmission medium may be based on one or more communication protocols. Examples of communication protocols include Ethernet protocol, Internet Protocol (IP), Voice over IP (VOIP), Peer-to-Peer (P2P) protocol, Hypertext Transfer Protocol (HTTP), Session Initiation Protocol (SIP), H.323, Media Gateway Control Protocol (MGCP), Signaling System #7 (SS7), Global System for Mobile Communications (GSM) protocol, and push-button Push-to-Talk (PTT) protocol, PTT over Cellular (POC) protocol, Universal Mobile Telecommunications System (UMTS), 3GPP Long-Term Evolution This may include Long Term Evolution (LTE) and / or other communication protocols.

[0146] Computing system devices may include, for example, computers, computers with browser devices, telephones, IP phones, mobile devices (e.g., mobile phones, personal digital assistant (PDA) devices, smartphones, tablets, laptop computers, email devices) and / or other communication devices. Browser devices include, for example, World Wide Web browsers (e.g., Chrome® from Google, Inc., Mi, available from Microsoft Corporation). Microsoft® Internet Explorer® and / or Mozilla® Firefox, available from Mozilla Corporation. Examples of computers having x) include desktop computers and / or laptop computers. Examples of mobile computing devices include Blackberry® from Research in Motion and Apple Corporation. Examples include iPhone® and / or Android®-based devices. IP phones include, for example, Cisco Systems, Inc. (Registered Trademark) Unified IP Phone 7985G and / or Cisco (Registered Trademark) One example is the Unified Wireless Phone 7920.

[0147] Additional definitions The terms “polypeptide” or “protein” refer to macromolecules having an amino acid sequence of a protein that includes one or more amino acid deletions, additions, and / or substitutions of the natural sequence, preferably with eight or fewer amino acid substitutions. Polypeptides or proteins are preferably isolated as specified herein. The term “polypeptide fragment” refers to an isolated polypeptide having amino-terminal deletions, carboxyl-terminal deletions, and / or internal deletions compared to a full-length natural protein. Such fragments may contain modified amino acids compared to the natural protein. Useful polypeptide fragments include immunofunctional fragments of antigen-binding molecules. Useful fragments include, but are not limited to, one or more CDR regions, variable domains of heavy and / or light chains, and parts of other parts of antibody chains.

[0148] The term “antibody” refers to any isotype of intact immunoglobulin, or a fragment thereof that can compete with an intact antibody for specific binding to a target antigen / molecule, including, for example, chimeric, humanized, fully human, and bispecific antibodies. “Antibody” is an antigen-binding molecular species as defined herein. Intact antibodies generally contain at least two full-length heavy chains and two full-length light chains, but may contain fewer chains, such as antibodies naturally occurring in camelids, which may contain only heavy chains. Antibodies may originate from a single source or may be chimeric, i.e., different parts of an antibody may originate from two different antibodies, as further described below. Antigen-binding molecules, antibodies, or binding fragments can be produced by recombinant DNA in a hybridoma, or by enzymatic or chemical cleavage of an intact antibody. Unless otherwise specified, the term “antibody” includes antibodies containing two full-length heavy chains and two full-length light chains, as well as their derivatives, variants, fragments, and mutant proteins, examples of which are listed below. Furthermore, unless explicitly excluded, antibodies include monoclonal antibodies, bispecific antibodies, minibodies, domain antibodies, synthetic antibodies (sometimes referred to herein as “antibody mimes”), chimeric antibodies, humanized antibodies, human antibodies, antibody fusions (sometimes referred to herein as “antibody conjugates”), and their fragments.

[0149] The variable regions typically exhibit the same overall structure of a relatively conserved framework region (FR) joined by three hypervariable regions (i.e., "CDRs"). The CDRs from each pair of chains are typically aligned by the framework region, enabling binding to specific epitopes. From the N-terminus to the C-terminus, both the light-chain and heavy-chain variable regions typically contain domains FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. By convention, the CDR regions of the heavy chain are typically referred to as HC CDR1, CDR2, and CDR3. The CDR regions of the light chain are typically referred to as LC CDR1, CDR2, and CDR3. The assignment of amino acids to each domain typically follows the Kabat, Chothia, or AbM definitions.

[0150] The term "Kabat numbering" and similar terms are recognized in the art and refer to a system for numbering amino acid residues in the variable regions of the heavy and light chains of antibodies, or in their antigen-binding regions. In certain embodiments, the CDR of an antibody can be identified according to the Kabat numbering system (see, for example, Kabat EA & Wu TT (1971) Ann NY Acad Sci 190: 382-391 and Kabat EA et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242). In antibody heavy chain molecules, CDRs are typically located at amino acid positions 31-35 (CDR1), 50-65 (CDR2), and 95-102 (CDR3), which may contain one or two additional amino acids following 35 (referred to as 35A and 35B in the Kabat numbering scheme). Using the Kabat numbering system, CDRs in antibody light chain molecules are typically located at amino acid positions 24-34 (CDR1), 50-56 (CDR2), and 89-97 (CDR3). In certain embodiments, the CDRs of the antibodies described herein were determined according to the Kabat numbering scheme.

[0151] In certain embodiments, the CDR of an antibody may be identified according to the Chothia numbering scheme, which refers to the position of the structural loop of the immunoglobulin (see, for example, Chothia C & Lesk AM, (1987), J Mol Biol 196: 901-917, Al-Lazikani B et al., (1997) J Mol Biol 273: 927-948, Chothia C et al., (1992) J Mol Biol 227: 799-817, Tramontano A et al., (1990) J Mol Biol 215(1): 175-82, and U.S. Patent No. 7,709,226). Typically, when using the Kabat numbering standard, the CDR-H1 loop of Chothia is located at amino acids 26-32, 33, or 34 of the heavy chain, the CDR-H2 loop of Chothia is located at amino acids 52-56 of the heavy chain, and the CDR-H3 loop of Chothia is located at amino acids 52-56 of the heavy chain. While the loops are located at amino acids 95-102 of the heavy chain, the Chothia CDR-L1 loop is located at amino acids 24-34 of the light chain, the Chothia CDR-L2 loop is located at amino acids 50-56 of the light chain, and the Chothia CDR-L3 loop is located at amino acids 89-97 of the light chain. The end of the Chothia CDR-HI loop, when numbered using the Kabat numbering scheme, varies between H32 and H34 depending on the length of the loop (this is because the Kabat numbering scheme places insertions at H35A and H35B, meaning the loop ends at 32 if neither 35A nor 35B exists, at 33 if only 35A exists, and at 34 if both 35A and 35B exist).

[0152] In certain embodiments, the CDRs of the antibodies described herein were identified according to the Chothia numbering scheme.

[0153] Several definitions of CDRs, namely Kabat numbering, Chothia numbering, AbM numbering, or contact numbering, are commonly used. The bM definition is a compromise between two approaches used by Oxford Molecular's AbM antibody modeling software. The contact definition is based on the analysis of available complex crystalline structures.

[0154] [Table 2]

[0155] As used herein, the term “heavy chain” may refer to any different types, such as alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), that give rise to the classes of antibodies IgA, IgD, IgE, IgG, and IgM, respectively, based on the amino acid sequence of the constant domain, including subclasses of IgG, such as IgG1, IgG2, IgG3, and IgG4.

[0156] As used herein, the term “light chain,” when used in reference to antibodies, may refer to any different type, such as kappa (κ) or lambda (λ), based on the amino acid sequence of the constant domain. Light chain amino acid sequences are known in the art. In certain embodiments, the light chain is a human light chain.

[0157] The term "variable region" or "variable domain" typically refers to a portion of an antibody's light and / or heavy chain, including approximately 120–130 amino acids at the amino terminus of the heavy chain and approximately 100–110 amino acids at the amino terminus of the light chain. The variable region of an antibody typically determines the specificity of a particular antibody to its target.

[0158] The variability is not evenly distributed throughout the variable domain of the antibody or antigen-binding molecule, but rather between the heavy and light chains. It is concentrated into each subdomain of the variable region. These subdomains These are called "hypervariable regions" or "complementarity-determining regions" (CDRs), as further described herein. The more conserved (i.e., non-hypervariable) portions of the variable domains are called "framework" regions (FRMs or FRs), which provide a scaffold for six CDRs in three-dimensional space and form an antigen-binding surface. The variable domains of the spontaneously occurring heavy and light chains each contain four FRM regions (FR1, FR2, FR3, and FR4) that primarily employ a β-sheet configuration, connected by three hypervariable regions that form loops connecting, and in some cases forming part of, a β-sheet structure. The hypervariable regions of each chain are linked very closely to the hypervariable regions of other chains by the FRMs, contributing to the formation of antigen-binding sites (see Kabat et al., further described herein).

[0159] Typically, CDRs form loop structures that can be classified as canonical structures. The term "canonical structure" refers to the main chain conformation adopted by the antigen-binding (CDR) loop. Comparative structural studies have shown that five of the six antigen-binding loops have only a limited repertoire of available conformations. Each canonical structure can be characterized by the twist angle of the polypeptide backbone. Therefore, the loops between corresponding antibodies can have very similar three-dimensional structures despite the high amino acid sequence variability of most of the loops (Chothia and Lesk, J. MoI. Biol., 1987, 196: 901, Chothia et al., Nature, (1989, 342: 877; Martin and Thornton, J. MoI. Biol, 1996, 263: 800). Furthermore, a correlation has been observed between the adopted loop structure and the surrounding amino acid sequence. The conformation of a particular canonical class is determined by the length of the loop, as well as the amino acid residues present at key positions within the loop and within the conserved framework (i.e., outside the loop). Therefore, assignment to a particular canonical class can be made based on the presence of these key amino acid residues.

[0160] The term "canonical structure" may also include considerations regarding the linear sequence of an antibody, as cataloged, for example, by Kabat (Kabat et al. as herein). The Kabat numbering scheme (system) is a widely adopted standard for consistently numbering amino acid residues in the antibody variable domain and is the preferred scheme applied to the present invention, as referred to elsewhere herein. Additional structural considerations may also be used in determining the canonical structure of an antibody. For example, differences not fully reflected in Kabat numbering can be explained by the numbering system of Chothia et al. and / or revealed by other techniques, such as crystallographic analysis and two- or three-dimensional computational modeling. Thus, a given antibody sequence can be classified into a canonical class that allows for the identification of a particularly appropriate chassis sequence (e.g., (For example, based on the desire to include various canonical structures in the library.) The Kabat numbering of antibody amino acid sequences and their influence on structural considerations described by Chothia et al. (hereinforced) and on the interpretation of the canonical configuration of antibody structures are described in the literature. The subunit structures and three-dimensional arrangements of various classes of immunoglobulins are known in the art. For an overview of antibody structures, see Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, eds. Harlow et al., 1988.

[0161] Light chain CDR3s, and especially heavy chain CDR3s, can be the most important determinants of antigen binding in the light and heavy chain variable regions. In some antibody constructs, heavy chain CDR3s appear to constitute the primary contact site between antigen and antibody. In vitro selection schemes that alter CDR3s alone can be used to change the binding properties of an antibody or to determine which residues contribute to antigen binding. Therefore, CDR3s are typically the greatest source of molecular diversity at antibody binding sites. For example, H3 can be short, consisting of two amino acid residues, or longer, consisting of more than 26 amino acids.

[0162] As used herein, the terms “constant region” and “constant domain” are synonymous and have the common meaning in the art. The constant region is the antibody portion of the light and / or heavy chain, such as the carboxyl terminus, which does not directly participate in the binding of antibodies to antigens but can exhibit various effector functions, such as interaction with Fc receptors. The constant region of immunoglobulin molecules generally has a more conserved amino acid sequence compared to the immunoglobulin variable domain.

[0163] The "Fc" region contains two heavy chain fragments, each containing the CH1 and CH2 domains of the antibody. The two heavy chain fragments are linked by two or more disulfide bonds and hydrophobic interactions of the CH3 domain.

[0164] A "Fab fragment" contains one light chain and one heavy chain containing the CH1 and variable regions. The heavy chain of a Fab molecule cannot form a disulfide bond with another heavy chain molecule. A "Fab'" fragment contains one light chain and a portion of a heavy chain containing the VH domain and CH1 domain, as well as the region between the CH1 and CH2 domains. Therefore, an interchain disulfide bond can be formed between the two heavy chains of the two Fab' fragments, forming an F(ab')2 molecule. An "F(ab')2 fragment" contains two light chains and two heavy chains containing a portion of the constant region between the CH1 and CH2 domains. Therefore, an interchain disulfide bond is formed between the two heavy chains. Thus, an F(ab')2 fragment is composed of two Fab' fragments linked by a disulfide bond between the two heavy chains.

[0165] The "Fv region" includes variable regions for both heavy and light chains, but does not have a steady-state region.

[0166] A "bivalent antigen-binding molecule" contains two antigen-binding sites. In some cases, the two binding sites have the same antigen specificity. A bivalent antigen-binding molecule may also be bispecific. A "multispecific antigen-binding molecule" targets two or more antigens or epitopes. "Bispecific," "dual-specific," or "bifunctional" The antigen-binding molecules of these molecules are hybrid antigen-binding molecules or antibodies, each having two different antigen-binding sites. The two binding sites of a bispecific antigen-binding molecule bind to two different epitopes that may be present on the same or different protein targets.

[0167] "Binding affinity" generally refers to the overall strength of the non-covalent interaction between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise specified, "binding affinity" as used herein refers to the intrinsic binding affinity that reflects the 1:1 interaction between the members of a binding pair (e.g., an antibody and an antigen). The affinity of molecule X to partner Y is generally expressed by the dissociation constant (K).D Affinity can be expressed by the equilibrium dissociation constant (K). D ) and equilibrium association constant (K A ) can be measured and / or represented by a number of methods known in the art, including but not limited to these. D is k off / k on It is calculated from the quotient of K A is k on / k off It is calculated from the quotient of k. on k refers to, for example, the association rate constant of an antibody against an antigen. off This refers, for example, to the dissociation of an antibody against an antigen. on and k off This can be determined by techniques known to those skilled in the art, such as BIAcore® or KinExA.

[0168] The term "neutralize" refers to an antigen-binding molecule, scFv, or antibody that binds to a ligand and inhibits or reduces the ligand's biological effect. This can be achieved, for example, by directly blocking the binding site on the ligand, or by binding to the ligand and altering its binding ability through indirect means (such as structural or energy changes in the ligand). In some embodiments, the term may also mean that the antigen-binding molecule prevents the protein to which it binds from performing its biological function.

[0169] When the term "competing" is used in relation to antigen-binding molecules competing for the same epitope, it means competition between antigen-binding molecules determined by an assay in which the antigen-binding molecule being tested (e.g., an antibody or its immune function fragment) blocks or inhibits (e.g., reduces) the specific binding of a reference antigen-binding molecule to the antigen. Numerous types of competitive binding assays, e.g., solid-phase direct or indirect radioimmunoassay (RIA), solid-phase direct or indirect enzyme immunoassay (EIA), sandwich competitive assays (Stahli et al., 1983, Methods in Enzymology 9:242-253); solid-phase direct biotin-avidin EIA (Kirkland et al., 1986, J. Immunol. 137:3614-3619), solid-phase direct labeling assays, solid-phase direct labeling sandwich assays (Harlow and Lane, 1988, Antibodies, A Laboratory Manual, Cold Spring Harbor Press); solid-phase direct labeling RIA using I-125 labeling (Morel et al., 1988, Molec. Immunol. 25:7-15); solid-phase direct biotin-avidin EIA (Cheung, et al., 1990, Virology Using 176:546-552 and directly labeled RIA (Moldenhauer et al., 1990, Scand. J. Immunol. 32:77-82), it is possible to determine whether one antigen-binding molecule competes with another.

[0170] As used herein, the term “epitope” refers to a localization region of an antigen to which an antibody can specifically bind. An epitope may be, for example, an adjacent amino acid of a polypeptide (linear or adjacent epitopes), or it may be, for example, derived from two or more non-adjacent regions of a polypeptide (contiguous, non-linear, discontinuous, or non-adjacent epitopes). In certain embodiments, the epitope to which the antibody binds may be identified, for example, by NMR spectroscopy, X-ray diffraction crystallography studies, ELISA assays, hydrogen / deuterium exchange coupled with mass spectrometry (e.g., liquid chromatography-electrospray mass spectrometry), array-based oligopeptide scanning assays, and / or mutagenic mapping (e.g., site-directed mutagenic mapping). For X-ray crystallography studies, crystallization may be performed using methods known in the art (e.g., Giege R. et al., (1994) Acta Crystallogr D Biol Crystallogr50(Pt 4): 339-350, McPherson A (1990) This can be carried out using any of the following methods: Eur J Biochem 189: 1-23, Chayen NE (1997) Structure 5: 1269-1274, McPherson A (1976) J Biol Chem 251: 6300-6303). Antibody: Antigen Crystals can be studied using well-known X-ray diffraction techniques, such as X-PLOR (Yale University, 1992, distributed by Molecular Simulations, Inc.; see, for example, Meth Enzymol (1985) volumes 114 & 115, eds Wyckoff HW et al.; U.S. Patent Application Publication) It can be refined using computer software such as (No. 2004 / 0014194) and BUSTER (Bricogne G (1993) Acta Crystallogr D Biol Crystallogr 49(Pt 1): 37-60, Bricogne G (1997) Meth Enzymol276A: 361-423, ed Carter CW, Roversi P et al., (2000) Acta Crystallogr D Biol Crystallogr 56(Pt 10): 1316-1323). Mutagenesis mapping studies can be carried out using any method known to those skilled in the art. For example, see Champe M et al., (1995) J Biol Chem 270: 1388-1394 and Cunningham BC & Wells JA (1989) Science 244: 1081-1085 for a description of mutagenesis techniques, including alanine scanning mutagenesis.

[0171] The terms “genetically modified” or “modified” refer to methods of modifying a cell’s genome, including but not limited to deletions of coding regions or non-coding regions, or parts thereof, or insertions of coding regions or parts thereof. In some embodiments, the cells being modified are lymphocytes, for example, T cells that can be obtained from either a patient or a donor. The cells may be modified to express exogenous constructs, such as chimeric antigen receptors (CARs) or T cell receptors (TCRs), which are incorporated into the cell’s genome, for example.

[0172] "Immune response" refers to an invading pathogen, cells or tissues infected with a pathogen, cancer or other This refers to the action of immune system cells (e.g., T lymphocytes, B lymphocytes, natural killer (NK) cells, macrophages, eosinophils, mast cells, dendritic cells, and neutrophils) and soluble macromolecules (including alpha, cytokines, and complement) produced by any of these cells or the liver, resulting in the selective targeting, binding to, damage to, destruction of, and / or elimination of abnormal cells or, in the case of autoimmune or pathological inflammation, normal human cells or tissues.

[0173] The term "immunotherapy" refers to the treatment of subjects affected by disease, or at risk of developing or relapsing from disease, by methods including inducing, enhancing, suppressing, or otherwise modifying the immune response. Examples of immunotherapy include, but are not limited to, T-cell therapy. T-cell therapy includes adoptive T-cell therapy. This may include tumor-infiltrating lymphocyte (TIL) immunotherapy, autologous cell therapy, engineered autologous cell therapy (eACT), and allogeneic T-cell transplantation. However, those skilled in the art will recognize that the conditioning methods disclosed herein may enhance the efficacy of any transplanted T-cell therapy. Examples of T-cell therapies are described in U.S. Patent Applications Publications 2014 / 0154228 and 2002 / 0006409, U.S. Patent No. 5,728,388, and International Publication No. 2008 / 081035.

[0174] T cells for immunotherapy may be derived from any source known in the art. For example, T cells may be differentiated in vitro from a hematopoietic stem cell population, or T cells may be obtained from a subject. T cells can be obtained, for example, from peripheral blood mononuclear cells (PBMCs), bone marrow, lymph node tissue, umbilical cord blood, thymic tissue, tissue from the site of infection, ascites, pleural fluid, splenic tissue, and tumors. In addition, T cells may be derived from one or more T cell lines available in the art. T cells may also be obtained from blood units collected from a subject using various techniques known to those skilled in the art, such as FICOLL® isolation and / or apheresis. Additional methods for isolating T cells for T cell therapy are disclosed in U.S. Patent Application Publication 2013 / 0287748, which is incorporated herein by reference in whole.

[0175] The term "engineered autologous cell therapy," which can be abbreviated as "eACT®," also known as adoptive cell transplantation, is a process in which a patient's own T cells are collected and subsequently genetically engineered to recognize and target one or more antigens expressed on the surface of one or more specific tumor cells or malignant tumor cells. T cells can be modified to express, for example, a chimeric antigen receptor (CAR) or a T cell receptor (TCR). CAR-positive (+) T cells are modified to express an extracellular single-strand variable fragment (scFv) with specificity to a particular tumor antigen, ligated to an intracellular signaling region containing at least one costimulatory domain and at least one activating domain. The costimulatory domain may be derived from (or corresponding to), for example, CD28, and the activating domain may be derived from (or corresponding to), for example, CD3-zeta. In certain embodiments, the CAR is designed to have two, three, four, or more costimulatory domains.

[0176] The term “autologous” refers to any substance derived from the same individual that is later reintroduced. For example, the method of engineered autologous cell therapy (eACT®) described herein includes collecting lymphocytes from a patient, subsequently modifying them to express, for example, a CAR construct, and then administering and returning them to the same patient.

[0177] The term "allogeneic" refers to any substance originating from one individual that is later introduced into another individual of the same species (for example, allogeneic T cell transplantation).

[0178] Standard techniques for recombinant DNA and oligonucleotide synthesis, as well as tissue culture and plasmapheresis. Alternatively, methods such as electroporation and lipofection may be used. Enzymatic reactions and purification methods may be carried out according to the manufacturer's specifications, as is commonly achieved in the art, or as described herein. The techniques and procedures described above may be carried out in general according to conventional methods known in the art, as described in the various general and more specific references cited and discussed throughout this specification. For example, see Sambrook et al. Molecular Cloning: A Laboratory Manual (2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989)) (which is incorporated herein by reference).

[0179] There are no restrictions on the plural forms of "comprise," "include," and / or. This includes the listed parts and may include additional parts not listed. "and / or" is unrestricted and includes one or more of the listed parts, and combinations of the listed parts.

[0180] It should be understood that the descriptions herein are illustrative and descriptive only and do not limit the claimed invention. In this application, unless otherwise specifically designated, the use of the singular includes the plural.

[0181] All documents or parts of documents cited herein, including but not limited to patents, patent applications, articles, books, and papers, expressly constitute part of this Specified

[0182] In this application, the use of "or" means "and / or" unless otherwise specified. Furthermore, the use of the term "including," as well as other forms such as "includes" and "included," is not limited. Also, "element" or "component" Unless otherwise specifically designated, terms such as these include both elements and components containing one unit, and elements and components containing two or more subunits.

[0183] References made herein should not be construed as recognition of prior art relating to the present invention. If any definition or term provided in a reference that, by means of reference, forms part of this specification differs from the terms and discussion provided herein, the terms and definitions of this invention shall prevail.

[0184] Those skilled in the art will recognize that the subject matter can be embodied in other specific forms without departing from the spirit or essential features of the present invention. Therefore, the embodiments described herein are intended to be illustrative in all respects and not limit the subject matter described herein.

[0185] The present invention is further illustrated by the following sequence.

[0186] CD28T DNA extracellular, transmembrane, intracellular CTTGATAATGAAAAGTCAAACGGAACAATCATTCACGTGAAGGGCAAGCACCTCTGTCCGTCACCCTTGTTCCCTGGTCCATCCAAGCCATTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTGCTCGTCACCGTG GCTTTTATAATCTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGC (SEQ ID NO: 1)

[0187] CD28T extracellular, transmembrane, intracellular AA LDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS(Sequence ID 2)

[0188] CD28T DNA-extracellular CTTGATAATGAAAAGTCAAACGGAACAATCATTCACGTGAAGGGCAAGCACCTCTGTCCGTCACCCTTGTTCCCTGGTCCATCCAAGCCA(Sequence ID 3)

[0189] CD28T AA-Extracellular LDNEKSNGTI IHVKGKHLCP SPLFPGPSKP(Sequence ID 4)

[0190] CD28 DNA transmembrane domain TTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTGCTCTCACCGTGGCTTTTATAATCTTCTGGGTT(Sequence No. 5)

[0191] CD28 AA transmembrane domain FWVLVVVGGV LACYSLLVTV AFIIFWV (Sequence ID 6)

[0192] CD28 DNA intracellular domain AGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGC (SEQ ID NO: 7)

[0193] CD28 AA intracellular domain RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS(Sequence ID 8)

[0194] CD3 Zeta DNA AGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCAGGAGGGT CTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG (SEQ ID NO: 9)

[0195] CD3 Zeta AA RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR(Sequence ID 10)

[0196] CD28 bit ATTGAGGTGATGTATCCACCGCCTTACCTGGATAACGAAAAGAGTAACGGTACCATCATTCACGTGAAAGGTAAACACCTGTGTCCTTCTCCCCTCTCCCCGGGCCATCAAAGCCC(array Number 11)

[0197] CD28 AA IEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP(Sequence ID 12)

[0198] CD8 DNA extracellular and transmembrane domains GCTGCAGCATTGAGCAACTCAATAATGTATTTTAGTCACTTTGTACCAGTGTTCTTGCCGGCTAAGCCTACTACCACACCCGCTCCACGGCCACCTACCCCAGCTCCTACCATCGCTTCACAGCCTCTGTCCCTGCGCCCAGAGGCTT GCCGACCGGCCGCAGGGGGCGCTGTTCATACCAGAGGACTGGATTTCGCCTGCGATATCTATATCTGGGCACCCCTGGCCGGAACCTGCGGCGTACTCCTGCTGTCCCTGGTCATCACGCTCTATTGTAATCACAGGAAC (SEQ ID NO: 13)

[0199] CD8 AA extracellular and transmembrane domains AAALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRN(Sequence ID 14)

[0200] Clone 24C1 HC DNA CAGGTGCAGCTGCAGGAATCCGGACCGGGGCTGGTGAAGCCCAGCGAGACTCTGAGTCTCACGTGTACAGTTTCTGGAGGTAGCATTAGCTCCTACTATTGGTCATGGATAAGGCAGCCCCCCGGGAAGGGATTGGAATGGATCGGCTATATTTACTACAGTGGGAGCACCAATTACAACCCCTCAC TGAAGTCTAGAGTTACAATCAGCGTTGACACCTCAAAGAATCAGTTCAGTTTGAAATTGTCTAGCGTCACAGCAGCTGATACAGCCGTCTATTATTGTGTTTCTCTGGTCTATTGCGGTGGGGATTGTTACAGTGGCTTTGACTATTGGGGGCAGGGTACTCTGGTTACAGTTTCTTCC (SEQ ID NO: 15)

[0201] Clone 24C1 HC AA (underlined in CDR) I YYSGS TNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCVS LVYCGGDCYSGFDY WGQGTLVTVSS (Sequence ID 16)

[0202] Clone 24C1 HC AA CDR1:GGSISSY (Sequence ID 17)

[0203] Clone 24C1 HC AA CDR2:YYSGS (Sequence ID 18)

[0204] Clone 24C1 HC AA CDR3:LVYCGGDCYS GFDY (Sequence ID 19)

[0205] Clone 24C1 LC DNA GACATCCAGTTGACACAGAGCCCGAGTTCCTTGTCCGCCTCCGTCGGGGATAGAGTGTCATTTACCTGTCAGGCCTCTCAGGATATTAATAACTTTCTGAATTGGTATCAGCAAAAGCCCGGAAAGGCACCCAAGCTGTTGATTTACGACGCCAGTAACCTGGAGACAGGCGTGCCCTCCCGGTTTAGTGGTAGCGGAAG CGGTACGGATTTTACCTTTACTATCAGCTCTCTCCAACCCGAAGACATTGCAACCTACTATTGTCAACAATATGGAAACCTGCCTTTTACATTTGGCGGCGGCACCAAGGTGGAGATTAAGCGG (SEQ ID NO: 20)

[0206] Clone 24C1 LC AA (CDR is underlined) DIQLTQSPSSLSASVGDRVSFTC QASQDINNFLN WYQQKPGKAPKLLIY DASNLET GVPSRFSGSGSGTTDFTFTISSLQPEDIATYYC QQYGNLPFT FGGGTKVEIKR (Sequence ID 21)

[0207] Clone 24C1 LC CDR1 AA:QASQDINNFLN (Sequence ID 22)

[0208] Clone 24C1 LC CDR2 AA:DASNLET (Sequence ID 23)

[0209] Clone 24C1 LC CDR3 AA:QQYGNLPFT (Sequence ID 24)

[0210] Clone 24C1 CD28T CD3 Zeta CAR DNA heavy and light strands ATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA (SEQ ID NO: 25)

[0211] Clone 24C1 CD28T CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold) TIFF2026082913000004.tif64168

[0212] Clone 24C1 CD28T CD3 Zeta CAR DNA heavy and light strands GCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTAT TCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG (SEQ ID NO: 27)

[0213] Clone 24C1 CD28T CD3 Zeta CAR AA Heavy and Light Chains QVQLQESGPGLVKPSETLSLTCTVSGGSISSYYWSWIRQPPGKGLEWIGYIYYSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCVSLVYCGGDCYSGFDYWGQGT LVTVSSGGGGSGGGGSGGGGSDIQLTQSPSSLSASVGDRVSFTCQASQDINNFLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYGNLPF TFGGGTKVEIKRAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR(Sequence ID 28)

[0214] Clone 24C1 CD28 CD3 Zeta CAR DNA heavy and light strands CACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAG ACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA (SEQ ID NO: 29)

[0215] Clone 24C1 CD28 CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold.) TIFF2026082913000005.tif64169

[0216] Clone 24C1 CD28 CD3 Zeta CAR DNA heavy and light strands CAGGTGCAGCTGCAGGAATCCGGACCGGGGCTGGTGAAGCCCAGCGAGACTCTGAGTCTCACGTGTACAGTTTCTGGAGGTAGCATTAGCTCCTACTATTGGTCATGGATAAGGCAGCCCCCCGGGAAGGGATTGGAATGGATCGGCTATATTTACTACAGTGGGAGCACCAATTACAACCCCTCACTGAAGTCTAGAGTTACAATCAGCGTTGACACCTCAAAGAATCAGTTCAGTTTGAAATTGTCTAGCGTCACAGCAGCTGATACAGCCGTCTATTATTGTGTTTCTCTGGTCTATTGCGGTGGGGATTGTTACAGTGGCTTTGACTATTGGGGGCAGGGTACTCTGGTTACAGTTTCTTCCGGGGGGGGAGGCTCTGGGGGCGGAGGCTCAGGTGGTGGAGGCAGCGACATCCAGTTGACACAGAGCCCGAGTTCCTTGTCCGCCTCCGTCGGGGATAGAGTGTCATTTACCTGTCAGGCCTCTCAGGATATTAATAACTTTCTGAATTGGTATCAGCAAAAGCCCGGAAAGGCACCCAAGCTGTTGATTTACGACGCCAGTAACCTGGAGACAGGCGTGCCCTCCCGGTTTAGTGGTAGCGGAAGCGGTACGGATTTTACCTTTACTATCAGCTCTCTCCAACCCGAAGACATTGCAACCTACTATTGTCAACAATATGGAAACCTGCCTTTTACATTTGGCGGCGGCACCAAGGTGGAGATTAAGCGGGCGGCAGCTATTGAGGTGATGTATCCACCGCCTTACCTGGATAACGAAAAGAGTAACGGTACCATCATTCACGTGAAAGGTAAACACCTGTGTCCTTCTCCCCTCTTCCCCGGGCCATCAAAGCCCTTCTGGGTTCTTGTGGTCG TGGGAGGCGTGCTTGCTTGTTATTCTCTGCTCGTTACCGTGGCGTTTATCATTTTTTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG(SEQ ID NO: 31)

[0217] Clone 24C1 CD28 CD3 zeta CAR AA heavy and light chains QVQLQESGPGLVKPSETLSLTCTVSGGSISSYYWSWIRQPPGKGLEWIGYIYYSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCVSLVYCGGDCYSGFDYW GQGTLVTVSSGGGGSGGGGGSGGGGSDIQLTQSPSSLSASVGDRVSFTCQASQDINNFLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYC QQYGNLPFTFGGGTKVEIKRAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLST ATKDTYDALHMQALPPR(Sequence ID 32)

[0218] Clone 24C1 CD8 CD3 Zeta CAR DNA heavy and light strands ATGGCACTCCCCGTAACTGCTCTGCTGCTGCCGTTGGCATTGCTCCTGCACGCCGCACGCCCGCAGGTGCAATTGCAAGAGTCCGGCCCCGGACTCGTTAAACCCAGTGAGACGCTTAGCCTGACCTGTACCGTCTCAGGGGGCAGCATCTCCTCTTATTACTGGAGCTGGATCAGGCAGCCTCCAGGAAAAGGCCTTGAATGGATTGGGTACATCTACTACTCTGGCTCAACAAATTATAATCCATCCCTGAAGTCCCGCGTGACTATCTCTGTGGACACCAGCAAGAATCAGTTTTCACTGAAGTTGTCTAGTGTTACCGCGGCCGACACCGCCGTATACTACTGTGTGTCTCTTGTGTACTGTGGCGGCGACTGCTATTCCGGGTTCGACTACTGGGGCCAAGGGACTCTGGTAACCGTGTCCTCAGGCGGCGGCGGGTCAGGAGGAGGCGGCAGTGGAGGTGGCGGCTCCGACATCCAGCTGACACAATCACCATCTTCCCTTTCAGCTTCAGTCGGGGACAGAGTGTCCTTCACATGCCAGGCCAGCCAGGATATCAATAACTTCCTGAACTGGTACCAACAGAAACCCGGAAAGGCTCCAAAGCTCCTGATCTATGATGCTTCCAACCTGGAGACCGGCGTGCCCTCCAGGTTCAGTGGTTCAGGATCAGGCACTGACTTTACGTTCACCATATCCAGTCTTCAGCCCGAAGACATTGCAACCTATTACTGCCAACAATACGGGAACCTTCCCTTTACATTCGGAGGCGGCACCAAGGTGGAAATCAAAAGGGC (Sequence ID 33)

[0219] Clone 24C1 CD8 CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold.) TIFF2026082913000006.tif70169

[0220] Clone 24C1 CD8 CD3 Zeta CAR DNA heavy and light strands CAGGTGCAATTGCAAGAGTCCGGCCCCGGACTCGTTAAACCCAGTGAGACGCTTAGCCTGACCTGTACCGTCTCAGGGGGCAGCATCTCCTCTTATTACTGGAGCTGGATCAGGCAGCCTCCAGGAAAAGGCCTTGAATGGATTGGGTACATCTACTACTCTGGCTCAACAAATTATAATCCATCCCTGAAGTCCCGCGTGACTATCTCTGTGGACACCAGCAAGAATCAGTTTTCACTGAAGTTGTCTAGTGTTACCGCGGCCGACACCGCCGTATACTACTGTGTGTCTCTTGTGTACTGTGGCGGCGACTGCTATTCCGGGTTCGACTACTGGGGCCAAGGGACTCTGGTAACCGTGTCCTCAGGCGGCGGCGGGTCAGGAGGAGGCGGCAGTGGAGGTGGCGGCTCCGACATCCAGCTGACACAATCACCATCTTCCCTTTCAGCTTCAGTCGGGGACAGAGTGTCCTTCACATGC

[0221] Clone 24C1 CD8 CD3 zeta CAR AA heavy and light chains QVQLQESGPGLVKPSETLSLTCTVSGGSISSYYWSWIRQPPGKGLEWIGYIYYSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCVSLVYCGGDCYSGFDYWGQGTLVTVSSGGGGSGGGGSGGGGSDIQLTQSPSSLSASVGDRVSFTCQASQDINNFLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYGNLPFTFGGGTKVEIKRAAALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATK DTYDALHMQALPPR (SEQ ID NO: 36)

[0222] Clone 24C1 CD28T CD3 zeta CAR DNA heavy and light chains ATGGCACTCCCCGTAACTGCTCTGCTGCTGCCGTTGGCATTGCTCCTGCACGCCGCACGCCCGGATATCCAGCTCACGCAATCCCCCTCAAGCTTGAGTGCCTCCGTGGGCGACCGGGTGTCCTTCACATGTCAGGCAAGCCAAGACATAAATAATTTCCTGAATTGGTACCAACAAAAACCCGGCAAGGCTCCCAAACTCCTGATTTATGATGCCTCCAATCTGGAGACCGGGGTCCCTTCTAGATTCAGCGGAAGTGGCAGCGGCACAGACTTTACAT<

[0223] Clone 24C1 CD28T CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold.) TIFF2026082913000007.tif64169

[0224] Clone 24C1 CD28T CD3 Zeta CAR DNA heavy and light strands GATATCCAGCTCACGCAATCCCCTCAAGCTTGAGTGCCTCCGTGGGCGACCGGGTGTCCTTCACATGTCAGGCAAGCCA

[0225] Clone 24C1 CD28T CD3 Zeta CAR AA Heavy and Light Chains DIQLTQSPSSLSASVGDRVSFTCQASQDINNFLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYGNLPFTFGGGGTKVEIKRGGGGSGG GSGGGGSQVQLQESGPGLVKPSETLSLTCTVSGGSISSYYWSWIRQPPGKGLEWIGYIYYSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCVSLVYCGGDCYSGFD YWGQGTLVTVSSAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (Sequence ID 40)

[0226] Clone 24C1 CD28 CD3 Zeta CAR DNA AA heavy and light strands ATGGCACTCCCCGTAACTGCTCTGCTGCTGCCGTTGGCAT

[0227] Clone 24C1 CD28 CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold.) TIFF2026082913000008.tif64168

[0228] Clone 24C1 CD28 CD3 Zeta CAR DNA heavy and light strands G (Sequence ID 43)

[0229] Clone 24C1 CD28 CD3 Zeta CAR AA Heavy and Light Chains DIQLTQSPSSLSASVGDRVSFTCQASQDINNFLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYGNLPFTFGGGGTKVEIKRGGGGSGGGGS GGGGSQVQLQESGPGLVKPSETLSLTCTVSGGSISSYYWSWIRQPPGKGLEWIGYIYYSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCVSLVYCGGDCYSGFDYWGQ GTLVTVSSAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR(Sequence ID 44)

[0230] Clone 24C1 CD8 CD3 Zeta CAR DNA heavy and light strands CGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA (SEQ ID NO: 45)

[0231] Clone 24C1 CD8 CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold.) TIFF2026082913000009.tif64168

[0232] Clone 24C1 CD8 CD3 Zeta CAR DNA heavy and light strands CCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG (SEQ ID NO: 47)

[0233] Clone 24C1 CD8 CD3 zeta CAR AA heavy and light chains DIQLTQSPSSLSASVGDRVSFTCQASQDINNFLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYGNLPFTFGGGTKVEIKRGGGGSGGGGSG GGGSQVQLQESGPGLVKPSETLSLTCTVSGGSISSYYWSWIRQPPGKGLEWIGYIYYSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTADTAVYYCVSLVYCGGDCYSGFDYWGQGTL VTVSSAAALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATK DTYDALHMQA LPPR (Sequence ID 48)

[0234] Clone 24C8 heavy chain (HC) DNA CAGGTACAGCTGCAGGAATCTGGGCCCGGACTTGTCAAGCCAAGTCAGACACTTTCTCTTACATGTACCGTGAGCGGCGGAAGTATAAGCAGTGGAGGCTTTTACTGGTCTTGGATACGGCAGCACCCAGGCAAAGGCTTGGAGTGGATTGGATACATTCATCATTCAGGATCTACACACTATAATCCAT CCCTTAAGTCCCGGGTCACCATTAGCATTGATACGTCTAAGAATCTGTTCAGTCTCAGGCTGTCCTCCGTCACTGCTGCCGACACAGCCGTGTACTACTGCGCCTCCTTGGTTTACTGCGGAGGCGACTGTTATAGCGGCTTTGATTATTGGGGGCAGGGGACCCTCGTAACCGTGAGCTCT (SEQ ID NO: 49)

[0235] Clone 24C8 AA HC (CDRs are underlined) QVQLQESGPGLVKPSQTLSLTCTVS GGSISSGGF YWSWIRQHPGKGLEWIGYI HHSGS THYNPSLKSRVTISIDTSKNLFSLRLSSVTAADTAVYYCAS LVYCGGDCYSGFDY WGQGTLVTVSS (SEQ ID NO: 50)

[0236] Clone 24C8 HC CDR1 AA: GGSISSGGF (SEQ ID NO: 51)

[0237] Clone 24C8 HC CDR2 AA: HHSGS (SEQ ID NO: 52)

[0238] Clone 24C8 HC CDR3 AA: LVYCGGDCYS GFDY (SEQ ID NO: 53)

[0239] Clone 24C8 light chain (LC) DNA GATATCCAGCTCACTCAAAGCCCCTCTAGTCTCTCTGCCTCAGTGGGGGATCGGGTCAGTTTTACTTGTCAAGCTTCACAGGATATCAACAACTTCCTTAATTGGTATCAGCAGAAGCCAGGAAAAGCACCCAAGCTGCTCATCTATGATGCCTCAAATTTGGAGACGGGTGTTCCCAGTCGATTCTCTGGGTCAGGGTCCGGGACCGACTTTACGTTTACGATCTCCTCTCTGCAGCCCGAAGACATCGCCACATACTATTGTCAACAGTACGGCAACTTGCCTTTCACATTTGGGGGCGGGACTAAGGTTGAAATCAAGAGG (SEQ ID NO: 54)

[0240] Clone 24C8 LC AA (CDRs are underlined) DIQLTQSPSSLSASVGDRVSFTC QASQDINNFLN WYQQKPGKAPKLLIY DASNLET GVPSRFSGSGSGTDFTFTISSLQPEDIATYYC QQYGNLPFTFGGGTKVEIKR (Sequence ID 55)

[0241] Clone 24C8 LC CDR1 AA:QASQDINNFLN (Sequence ID 56)

[0242] Clone 24C8 LC CDR2 AA:DASNLET (Sequence ID 57)

[0243] Clone 24C8 LC CDR3 AA:QQYGNLPFT (Sequence ID 58)

[0244] Clone 24C8 CD28T CD3 Zeta CAR DNA heavy and light strands ATGGCACTCCCCGTAACTGCTCTGCTGCTGCCGTTGGCATTGCTCCTGCACGCCGCACGCCCGCAGGTACAGCTGCAGGAATCTGGGCCCGGACTTGTCAAGCCAAGTCAGACACTTTCTCTTACATGTACCGTGAGCGGCGGAAGTATAAGCAGTGGAGGCTTTTACTGGTCTTGGATACGGCAGCACCCAGGCAAAGGCTTGGAGTGGATTGGATACATTCATCATTCAGGATCTACACACTATAATCCATCCCTTAAGTCCCGGGTCACCATTAGCATTGATACGTCTAAGAATCTGTTCAGTCTCAGGCTGTCCTCCGTCACTGCTGCCGACACAGCCGTGTACTACTGCGCCTCCTTGGTTTACTGCGGAGGCGACTGTTATAGCGGCTTTGATTATTGGGGGCAGGGGACCCTCGTAACCGTGAGCTCTGGAGGGGGTGGGAGCGGGGGAGGAGGTTCAGGGGGGGGCGGCTCCGATATCCAGCTCACTCAAAGCCCCTCTAGTCTCTCTGCCTCAGTGGGGGATCGGGTCAGTTTTACTTGTCAAGCTTCACAGGATATCAACAACTTCCTTAATTGGTATCAGCAGAAGCCAGGAAAAGCACCCAAGCTGCTCATCTATGATGCCTCAAATTTGGAGACGGGTGTTCCCAGTCGATTCTCTGGGTCAGGGTCCGGGACCGACTTTACGTTTACGATCTCCTCTCTGCAGCCCGAAGACATCGCCACATACTATTGTCAACAGTACGGCAACTTGCCTTTCACATTTGGGGGCGGGACTAAGGTTGAAATCAAGAGGGCCGCTGCACTGGACAATGAGAAGTCCAACGGCACCATCATCCACGTGAAGGGCAAGCACCTGTGCCCTAGTCCTCTGTTCCCAGGCCCATCCAAACCTTTTTGGGTTCTTGTTGTGGTCGGGGGGGTGCTGGCCTGCTATTCTCTGCTGGTCACG GTGGCCTTCATAATTTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCT AGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACG TTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAG GCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA (SEQ ID NO: 59)

[0245] Clone 24C8 CD28T CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold.) TIFF2026082913000010.tif64168

[0246] Clone 24C8 CD28T CD3 Zeta CAR DNA heavy and light strands CAGGTACAGCTGCAGGAATCTGGGCCCGGACTTGTCAAGCCAAGTCAGACACTTTCTCTTACATGTACCGTGAGCGGCGGAAGTATAAGCAGTGGAGGCTTTTACTGGTCTTGGATACGGCAGCACCCAGGCAAAGGCTTGGAGTGGATTGGATACATTCATCATTCAGGATCTACACACTATAATCCATCCCTTAAGTCCCGGGTCACCATTAGCATTGATACGTCTAAGAATCTGTTCAGTCTCAGGCTGTCCTCCGTCACTGCTGCCGACACAGCCGTGTACTACTGCGCCTCCTTGGTTTACTGCGGAGGCGACTGTTATAGCGGCTTTGATTATTGGGGGCAGGGGACCCTCGTAACCGTGAGCTCTGGAGGGGGTGGGAGCGGGGGAGGAGGTTCAGGGGGGGGCGGCTCCGATATCCAGCTCACTCAAAGCCCCTCTAGTCTCTCTGCCTCAGTGGGGGATCGGGTCAGTTTTACTTGTCAAGCTTCACAGGATATCAACAACTTCCTTAATTGGTATCAGCAGAAGCCAGGAAAAGCACCCAAGCTGCTCATCTATGATGCCTCAAATTTGGAGACGGGTGTTCCCAGTCGATTCTCTGGGTCAGGGTCCGGGACCGACTTTACGTTTACGATCTCCTCTCTGCAGCCCGAAGACATCGCCACATACTATTGTCAACAGTACGGCAACTTGCCTTTCACATTTGGGGGCGGGACTAAGGTTGAAATCAAGAGGGCCGCTGCACTGGACAATG AGAAGTCCAACGGCACCATCATCCACGTGAAGGGCAAGCACCTGTGCCCTAGTCCTCTGTTCCCAGGCCCATCCAAACCTTTTTGGGTTCTTGTTGTGGTCGGGGGGGTGCTGGCCTGCTATTCTCTGCTGGTCACGGTGGCCTTCATAATTTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG (SEQ ID NO: 61)

[0247] Clone 24C8 CD28T CD3 zeta CAR AA heavy and light chains QVQLQESGPGLVKPSQTLSLTCTVSGGSISSGGFYWSWIRQHPGKGLEWIGYIHHSGSTHYNPSLKSRVTISIDTSKNLFSLRLSSVTAADTAVYYCASLVYCGGDCYSGFDYWGQG TLVTVSSGGGGGSGGGGSGGGGSDIQLTQSPSSLSASVGDRVSFTCQASQDINNFLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTFTFTISSLQPEDIATYYCQQYGNLP FTFGGGTKVEIKRAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (Sequence ID 62)

[0248] Clone 24C8 CD28 CD3 Zeta CAR DNA heavy and light strands ATGGCACTCCCCGTAACTGCTCTGCTGCTGCCGTTGGCATTGCTCCTGCACGCCGCACGCCCGCAGGTGCAGCTGCAGGAAAGCGGTCCGGGACTTGTCAAGCCGTCCCAAACGCTGAGTCTGACGTGTACTGTCTCTGGTGGCTCTATTTCTTCCGGGGGCTTTTATTGGTCTTGGATCAGACAACACCCTGGCAAAGGGCTGGAGTGGATAGGGTATATTCACCACTCTGGGTCCACTCACTACAACCCATCATTGAAATCCAGAGTGACTATCTCAATCGACACATCCAAGAACCTTTTCAGCCTGAGGTTGTCATCAGTTACCGCCGCTGACACCGCGGTGTATTATTGCGCCTCTCTCGTGTACTGCGGTGGCGATTGTTATAGTGGCTTTGACTACTGGGGGCAGGGGACATTGGTTACCGTTTCAAGTGGAGGCGGTGGGTCTGGCGGGGGCGGTAGCGGAGGTGGGGGGAGCGACATACAGCTTACGCAGAGCCCCTCCAGCCTTTCAGCCTCCGTGGGGGATAGGGTGTCCTTTACCTGCCAGGCTTCCCAGGACATAAACAACTTCCTCAATTGGTATCAGCAAAAGCCCGGGAAAGCACCAAAGCTGCTCATCTACGATGCCAGCAACCTGGAAACCGGAGTGCCGTCTCGCTTCTCTGGAAGTGGCAGTGGGACCGATTTCACTTTTACAATCTCAAGTTTGCAGCCA (Sequence No. 63)

[0249] Clone 24C8 CD28 CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold.) TIFF2026082913000011.tif64169

[0250] Clone 24C8 CD28 CD3 Zeta CAR DNA heavy and light strands CAGGTGCAGCTGCAGGAAAGCGGTCCGGGACTTGTCAAGCCGTCCCAAACGCTGAGTCTGACGTGTACTGTCTCTGGTGGCTCTATTTCTTCCGGGGGCTTTTATTGGTCTTGGATCAGACAACACCCTGGCAAAGGGCTGGAGTGGATAGGGTATATTCACCACTCTGGGTCCACTCACTACAACCCATCATTGAAATCCAGAGTGACTATCTCAATCGACACATCCAAGAACCTTTTCAGCCTGAGGTTGTCATCAGTTACCGCCGCTGACACCGCGGTGTATTATTGCGCCTCTCTCGTGTACTGCGGTGGCGATTGTTATAGTGGCTTTGACTACTGGGGGCAGGGGACATTGGTTACCGTTTCAAGTGGAGGCGGTGGGTCTGGCGGGGGCGGTAGCGGAGGTGGGGGGAGCGACATACAGCTTACGCAGAGCCCCTCCAGCCTTTCAGCCTCCGTGGGGGATAGGGTGTCCTTT ACCTGCCAGGCTTCCCAGGACATAAACAACTTCCTCAATTGGTATCAGCAAAAGCCCGGGAAAGCACCAAAGCTGCTCATCTACGATGCCAGCAACCTGGAAACCGGAGTGCCGTCTCGCTTCTCTGGAAGTGGCAGTGGGACCGATTTCACTTTTACAATCTCAAGTTTGCAGCCAGAAGACATTGCAACATACTACTGTCAACAGTACGGCAATCTCCCCTTTACATTTGGGGGGGGAACTAAAGTGGAGATTAAGCGCGCTGCAGCCATTGAAGTTATGTATCCGCCCCCGTATCTGGATAACGAGAAATCTAATGGTACCATAATACATGTGAAGGGGAAGCACCTCTGTCCATCACCGCTGTTCCCCGGCCCTTCAAAACCTTTCTGGGTACTCGTTGTCGTGGGTGGAGTTCTGGCCTGCTATAGTCTGCTGGTGACCGTGGCGTTTATCATCTTCTGGGTAAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG(SEQ ID NO: 65)

[0251] Clone 24C8 CD28 CD3 Zeta CAR AA Heavy and Light Chains QVQLQESGPGLVKPSQTLSLTCTVSGGSISSGGFYWSWIRQHPGKGLEWIGYIHHSGSTHYNPSLKSRVTISIDTSKNLFSLRLSSVTAADTAVYYCASLVYCGGDCYSGFDYWGQGTL VTVSSGGGGGSGGGGSGGGGSDIQLTQSPSSLSASVGDRVSFTCQASQDINNFLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYGNLPFTFG GGTKVEIKRAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (Sequence ID 66)

[0252] Clone 24C8 CD8 CD3 Zeta CAR DNA heavy and light strands ATGGCACTCCCCGTAACTGCTCTGCTGCTGCCGTTGGCATTGCTCCTGCACGCCGCACGCCCGCAGGTGCAGTTGCAGGAAAGCGGGCCTGGCCTTGTGAAACCAAGCCAGACACTGAGCCTGACATGCACTGTGTCCGGCGGGTCCATATCTTCCGGGGGTTTTTATTGGTCCTGGATACGCCAGCATCCCGGGAAAGGACTTGAATGGATTGGATATATCCACCATTCCGGAAGCACCCACTACAATCCAAGCCTTAAATCCCGGGTGACAATCTCCATCGACACCTCAAAGAATCTTTTTTCCCTGCGGTTGTCTTCAGTAACTGCCGCCGATACCGCTGTGTACTACTGTGCCAGCCTCGTCTATTGCGGCGGAGATTGTTATTCTGGGTTCGATT

[0253] Clone 24C8 CD8 CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold.) TIFF2026082913000012.tif64169

[0254] Clone 24C8 CD8 CD3 Zeta CAR DNA heavy and light strands CAGGTGCAGTTGCAGGAAAGCGGGCCTGGCCTTGTGAAACCAAGCCAGACACTGAGCCTGACATGCACTGTGTCCGGCGG

[0255] Clone 24C8 CD8 CD3 Zeta CAR AA Heavy and Light Chains QVQLQESGPGLVKPSQTLSLTCTVSGGSISSGGFYWSWIRQHPGKGLEWIGYIHHSGSTHYNPSLKSRVTISIDTSKNLFSLRLSSVTAADTAVYYCASLVYCGGDCYSGFDYWGQGTLV TVSSGGGGSGGGSGGGGSDIQLTQSPSSLSASVGDRVSFTCQASQDINNFLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYGNLPFTFGGG TKVEIKRAAALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNRSKRSRLLHSDYMNMTP RRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTA TKDTYDALHM QALPPR (Sequence ID 70)

[0256] Clone 20C5.1 HC DNA CAGGTCCAACTGGTGCAGTCCGGAGCCGAAGTCAAGAAACCAGGTGCCTCCGTTAAAGTGAGTTGCAAAGTCTCTGGATACACTCTGACCGAGCTCTCTATGCACTGGGTCCGGCAGGCCCCCGGCAAGGGATTGGAATGGATGGGCGGGTTCGATCCTGAGGACGGAGAGACTATCTACGCTCA AAAATTCCAGGGACGAGTGACTGTGACCGAAGACACTAGTACCGACACTGCCTACATGGAACTTTCCTCTCTGCGATCAGAAGATACCGCAGTGTACTACTGTGCTACTGAATCTAGGGGCATTGGATGGCCCTACTTCGATTACTGGGGTCAGGGAACTCTGGTGACTGTCTCCAGC (SEQ ID NO: 71)

[0257] Clone 20C5.1 AA HC (CDR is underlined) QVQLVQSGAEVKKPGASVKVSCKVS GYTLTEL SMHWVRQAPGKGLEWMGGF DPEDGE TIYAQKFQGRVTVTEDTSTDTAYMELSSLRSEDTAVYYCAT ESRGIGWPYFDY WGQGTLVTVSS (Sequence ID 72)

[0258] Clone 20C5.1 HC AA CDR1:GYTLTEL (Sequence ID 73)

[0259] Clone 20C5.1 HC AA CDR2:DPEDGE (Sequence ID 74)

[0260] Clone 20C5.1 HC AA CDR3:ESRGIGWPYFDY (Sequence ID 75)

[0261] Clone 20C5.1 LC DNA GATATTCAGATGACTCAATCTCCTTCTTCTCTGTCCGCTTCCGTGGGCGATAGAGTGACCATTACTTGTAGGGCGTCCCAGTCAATCTCCAGTTATTTGAATTGGTATCAGCAGAAGCCCGGGAAAGCACCTAAGCTGTTGATCAGCGGGGCTTCTAGCCTGAAGA GTGGGGTACCTTCACGGTTCAGCGGAAGCGGAAGCGGAACCGATTTCACCCTGACTATCAGCAGCCTGCCACCTGAGGACTTTGCAACTTACTACTGCCAACAGTCATACAGCACTCCGATCACTTTCGGCCAGGGCACCCGGCTCGAAATCAAGCGC (SEQ ID NO: 76)

[0262] Clone 20C5.1 AA LC (CDR is underlined) DIQMTQSPSSLSASVGDRVTITC RASQSISSYLN WYQQKPGKAPKLLIS GASSLKS GVPSRFSGSGSGTDFLTISSLPPEDFATYYC QQSYSTPIT FGQGTRLEIKR (Sequence ID 77)

[0263] Clone 20C5.1 AA LC CDR1:RASQSISSYLN (Sequence ID 78)

[0264] Clone 20C5.1 AA LC CDR2:GASSLKS (Sequence ID 79)

[0265] Clone 20C5.1 AA LC CDR3:QQSYSTPIT (Sequence ID 80)

[0266] Clone 20C5.1 CD28T CD3 Zeta CAR DNA heavy and light strands

[0267] Clone 20C5.1 CD28T CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold.) TIFF2026082913000013.tif64169

[0268] Clone 20C5.1 CD28T CD3 Zeta CAR DNA heavy and light strands

[0269] Clone 20C5.1 CD28T CD3 Zeta CAR AA Heavy and Light Chains QVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMGGFDPEDGETIYAQKFQGRVTVTEDTSTDTAYMELSSLRSEDTAVYYCATESRGIGWPYFDYWGQGTL VTVSSGGGGSGGGGSGGGSDIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLISGASSLKSGVPSRFSGSGSGTDFTLTISSLPPEDFATYYCQQSYSTPI TFGQGTRLEIKRAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR(Sequence ID 84)

[0270] Clone 20C5.1 CD28 CD3 Zeta CAR DNA heavy and light strands AGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA (SEQ ID NO: 85)

[0271] Clone 20C5.1 CD28 CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold.) TIFF2026082913000014.tif64169

[0272] Clone 20C5.1 CD28 CD3 Zeta CAR DNA heavy and light strands CGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAAT AGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG (SEQ ID NO: 87)

[0273] Clone 20C5.1 CD28 CD3 Zeta CAR AA Heavy and Light Chains QVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMGGFDPEDGETIYAQKFQGRVTVTEDTSTDTAYMELSSLRSEDTAVYYCATESRGIGWPYFDYWGQGT LVTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLISGASSLKSGVPSRFSGSGSGTDFTLTISSLPPEDFATYYCQQSYST PITFGQGTRLEIKRAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHM QALPPR (Sequence ID 88)

[0274] Clone 20C5.1 CD8 CD3 Zeta CAR DNA heavy and light strands AACAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAA GCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA (SEQ ID NO: 89)

[0275] Clone 20C5.1 CD8 CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold.) TIFF2026082913000015.tif64169

[0276] Clone 20C5.1 CD8 CD3 Zeta CAR DNA heavy and light strands CAGGTGCAGTTGGTGCAAAGCGGCGCAGAAGTTAAGAAACCTGGGGCGTCAGTTAAGGTGTCTTGCAAAGTATCTGGCTATACCCTCACTGAGCTGTCCATGCATTGGGTAAGGCAGGCTCCTGGAAAGGGGCTCGAATGGATGGGAGGATTTGACCCTGAAGACGGAGAGACCATCTACGCCCAGAAATTCCAGGGTAGAGTAACAGTGACTGAGGACACTAGCACTGACACAGCGTACATGGAGCTGAGTTCTCTGAGAAGTGAGGACACAGCCGTTTACTACTGCGCTACCGAGTCCAGAGGTATTGGCTGGCCATACTTCGACTATTGGGGTCAGGGCACCCTGGTTACAGTGAGTTCAGGAGGCGGGGGCTCTGGGGGGGGCGGTTCCGGAGGGGGGGGCTCAGATATACAGATGACGCAGAGTCCATCAAGTCTCTCAGCCAGCGTGGGAGATCGCGTGACTATTACTTGCCGCGCCAGCCAGAGTATTAGCTCCTATCTGAATTGGTACCAGCAAAAGCCCGGGAAGGCCCCTAAGCTTCTGATTTCTGGCGCCTCCTCTTTGAAGTCAGGTGTGCCAAGCAGATTTAGCGGGTCTGGAAGTGGCACTGACTTTACACTTACTATCTCCAGCCTGCCCCCAGAGGATTTTGCCACATATTACTGTCAGCAAAGCTACTCTACTCCAATCACTTTCGGCCAGGGCACAAGATTGGAGATTAAGAGGGCTGCCGCACTTTCAAATTCCATCATGTATTTCAGCCATTTTGTGCCTGTTTTTCTTCCGGCCAAACC TACAACCACTCCCGCCCCACGCCCACCTACTCCCGCCCCTACCATTGCCTCCCAGCCTCTGTCTCTTAGACCTGAGGCTTGTAGACCTGCTGCCGGCGGAGCCGTGCACACTCGCGGTCTGGACTTCGCCTGCGACATCTATATCTGGGCCCCTCTGGCCGGCACCTGCGGCGTTCTCCTTCTCTCACTCGTAATCACACTCTATTGCAATCACAGGAACAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG (SEQ ID NO: 91)

[0277] Clone 20C5.1 CD8 CD3 zeta CAR AA heavy and light chains QVQLVQSGAEVKKPGASVKVSCKVSGYTLTELSMHWVRQAPGKGLEWMGGFDPEDGETIYAQKFQGRVTVTEDTSTDTAYMELSSLRSEDTAVYYCATESRGIGWPYFDYWGQGTLVTVSSGGGG SGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLISGASSLKSGVPSRFSGSGSGTDFTLTISSLPPEDFATYYCQQSYSTPITFGQGTRLEIKRAAALSN SIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR(Sequence ID 92)

[0278] Clone 20C5.2 HC DNA CAGGTCCAGTTGGTCGAAAGTGGCGGTGGTGTAGTGCAGCCGGGCCGCAGTTTGAGGCTTTCCTGTGCGGCTTCAGGCTTTACTTTTTCCAGCTATGGAATGCACTGGGTGCGGCAGGCCCCCGGCAAAGGACTTGAGTGGGTGGCCGTCATTTCTTATGACGGATCAGATAAGTACTA CGTGGACAGCGTCAAGGGCAGATTCACCATCTCTAGGGACAACAGTAAAAATAGACTCTACCTCCAGATGAATAGCCTCAGAGCTGAAGACACGGCCGTCTACTATTGTGCTCGGGAGCGGTATAGTGGCAGAGACTACTGGGGGCAGGGCACACTCGTTACAGTGAGTAGC (SEQ ID NO: 93)

[0279] Clone 20C5.2 AA HC (CDR is underlined) QVQLVESGGGVVQPGRSLRLSCAAS GFTFSSY GMHWVRQAPGKGLEWVAVI SYDGSD KYYVDSVKGRFTISRDNSKNRLYLQMNSLRAEDTAVYYCAR ERYSGRDY WGQGTLVTVSS (Sequence ID 94)

[0280] Clone 20C5.2 HC AA CDR1:GFTFSSY (Sequence ID 95)

[0281] Clone 20C5.2 HC AA CDR2:SYDGSD (Sequence ID 96)

[0282] Clone 20C5.2 HC AA CDR3:ERYSGRDY (Sequence ID 97)

[0283] Clone 20C5.2 LC DNA GAGATTGTTATGACCCAGAGTCCTGCGACCCTCTCAGTCAGCCCCGGGGAGCGCGCAACTTTGTCTTGCAGAGCTAGTCAGTCCGTGTCCTCTCTTCTGACATGGTACCAGCAAAAGCCCGGGCAGGCTCCGCGCCTTTTGATCTTTGGGGCTTCAACAAGAGCCA CTGGGATTCCCGCACGATTCTCTGGCTCCGGGAGCGGTACTGGTTTCACCCTGACGATTAGCAGTCTCCAGAGCGAGGACTTCGCCGTATACTACTGCCAGCAGTACGATACGTGGCCATTCACTTTTGGACCAGGGACTAAAGTGGATTTTAAGCGC (SEQ ID NO: 98)

[0284] Clone 20C5.2 AA LC (CDR is underlined) EIVMTQSPATLSVSPGERATLSC RASQSVSSLLT WYQQKPGQAPRLLIF GASTRAT GIPARFSGSGSGTGFTLTISSLQSEDFAVYYC QQYDTWPFT FGPGTKVDFKR (Sequence ID 99)

[0285] Clone 20C5.2 AA LC CDR1:RASQSVSSLLT (Sequence ID 100)

[0286] Clone 20C5.2 AA LC CDR2:GASTRAT (Sequence ID 101)

[0287] Clone 20C5.2 AA LC CDR3:QQYDTWPFT (Sequence ID 102)

[0288] Clone 20C5.2 CD28T CD3 Zeta CAR DNA heavy and light strands ATGGCACTCCCCGTAACTGCTCTGCTGCTGCCGTTGGCATTGCTCCTGCACGCCGCACGCCCGCAGGTCCAGTTGGTCGAAAGTGGCGGTGGTGTAGTGCAGCCGGGCCGCAGTTTGAGGCTTTCCTGTGCGGCTTCAGGCTTTACTTTTTCCAGCTATGGAATGCACTGGGTGCGGCAGGCCCCCGGCAAAGGACTTGAGTGGGTGGCCGTCATTTCTTATGACGGATCAGATAAGTACTACGTGGACAGCGTCAAGGGCAGATTCACCATCTCTAGGGACAACAGTAAAAATAGACTCTACCTCCAGATGAATAGCCTCAGAGCTGAAGACACGGCCGTCTACTATTGTGCTCGGGAGCGGTATAGTGGCAGAGACTACTGGGGGCAGGGCACACTCGTTACAGTGAGTAGCGGCGGAGGAGGGAGTGGGGGCGGTGGCTCCGGTGGAGGAGGTTCTGAGATTGTTATGACCCAGAGTCCTGCGACCCTCTCAGTCAGCCCCGGGGAGCGCGCAACTTTGTCTTGCAGAGCTAGTCAGTCCGTGTCCTCTCTTCTGACATGGTACCAGCAAAAGCCCGGGCAGGCTCCGCGCCTTTTGATCTTTGGGGCTTCAACAAGAGCCACTGGGATTCCCGCACGATTCTCTGGCTCCGGGAGCGGTACTGGTTTCACCCTGACGATTAGCAGTCTCCAGAGCGAGGACTTCGCCGTATACTAC (Sequence ID 103)

[0289] Clone 20C5.2 CD28T CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold.) TIFF2026082913000016.tif64169

[0290] Clone 20C5.2 CD28T CD3 Zeta CAR DNA heavy and light strands CAGGTCCAGTTGGTCGAAAGTGGCGGTGGTGTAGTGCAGCCGGGCCGCAGTTTGAGGCTTTCCTGTGCGGCTTCAGGCTTTACTTTTTCCAGCTATGGAATGCACTGGGTGCGGCAGGCCCCCGGCAAAGGACTTGAGTGGGTGGCCGTCATTTCTTATGACGGATCAGATAAGTACTACGTGGACAGCGTCAAGGGCAGATTCACCATCTCTAGGGACAACAGTAAAAATAGACTCTACCTCCAGATGAATAGCCTCAGAGCTGAAGACACGGCCGTCTACTATTGTGCTCGGGAGCGGTATAGTGGCAGAGACTACTGGGGGCAGGGCACACTCGTTACAGTGAGTAGCGGCGGAGGAGGGAGTGGGGGCGGTGGCTCCGGTGGAGGAGGTTCTGAGATTGTTATGACCCAGAGTCCTGCGACCCTCTCAGTCAGCCCCGGGGAGCGCGCAACTTTGTCTTGCAGAGCTAGTCAGTCCGTGTCCTCTCTTCTGACATGGTACCAGCAAAAGCCCGGGCAGGCTCCGCGCCTTTTGATCTTTGGGGCTTCAACAAGAGC CACTGGGATTCCCGCACGATTCTCTGGCTCCGGGAGCGGTACTGGTTTCACCCTGACGATTAGCAGTCTCCAGAGCGAGGACTTCGCCGTATACTACTGCCAGCAGTACGATACGTGGCCATTCACTTTTGGACCAGGGACTAAAGTGGATTTTAAGCGCGCCGCCGCTCTCGATAACGAAAAGTCAAATGGCACCATAATCCACGTCAAAGGCAAGCACCTGTGCCCTTCCCCGCTCTTCCCCGGACCCAGTAAACCATTTTGGGTGCTGGTTGTTGTGGGGGGCGTGCTGGCCTGCTATAGCCTTTTGGTCACTGTAGCCTTCATTATTTTTTGGGTCAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG(SEQ ID NO: 105)

[0291] Clone 20C5.2 CD28T CD3 zeta CAR AA heavy and light chains QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSDKYYVDSVKGRFTISRDNSKNRLYLQMNSLRAEDTAVYYCARERYSGRDYWGQGTLVTV SSGGGGSGGGGSGGGGSEIVMTQSPATLSVSPGERATLSCRASQSVSSLLTWYQQKPGQAPRLLIFGASTRATGIPARFSGSGSGTGFTLTISSLQSEDFAVYYCQQYDTWPFTFG PGTKVDFKRAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (Sequence ID 106)

[0292] Clone 20C5.2 CD28 CD3 Zeta CAR DNA heavy and light strands ATGGCACTCCCCGTAACTGCTCTGCTGCTGCCGTTGGCATTGCTCCTGCACGCCGCACGCCCGCAGGTGCAGCTCGTGGAGTCTGGCGGCGGCGTGGTCCAGCCCGGCCGGTCCCTGCGCCTGTCCTGCGCCGCCAGCGGGTTTACTTTTTCCTCCTACGGCATGCACTGGGTGCGCCAGGCTCCCGGCAAGGGCCTCGAGTGGGTCGCCGTGATCTCATACGATGGGTCAGACAAATACTATGTCGATTCTGTTAAAGGGCGGTTTACCATTTCAAGAGATAACTCTAAGAATAGGCTGTATTTGCAGATGAACAGCCTGAGGGCTGAAGATACCGCAGTGTACTATTGCGCTAGGGAGCGGTATAGTGGCCGCGATTACTGGGGACAGGGTACACTGGTGACCGTGAGCTCTGGGGGTGGCGGAAGCGGGGGTGGCGGAAGCGGCGGAGGGGGTAGTGAAATTGTGATGACCCAGTCTCCGGCTACACTTTCAGTCTCCCCTGGGGAGAGAGCTACAC TGTCATGCAGAGCGTCCCAGTCCGTCTCTTCTCTCCTTACCTGGTATCAGCAGAAGCCCGGCCAGGCTCCTCGACTGCTGATCTTCGGTGCCTCCACAAGGGCGACCGGGATTCCAGCCCGCTTCTCAGGTTCTGGGAGCGGAACTGGTTTCACTTTGACAATCAGTTCACTGCAGTCAGAGGATTTCGCCGTGTACTACTGCCAGCAATACGACACATGGCCATTCACTTTCGGACCCGGTACCAAAGTCGATTTCAAGAGAGCCGCGGCCATCGAGGTTATGTACCCACCACCATATCTGGACAATGAAAAAAGCAATGGAACCATTATCCATGTGAAGGGTAAACACCTCTGCCCTAGCCCACTTTTCCCTGGCCCATCAAAGCCCTTCTGGGTCTTGGTGGTCGTGGGGGGTGTGCTGGCCTGTTACAGCCTTCTGGTGACGGTTGCTTTCATTATCTTCTGGGTTAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA(SEQ ID NO: 107)

[0293] Clone 20C5.2 CD28 CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold.) TIFF2026082913000017.tif64169

[0294] Clone 20C5.2 CD28 CD3 Zeta CAR DNA heavy and light strands CAGGTGCAGCTCGTGGAGTCTGGCGGCGGCGTGGTCCAGCCCGGCCGGTCCCTGCGCCTGTCCTGCGCGCCAGCGGGTTTACTTTTTCCTCCTACGGCATGCACTGGGTGCGCCAGGCTCCCGGCAAGGGCCTCGAGTGGGTCGCCGTGATCTCATACG ATGGGTCAGACAAATACTATGTCGATTCTGTTAAAGGGCGGTTTACCATTTCAAGAGATAACTCTAAGAATAGGCTGTATTTGCAGATGAACAGCCTGAGGGCTGAAGATACCGCAGTGTACTATTGCGCTAGGAGCGGTATAGTGGCCGCGATTACTG

[0295] Clone 20C5.2 CD28 CD3 Zeta CAR AA Heavy and Light Chains QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSDKYYVDSVKGRFTISRDNSKNRLYLQMNSLRAEDTAVYYCARERYSGRDYWGQGTLVTVSS GGGGSGGGGSGGGGSEIVMTQSPATLSVSPGERATLSCRASQSVSSLLTWYQQKPGQAPRLLIFGASTRATGIPARFSGSGSGTGFTLTISSLQSEDFAVYYCQQYDTWPFTFGPGTK VDFKRAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (Sequence ID 110)

[0296] Clone 20C5.2 CD8 CD3 Zeta CAR DNA heavy and light strands ATGGCACTCCCCGTAACTGCTCTGCTGCTGCCGTTGGCATTGCTCCTGCACGCCGCACGCCCGCAGGTGCAGTTGGTTGAATCAGGAGGGGGTGTGGTGCAACCCGGTCGGTCACTGCGCCTCAGTTGTGCTGCTTCCGG GTTTACTTTCAGCTCATATGGGATGCACTGGGTACGGCAGGCTCCAGGTAAAGGCTTGGAATGGGTGGCGGTGATCAGCTATGACGGCTCTGACAAATATTATGTGGACTCCGTGAAAGGCAGATTCACCATCAGTCGAG

[0297] Clone 20C5.2 CD8 CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold.) TIFF2026082913000018.tif64169

[0298] Clone 20C5.2 CD8 CD3 Zeta CAR DNA heavy and light strands

[0299] Clone 20C5.2 CD8 CD3 Zeta CAR AA Heavy and Light Chains QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSDKYYVDSVKGRFTISRDNSKNRLYLQMNSLRAEDTAVYYCARERYSGRDYWGQG TLVTVSSGGGGSGGGGSGGGGSEIVMTQSPATLSVSPGERATLSCRASQSVSSLLTWYQQKPGQAPRLLIFGASTRATGIPARFSGSGSGTGFTLTISSLQSEDFAVYYC QQYDTWPTFGPGTKVDFKRAAALSNSIMYFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITL YCNHRNRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQ KDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (Sequence ID 114)

[0300] Clone 20C5.2 CD28T CD3 Zeta CAR DNA heavy and light strands

[0301] Clone 20C5.2 CD28T CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold.) TIFF2026082913000019.tif64169

[0302] Clone 20C5.2 CD28T CD3 Zeta CAR DNA heavy and light strands

[0303] Clone 20C5.2 CD28T CD3 Zeta CAR AA Heavy and Light Chains EIVMTQSPATLSVSPGERATLSCRASQSVSSLLTWYQQKPGQAPRLLIFGASTRATGIPARFSGSGSGTGFTLTISSLQSEDFAVYYCQQYDTWPTFTFGPGTKVDFKRGGGGSGG GGSGGGGSQVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSDKYYVDSVKGRFTISRDNSKNRLYLQMNSLRAEDTAVYYCARERYSGRDYWG QGTLVTVSSAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (Sequence ID 118)

[0304] Clone 20C5.2 CD28 CD3 Zeta CAR DNA heavy and light strands GAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA (SEQ ID NO: 119)

[0305] Clone 20C5.2 CD28 CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold.) TIFF2026082913000020.tif64168

[0306] Clone 20C5.2 CD28 CD3 Zeta CAR DNA heavy and light strands GCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG (SEQ ID NO: 121)

[0307] Clone 20C5.2 CD28 CD3 Zeta CAR AA Heavy and Light Chains EIVMTQSPATLSVSPGERATLSCRASQSVSSLLTWYQQKPGQAPRLLIFGASTRATGIPARFSGSGSGTGFTLTISSLQSEDFAVYYCQQYDTWPTFFGGPGTKVDFKRGGGGSGGGG SGGGGSQVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSDKYYVDSVKGRFTISRDNSKNRLYLQMNSLRAEDTAVYYCARERYSGRDYWGQGTL VTVSSAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (Sequence ID 122)

[0308] Clone 20C5.2 CD8 CD3 Zeta CAR DNA heavy and light strands CACGCCGCCCTGGCCCACAAGGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGG TGGCAAACCAAGACGAAAAAACCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGGTAA (SEQ ID NO: 123)

[0309] Clone 20C5.2 CD8 CD3 Zeta CAR AA Heavy and Light Chains (Signal peptides are in bold.) TIFF2026082913000021.tif64169

[0310] Clone 20C5.2 CD8 CD3 Zeta CAR DNA heavy and light strands GAAATAGTGATGACTCAGTCCCCGGCCACCCTCAGCGTGTCCCCCGGGGAGCGAGCGACCCTGTCATGCAGGGCTTCCCAGAGTGTCAGCTCCCTGCTCACTTGGTATCAGCAAAAGCCGGGGCAGGCTCCCCGCCTCCTCATCTTCGGGGCATCAACTAGGGCCACCGGCATTCCTGCAAGATTTTCCGGGTCTGGCAGCGGCACCGGCTTCACCCTTACCATTAGCTCTCTGCAGTCTGAGGACTTCGCCGTTTACTATTGTCAGCAGTATGATACTTGGCCCTTTACCTTCGGTCCCGGAACTAAGGTGGACTTCAAGCGCGGGGGGGGTGGATCTGGAGGTGGTGGCTCCGGGGGCGGTGGAAGCCAGGTCCAGTTGGTTGAGAGCGGCGGCGGAGTGGTGCAGCCCGGGAGGTCCTTGCGGCTGAGCTGTGCAGCCTCCGGTTTTACTTTTTCTAGCTATGGAATGCATTGGGTAAGACAGGCTCCCGGAAAAGGCCTCGAGTGGGTGGCGGTCATTAGCTATGATGGATCTGATAAATACTATGTGGACTCAGTTAAGGGGCGCTTCACAATCTCAAGAGACAATAGCAAAAATAGACTGTACCTGCAGATGAATAGTCTGCGCGCCGAGGACACTGCCGTGTACTACTGCGCCCGCGAGAGATACAGCGGACGGGATTACTGGGGCCAGGGTACCCTCGTAACGGTGTCCTCCGCTGCCGCCCTTAGCAACAGCATTATGTACTTTTCTCATTTCGTGCCAGTCTTTCTCCCAGCAAAGCCCACCACTACCCCGGCCCCCAGGCCGCCTACTCCTGCCCCCACTATCGCGTCT CAGCCTCTCTCCTTGCGGCCCGAGGCCTGCCGGCCAGCCGCAGGGGGCGCCGTACATACTCGGGGTTTGGATTTCGCTTGCGACATATATATTTGGGCCCCCCTCGCCGGCACATGTGGAGTGCTGCTCCTGAGTCTCGTTATAACCCTCTATTGCAACCATAGAAACAGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGCAGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAGGGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG (SEQ ID NO: 125)

[0311] Clone 20C5.2 CD8 CD3 zeta CAR AA heavy and light chains EIVMTQSPATLSVSPGERATLSCRASQSVSSLLTWYQQKPGQAPRLLIFGASTRATGIPARFSGSGSGTGFTLTISSLQSEDFAVYYCQQYDTWPFTFGPGTKVDFKRGGGGSGGGGSGGGGSQ VQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSDKYYVDSVKGRFTISRDNSKNRLYLQMNSLRAEDTAVYYCARERYSGRDYWGQGTLVTVSSAAALSNSIM YFSHFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (Sequence ID 126)

[0312] CAR signal peptide DNA ATGGCACTCCCCGTAACTGCTCTGCTGCTGCCGTTGGCATTGCTCCTGCACGCCGCACGCCCG (Sequence ID 127)

[0313] CAR signaling peptide: MALPVTALLLPLALLLHAARP (SEQ ID NO: 128)

[0314] scFv G4S Linker DNA GGCGGTGGAGGCTCCGGAGGGGGGGGCTCTGGCGGAGGGGGCTCC(Sequence 129)

[0315] scFv G4s linker:GGGGSGGGGSGGGGS (Sequence ID 130)

[0316] scFv Whitlow Linker DNA GGGTCTACATCCGGCTCCGGGAAGCCCGGAAGTGGCGAAGGTAGTACAAAGGGG (Sequence No. 131)

[0317] scFv Whitlow linker: GSTGSSGKPGSGEGSTKG (sequence number) No. 132)

[0318] CD28 AA extracellular domain MLRLLLALNLFPSIQVTGNKILVKQSPMLVAYDNAVNLSCKYSYNLFSREFRASLHKGLDSAVEVCVVYGNYSQQLQVYSKTGFNCDGKLGNESVTFYLQNLYVNQTDIYFCKIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP(Sequence ID 133)

[0319] GX2X3X4X5X6X7X8X9 (Sequence number 134)

[0320] X1X2X3X4X5X6 (Sequence number 135)

[0321] X1X2X3X4X5X6X7X8X9X 10 X 11 X 12 DY (Sequence No. 136)

[0322] X1ASQX5X6X7X8X9LX 11 (Sequence ID 137)

[0323] X1ASX4X5X6X7 (Sequence ID 138)

[0324] QQX3X4X5X6PX8T (Sequence ID 139)

[0325] 4-1BB nucleic acid sequence (intracellular domain) AAGCGCGGCAGGAAGAAGCTCCTCTACATTTTTAAGCAGCCTTTTATGAGGCCCGTACAGACAACACAGGAGGAAGATGGCTGTAGCTGCAGATTTCCCGAGGAGGAGGAAGGTGGGTGCGAGCTG (SEQ ID NO: 140)

[0326] 4-1BB AA (intracellular domain) KRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL (Sequence ID 141)

[0327] OX40 AA RRDQRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI (Sequence ID 142)

[0328] Leader array AA MALPVTALLLPLALLLHAARP (Sequence ID 143)

[0329] Additional G4S linker: GGGGSGGGGSGGGGSGGGGS (Sequence ID 144)

[0330] CD3 zeta variant AA RVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR(Sequence ID 145)

[0331] Axi Cel(KTE-C19) DNA(SEQ ID NO: 146) Atgcttctcctggtgacaagccttctgctctgtgagttaccacacccagcattcctcctgatcccagacatccagatgac

[0332] Axi Cel(KTE-C19) AA(Sequence ID 147) MLLLVTSLLLCELPHPAFLLIPDIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGG GTKLEITGSTSGSGKPGSGEGSTKGEVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAK HYYYGGSYAMDYWGQGTSVTVSSAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPY APPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR

[0333] Humanized anti-CD19 CAR DNA (SEQ ID NO: 148)

[0334] Humanized anti-CD19 CAR AA (SEQ ID NO: 149) MALPVTALLLPLALLLHAARPDIQMTQSPSSLSASVGDRVTITCRASQDISKYLNWYQQKPDQAPKLLIKHTSRLHSGVPSRFSGSGSGTDYTLTISSLQ PEDFATYYCQQGNTLPYTFGQGTKLEIKGSTSGSGKPGSGEGSTKGEVQLVESGGGLVQPGRSLRLSCTASGVSLPDYGVSWIRQPPGKGLEWIGVIWGS ETTYYNSALKSRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKHYYYGGSYAMDYWGQGTLVTVSSAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYD VLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR

[0335] Fully human anti-CD19 CAR DNA (SEQ ID NO: 150)

[0336] Fully human anti-CD19 CAR AA (SEQ ID NO: 151) MALPVTALLLPLALLLHAARPEIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSRFTF GPGTKVDIKGSTSGSGKPGSGEGSTKGQVQLVQSGAEVKKPGSSVKVSCKDSGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTTNYAQQFQGRVTITADESTSTAYMELSSLRSEDTAV YYCAREAVAADWLDPWGQGTLVTVSSFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNRSKRSRLLHSDYMNMTPRRPGP TRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR

[0337] MAGE A3 / A6 TCR DNA (SEQ ID NO: 152) cacacagaacatcagtgcagaggcctggggccgagcagactgtggaatcacttcagcatcctatcatcagggggttctgtctgcaaccatcctctatgagatcctactggggaaggccaccctatatgctgtgctggtcagtggcctggtgctgatggctatggtcaaaagaaagaactcatga

[0338] MAGE A3 / A6 TCR AA (SEQ ID NO: 153) M A C P G F L W A L V I S T C L E F S M A Q T V T Q S Q P E M S V Q E A E T V T L S C T Y D T S E S D Y Y L F W Y K Q P P S R Q M I L V I R Q E A Y K Q Q N A T E N R F S V N F Q K A A K S F S L K I S D S Q L G D A A M Y F C A L R S S G T Y K Y I F G T G T R L K V L A N I Q N P E P A V Y Q L K D P R S Q D S T L C L F T D F D S Q I N V P K T M E S G T F I T D K T V L D M K A M D S K S N G A I A W S N Q T S F T C Q D I F K E T N A T Y P S S D V P C D A T L T E K S F E T D M N L N F Q N L S V M G L R I L L L K V A G F N L L M T L R L W S S R A K R S G S G A T N F S L L K Q A G D V E E N P G PM G T R L L F W V A F C L L G A D H T G A G V S Q S P S N K V T E K G K D V E L R C D P I S G H T A L Y W Y R Q S L G Q G L E F L I Y F Q G N S A P D K S G L P S D R F S A E R T G G S V S T L T I Q R T Q Q E D S A V Y L C A SI R T G P F F S G N T I Y F G E G S W L T V V E D L R N V T P P K V S L F E P S K A E I A N K Q K A T L V C L A R G F F P D H V E L S W W V N G K E V H S G V S T D P Q A Y K E S N Y S Y C L S S R L R V S A T F W H N P R N H F R C Q V Q F H G L S E E D K W P E G S P K P V T Q N I S A E A W G R A D C G I T S A S Y H Q G V L S A T I L Y E I L L G K A T L Y A V L V S G L V L M A M V K R K N S Stop

[0339] Anti-CLL-1 CAR DNA (SEQ ID NO: 154) ATGGCACTCCCCGTAACTGCTCTGCTGCTGCCGTTGGCATTGCTCCTGCACGCCGCACGCCCGCAGGTCCAACTGCAAGAAAGCGGACCCGGACTGGTGAAGCCTTCTGAGACACTTAGTCTGACGTGCACGGTCAGTGGCGGCTCCATCTCCTCCTATTATTGGTCATGGATACGACAACCCCCAGGTAAGGGCCTGGAATGGATTGGCTATATCTACTATTCAGGAAGCACGAACTACAATCCCAGCCTGAAGTCCCGAGTGACAATTTCAGTAGATACCAGTAAAAACCAGTTCAGTCTTAAACTGTCAAGCGTGACAGCTGCCGACACCGCTGTGTATTACTGCGTCTCACTGGTGTATTGTGGAGGGGATTGTTATAGCGGGTTCGATTATTGGG

[0340] Anti-CLL-1 CAR AA MALPVTALLLPLALLLHAARPQVQLQESGPGLVKPSETLSLTCTVSGGSISSYYWSWIRQPPGKGLEWIGYIYYSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYCVSLVYCGGDCYSGFDYWGQGTLVTVSSGGGGSGGGGSGGGGSDIQLTQSPSSLSASVGDRVSFTCQASQDINNFLNWYQQKPGKAPKLIYDASNLETGVPSRFSGSGTDFFTISSLQPE DIATYYCQQYGNLPFTFGGGTKVEIKRAAALDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR

[0341] Anti-BCMA CAR DNA ATGGCACTCCCCGTAACTGCTCTGCTGCTGCCGTTGGCATTGCTCCTGCACGCCGCACGCCCGCAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTAGCTATGGCATGCACTGGGTCCGCCAGGCTCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCGTATGATGGAAGTAATAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCT

[0342] Anti-BCMA CAR AA MALPVTALLLPLALLLHAARPQVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYVKGPLQEPPYDYGMDVWGQGTTVTVSSGSTSGSGKPGSGEGSTKGEIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYSASTRATGIPARFSGSGSGTEFTLTISSLQ SEDFAVYCQQHHVWPLTFGGGTKVEIKRAAALDNEKSNGTIIHVKGKHLCPSPLFGPSPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR

[0343] C185 E7 11-19 / HLA-A*02:01 specificTCR DNA(SEQ ID NO:158) atgggcaccagactgttcttttatgtggccctgtgtctgctgtggaccggccacatggatgccggaattacacagagccccagacacaaagtgaccgagacaggcacccctgtgacactg

[0344] C185 E7 11-19 / HLA - A*02:01 specific TCR AA (SEQ ID NO: 159) MGTRLFFYVALCLLWTGHMDAGITQSPRHKVTETGTPVTLRCHQTENHRYMYWYRQDPGHGLRLIHYSYGVKDTDKGEVS DGYSVSRSKTEDFLLTLESATSSQTSVYFCAISGYKNTEAFFGQGTRLTVVEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRGRAKRSGSGATNFSLLKQAGDVEENPGPMKSLRVLLVILWLQLSWVWSQGQQVMQIPQYQHVQEGEDFTTYCNSSTTLSNIQWYKQRPGGHPVFLIQLVKSGEVKKQKRLTFQFGEAKKNSSLHITATQTTDVGTYFCAGREGGSEKLVFGKGTKLTVNPDIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS

[0345] E7 11-19 atggaaacaacagactggccttcggcaagggcaaccaggtggtggtcatccccgacattcagaaccccgatccagccgtgtaccagctgagagacagcaagagcagcgacaagagcgtgtgtctgttcaccgacttcgactcccagaccaatgtgtcccagagcaaggactccgacgtgtacatcaccgataagaccgtgctggacatgcggagcatggacttcaagagcaatagcgccgtggcttggagcaacaagagcgactttgcctgcgccaacgccttcaacaacagcatcatccccgaggacacattcttcccaagtcctgagagcagctgcgacgtgaagctggtggaaaagagcttcgagacagacaccaacctgaacttccagaacctgagcgtgatcggcttcagaatcctgctgctgaaggtggccggcttcaacctgctgatgactctgagactgtggtccagctgaattcggatccaagcttaggcctgctcgctttcttgctgtcccatttctattaaaggttcctttgttc

[0346] E7 11-19 / HLA-A*02:01 specific TCR AA (SEQ ID NO: 161) MGPGLLCWALLCLLGAGLVDAGVTQSPTHLIKTRGQQVTLRCSPKSGHDTVSWYQQALGQGPQFIFQYYEEEERQRGNFPDRFSGHQFPNYSSELNVNALLLGDSALYLCASSLGWRGGRYNEQFFGPGTRLTVLEDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRGRAKRSGSGATNFSLLKQAGDVEENPGPMKSLRVLLVILWLQLSWVWSQGQNIDQPTEMTATEGAIVQINCTYQTSGFNGLFWYQQHAGEAPTFLSYNVLDGLEEKGRFSSFLSRSKGYSYLLLKELQMKDSASYLCASVDGNNRLAFGKGNQVVVIPDIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS

[0347] Axi Cel (KTE-C19) leader (CSF2RA) DNA (SEQ ID NO: 162) Atgcttctcctggtgacaagccttctgctctgtgagttaccacacccagcattcctcctgatccca

[0348] Axi Cel (KTE-C19) leader (CSF2RA) AA (SEQ ID NO: 163) MLLLVTSLLLCELPHPAFLLIP

[0349] Axi Cel (KTE-C19) scFv heavy chain DNA (SEQ ID NO: 164) gacatccagatgacacagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggcaagtcaggacattagtaaatatttaaattggtatcagcagaaaccagatggaactgttaaactcctgatctaccatacatcaagattacactcaggagtcccatcaaggttcagtggcagtgggtctggaacagattattctctcaccattagcaacctggagcaagaagatattgccacttacttttgccaacagggtaatacgc ttccgtacacgttcggaggggggactaagttggaaataaca

[0350] Axi Cel (KTE-C19) scFv heavy chain AA (SEQ ID NO: 165) DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT

[0351] Axi Cel (KTE-C19) linker (Whitlow) DNA (SEQ ID NO: 166) ggctccacctctggatccggcaagcccggatctggcgagggatccaccaagggc

[0352] Axi Cel (KTE-C19) linker (Whitlow) AA (SEQ ID NO: 167) GSTSGSGKPGSGEGSTKG

[0353] Axi Cel (KTE-C19) scFv light chain DNA (SEQ ID NO: 168) gaggtgaaactgcaggagtcaggacctggcctggtggcgccctcacagagcctgtccgtcacatgcactgtctcaggggtctcattacccgactatggtgtaagctggattcgccagcctccacgaaagggtctggagtggctgggagtaatatggggtagtgaaaccacatactataat tcagctctcaaatccagactgaccatcatcaaggacaactccaagagccaagttttcttaaaaatgaacagtctgcaaactgatgacacagccatttactactgtgccaaacattattactacggtggtagctatgctatggactactggggtcaaggaacctcagtcaccgtctcctca

[0354] Axi Cel(KTE-C19) scFv Light Chain AA (Sequence ID 169) EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS

[0355] Axi Cel (KTE-C19) Mini Spacer DNA (SEQ ID NO: 170) gcggccgca

[0356] Axi Cel (KTE-C19) Mini Spacer AA (Sequence ID 171) AAA

[0357] Axi Cel(KTE-C19) CD28 Spacer (Extracellular / TM region of CD28) DNA (SEQ ID NO: 172) attgaagttatgtatcctcctccttacctagacaatgagaagagcaatggaaccattatccatgtgaaagggaaacacctttgtccaagtcccctattt cccggaccttctaagcccttttgggtgctggtggtggttgggggagtcctggcttgctatagcttgctagtaacagtggcctttattattttctgggtg

[0358] Axi Cel(KTE-C19)CD28 Spacer (Extracellular / TM region of CD28) AA (SEQ ID NO: 173) IEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPF WVLVVVGGVLACYSLLVTVAFIIFWV

[0359] Axi Cel(KTE-C19)CD28 co-stimulation (intracellular region of CD28) DNA (SEQ ID NO: 174) aggagtaagaggagcaggctcctgcacagtgactacatgaacatgactccccgccgccccggggcccacccgcaagcattaccagccctatgccccaccacgcgacttcgcagcctatcgctcc

[0360] Axi Cel(KTE-C19)CD28 co-stimulation (intracellular region of CD28) AA (SEQ ID NO: 175) RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS

[0361] Axi Cel(KTE-C19) CD3 Zeta DNA (SEQ ID NO: 176) agagtgaagttcagcaggagcgcagacgcccccgcgtaccagcagggccagaaccagctctataacgagctcaatctaggacgaagagaggagtacgatgttttggacaagagacgtggccgggaccctgagatggggggaaagccgagaaggaagaaccctcaggaaggcctgtacaatgaactgcagaaagataagatggcggaggcctacagtgagattgggatgaaaggcgagcgccggaggggcaaggggcacgatggcctttaccagggtctcagtacagccaccaaggacacctacgacgcccttcacatgcaggccctgccccctcgc

[0362] Axi Cel (KTE-C19) CD3 zeta AA (SEQ ID NO: 177) RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR

[0363] Axi Cel (KTE-C19) VL CDR1 (Chothia) (SEQ ID NO: 178) RASQDISKYLN

[0364] Axi Cel (KTE-C19) VL CDR2 (Chothia) (SEQ ID NO: 179) HTSRLHS

[0365] Axi Cel (KTE-C19) VL CDR3 (Chothia) (SEQ ID NO: 180) QQGNTLPYT <(

[0366] Axi Cel (KTE-C19) VH CDR1 (Chothia) (SEQ ID NO: 181) GVSLPDY

[0367] Axi Cel(KTE-C19) VH CDR2(Chothia)(Sequence ID 182) WGSET

[0368] Axi Cel(KTE-C19) VH CDR3(Chothia)(Sequence ID 183) HYYYGG Go AMDY

[0369] Axi Cel(KTE-C19) VL CDR1(Kabat)(Sequence ID 184) RASQDISKYLN

[0370] Axi Cel(KTE-C19) VL CDR2(Kabat)(Sequence ID 185) HTSRLHS

[0371] Axi Cel(KTE-C19) VL CDR3(Kabat)(Sequence ID 186) QQGNTLPYT

[0372] Axi Cel(KTE-C19) VH CDR1(Kabat)(Sequence ID 187) DYGVS

[0373] Axi Cel(KTE-C19) VH CDR2(Kabat)(Sequence ID 188) VIWGSETTYYNSALKS

[0374] Axi Cel(KTE-C19) VH CDR3(Kabat)(Sequence ID 189) HYYYGG Go AMDY

[0375] Humanized anti-CD19 CAR CD8 leader DNA (SEQ ID NO: 190) ATGGCACTCCCCGTAACTGCTCTGCTGCTGCCGTTGGCATTGCTCCTGCACGCCGCACGCCCG

[0376] Humanized anti-CD19 CAR CD8 leader AA (SEQ ID NO: 191) MALPVTALLLPLALLLHAARP

[0377] Humanized anti-CD19 CAR scFv heavy chain DNA (SEQ ID NO: 192) GATATTCAAATGACCCAGTCCCCGTCCTCCCTGAGTGCCTCCGTCGGTGACCGTGTTACGATTACCTGCCGTGCGAGCCAAGACATCTCTAAATACCTGAACTGGTATCAGCAAAAACCGGATCAGGCACCGAAACTGCTGATCAAACATACCTCACGTC TGCACTCGGGTGTGCCGAGCCGCTTTAGTGGTTCCGGCTCAGGTACCGATTACACCCTGACGATCAGCTCTCTGCAGCCGGAAGACTTTGCCACGTATTACTGCCAGCAAGGTAATACCCTGCCGTATACGTTCGGCCAAGGTACCAAACTGGAAATCAAA

[0378] Humanized anti-CD19 CAR scFv heavy chain AA (SEQ ID NO: 193) DIQMTQSPSSLSASVGDRVTITCRASQDISKYLNWYQQKPDQAPKLLIKHTSRLHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPYTFGQGTKLEIK

[0379] Humanized anti-CD19 CAR linker (Whitlow) DNA (SEQ ID NO: 194) GGCTCGACGAGCGGCTCTGGTAAACCGGGCTCTGGTGAAGGCAGTACCAAAGGT

[0380] Humanized anti-CD19 CAR linker (Whitlow) AA (SEQ ID NO: 195) GSTSGSGKPGSGEGSTKG

[0381] Humanized anti-CD19 CAR scFv light chain DNA (SEQ ID NO: 196) GAAGTGCAGCTGGTTGAAAGCGGTGGTGGTCTGGTTCAACCGGGTCGTTCCCTGCGTCTGTCATGTACGGCGAGTGGTGTCTCCCTGCCGGACTATGGCGTGTCCTGGATTCGTCAGCCGCCGGTAAAGGCCTGGAATGGATTGGTGTCATCTGGGGCAGTGAAACCACGTATTACAAC TCGGCCCTGAAAAGCCGTTTCACCATCTCTCGCGATAACAGTAAAAATACGCTGTACCTGCAGATGAATAGCCTGCGCGCGGAAGACACCGCCGTTTACTACTGCGCAAAACATTACTACTACGGTGGCAGCTATGCTATGGATTACTGGGGTCAAGGCACGCTGGTCACCGTTTCGTCA

[0382] Humanized anti-CD19 CAR scFv light chain AA (SEQ ID NO: 197) EVQLVESGGGLVQPGRSLRLSCTASGVSLPDYGVSWIRQPPGKGLEWIGVIWGSETTYYNSALKSRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKHYYYGGSYAMDYWGQGTLVTVSS

[0383] Humanized anti-CD19 CAR mini-spacer DNA (SEQ ID NO: 198) GCCGCTGCC

[0384] Humanized anti-CD19 CAR mini spacer AA (SEQ ID NO: 199) AAA

[0385] Humanized anti-CD19 CAR CD28T spacer (extracellular / TM region of CD28) DNA (SEQ ID NO: 200) CTTGATAATGAAAAGTCAAACGGAACAATCATTCACGTGAAGGGCAAGCACCTCTGTCCGTCACCCTTGTTCCCTGGTCCATCCAAGCCATTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTGCTCGTCACCGTGGCTTTTATAATCTTCTGGGTT

[0386] Humanized anti-CD19 CAR CD28T spacer (extracellular / TM region of CD28) AA (SEQ ID NO: 201) LDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWV

[0387] Humanized anti-CD19 CAR CD28 co-stimulation (intracellular region of CD28) DNA (SEQ ID NO: 202) AGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGC

[0388] Humanized anti-CD19 CAR co-stimulation (intracellular region of CD28) AA (SEQ ID NO: 203) RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS

[0389] Humanized anti-CD19 CAR CD3 zeta DNA (SEQ ID NO: 204) AGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAG GGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCAcctagg

[0390] Humanized anti-CD19 CAR CD3 zeta AA (SEQ ID NO: 205) RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR

[0391] Humanized anti-CD19 CAR VL, fully human anti-CD19 CDR1(Chothia) (SEQ ID NO: 206) RASQSVSSSYLA

[0392] Humanized anti-CD19 CAR VL, fully human anti-CD19 CDR2 (Chothia) (SEQ ID NO: 207) GASSRAT

[0393] Humanized anti-CD19 CAR VL, fully human anti-CD19 CDR3 (Chothia) (SEQ ID NO: 208) QQYGSSRFT

[0394] Humanized anti-CD19 CAR VH, fully human anti-CD19 CDR1(Chothia) (SEQ ID NO: 209) GGTFSSY

[0395] Humanized anti-CD19 CAR VH, fully human anti-CD19 CDR2 (Chothia) (SEQ ID NO: 210) IPIFGT

[0396] Humanized anti-CD19 CAR VH, fully human anti-CD19 CDR3 (Chothia) (SEQ ID NO: 211) EAVAADWLDP

[0397] Humanized anti-CD19 CAR VL, fully human anti-CD19 CDR1 (Kabat) (SEQ ID NO: 212) RASQSVSSSYLA

[0398] Humanized anti-CD19 CAR VL, fully human anti-CD19 CDR2 (Kabat) (SEQ ID NO: 213) GASSRAT

[0399] Humanized anti-CD19 CAR VL, fully human anti-CD19 CDR3 (Kabat) (SEQ ID NO: 214) QQYGSSRFT

[0400] Humanized anti-CD19 CAR VH, fully human anti-CD19 CDR1 (Kabat) (SEQ ID NO: 215) SYAIS

[0401] Humanized anti-CD19 CAR VH, fully human anti-CD19 CDR2 (Kabat) (SEQ ID NO: 216) GIIPIFGTTNYAQQFQG

[0402] Humanized anti-CD19 CAR VH, fully human anti-CD19 CDR3 (Kabat) (SEQ ID NO: 217) EAVAADWLDP

[0403] Fully human anti-CD19 CAR leader (CD8a) DNA (SEQ ID NO: 218) Atggccctgcctgtgacagctctgctgctgcccctggccctgctgctgcatgccgccagacct

[0404] Fully human anti-CD19 CAR leader (CD8a) AA (Sequence ID 219) MALPVTALLLPLALLLHAARP

[0405] Fully human anti-CD19 CAR scFv light chain DNA (SEQ ID NO: 220) gagatcgtgctgacccagtctcccggcaccctgtctctcagcccaggagagagagccaccctgagctgcagagccagccagagcgtgtccagcagctacctggcctggtatcagcagaagcccggacaggcccccagactgctgatctacggcgccagctct agagccaccggcatccccgacagattcagcggcagcggcagtggcaccgacttcaccctgaccatcagcagactggaacccgaggacttcgccgtgtactactgccagcagtacggcagcagccggttcaccttcggccctggcaccaaggtggacatcaag

[0406] Fully human anti-CD19 CAR scFv light chain AA (SEQ ID NO: 221) EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSRFTFGPGTKVDIK

[0407] Fully human anti-CD19 CAR linker (Whitlow) DNA (SEQ ID NO: 222) ggcagcacctccggcagcggcaagcctggctctggcgagggctctaccaagggc

[0408] Fully human anti-CD19 CAR linker (Whitlow) AA (SEQ ID NO: 223) GSTGSGKPGSGEGSTKG

[0409] Fully human anti-CD19 CAR scFv heavy-chain DNA (SEQ ID NO: 224) caggtgcagctggtgcagtctggcgccgaagtgaagaaacccggctctagcgtgaaggtgtcctgcaaggacagcggcggcaccttcagcagctacgccatcagctgggtgcgccaggccccaggacaggggctggaatggatgggcggcatcatccccatcttcggcaccaccaact acgcccagcagttccagggcagagtgaccatcaccgccgacgagagcaccagcaccgcctacatggaactgagcagcctgcggagcgaggacacagccgtgtattactgtgcccgcgaggccgtggccgccgactggctggatccttggggacagggcaccctggtgacagtgtccagc

[0410] Fully human anti-CD19 CAR scFv heavy chain AA (SEQ ID NO: 225) QVQLVQSGAEVKKPGSSVKVSCKDSGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTTNYAQQFQGRVTITADESTSTAYMELSSLRSEDTAVYYCAREAVAADWLDPWGQGTLVTVSS

[0411] Fully human anti-CD19 CAR CD8a spacer and TM region DNA (SEQ ID NO: 226) ttcgtgcccgtgttcctgcccgccaagcctaccaccacccctgcccctagacctcccaccccagccccaacaatcgccagccagcctctgtccctgcggcccgaagcctgtagacctgctgccg gcggagccgtgcacaccagaggcctggacttcgcctgcgatatctacatctgggcccctctggccggcacctgtggcgtgctgctgctgagcctggtgatcaccctgtactgcaaccaccggaac

[0412] Fully human anti-CD19 CAR CD8a spacer and TM region AA (SEQ ID NO: 227) FVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRN

[0413] Fully human anti-CD19 CAR CD28 costim. DNA (SEQ ID NO: 228)agaagcaagcggagccggctgctgcacagcgactacatgaacatgaccccaagacggcctggccccacccggaagcactaccagccttacgccctcccagagacttcgccgcctaccggtcc

[0414] Fully human anti-CD19 CAR CD28 costim. AA (Sequence ID 229) RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS

[0415] Fully human anti-CD19 CAR CD3 zeta DNA (SEQ ID NO: 230) agagtgaagttcagcagatccgccgacgcccctgcctaccagcagggacagaaccagctgtacaacgagctgaacctgggcagacgggaagagtacgacgtgctggacaagcggagaggccgggaccccgagatgggcggaaagcccagacggaagaacc cccaggaaggcctgtataacgaactgcagaaagacaagatggccgaggcctacagcgagatcggcatgaagggcgagcggaggcgcggcaagggccacgatggcctgtaccagggcctgagcaccgccaccaaggacacctacgacgccctgcacatgca ggccctgccccccaga

[0416] Fully human anti-CD19 CAR CD3 zeta AA (SEQ ID NO: 231) RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR

[0417] MAGE A3 / A6 TCR construct - variable alpha chain - AV38 - 2 DNA (SEQ ID NO: 232) atggcatgccctggcttcctgtgggcacttgtgatctccacctgtcttgaatttagcatggctcagacagtcactcagtctcaaccagagatgtctgtgcaggaggcagagaccgtgaccctgagctgcacatatgacaccagtgagagtgattattatttattctggtacaagcagcctcccagcaggcagatgattctcgttattcgccaagaagcttataagcaacagaatgcaacagagaatcgtttctctgtgaacttccagaaagcagccaaatccttcagtctcaagatctcagactcacagctgggggatgccgcgatgtatttctgtgct

[0418] MAGE A3 / A6 TCR construct - variable alpha chain - AV38 - 2 AA (SEQ ID NO: 233) M A C P G F L W A L V I S T C L E F S M A Q T V T Q S Q P E M S V Q E A E T V T L S C T Y D T S E S D Y Y L F W Y K Q P P S R Q M I L V I R Q E A Y K Q Q N A T E N R F S V N F Q K A A K S F S L K I S D S Q L G D A A M Y F C A L R S S G T Y K Y I F G T G T R L K V L A N

[0419] MAGE A3 / A6 TCR construct - TRAJ40 DNA (SEQ ID NO: 234) ctccggagctcaggaacctacaaatacatctttggaacaggcaccaggctgaaggttttagcaaat

[0420] MAGE A3 / A6 TCR construct - murine constant α - murine CA DNA (SEQ ID NO: 235) atccagaaccctgaacctgctgtgtaccagttaaaagatcctcggtctcaggacagcaccctctgcctgttcaccgactttgactcccaaatcaatgtgccgaaaaccatggaatctggaacgttcatcactgacaaaactgtgctggacatgaaagctatggattccaagagcaatggggccattgcctggagcaaccagacaagcttcacctgccaagatatcttcaaagagaccaacgccacctaccccagttcagacgttccctgtgatgccacgttgactgagaaaagctttgaaacagatatgaacctaaactttcaaaacctgtcagttatgggactccgaatcctcctgctgaaagtagccggatttaacctgctcatgacgctgaggctgtggtccagt

[0421] MAGE A3 / A6 TCR construct - murine constant α - murine CA AA (SEQ ID NO: 236) I Q N P E P A V Y Q L K D P R S Q D S T L C L F T D F D S Q I N V P K T M E S G T F I T D K T V L D M K A M D S K S N G A I A W S N Q T S F T C Q D I F K E T N A T Y P S S D V P C D A T L T E K S F E T D M N L N F Q N L S V M G L R I L L L K V A G F N L L M T L R L W S S

[0422] MAGE A3 / A6 TCR construct-furin-SG SG-P2A DNA (SEQ ID NO: 237) cgggccaagcggtccggatccggagccaccaacttcagcctgctgaagcaggccggcgacgtggaggagaaccccggcccc

[0423] MAGE A3 / A6 TCR Construct-Furin-SG SG-P2A AA (Sequence ID 238) RAKRSGSGATNFSLLKQAGDV EENPGP

[0424] MAGE A3 / A6 TCR construct-variable β-chain-BV7-2 DNA (SEQ ID NO: 239) atgggcaccaggctcctcttctgggtggccttctgtctcctgggggcagatcacacaggagctggagtctcccagtcccccagtaacaaggtcacagagaagggaaaggatgtagagctcaggtgtgatccaatttcaggtcatactgccctttactggtaccgacagag cctggggcagggcctggagttttaatttacttccaaggcaacagtgcaccagacaaatcagggctgcccagtgatcgcttctctgcagagaggactgggggatccgtctccactctgacgatccagcgcacacagcaggaggactcggccgtgtatctctgtgccagca

[0425] MAGE A3 / A6 TCR construct-variable β-chain-BV7-2 AA (SEQ ID NO: 240) MGTRLLFWVAFCLLGADHTGA GVSQSPSNKVTEKGKDVELRC DPISGHTALYWYRQSLGQGLE FLIYFQGNSAPDKSGLPSDRF SAERTGGSVSTLTIQRTQQED SAVYLCA SI RTGPFFS GNTIYFGEGSWLTVVE

[0426] MAGE A3 / A6 TCR fragment-TRBJ1-3 DNA(protein complex 241) . tccggacaggggccttttttctctggaacaccatatattttggagaggggagttggctcactgttgtag

[0427] MAGE A3 / A6 TCR β-proliferase β-protein CB1 DNA(free residue 242) . gacctgagaaacgtgaccccacccaaggtctccttgtttgagccatcaaaagcagagattgcaaacaaacaaaaggctac cctcgtgtgcttggccaggggcttcttccctgaccacgtggagctgagctggtgggtgaatggcaaggaggtccacagtggggtcagcacggaccctcaggcctacaaggagagcaattatagctactgcctgagcagccgcctgagggtctctgctaccttctggcacaatcctcgaaaccacttccgctgccaagtgcagttccatgggctttcagaggaggacaagtggccagagggctcacccaaacctgtcacacagaacatcagtgcagaggcctggggccgagcagactgtggaatcacttcagcatcctatcatcagggggttctgtctgcaaccatcctctatgagatcctactggggaaggccaccctatatgctgtgctggtcagtggcctggtgctgatggctatggtcaaaagaaagaactcatga

[0428] MAGE A3 / A6 TCR construct - murine constant β - murine CB1 AA (SEQ ID NO: 243) D L R N V T P P K V S L F E P S K A E I A N K Q K A T L V C L A R G F F P D H V E L S W W V N G K E V H S G V S T D P Q A Y K E S N Y S Y C L S S R L R V S A T F W H N P R N H F R C Q V Q F H G L S E E D K W P E G S P K P V T Q N I S A E A W G R A D C G I T S A S Y H Q G V L S A T I L Y E I L L G K A T L Y A V L V S G L V L M A M V K R K N S Stop

[0429] MAGE A3 / A6 TCR Va CDR1 (SEQ ID NO: 244) TYDTSESDYYLF

[0430] MAGE A3 / A6 TCR Va CDR2 (Sequence ID 245) QEAYKQQ

[0431] MAGE A3 / A6 TCR Va CDR3 (Sequence ID 246) ALRSSGTYKYI

[0432] MAGE A3 / A6 TCR Vb CDR1 (Sequence ID 247) DPISGHTALY

[0433] MAGE A3 / A6 TCR Vb CDR2 (Sequence ID 248) FQGNSAPDKSG

[0434] MAGE A3 / A6 TCR Vb CDR3 (Sequence ID 249) ASIRTGPFFSGNTIY

[0435] Anti-CLL-1 CAR CD8 Leader DNA (SEQ ID NO: 250) ATGGCACTCCCCGTAACTGCTCTGCTGCTGCCGTTGGCATTGCTCCTGCACGCCGCACGCCCG

[0436] Anti-CLL-1 CAR CD8 Leader AA (SEQ ID NO: 251) MALPVTALLLPLALLLHAARP

[0437] Anti-CLL-1 CAR scFv heavy-chain DNA (SEQ ID NO: 252) CAGGTCCAACTGCAAGAAAGCGGACCCGGACTGGTGAAGCCTTCTGAGACACTTAGTCTGACGTGCACGGTCAGTGGCGGCTCCATCTCCTCCTATTATTGGTCATGGATACGACAACCCCCAGGTAAGGGCCTGGAATGGATTGGCTATATCTACTATTCAGGAAGCACGAACTACAATCCC AGCCTGAAGTCCCGAGTGACAATTTCAGTAGATACCAGTAAAAACCAGTTCAGTCTTAAACTGTCAAGCGTGACAGCTGCCGACACCGCTGTGTATTACTGCGTCTCACTGGTGTATTGTGGAGGGGATTGTTATAGCGGGTTCGATTATTGGGGACAGGGAACCCTGGTGACTGTATCTTCC

[0438] Anti-CLL-1 CAR scFv heavy chain AA (SEQ ID NO: 253) QVQLQESGPGLVKPSETLSLTCTVSGGSISSYYWSWIRQPPGKGLEWIGYIYYSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCVSLVYCGGDCYSGFDYWGQGTLVTVSS

[0439] Anti-CLL-1 CAR G4S linker DNA (SEQ ID NO: 254) GGCGGCGGCGGCTCAGGGGGTGGCGGTAGTGGCGGTGGGGGTTCC

[0440] Anti-CLL-1 CAR G4S Linker AA (Sequence ID 255) GGGGSGGGGSGGGGS

[0441] Anti-CLL-1 CAR scFv light chain DNA (SEQ ID NO: 256) GATATTCAACTGACACAATCCCCCAGCTCACTCAGCGCCAGCGTGGGGACAGGGTTAGCTTTACCTGTCAAGCCTCTCAGGATATAAATAACTTTCTGAACTGGTATCAACAGAAGCCTGGGAAGGCGCCCAAACTCCTGATCTATGATGCGTCCAACCTG GAAACTGGCGTGCCTTCACGCTTTAGCGGCTCTGGCAGTGGTACAGACTTCACTTTTACCATCTCTTCACTTCAGCCGGAGGACATCGCCACATATTACTGTCAACAGTACGGAAACTTGCCCTTTACTTTTGGAGGCGGCACCAAAGTTGAAATCAAAAGG

[0442] Anti-CLL-1 CAR scFv light chain AA (SEQ ID NO: 257) DIQLTQSPSSLSASVGDRVSFTCQASQDINNFLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYGNLPFTFGGGTKVEIKR

[0443] Anti-CLL-1 CAR mini spacer DNA (SEQ ID NO: 258) GCCGCTGCC

[0444] Anti-CLL-1 CAR Mini Spacer AA (Sequence ID 259) AAA

[0445] Anti-CLL-1 CAR CD28T (extracellular / TM region of CD28) DNA (SEQ ID NO: 260) CTGGATAACGAAAAGAGCAATGGGACTATAATACATGTTAAAGGAAAACACCTGTGTCCATCTCCCCTGTTCCCTGGACC GTCAAAGCCATTTTGGGTGCTCGTGGTTGTCGGTGGCGTTCTCGCCTGTTATAGCTTGCTGGTGACAGTAGCCTTCATTATCTTTTGGGTG

[0446] Anti-CLL-1 CAR CD28T (extracellular / TM region of CD28) AA (SEQ ID NO: 261) LDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWV

[0447] Anti-CLL-1 CAR CD28 (intracellular co-stimulatory region of CD28) DNA (SEQ ID NO: 262) AGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGC

[0448] Anti-CLL-1 CAR CD28 (intracellular co-stimulatory region of CD28) AA (SEQ ID NO: 263) RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS

[0449] Anti-CLL-1 CAR CD3 zeta DNA (SEQ ID NO: 264) CGAGTGAAATTTTCTAGATCAGCTGATGCTCCCGCCTATCAGCAGGGACAGAATCAACTTTACAATGAGCTGAACCTGGGTCGCAGAGAAGAGTACGACGTTTTGGACAAACGCCGGGGCCGAGATCCTGAGATGGGGGGAAGCCGAGAAGGAAGAATCCTCAAGAA GGCCTGTACAACGAGCTTCAAAAAGACAAAATGGCTGAGGCGTACTCTGAGATCGGCATGAAGGGCGAGCGGAGACGAGGCAAGGGTCACGATGGCTTGTATCAGGGCCTGAGTACAGCCACAAAGGACACCTATGACGCCCTCCACATGCAGGCACTGCCCCCACGC

[0450] Anti-CLL-1 CAR CD3 Zeta AA (SEQ ID NO: 265) RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR

[0451] CLL scFv(24C1) VL CDR1(Sequence ID 266) QASQDINNFLN

[0452] CLL scFv(24C1) VL CDR2(Sequence ID 267) DASNLET

[0453] CLL scFv(24C1) VL CDR3(SEQ ID NO: 268) QQYGNLPFT

[0454] CLL scFv(24C1) VH CDR1(SEQ ID NO: 269) GGSISSY

[0455] CLL scFv(24C1) VH CDR2(Sequence ID 270) YYSGS

[0456] CLL scFv(24C1) VH CDR3(SEQ ID NO: 271) LVYCGGDCYSGFDY

[0457] Anti-BCMA CAR leader (CD8a) DNA (SEQ ID NO: 272) ATGGCACTCCCCGTAACTGCTCTGCTGCTGCCGTTGGCATTGCTCCTGCACGCCGCACGCCCG

[0458] Anti-BCMA CAR leader (CD8a) AA (Sequence ID 273) MALPVTALLLPLALLLHAARP

[0459] Anti-BCMA CAR scFv heavy-chain DNA (SEQ ID NO: 274) CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTAGCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCGTATGATGGAAGTAATAAATACTATGCAG ACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCGGTGTACTACTGCGTCAAGGGGCCGTTGCAGGAGCCGCCATACGATTATGGAATGGACGTATGGGGCCAGGGAACAACTGTCACCGTCTCCTCA

[0460] Anti-BCMA CAR scFv heavy chain AA (SEQ ID NO: 275) QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVKGPLQEPPYDYGMDVWGQGTTVTVSS

[0461] Anti-BCMA CAR linker (Whitlow) DNA (SEQ ID NO: 276) GGGTCTACATCCGGCTCCGGGAAGCCCGGAAGTGGCGAAGGTAGTACAAAGGGG

[0462] Anti-BCMA CAR Linker (Whitlow) AA (SEQ ID NO: 277) GSTSGSGKPGSGEGSTKG

[0463] Anti-BCMA CAR scFv light chain DNA (SEQ ID NO: 278) GAAATAGTGATGACGCAGTCTCCAGCCACCCTGTCTGTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGCAGCAACTTAGCCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATAGCGCATCCACCAGG GCCACTGGTATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGAGTTCACTCTCACCATCAGCAGCCTGCAGTCTGAAGATTTTGCAGTTTATTACTGTCAGCAGCACCACGTCTGGCCTCTCACTTTTGGCGGAGGGACCAAGGTTGAGATCAAACGG

[0464] Anti-BCMA CAR scFv light chain AA (Sequence ID 279) EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKP GQAPRLLIYSASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQHHVWPLTFGGGTKVEIKR

[0465] Anti-BCMA CAR Mini Spacer DNA (SEQ ID NO: 280) GCCGCTGCC

[0466] Anti-BCMA CAR Mini Spacer AA (Sequence ID 281) AAA

[0467] Anti-BCMA CAR CD28T spacer DNA (SEQ ID NO: 282) CTTGATAATGAAAAGTCAAACGGAACAATCATTCACGTGAAGGGCAAGCACCTCTGTCCGTCACCCTTGTTCCCTGGTCCATCCAAGCCATTCTGGGTGTTGGTCGTAGTGGGTGGAGTCCTCGCTTGTTACTCTCTGCTCGTCACCGTGGCTTTTATAATCTTCTGGGTT

[0468] Anti-BCMA CAR CD28T Spacer AA (Sequence No. 283) LDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWV

[0469] Anti-BCMA CAR CD28 co-stimulatory region DNA (SEQ ID NO: 284) AGATCCAAAAGAAGCCGCCTGCTCCATAGCGATTACATGAATATGACTCCACGCCGCCCTGGCCCCACAAGGAAACACTACCAGCCTTACGCACCACCTAGAGATTTCGCTGCCTATCGGAGC

[0470] Anti-BCMA CAR CD28 co-stimulatory region AA (SEQ ID NO: 285) RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS

[0471] Anti-BCMA CAR CD3 zeta DNA (SEQ ID NO: 286) AGGGTGAAGTTTTCCAGATCTGCAGATGCACCAGCGTATCAGCAGGGCCAGAACCAACTGTATAACGAGCTCAACCTGGGACGCAGGGAAGAGTATGACGTTTTGGACAAGCGCAGAGGACGGGACCCTGAGATGGGTGGCAAACCAAGACGAAAAAACCCCCAGGAG GGTCTCTATAATGAGCTGCAGAAGGATAAGATGGCTGAAGCCTATTCTGAAATAGGCATGAAAGGAGAGCGGAGAAGGGGAAAAGGGCACGACGGTTTGTACCAGGGACTCAGCACTGCTACGAAGGATACTTATGACGCTCTCCACATGCAAGCCCTGCCACCTAGG

[0472] Anti-BCMA CAR CD3 Zeta AA (SEQ ID NO: 287) RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR

[0473] Anti-BCMA CAR VL CDR1 (SEQ ID NO: 288) RASQSVSSNLA

[0474] Anti-BCMA CAR VL CDR2 (SEQ ID NO: 289) SASTRAT

[0475] Anti-BCMA CAR VL CDR3 (SEQ ID NO: 290) QQHHVWPLTF

[0476] Anti-BCMA CAR VH CDR1 (SEQ ID NO: 291) GFTFSSY

[0477] Anti-BCMA CAR VH CDR2 (SEQ ID NO: 292) VISYDGSNKYYADSVKG

[0478] Anti-BCMA CAR VH CDR3 (SEQ ID NO: 293) VKGPLQEPPYDYGMDV

[0479] C185 E7 11-19 / HLA-A*02:01 specific TCR-TCR β chain variable region DNA (SEQ ID NO: 294) atgggcaccagactgttcttttatgtggccctgtgtctgctgtggaccggccacatggatgccggaattacacagagccccagacacaaagtgaccgagacaggcacccctgtgacactgagatgccaccagaccgagaaccaccgctacatgtactggtacagacaggatccaggccacggcctgagactgatccactacagctacggcgtgaaggacaccgacaagggcgaagtgtctgacggctacagcgtgtccagaagcaagaccgaggatttcctgctgaccctggaaagcgccacaagcagccagaccagcgtgtacttttgtgccatcagcggctacaagaacaccgaggcctttttcggccaaggcaccaggctgacagtggtg

[0480] C185 E7 11-19 / HLA-A*02:01 specific TCR - TCR β-chain variable region AA (SEQ ID NO: 295) MGTRLFFYVALCLLWTGHMDAGITQSPRHKVTETGTPVTLRCHQTENHRYMYWYRQDPGHGLRLIHYSYGVKDTDKGEVSDGYSVSRSKTEDFLLTLESATSSQTSVYFCAISGYKNTEAFFGQGTRLTVV

[0481] C185 E7 11-19 / HLA-A*02:01 specific TCR - TCR β-chain constant region DNA (SEQ ID NO: 296) gaagatctgaagaacgtgttcccacctgaggtggccgtgtttgagccttctgaggccgagatcagccacacacagaaagccacactcgtgtgcctggccaccggcttttatcccgatcacgtggaactgtcttggtgggtcaacggcaaagaggtgcacagcggcgttagcacagaccctcagcctctgaaagagcagcccgctctgaacgacagcagatactgtctgagcagcagactgagagtgtccgccaccttctggcagaaccccagaaaccacttcagatgccaggtgcagttctacggcctgtccgagaatgacgagtggacccaggatagagccaagccagtgacacagattgtgtctgccgaagcctggggcagagccgattgtggctttacaagcgagagctaccagcagggcgtgctgtctgccacaatcctgtatgagatcctgctgggcaaagccactctgtacgctgtgctggtgtctgccctggtgctgatggccatggtcaagagaaaggacagcagaggc

[0482] C185 E7 11-19 / HLA - A*02:01 specific TCR - TCR β - chain constant region AA (SEQ ID NO: 297) EDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG

[0483] C185 E7 11-19 / HLA - A*02:01 specific TCR - P2A peptide (with furin cleavage site and linker) DNA (SEQ ID NO: 298) agagccaagagatctggcagcggcgccacaaactttagcctgctgaaacaggccggcgacgtggaagagaaccctggacct

[0484] C185 E7 11-19 / HLA-A*02:01 Specific TCR-P2A peptide (with furin cleavage site and linker) AA (SEQ ID NO: 299) RAKRSGSGATNFSLLKQAGDVEENPGP

[0485] C185 E7 11-19 / HLA-A*02:01 specific TCR-TCR α chain variable region DNA (SEQ ID NO: 300) atgaagtccctgcgggtgctgctggttattctgtggctgcagctgagctgggtttggagccagggacagcaagtgatgcagatccctcagtaccagcac gtgcaagaaggcgaggacttcaccacctactgcaacagcagcacaaccctgagcaacatccagtggtacaagcagaggcctggcgggcaccctgtgttt ctgatccagctggttaagagcggcgaagtgaagaagcagaagcggctgacctttcagttcggcgaggccaagaagaacagcagcctgcacattaccgcc acacagaccacgacgtgggcacatatttttgcgctggcagagaaggcggcagcgagaagctggtttttggcaagggcaccaaactgaccgtgaatccc

[0486] C185 E7 11-19 / HLA-A*02:01 Specific TCR-TCR α chain variable region AA (SEQ ID NO: 301) MKSLRVLLVILWLQLSWVWSQGQQVMQIPQYQHVQEGEDFTTYCNSSTTLSNIQWYKQRPGGHPVFLIQLVKSGEVKKQKRLTFQFGEAKKNSSLHITATQTTDVGTYFCAGREGGSEKLVFGKGTKLTVNP

[0487] C185 E7 11-19 / HLA-A*02:01 specific TCR-TCR α-chain constant region DNA (SEQ ID NO: 302) gacattcagaaccccgatccagccgtgtaccagctgagagacagcaagagcagcgacaagagcgtgtgtctgttcaccgacttcgactcccagaccaatgtgtcccagagcaaggactccgacgtgtacatcaccgataagaccgtgctggacatgcggagcatggacttcaagagcaatagcgccgtggcttggagcaacaagagcgactttgcctgcgccaacgccttcaacaacagcatcatccccgaggacacattcttcccaagtcctgagagcagctgcgacgtgaagctggtggaaaagagcttcgagacagacaccaacctgaacttccagaacctgagcgtgatcggcttc agaatcctgctgctgaaggtggccggcttcaacctgctgatgactctgagactgtggtccagctgaattcggatccaagcttaggcctgctcgctttcttgctgtcccatttctattaaaggttcctttgttc [[ID=eleven]]

[0488] C185 E7 11-19 / HLA-A*02:01 specific TCR-TCR α-chain constant region AA (SEQ ID NO: 303) DIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS

[0489] C185 E7 11-19 / HLA-A*02:01 specific TCR-α CDR1 (SEQ ID NO: 304) TTLSN

[0490] C185 E7 11-19 / HLA-A*02:01 Specific TCR-α CDR2 (Layout No. 305) LVKSGEV

[0491] C185 E7 11-19 / HLA-A*02:01 Specific TCR-α CDR3 (Layout No. 306) AGREGGSEKLV

[0492] C185 E7 11-19 / HLA-A*02:01 Specific TCR-β CDR1 (Layout No. 307) ENHRY

[0493] C185 E7 11-19 / HLA-A*02:01 Specific TCR-β CDR2 (Layout No. 308) SYGVKD

[0494] C185 E7 11-19 / HLA-A*02:01 Specific TCR-β CDR3 (Layout No. 309) AISGYKNTEAF

[0495] E7 11-19 / HLA-A*02:01 Specific TCR-TCR β-lock can damage domain DNA (allocation number 310) atgggacctggattgctttgttgggccctgctgtgtctgcttggagctggacttgtggatgccggcgtgacacagtctcccacacacctgatcaagaccagaggccagcaagtgaccctgagatgtagccctaagagcggccacgacaccgtgtcttggtatcagcaggctcttggccagggacctcagttcatcttccagtactacgaggaagaggaacggcagcggggcaacttccctgatagattctctggccatcagttccccaactacagcagcgagctgaacgtgaacgctctgctgctgggcgatagcgccctgtatctgtgtgccagttctcttggttggagaggcggcagatacaacgagcagttctttggccctggcaccagactgaccgtgctg

[0496] E7 11-19 / HLA - A*02:01 specific TCR - TCR β - chain variable region AA (SEQ ID NO: 311) MGPGLLCWALLCLLGAGLVDAGVTQSPTHLIKTRGQQVTLRCSPKSGHDTVSWYQQALGQGPQFIFQYYEEEERQRGNFPDRFSGHQFPNYSSELNVNALLLGDSALYLCASSLGWRGGRYNEQFFGPGTRLTVL

[0497] E7 11-19 / HLA - A*02:01 specific TCR - TCR β - chain constant region DNA (SEQ ID NO: 312) gaagatctgaagaacgtgttcccacctgaggtggccgtgtttgagccttctgaggccgagatcagccacacacagaaagccacactcgtgtgcctggccaccggcttttatcccgatcacgtggaactgtcttggtgggtcaacggcaaagaggtgcacagcggcgttagcacagaccctcagcctctgaaagagcagcccgctctgaacgacagcagatactgtctgagcagcagactgagagtgtccgccaccttctggcagaaccccagaaaccacttcagatgccaggtgcagttctacggcctgtccgagaatgacgagtggacccaggatagagccaagccagtgacacagattgtgtctgccgaagcctggggcagagccgattgtggctttacaagcgagagctaccagcagggcgtgctgtctgccacaatcctgtatgagatcctgctgggcaaagccactctgtacgctgtgctggtgtctgccctggtgctgatggccatggtcaagagaaaggacagcagaggc

[0498] E7 11-19 / HLA-A*02:01 specific TCR - TCR β chain constant region AA (SEQ ID NO: 313) EDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVSTDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMVKRKDSRG

[0499] E7 11-19 / HLA-A*02:01 specific TCR - P2A (with furin cleavage site and linker) DNA (SEQ ID NO: 314) agagccaagagatctggcagcggcgccacaaactttagcctgctgaaacaggccggcgacgtggaagagaaccctggacct

[0500] E7 11-19 / HLA - A*02:01 specific TCR - P2A (with furin cleavage site and linker) AA (SEQ ID NO: 315) RAKRSGSGATNFSLLKQAGDVEENPGP

[0501] E7 11-19 / HLA - A*02:01 specific TCR - TCR α - chain variable region DNA (SEQ ID NO: 316) atgaagtccctgcgggtgctgctggttattctgtggctgcagctgagctgggtttggagccagggacagaacatcgaccagcctaccgagatgacagccaccgaaggcgccatcgtgcagatcaattgcacctaccagaccagcggcttcaacggcctgttctggtatcaacagcatgccggcgaggcccctaccttcctgagctataatgtgctggacggcctggaagaaaagggcaga ttcagcagcttcctgtccagaagcaagggctacagctacctgctgctgaaagaactccagatgaaggacagcgcctcctacctgtgtgcctccgtggatggaaacaacagactggccttcggcaagggcaaccaggtggtggtcatcccc

[0502] E7 11-19 / HLA - A*02:01 specific TCR - TCR α - chain variable region AA (SEQ ID NO: 317) MKSLRVLLVILWLQLSWVWSQGQNIDQPTEMTATEGAIVQINCTYQTSGFNGLFWYQQHAGEAPTFLSYNVLDGLEEKGRFSSFLSRSKGYSYLLLKELQMKDSASYLCASVDGNNRLAFGKGNQVVVIP

[0503] E7 11-19 / HLA-A*02:01 specific TCR-TCR α-chain constant region DNA (SEQ ID NO: 318) gacattcagaaccccgatccagccgtgtaccagctgagagacagcaagagcagcgacaagagcgtgtgtctgttcaccgacttcgactcccagaccaatgtgtcccagagcaaggactccgacgtgtacatcaccgataagaccgtgctggacatgcggagcatggacttcaagagcaatagcgccgtggcttggagcaacaagagcgactttgcctgcgccaacgccttcaacaacagcatcatccccgaggacacattcttcccaagtcctgagagcagctgcgacgtgaagctggtggaaaagagcttcgagacagacaccaacctgaacttccagaacctgagcgtgatcggcttcagaatcctgctgctgaaggtggccggcttcaacctgctgatgactctgagactgtggtccagctgaattcggatccaagcttaggcctgctcgctttcttgctgtcccatttctattaaaggttcctttgttc

[0504] E7 11-19 / HLA-A*02:01 specific TCR-TCR α-chain constant region AA (SEQ ID NO: 319) DIQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFRILLLKVAGFNLLMTLRLWSS

[0505] E7 11-19 / HLA-A*02:01 specific TCR-α CDR1 (SEQ ID NO: 320) TSGFNG

[0506] E7 11-19 / HLA-A*02:01 specific TCR-α CDR2 (SEQ ID NO: 321) NVLDGL

[0507] E7 11-19 / HLA-A*02:01 specific TCR-α CDR3 (SEQ ID NO: 322) ASVDGNNRLA

[0508] E7 11-19 / HLA-A*02:01 specific TCR-β CDR1 (SEQ ID NO: 323) SGHDT

[0509] E7 11-19 / HLA-A*02:01 specific TCR-β CDR2 (SEQ ID NO: 324) YYEEEE

[0510] E7 11-19 / HLA-A*02:01 specific TCR-β CDR3 (SEQ ID NO: 325) ASSLGWRGGRYNEQF

Claims

1. A method for implementing patient-specific immunotherapy procedures, The computing device receives cell order requests to produce transfected T cells for patients. The computing device generates a patient-specific identifier associated with the cell order request (the patient-specific identifier includes a patient identification element, a sales order identifier, and a cell order lot number), The computing device initiates a process to produce transfected T cells for injection into the patient's bloodstream. Includes, The process is, A leukocyte apheresis procedure is performed on the blood sample of the aforementioned patient to collect T cells from the sample. Transferring the collected T cells to a container, Labeling the container with the aforementioned patient-specific identifier, Transferring the collected T cells to a manufacturing facility, Using cell modification technology, transfected T cells are produced from the collected T cells. Receiving the transfected T cells from the manufacturing facility, and Injecting the transfected T cells into the patient's bloodstream, Includes, The computing device records a tracking event for each step of the process, and each tracking event includes the patient-specific identifier. A method wherein the tracking event includes the continued management of the patient's T cells during the process.

2. The method according to claim 1, wherein the transfected T cells are produced by transfecting the collected T cells with a polynucleotide encoding a chimeric antigen receptor (CAR), and the CAR comprises an antigen-binding molecule that specifically binds to a target molecule.

3. The method according to claim 2, wherein the antigen-binding molecule is a single-stranded variable fragment.

4. The method according to claim 2, wherein the target molecule is a cancer-related antigen derived from blood.

5. The method according to claim 2, wherein the target molecule is a viral infection-associated antigen.

6. The method according to claim 2, wherein the chimeric antigen receptor (CAR) further comprises at least one costimulatory domain.

7. The method according to claim 2, wherein the chimeric antigen receptor (CAR) further comprises at least one activation domain.

8. The method according to claim 2, wherein the polynucleotide is a component of the vector.

9. The method according to claim 1, wherein the transfected T cells are produced by transfecting the collected T cells with a polynucleotide encoding a T cell receptor (TCR).

10. The method according to claim 9, wherein the TCR binds to a tumor-associated antigen.

11. The method according to claim 9, wherein the TCR binds to a viral infection-associated antigen.

12. The method according to claim 9, wherein the polynucleotide is a component of the vector.

13. Initiating the process of creating transfected T cells is The computing device receives indicia that the transfected T cells were shipped from the first site after they were produced, and The transfected T cells receive indicia that was received at a second site before injection. It further includes, When the computing device receives an Indicia indicating that the transfected T cells have been shipped, it records a tracking event. The method according to claim 1, wherein when the computing device receives an indicia indicating that the transfected T cells have been received at a second site, it records a tracking event.

14. The method according to claim 1, wherein the patient identification element includes a first patient ID associated with the immunotherapy procedure and a second patient ID associated with a facility that performs one or more of the leukocyte apheresis procedure or the transfected T cell infusion.

15. The method according to claim 1, wherein the computing device stores the tracking events in a regular order.

16. A method for tracking cell orders during an immunotherapy procedure: A computing device receives a cell order request to produce transfected T cells for a patient; The computing device generates a patient-specific identifier associated with the cell order request (the patient-specific identifier includes a patient identification element, a sales order identifier, and a cell order lot number); The computing device monitors the process of producing transfected T cells to be injected into the patient's bloodstream. (The process is, To collect T cells from the aforementioned sample, an indicia is received indicating that a leukocyte apheresis procedure has been performed on the patient's blood sample. Receiving an indicia that the collected T cells have been transferred to a container, Receiving information that the container has been labeled with the patient-specific identifier, Receiving an Indicia that the collected T cells have been sent to the manufacturing facility, Using cell modification technology, receive indicia indicating that transfected T cells have been produced from the collected T cells. The transfected T cells receive indicia which they received from the manufacturing facility, and The transfected T cells receive an indication that they have been injected into the patient's bloodstream. (including); The computing device records a tracking event when an Indicia is received (each tracking event includes the patient-specific identifier); and The computing device maintains continuous management of the patient's T cells by saving the tracking events during the process. Methods that include...

17. The method according to claim 16, wherein the transfected T cells are produced by transfecting the collected T cells with a polynucleotide encoding a chimeric antigen receptor (CAR), and the CAR comprises an antigen-binding molecule that specifically binds to a target molecule.

18. The method according to claim 17, wherein the antigen-binding molecule is a single-stranded variable fragment.

19. The method according to claim 17, wherein the target molecule is a cancer-related antigen derived from blood.

20. The method according to claim 17, wherein the target molecule is a viral infection-associated antigen.

21. The method according to claim 17, wherein the chimeric antigen receptor (CAR) further comprises at least one costimulatory domain.

22. The method according to claim 17, wherein the chimeric antigen receptor (CAR) further comprises at least one activation domain.

23. The method according to claim 17, wherein the polynucleotide is a component of the vector.

24. The method according to claim 16, wherein the transfected T cells are produced by transfecting the collected T cells with a polynucleotide encoding a T cell receptor (TCR).

25. The method according to claim 24, wherein the TCR binds to a tumor-associated antigen.

26. The method according to claim 24, wherein the TCR binds to a viral infection-associated antigen.

27. The method according to claim 24, wherein the polynucleotide is a component of the vector.

28. Initiating the process of creating transfected T cells is The computing device receives indicia that the transfected T cells were shipped from the first site after they were produced, and The transfected T cells receive indicia that was received at a second site before injection. It further includes, When the computing device receives an Indicia indicating that the transfected T cells have been shipped, it records a tracking event. The method according to claim 16, wherein when the computing device receives an indicia indicating that the transfected T cells have been received at a second site, it records a tracking event.

29. The method according to claim 16, wherein the patient identification element includes a first patient ID associated with the immunotherapy procedure and a second patient ID associated with a facility that performs one or more of the leukocyte apheresis procedure or the transfected T cell infusion.

30. The method according to claim 16, wherein the computing device stores the tracking events in a regular order.

31. A method for implementing patient-specific immunotherapy procedures: Receiving cell order requests to produce transfected T cells for patients; An event tracking module running on the processor generates a patient-specific identifier associated with the cell order request; To initiate the process of producing transfected T cells to be injected into the bloodstream of the aforementioned patient. (The process is, The procedure involves performing a leukocyte apheresis procedure on the patient's blood sample to collect T cells from the sample, Transferring the collected T cells to a container, Labeling the container with the aforementioned patient-specific identifier, Sending the collected T cells to a manufacturing facility, Using cell modification technology, transfected T cells are produced from the collected T cells, Receiving transfected T cells from the aforementioned manufacturing facility, and Injecting the transfected T cells into the patient's bloodstream. (including); From a first client device located at the site of the leukocyte apheresis procedure, the leukocyte apheresis procedure is confirmed, and a first tracking event including the patient-specific identifier is received by the event tracking module; The event tracking module integrates the first tracking event into a data structure relating to the patient-specific identifier (the data structure is stored in a database, and the integration process records a first timestamp along with the first tracking event); From a second client device located in the manufacturing facility, confirmation is received of the collected T cells at the manufacturing facility, and a second tracking event including a patient-specific identifier is received by the event tracking module; and, The event tracking module integrates the second tracking event into the data structure relating to the patient-specific identifier (the integration step includes recording a second timestamp along with the second tracking event); Methods that include...

32. A method for implementing patient-specific immunotherapy procedures: A tracking module running on the processor receives a cell order request to produce transfected T cells for the patient; The tracking module generates a patient-specific identifier that identifies the patient, and a cell order lot, associated with the cell order request; To generate data records in a database that track the orders for the aforementioned cells (these data records are identified in the database according to the patient-specific identifier); The tracking module receives a first tracking event indicating that the collected T cells are ready for shipment to the manufacturing facility; Updating the data record corresponding to the patient-specific identifier in accordance with the first tracking event; The tracking module, based on the container received at the manufacturing facility, receives a second tracking event indicating that the collected T cells were received at the manufacturing facility; Updating the data record corresponding to the patient-specific identifier in accordance with the second tracking event; The tracking module, based on a manufacturing facility that produced transfected T cells from the collected T cells using cell modification technology, receives a third tracking event indicating that the transfected T cells were produced; Updating the data record corresponding to the patient-specific identifier in accordance with the third tracking event; The tracking module based on transfected T cells received from the manufacturing facility The recipient receives a fourth follow-up event indicating that the transfected T cells have been received; Updating the data record corresponding to the patient-specific identifier in accordance with the fourth tracking event; A tracking module based on transfected T cells injected into the patient's bloodstream receives a fifth tracking event indicating that the transfected T cells were injected into the patient's bloodstream; and Updating the data record corresponding to the patient-specific identifier in accordance with the fifth tracking event; Includes, Each of the first, second, third, fourth, and fifth tracking events includes the patient-specific identifier, timestamp, and event identifier. The data record corresponding to the patient-specific identifier stores the first, second, third, fourth, and fifth tracking events in a regular order as the data record is updated according to each event. method.

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