Molecules and methods for activating CAR-T cells

Description

Molecules and methods for activating CAR-T cells Field of invention

[001] The present invention relates to molecules and methods for activating immune cells expressing an exogenous cell surface receptor comprising an intracellular signaling domain (e.g., chimeric antigen receptors), for the prevention and / or treatment of various conditions, including cancer. Related request

[002] This application claims priority over Australian provisional application AU 2023902090, the full content of which is incorporated herein by reference. Background of the invention

[003] Cancer immunotherapy is a rapidly growing field. The development of T cells expressing chimeric antigen receptors (CARs) has revolutionized adoptive cell therapies.

[004] The potential of this approach has been demonstrated in clinical trials in which CAR-T cells were infused into adult and pediatric patients with B-cell malignancies, neuroblastoma, and sarcoma. To date, more than 500 clinical trials have emerged worldwide, designed to test the efficacy of CAR-T cells targeted for binding to 64 different tumor-associated antigens. Among these, three CD19-specific CAR-T cell products have been approved for the treatment of acute lymphoblastic leukemia (ALL), large B-cell lymphoma, and mantle cell lymphoma. To date, most success with CAR-T cell therapies has been observed in the context of so-called liquid tumors, or where CARs are targeted to CD19, CD22, or the B-cell maturation antigen (BCMA).

[005] Several challenges remain in the clinical application of CAR-T cell therapies. These include difficulties in achieving therapeutic responses. Petition 870250089762, dated 02 / 10 / 2025, pp. 362 / 580 2 / 176 sufficient in the context of solid tumors, which may be due to insufficient activation, expansion, and persistence of CAR-T cells and / or the immunosuppressive tumor microenvironment. Furthermore, the long-term clinical application of CAR-T cell therapies can be negatively impacted by selective pressure from monospecific CAR-T cells, negative antigen escape variants, and antigen depletion. Additionally, low levels of target antigen expression in healthy tissues can result in severe toxicities at the target, outside the tumor. Finally, cytokine release syndrome (CRS) and CAR-T cell-related encephalopathy syndrome (CRES) are frequently observed side effects of CAR-T cell therapy.

[006] Consequently, there is a need for new and improved approaches to CAR therapies, preferably CAR-T.

[007] Reference to any prior art in the descriptive report is not an acknowledgment or suggestion that such prior art is part of the common general knowledge in any jurisdiction or that such prior art could reasonably be expected to be understood, considered relevant and / or combined with other pieces of prior art by a person skilled in the art. Summary of the invention

[008] The present invention is based on the inventors' surprising discovery that molecules having a particular architecture are especially useful for activating immune cells expressing an exogenous cell surface receptor comprising an intracellular signaling domain (e.g., chimeric antigen receptors, including those expressed on T cells). These molecules may be referred to in the present invention as "bridge molecules" (or may also be referred to as adapter molecules). Petition 870250089762, dated 02 / 10 / 2025, pp. 363 / 580 3 / 176

[009] Therefore, in a first aspect, the invention provides a molecule capable of simultaneously binding to a target cell (such as a tumor cell) and to an immune cell expressing an exogenous cell surface receptor comprising an intracellular signaling domain, the molecule comprising: a heterodimer consisting of a first portion comprising a first Fab arm of an antibody and a second portion comprising a second Fab arm of an antibody, wherein the first and second Fab arms each comprise an antigen-binding domain for binding to a cell surface molecule on a target cell, wherein the first Fab arm comprises at least a fraction capable of being bound by the exogenous immune cell receptor, wherein the second Fab arm does not comprise any fractions capable of being bound by the exogenous immune cell receptor, and wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains.

[010] In a further aspect, the invention provides a molecule capable of simultaneously binding to a target cell (such as a tumor cell) and to an immune cell expressing an exogenous cell surface receptor comprising an intracellular signaling domain, the molecule comprising: a heterodimer consisting of a first portion comprising a first Fab arm of an antibody and a second portion comprising a second Fab arm of an antibody, Petition 870250089762, dated 02 / 10 / 2025, pp. 364 / 580 4 / 176 wherein the first and second Fab arms each comprise an antigen-binding domain for binding to a cell surface molecule on a target cell, wherein the first Fab arm comprises at least two fragments capable of being bound by the exogenous immune cell receptor, and wherein the second Fab arm does not comprise any fragments capable of being bound by the exogenous immune cell receptor. Preferably, wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains.

[011] In a third aspect, a two-component therapeutic approach is provided comprising: (a) an immune cell or progenitor thereof expressing an exogenous receptor comprising an intracellular signaling domain; and (b) a bridging molecule comprising: a heterodimer consisting of a first portion comprising a first Fab arm of an antibody and a second portion comprising a second Fab arm of an antibody, wherein the first and second Fab arms each comprise an antigen-binding domain for binding to a cell surface molecule on a target cell, wherein the first Fab arm comprises at least a fraction capable of being bound by the exogenous immune cell receptor, wherein the second Fab arm does not comprise any fractions capable of being bound by the exogenous immune cell receptor, and wherein the first and second portions of the heterodimer Petition 870250089762, dated 02 / 10 / 2025, pp. 365 / 580 5 / 176 comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains.

[012] In a fourth aspect, a composition is provided comprising: (a) an immune cell or progenitor thereof expressing an exogenous receptor comprising an intracellular signaling domain; and (b) a bridging molecule comprising: a heterodimer consisting of a first portion comprising a first Fab arm and a second portion comprising a second Fab arm, wherein the first and second Fab arms comprise an antigen-binding domain for binding to a cell surface molecule on a target cell, wherein the first Fab arm comprises at least a fraction capable of being bound by the exogenous immune cell receptor, wherein the second Fab arm does not comprise any fractions capable of being bound by the exogenous immune cell receptor, and wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains.

[013] The invention also provides a molecule capable of simultaneously binding to a target cell (such as a tumor cell) and to an immune cell expressing an exogenous cell surface receptor comprising an intracellular signaling domain, the molecule comprising: a heterodimer consisting of a first portion comprising Petition 870250089762, dated 02 / 10 / 2025, pp. 366 / 580 6 / 176 a first Fab arm of an antibody and a second portion comprising a second Fab arm of an antibody, wherein the first and second Fab arms each comprise means for binding to a cell surface molecule on a target cell, wherein the first Fab arm comprises at least a fraction capable of being bound by the exogenous immune cell receptor, wherein the second Fab arm does not comprise any fractions capable of being bound by the exogenous immune cell receptor, and wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains.

[014] In a further aspect, the invention provides a molecule capable of simultaneously binding to a target cell (such as a tumor cell) and to an immune cell expressing an exogenous cell surface receptor comprising an intracellular signaling domain, the molecule comprising: A heterodimer consisting of a first portion comprising a first Fab arm of an antibody and a second portion comprising a second Fab arm of an antibody, wherein the first and second Fab arms each comprise means for binding to a cell surface molecule on a target cell, wherein the first Fab arm comprises at least two fragments capable of being bound by the exogenous immune cell receptor, and wherein the second Fab arm does not comprise any fragments capable of being bound by the exogenous immune cell receptor. Preferably, wherein the first and second portions of the heterodimer comprise sequences Petition 870250089762, dated 02 / 10 / 2025, pp. 367 / 580 7 / 176 of amino acids to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains.

[015] Additionally, a two-component therapeutic is provided comprising: (a) an immune cell or progenitor thereof expressing an exogenous receptor comprising an intracellular signaling domain; and (b) a bridging molecule comprising: a heterodimer consisting of a first portion comprising a first Fab arm of an antibody and a second portion comprising a second Fab arm of an antibody, wherein the first and second Fab arms each comprise means for binding to a cell surface molecule on a target cell, wherein the first Fab arm comprises at least a fraction capable of being bound by the exogenous immune cell receptor, wherein the second Fab arm does not comprise any fractions capable of being bound by the exogenous immune cell receptor, and wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains.

[016] In an additional aspect, a composition comprising is provided: (a) an immune cell or progenitor thereof expressing an exogenous receptor comprising an intracellular signaling domain; and (b) a bridging molecule comprising: a heterodimer consisting of a first portion Petition 870250089762, dated 02 / 10 / 2025, pp. 368 / 580 8 / 176 comprising a first Fab arm and a second portion comprising a second Fab arm, wherein the first and second Fab arms comprise means for binding to a cell surface molecule on a target cell, wherein the first Fab arm comprises at least a fraction capable of being bound by the exogenous immune cell receptor, wherein the second Fab arm does not comprise any fractions capable of being bound by the exogenous immune cell receptor, and wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains.

[017] In any embodiment, a means for binding to a cell surface molecule on a target cell comprises or consists of an antigen-binding domain for binding to a cell surface molecule on a target cell.

[018] Additionally, a kit is also provided comprising: (a) an immune cell or progenitor thereof expressing an exogenous receptor comprising an intracellular signaling domain; and (b) a bridging molecule comprising: a heterodimer consisting of a first portion comprising a first Fab arm and a second portion comprising a second Fab arm, wherein the first and second Fab arms comprise an antigen-binding domain for binding to a cell surface molecule on a target cell, wherein the first Fab arm comprises at least a fraction Petition 870250089762, dated 02 / 10 / 2025, pp. 369 / 580 9 / 176 capable of being bound by the exogenous immune cell receptor, wherein the second Fab arm does not comprise any fractions capable of being bound by the exogenous immune cell receptor, wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains.

[019] In another aspect, the present invention provides a nucleic acid comprising a nucleotide sequence encoding a heterodimer (bridge molecule) as described in the present invention. Preferably, the nucleic acid comprises a first nucleotide sequence encoding the first portion of the molecule and a second nucleotide sequence encoding the second portion of the molecule.

[020] In another aspect, the present invention provides a method of treating a disease or disorder in an individual, the method comprising administering to the individual: (a) an immune cell or progenitor thereof expressing an exogenous receptor comprising an intracellular signaling domain; and (b) a bridging molecule comprising: a heterodimer consisting of a first portion comprising a first Fab arm and a second portion comprising a second Fab arm, wherein the first and second Fab arms comprise an antigen-binding domain for binding to a cell surface molecule on a target cell, wherein the first Fab arm comprises at least a portion capable of being bound by the exogenous immune cell receptor, Petition 870250089762, dated 02 / 10 / 2025, pages 370 / 580 10 / 176 wherein the second Fab arm does not comprise any fractions capable of being bound by the exogenous immune cell receptor, and wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains, thereby treating the disease or disorder in the individual.

[021] In another aspect, the present invention provides a method of treating cancer in an individual, the method comprising administering to the individual: (a) an immune cell or progenitor thereof expressing an exogenous receptor comprising an intracellular signaling domain; and (b) a bridging molecule comprising: a heterodimer consisting of a first portion comprising a first Fab arm and a second portion comprising a second Fab arm, wherein the first and second Fab arms comprise an antigen-binding domain for binding to a cell surface molecule on a tumor cell, wherein the first Fab arm comprises at least a fraction capable of being bound by an exogenous immune cell receptor, wherein the second Fab arm does not comprise any fractions capable of being bound by an exogenous immune cell receptor, and wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains, thus Petition 870250089762, dated 02 / 10 / 2025, pp. 371 / 580 11 / 176 treating cancer in the individual.

[022] In another aspect, the present invention provides a method for killing a target cell, the method comprising exposing or bringing the target cell into contact with: (a) an immune cell or progenitor thereof expressing an exogenous receptor comprising an intracellular signaling domain; and (b) a bridging molecule comprising: a heterodimer consisting of a first portion comprising a first Fab arm and a second portion comprising a second Fab arm, wherein the first and second Fab arms comprise an antigen-binding domain for binding to a cell surface molecule on a tumor cell, wherein the first Fab arm comprises at least a fraction capable of being bound by the exogenous immune cell receptor, wherein the second portion of the Fab arm does not comprise any fractions capable of being bound by the exogenous immune cell receptor, and wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains, thereby killing the target cell.

[023] Additionally, the use of a heterodimer as described in the present invention is also provided for in the manufacture of a medicament to treat a disorder, wherein optionally the disorder is cancer or a disease or condition requiring depletion of T cells or B cells (such as an autoimmune disease or condition or a transplant-related condition, such Petition 870250089762, dated 02 / 10 / 2025, pp. 372 / 580 12 / 176 such as graft-versus-host disease).

[024] Preferably, the use comprises administering the drug comprising the heterodimer and administering an immune cell or progenitor thereof expressing an exogenous receptor comprising an intracellular signaling domain, wherein the binding domain is capable of binding to the heterodimer.

[025] In yet another aspect, a pharmaceutical composition is provided comprising a heterodimer as described in the present invention, optionally comprising a pharmaceutically acceptable excipient. The pharmaceutical composition may optionally further comprise an immune cell or progenitor thereof expressing an exogenous receptor comprising an intracellular signaling domain.

[026] In a further aspect, a heterodimer as described in the present invention is provided, optionally in combination with an immune cell or progenitor thereof expressing an exogenous receptor comprising an intracellular signaling domain; or a pharmaceutical composition as described in the present invention, for use in the treatment of a disease or disorder in an individual. Preferably the disease is cancer. Alternatively, the disease or condition may be one that requires depletion of T cells or B cells (such as an autoimmune disease or condition or a transplant-related condition, such as graft-versus-host disease).

[027] Furthermore, the invention provides methods in which the bridging molecules described in the present invention can be administered synchronously to an individual in need of them. This allows for fine-tuning of the therapeutic approach, so that a therapeutic immune cell can be directed to bind to target cells (such as cells Petition 870250089762, dated 02 / 10 / 2025, pp. 373 / 580 13 / 176 cancerous) via different antigens (including cancer-associated antigens), at different times during the course of the patient's therapeutic regimen. In other words, the molecules can be administered sequentially or at the same time (simultaneously) or separately from the administration of therapeutic immune cells.

[028] In any embodiment of the aspects directed to the treatment methods in the present invention, the bridge molecule can be delivered via infusion to the individual or can be expressed by the immune cell expressing the receptor or an immune cell designed for expression of the molecule with or without an inducible expression system. The bridge molecule can be a polypeptide, which is encoded in an inducible or constitutive expression construct contained in the immune cell.

[029] As used in the present invention, the term Fab arm (or Fab region) will be understood as referring to an antigen-binding region of the Fab fragment or region of an antibody. Those skilled in the art will be familiar with the use of this term to describe a region of an antibody comprising an antigen-binding domain and consisting of a constant and a variable domain of each of the heavy and light chains of an antibody.

[030] In preferred embodiments of any aspect of the present invention, the first and second portions comprise an Fc region of an antibody, such as a CH2 region and / or a CH3 region of an antibody, wherein the Fc region is linked to the Fab arms of the molecule. Therefore, in preferred embodiments of any aspect, the molecule is in the form of a heterodimeric Fab-Fc fusion protein (such as a conventional antibody structure) comprising at least one portion capable of being bound by an exogenous immune cell receptor (such as a receptor of Petition 870250089762, dated 02 / 10 / 2025, pp. 374 / 580 14 / 176 chimeric antigen) bound to the first Fab region / arm of the antibody.

[031] It will be appreciated that the first Fab region / arm of the heterodimeric molecule of the invention can be oriented to the left or right side of the molecule, as represented in Figure 2B in the present invention.

[032] A person skilled in the art will be familiar with routine techniques for designing a heterodimeric (asymmetric) molecule as described in the present invention, including the use of recombinant techniques to ensure proper dimerization of non-identical Fab and Fab-Fc fusion proteins (i.e., to force the pairing of a light chain polypeptide with its correct heavy chain counterpart). More specifically, a person skilled in the art will appreciate that the correct heavy chain / light chain assembly to achieve the asymmetric design of the molecules required the following heavy chain / light chain pairing: • A. Heterodimeric heavy chain pairing techniques and • B. Homologous light chain / heavy chain pairing techniques.

[033] Examples of such techniques include the use of so-called CrossMab and DuetMab systems, as described in Mazor et al., (2015) mAbs, 7: 377-389 and Klein et al., (2016) mAbs, 8: 1010-1020, incorporated into the present invention by reference. These references discuss techniques for enabling the correct association of generic light chains using immunoglobulin domain crossing (CrossMAb technology), which can be combined with approaches that enable the correct association of heavy chains, such as knob-into-holes (KIH) technology (Ridgway et al., (1996), Protein Eng, 9: 617-621) or electrostatic targeting. WO 2009 / 080251, WO 2009 / 080252, WO 2009 / 080253, WO 2009 / 080254 and Schaefer, W. et al, PNAS, 108 (2011) 111871191 disclose methods for obtaining bivalent and bispecific IgG antibodies with Petition 870250089762, dated 02 / 10 / 2025, pp. 375 / 580 15 / 176 a domain crossover. WO 2010 / 145792 discloses methods for obtaining tetravalent antigen-binding proteins with a domain crossover. Multispecific antibodies with a VH / VL substitution / exchange at a binding site to prevent light chain mismatch (CrossMab VHA L), which are described in WO 2009 / 080252 (see also Schaefer, W. et al, PNAS, 108 (2011) 11: 187-1191), clearly reduce the production of mismatched variants caused by the mismatch of a light chain against a first antigen with the wrong heavy chain against a second antigen (compared to approaches without such a domain crossover).The methods disclosed in the aforementioned documents for preventing mismatch between light and heavy chains (and thus ensuring that the fraction capable of being bound by the exogenous immune cell receptor is present in only a portion of the heterodimeric molecules of the invention) are incorporated into the present invention by reference.

[034] Similarly, DuetMab technology uses knobsinto-holes (KIH) technology for the heterodimerization of two distinct heavy chains and increases the pairing efficiency of cognate heavy and light chains by replacing the native disulfide bond at one of the CH1-CL interfaces with an engineered disulfide bond.

[035] The person skilled in the art will also be familiar with other technologies that can be used to achieve the same result. Some examples include, but are not limited to: A) for heterodimeric heavy chain technology: KiHS-S: T366S / L368A / Y407V / Y349C KiH mutations + SS interdomain CH3 linkage (Carter P. Bispecific human IgG by design. J Immunol Methods (2001) 248(1-2):7-15); EW-RVTS-S: K360E / K409W / Y349C Q347R / D399V / F405T / S354C EW-RVT mutations + SS interdomain CH3 linkage (Choi HJ, Seok SH, Kim YJ, Seo MD, Kim YS. Fc immunoglobulin crystal structures Petition 870250089762, dated 02 / 10 / 2025, pp. 376 / 580 16 / 176 heterodimers reveal the molecular basis for heterodimer formation. Mol Immunol (2015) 65(2):377-83); B) para pareamento homólogo de cadeia leve / cadeia pesada: Cadeia leve comum (Merchant AM, Zhu Z, Yuan JQ, Goddard A, Adams CW, Presta LG, et al. An efficient route to human bispecific IgG. Nat Biotechnol (1998) 16(7):677-81); tecnologia Ortho-Fab IgG (Lewis SM, Wu X, Pustilnik A, Sereno A, Huang F, Rick HL, et al. Generation of bispecific IgG antibodies by structure-based design of an orthogonal Fab interface. Nat Biotechnol (2014) 32(2):191-8).

[036] It is well within the competence of a person skilled in the art to be able to determine which technology among KIH, DuetMab, or CrossMab or another to employ in order to generate molecules that fall within the scope of the invention. More specifically, a person skilled in the art will recognize that, in order to generate a molecule mono-tagged on the heavy chain of the molecule, a knob-in-hole technology would be sufficient to achieve such a molecule, since the light chains would be identical and no DuetMab or CrossMab design would be necessary. The isolated use of DuetMab or CrossMab could also be used to provide such a molecule (such as XmAb1).

[037] For the generation of mono-tagged antibodies or molecules with two, three, four, five, etc. tags that have at least one light chain tag (N-TERMINAL, between VL and CL, or C-TERMINAL), it is necessary that DuetMab or CrossMab technologies be used with a knob-inhole (KIH) technology. Otherwise, the molecule cannot be intentionally generated and will be subject to a stochastic distribution of expression and protein assembly. The combination of KIH and DuetMab or CrossMab reliably facilitates the generation of molecules that exhibit the desired properties of having the tag only on one side of the molecule.

[038] In any respect, the antigen-binding domains of Petition 870250089762, dated 02 / 10 / 2025, pp. 377 / 580 17 / 176 The first and second Fab arms of the heterodimeric molecule may be identical in amino acid sequence. Alternatively, the antigen-binding domains may be non-identical but bind to the same antigen epitope or may bind to different epitopes of the same antigen. In further embodiments, the antigen-binding domains may be non-identical and may bind to distinct (different) antigens.

[039] As will be understood in the present invention, the first and second Fab arms are preferably for binding to an antigen that is an antigen of a target cell. In other words, the antigen-binding domains of the Fabs are capable of specifically binding to an antigen of a target cell.

[040] In certain modalities, the target cell is a cancerous cell, or a cell capable of presenting a peptide from an infectious agent to an MHC class receptor.

[041] In particularly preferred embodiments, the target cell is a cancer cell, so that the first and second Fab arms therefore comprise antigen-binding domains for binding to antigens on the surface of cancer cells. The antigens referred to are further described in the present invention.

[042] The heterodimeric molecule comprises at least one moiety capable of being bound by an exogenous immune cell receptor (such as a chimeric antigen receptor or universal CAR system receptor). The use of appropriate heavy and / or light chain pairing approaches, such as those discussed above, allows the design of bridging molecules comprising a single moiety (tag) in a heterodimeric molecule. Those skilled in the art will appreciate that, without using this approach, the fabrication of a dimeric molecule would result in the generation of homodimeric species. Petition 870250089762, dated 02 / 10 / 2025, pp. 378 / 580 18 / 176 resulting in molecules comprising an epitope fraction in each Fab arm. Thus, the use of appropriate heavy and / or light chain pairing (such as the use of KIH in combination with Cross-Mab and DuetMab) ensures that only one Fab arm in the dimeric molecule comprises one or more fractions capable of being bound by the exogenous immune cell receptor.

[043] Furthermore, the location of one or more fragments on a Fab arm of the heterodimeric molecule ensures binding to a single CAR molecule on an immune cell only and, in this way, reduces the likelihood of transactivation of different CAR receptors on individual CAR-T cells.

[044] In further embodiments, the heterodimeric molecule comprises 2, 3, or 4 or more fractions capable of being bound by an exogenous immune cell receptor (such as a chimeric antigen receptor or universal CAR system receptor), wherein the fractions are present in one Fab arm but not in the other Fab arm. Preferably, the heterodimeric molecule comprises no more than 5, no more than 3, or no more than 2 fractions capable of being bound by an exogenous immune cell receptor as described in the present invention, wherein the fractions are present in one Fab arm of the heterodimeric molecule but not in the other Fab arm.

[045] In any aspect, the heterodimeric molecule may comprise 3 or more fractions capable of being bound by the exogenous immune cell receptor.

[046] In any aspect, the heterodimeric molecule may comprise 4 or more fractions capable of being bound by the exogenous immune cell receptor.

[047] At least 2 fractions (or 3 or 4 or more fractions) may comprise or consist of identical sequences, or of sequences that are not identical. Petition 870250089762, dated 02 / 10 / 2025, pp. 379 / 580 19 / 176 identical. Non-identical sequences may comprise overlapping identical amino acid sequences. Non-identical sequences may comprise amino acid sequences of a single epitope capable of being bound by an immune cell receptor. Alternatively, non-identical sequences may comprise amino acid sequences of more than one epitope capable of being bound by an immune cell receptor. For example, in either aspect, a bridge molecule may comprise a fraction comprising an amino acid sequence of a dysfunctional P2X7 receptor epitope in the form of the E200 epitope and an additional fraction comprising an amino acid sequence of a dysfunctional P2X7 receptor epitope in the form of the E300 epitope.Alternatively, in any aspect, a bridge molecule may comprise a fraction of dysfunctional P2X7 receptor epitope in the form of the E200 epitope and an additional fraction of dysfunctional P2X7 receptor epitope in the form of the compound epitope. Further, in any aspect, a bridge molecule may comprise a first fraction of dysfunctional P2X7 receptor epitope in the form of the E200 epitope and an additional fraction of dysfunctional P2X7 receptor epitope in the form of the E200 epitope.

[048] It will be appreciated by those skilled in the art that the need to ensure proper pairing of the heavy and light chains of the bridge molecule will be dictated, to some extent, by the location of the epitope fragment in the Fab and by the number of fragments. For example, where the first and second Fab arms comprise identical antigen-binding domain sequences (e.g., in the scenario where the bridge molecule is bivalent and monospecific) and the first Fab arm comprises a single fragment (or indeed additional fragments) capable of being bound by the exogenous immune cell receptor, it is necessary to ensure proper pairing only of the heavy chains, but Petition 870250089762, dated 02 / 10 / 2025, pages 380 / 580 20 / 176 not from the light chains, if the fraction is attached to the heavy chain of the first Fab.

[049] On the other hand, if the first and second Fab arms comprise non-identical antigen-binding sequences (for example, in the scenario where the bridge molecule is bispecific), then it is necessary to ensure proper pairing of distinct heavy chains and correct pairing of cognate light chains (i.e., pairing of cognate light chains with the respective pair of heavy chains).

[050] Similarly, if the first and second Fab arms comprise identical antigen-binding domain sequences (e.g., the molecule is bivalent and monospecific), and the first Fab arm comprises a single fragment (or indeed additional fragments) capable of being bound by the exogenous immune cell receptor, it is necessary to ensure proper pairing of distinct heavy chains and proper pairing of cognate light chains if the fragment is bound to the Fab light chain.

[051] In any aspect, the fraction may comprise or consist of a peptide or antibody or antibody fragment. Alternatively, the targeting fraction may comprise a ligand or binding partner for a protein or receptor present on the surface of the target cell.

[052] In any respect, the fraction capable of being bound by the exogenous immune cell receptor may be any peptide, polysaccharide or lipid or other fraction capable of being bound by a receptor that is useful in a universal CAR or switchable CAR system or similar. Such systems are known to those skilled in the art and are described, for example, in Wermke et al., (2021), Blood, 137: 3145-3148; Lin et al., (2021) Frontiers in Immunology, 12: https: / / doi.org / 10.3389 / fimmu.2021.744823 and Liu et al., (2019) J. Hematol. & Oncol. 12:69; and WO 2019 / 238722, all incorporated herein by reference. In certain embodiments, the receptor is capable of binding to a Petition 870250089762, dated 02 / 10 / 2025, pp. 381 / 580 21 / 176 neoepitope or exogenous epitope or peptide tag, such as a dye (e.g., fluorescein isothiocyanate, FITC), biotin fraction, AZip tag (or leucine zipper fraction), Blm peptide Latch, SpyTag, the 5B9 tag (i.e., comprising 10 amino acids derived from the human nuclear autoantigen La / SS-B), the PNE tag (i.e., a 14-amino acid peptide neoepitope derived from a yeast transcription factor), or any peptide or tag capable of being used and bound by an extracellular receptor for use in a UniCAR, OmniCAR (SpyCatcher), Domain-D-based CAR-T cells (ARC-SparX), or other similar universal CAR system. In the art, such neoepitopes or tags may also be referred to as switching elements in the context of so-called Universal CAR systems. Those skilled in the art will appreciate that such neoepitopes are not typically epitopes associated with cancer antigens.

[053] In any aspect, in scenarios where the exogenous immune cell receptor is a chimeric antigen receptor, the fraction capable of being bound by the exogenous immune cell receptor is a fraction capable of being bound by the binding domain or other extracellular domain of the chimeric antigen receptor (CAR). Typically, CARs are designed for binding to tumor antigens and, therefore, the fraction capable of being bound by the CAR is preferentially a tumor antigen or a sequence derived from a tumor antigen. The tumor antigen may be expressed in a solid or liquid tumor. Therefore, in such embodiments, the fractions in the heterodimeric molecule comprise an amino acid sequence of an epitope of an antigen from a solid or liquid tumor.

[054] In any aspect, the CAR recognizes an epitope of a tumor-specific or tumor-associated antigen and the fraction capable of being bound by the exogenous immune cell is the epitope of the tumor-specific or tumor-associated antigen. Petition 870250089762, dated 02 / 10 / 2025, pp. 382 / 580 22 / 176 a tumor. Therefore, in such embodiments, the fractions in the heterodimeric molecule comprise an amino acid sequence of an epitope of a tumor-specific antigen or tumor-associated antigen, or a neoepitope or other peptide sequence (as described in the present invention in the context of universal CAR systems).

[055] In one embodiment, the tumor-associated or tumor-specific antigen is an epitope derived from the P2X7, EGFRvIII, or CLDN6 antigens. Therefore, in such embodiments, the fractions in the heterodimeric molecule comprise an amino acid sequence of an epitope derived from the P2X7, EGFRvIII, or CLDN6 antigens. Examples of such amino acid sequences are provided in the Tables in the present invention.

[056] In one example, the CAR recognizes a P2X7 receptor epitope and the fractions present in the heterodimeric bridge molecule of the invention therefore comprise at least the amino acid sequence as set out in SEQ ID NO: 162. In any respect, the P2X7 receptor epitope fraction comprises or consists of a fragment of a dysfunctional P2X7 receptor. Exemplary fragments include GHNYTTRNILPGLNITC (SEQ ID NO: 153; also referred to in the present invention as the E200 epitope) and variants thereof (exemplary variants are provided in SEQ ID NOs: 154 to 162 and 166 to 218); KYYKENNVEKRTLIK and variants thereof (SEQ ID NO: 163 and 164; also referred to in the present invention as the E300 epitope); or GHNYTTRNILPGAGAKYYKENNVEK (SEQ ID NO: 165; also referred to in the present invention as the E200 / E300 epitope or composite).

[057] In further preferred embodiments, the CAR recognizes an epitope of the EGFRvIII receptor and the fractions present in the heterodimeric bridge molecule of the invention therefore comprise at least the sequence Petition 870250089762, dated 02 / 10 / 2025, pp. 383 / 580 23 / 176 amino acids as established in SEQ ID NO: 134.

[058] In further preferred embodiments, CAR recognizes an epitope of CLDN6 and the fractions present in the heterodimeric bridge molecule of the invention therefore comprise at least the amino acid sequence as set forth in SEQ ID NO: 143.

[059] In certain embodiments, the heterodimeric bridge molecule of the invention is bivalent and bispecific. In such embodiments, the epitope moieties located on the molecule are preferably located on a Fab portion that is capable of binding to the target cell.

[060] In particularly preferred embodiments, the heterodimeric bridge molecule of the invention is bivalent and monospecific. In other words, the molecule preferably comprises two binding sites, each for binding to the same target antigen.

[061] Exemplary sequences of certain heterodimeric bridge molecules of the invention are bivalent and monospecific and are provided in Tables 1 and 2. Table 1 provides examples of molecule sequences comprising fragments capable of being linked by an anti-E200 CAR (i.e., comprising peptide fragment tags derived from the E200 epitope of the P2X7 receptor, to be linked by a CAR capable of binding to the E200 epitope). The molecules defined in Table 1 include various molecules capable of binding to a wide range of different cancer antigens, including CD19, CD33 and others.

[062] Table 2 provides alternative tag sequences and alternative CAR / bridge molecule systems, such as bridge molecules capable of binding to CD19 or CD33 and comprising: epitope fractions comprising the amino acid sequence of PNE (PNE epitope portions), epitope fractions comprising the amino acid sequence of a Petition 870250089762, dated 02 / 10 / 2025, pp. 384 / 580 24 / 176 EGFRvIII fragment or epitope fractions comprising the amino acid sequence of a CLDN6 fragment.

[063] It will be understood that, according to the molecules and methods of the invention, the exogenous immune cell receptor is a cell surface receptor comprising an extracellular domain capable of binding to or being bound by a bridge molecule as described in the present invention, and wherein the receptor further comprises an intracellular signaling domain. Typically, wherein the extracellular domain is an antigen-binding domain, cell surface receptors may be referred to as chimeric antigen receptors (or CARs). In the scenario wherein the extracellular domain is one comprising a peptide or other capable-of-binding fragment (e.g., a so-called neoepitope), the receptor may instead be referred to as a universal CAR (meaning that the CAR is not one that is limited to recognizing and binding to a specific antigen via an antigen-binding domain).

[064] In certain modalities, where the immune cell receptor is a CAR, the binding domain may be able to bind to a tumor-specific or tumor-associated antigen. The immune cell comprising the exogenous immune cell surface receptor (such as a CAR or universal CAR) can be any immune cell capable of eliciting a cell death response, such as a T cell, NK cell, NKT cell, B cell, cytokine-induced killer (CIK) cell, lymphokine-activated killer (LAK) cell, or gamma-delta T cell. Other examples of immune cells are described further in the present invention. Preferably, the immune cell is a CAR-T cell or a universal CAR-T cell.

[065] In any aspect, the immune cell or a cell that is capable of Petition 870250089762, dated 02 / 10 / 2025, pages 385 / 580 25 / 176 to differentiate into an immune cell (e.g., an immune cell progenitor) is a cell that is capable of differentiating into an immune cell (e.g., a T cell that will express the receptor) and may be a stem cell, multilineage progenitor cell, or induced pluripotent stem cell.

[066] In any embodiment, the exogenous immune cell receptor may be a CAR comprising an antigen recognition domain for binding to a tumor-associated or tumor-specific antigen. Examples of such tumor-associated and tumor-specific antigens are generally known in the art, as are the antigen recognition domains for binding to them in the context of a CAR. Some examples of tumor-associated and tumor-specific antigens are described further in the present invention, although those skilled in the art will appreciate that the present invention finds application in improving the usability and reducing undesirable toxicities for any CAR construct.

[067] An antigen recognition domain of a CAR typically comprises an antigen-binding domain or a fragment derived from, or that is, an antibody.

[068] In non-limiting examples, which exemplify the usefulness of the invention, the exogenous immune cell receptor may be a CAR comprising an antigen recognition domain for binding to a P2X7 receptor epitope (such as dysfunctional or non-functional forms of the receptor as defined further in the present invention). In any embodiment, the binding domain (antigen recognition domain) of the exogenous receptor binds to an epitope associated with an adenosine triphosphate (ATP) binding site of the P2X7 receptor. In particularly preferred embodiments, the P2X7 epitope comprises or is a derivative of the E200 epitope as defined in this invention, including in Table 3. In embodiments Petition 870250089762, dated 02 / 10 / 2025, pp. 386 / 580 26 / 176, particularly preferred, the P2X7 receptor epitope comprises at least the amino acid sequence as set forth in SEQ ID NO: 162.

[069] In any embodiment, the antigen recognition domain binds to an epitope that includes proline at amino acid position 210 of the P2X7 receptor. In some embodiments, the antigen recognition domain binds to an epitope that includes one or more amino acid residues extending from glycine at amino acid position 200 to cysteine ​​at amino acid position 216 inclusive of the P2X7 receptor, or a variant thereof.

[070] The receptor antigen recognition domain can be any suitable molecule that can interact with and bind specifically to a P2X7 receptor. However, in some embodiments, the antigen recognition domain includes amino acid sequence homology with the amino acid sequence of an antibody, or a fragment thereof, that binds to the dysfunctional P2X7 receptor. In some embodiments, the antigen recognition domain includes amino acid sequence homology with the amino acid sequence of an antigen-binding fragment (Fab) portion of an antibody that binds to a dysfunctional P2X7 receptor. In some embodiments, the antibody is a humanized antibody.

[071] In any embodiment, the antigen recognition domain includes amino acid sequence homology with the amino acid sequence of a VHH (i.e., a heavy chain antibody or nanobody), a d-domain-based recognition protein, a single-chain variable fragment (scFv), or a multivalent scFv that binds to the E200 epitope (or variant thereof) or another P2X7 receptor epitope (dysfunctional). In some embodiments, the multivalent scFv is a divalent or trivalent scFv.

[072] In any modality, the domain of recognition of Petition 870250089762, dated 02 / 10 / 2025, pp. 387 / 580 The 27 / 176 antigen includes amino acid sequence homology with a single antibody domain (sdAb) that binds to a dysfunctional P2X7 receptor.

[073] In any embodiment, the antigen recognition domain includes a binding polypeptide that includes amino acid sequence homology with one or more complementarity-determining regions (CDRs) of an antibody that binds to the E200 epitope (or variant thereof) or another epitope of a (dysfunctional) P2X7 receptor. In any embodiment, the antigen recognition domain includes amino acid sequence homology with the CDR1, 2, and 3 domains of the Vh and / or Vl chain of an antibody that binds to a dysfunctional P2X7 receptor.In preferred embodiments, the antigen recognition domain comprises the amino acid sequence of the CDRs of the Vh and / or Vl chains of an antibody, or the amino acid sequence of the Vh and / or Vl chains of an antibody, or the amino acid sequence of an antibody or fragment thereof, wherein the antibody or fragment thereof comprises the amino acid sequences of any antibody described in PCT / AU2002 / 000061 or PCT / AU2002 / 001204 (or in any of the corresponding US patents US 7,326,415, US 7,888,473, US 7,531,171, US 8,080,635, US 8,399,617, US 8,709,425, US 9,663,584 or US 10,450,380), PCT / AU2007 / 001540 (or in the corresponding US patent). US. 8,067,550), PCT / AU2007 / 001541 (or in the corresponding US patent publication US 2010-0036101), PCT / AU2008 / 001364 (or in any of the corresponding US patents US 8,440,186, US 9,181,320, US 9,944,701 or US 10,597,451), PCT / AU2008 / 001365 (or in any of the corresponding US patents US 8,293,491 or US 8,658,385), PCT / AU2009 / 000869 (or in any of the corresponding US patents US 8,597,643, US 9,328,155 or US 10,238,716), PCT / AU2010 / 001070 (or in any of the corresponding publications WO / 2011 / 020155, US 9,127,059, US 9,688,771, or US Petition 870250089762, dated 02 / 10 / 2025, pp. 388 / 580 28 / 176 10,053,508), and PCT / AU2010 / 001741 (or in any of the corresponding publications WO 2011 / 075789 or US 8,835,609), the full content of which is incorporated herein by reference.

[074] Preferably the antigen recognition domain comprises the CDR amino acid sequences of the 2-2-1 antigen-binding domain described in PCT / AU2010 / 001070 (or in any of the corresponding US patents US 9,127,059, US 9,688,771 or US 10,053,508) or BPM09 described in PCT / AU2007 / 001541 (or in the corresponding US patent publication US 2010-0036101) and produced by the AB253 hybridoma deposited in the European Collection of Cultures (ECACC) under Accession number 06080101.

[075] In additional preferred embodiments, the antigen recognition domain is a binding domain as described in WO 2023 / 028653.

[076] In some embodiments, the immune cell receptor signaling domain (such as a CAR) includes a portion derived from an activation receptor. In some embodiments, the activation receptor is a member of the CD3 coreceptor complex or is an Fc receptor. In some embodiments, the portion derived from the CD3 coreceptor complex is CD3-Z. In some embodiments, the portion derived from the Fc receptor is FcεRI or FcyRI.

[077] In some modalities, the signaling domain includes a portion derived from a co-stimulatory receptor. In some modalities, the signaling domain includes a portion derived from an activating receptor and a portion derived from a co-stimulatory receptor. In some modalities, the co-stimulatory receptor is selected from the group consisting of CD27, CD28, CD30, CD40, DAP10, OX40, 4-1BB (CD137), and ICOS.

[078] As used in the present invention, except where the context Petition 870250089762, dated 02 / 10 / 2025, pp. 389 / 580 29 / 176 requires otherwise, the term "comprise" and variations of the term, such as "comprise," "comprise," and "comprising," are not intended to exclude additives, components, whole items, or additional steps.

[079] Additional aspects of the present invention and further embodiments of the aspects described in the preceding paragraphs will become apparent from the following description, provided by way of example and with reference to the accompanying drawings. Brief description of the drawings

[080] Figure 1: Diagrams showing examples of target antigen-independent toxicity resulting from a suboptimal bridge molecule design (i.e., comprising epitope tags capable of being bound by CAR-T cells on both arms of the bridge molecule). A: Chronic activation, T cell exhaustion, and induction of apoptosis after binding of a suboptimal bridge molecule to a CAR-T cell; B: Fratricide resulting from toxic events between two CAR-expressing T cells.

[081] Figure 2: Proposed solution to the problem: design of a mono-tagged and doubly-tagged heterodimeric antibody molecule, where the epitope (tag) bound by the CAR is present only on one arm of the molecule.

[082] Figure 3: Exemplary formats of heterodimeric molecules of the invention showing the location of the CAR-bound epitope (label) on antibody molecules. Dark green and light green coloring denotes different antibody chains. The figure indicates possible combinations of labels and label locations, however, it does not show all possible combinations. The main sites for peptide label placement are indicated in the figure and are: N-terminal light chain, N-terminal heavy chain, C-terminal light chain, C-terminal heavy chain, between VL and CL, between Petition 870250089762, dated 02 / 10 / 2025, pages 390 / 580 30 / 176 VH and CH1. All label locations and all combinations can be used and individually optimized for the target antigen.

[083] Figure 4: Exemplary data showing cell death of U937 cells (expressing CD33) by anti-E200 P2X7 CAR-T cells and a bridging molecule comprising Fabs for CD33 binding and the E200 peptide tag. Mono-tagged (i.e., comprising a single copy of the E200 epitope) and doubly-tagged (comprising two copies of the E200 epitope) bridging molecules were tested. Doubly-tagged bridging molecules provide superior cell death in combination with anti-E200 P2X7 CAR-T cells compared to bridging molecules that have only one tag for binding by CAR-T cells. Concentration of bridging molecules used (y-axis) Concentration of bridging molecules used (y-axis) 0, 6, 4, 15, 40, 100 ng / ml or 1 pg / ml.

[084] Figure 5: Exemplary data showing that Jeko-1 cells are not killed by anti-E200 P2X7 CAR-T cells when combined with a bridging molecule comprising Fabs for CD33 binding and E200 peptide tag. These data indicate that there is no non-specific activation of CAR-T cells by the bridging molecule. Concentration of bridging molecules used (y-axis) 0, 6, 4, 15, 40, or 100 ng / ml.

[085] Figure 6: Other exemplary data showing cell death of U937 cells (expressing CD33) by a second anti-E200 P2X7 CAR-T cell line and a bridging molecule comprising Fabs for CD33 binding and E200 peptide tag. Mono-tagged (i.e., comprising a single copy of the E200 epitope) and doubly-tagged (comprising two copies of the E200 epitope) bridging molecules were tested. Doubly-tagged bridging molecules provide superior cell death in combination with anti-E200 P2X7 CAR-T cells compared to Petition 870250089762, dated 02 / 10 / 2025, pp. 391 / 580 31 / 176 bridge molecules that have only one tag for binding by CAR-T cells. Concentration of bridge molecules used (y-axis) 100 ng / ml or 1 ng / ml.

[086] Figure 7: Diagram showing A) ideal tag presentation (orthogonal tag presentation) on bridging molecules to enable engagement of CAR-T cells and APCs; B) suboptimal tag presentation (non-orthogonal) on bridging molecules.

[087] Figure 8: Cell death of U937 cells (expressing CD33) or Jeko-1 cells (expressing CD19) by anti-E200 P2X7 CAR-T cells and various bridging molecules at 24 and 48 hours. The bridging molecules are bivalent and can be mono- or bispecific for binding to CD33 (green) and / or CD19 (red) and one or more E200 peptide tags. Concentration of bridging molecules used (y-axis) 0, 1, 10, 100 ng / ml or 1 pg / ml.

[088] Figure 9: Undesired activation induced by the presence of a doubly labeled bridge molecule (where the labels are present on each of the antibody's Fab arms). The different antibodies used in this assay comprised symmetrical antibodies with 2 epitope fractions (peptide labels) on the light chains (mAb2) or heavy chains (mAb2), as well as a symmetrical antibody with 4 labels (mAb4) [two on the light chain and 2 on the heavy chain]. In addition, the assay comprised Cross-MAB antibody variants with a single label on the light chain (XmAb1) of an antibody's Fab arm or on the heavy chain (XmAb1) of an antibody's Fab arm, as well as with two labels on the light chain and on the heavy chain of an antibody's Fab arm (XmAb2). All labels tested were positioned at the N-terminal end of the molecules. Concentration of bridge molecules used (y-axis) 10, 40, 100 or 400 ng / ml.

[089] Figure 10: A. Cytolysis of the AML U937 cell line at 24 hours, Petition 870250089762, dated 02 / 10 / 2025, pp. 392 / 580 32 / 176 B - at 48 hours and C - at 72 hours. Ai, Bi and Ci: cytolysis of the AML U937 cell line by an anti-E200 CAR was measured using a luciferase-based cell death assay at an ET ratio of 0.5:1 (CAR-T to target ratio) in conjunction with and without the represented bridging molecule variants (comprising anti-CD33 binding domains and E200 tags at the indicated concentrations). The second condition (Aii, Bii and Cii) makes use of a CAR-T cell product that is unable to bind to the E200 peptide (via a mutated antigen-binding domain in the CAR) and serves as a negative control. Concentration of bridging molecules used (y-axis) 0, 1, 2, 6, 6, 4, 16, 40 or 100 ng / ml.

[090] Figure 11: Comparison of CAR anti-E200 cytolysis versus AML U937 cell line at 1 ng / mL of the corresponding adapter molecule formats at 24 hours, 48 ​​hours, and 72 hours. Statistical analysis of the various antibodies demonstrated superior function of the symmetric antibodies mAb2, mAb4, and XmAb2 compared to XmAb1 and Fab1.

[091] Figure 12: Kaplan-Meier plot showing the overall survival of mice treated with anti-E200 CAR-T cells + doubly tagged bridge molecule (XmAb2) comprising anti-CD33 binding domains (lintuzumab-based) + an E200 peptide on the N-terminal light chain and an E200 peptide on the N-terminal heavy chain of one Fab arm of the antibody (the other Fab arm not having E200 peptide tags), compared with untreated mice, or mice treated with non-transduced activated and expanded T cells, or mice receiving anti-CD33 CAR-T cell treatment (lintuzumab-based).

[092] Figure 13: Cell death of U937 cells by a universal CAR targeting PNE, using bridging molecules (0 ng / ml or 100 ng / ml) comprising a PNE peptide and antigen-binding domains based Petition 870250089762, dated 02 / 10 / 2025, pp. 393 / 580 33 / 176 in lintuzumab. Similar to the results shown using E200-based CARs and bridging molecules comprising E200 peptides, PNE-based bridging molecules and CAR systems are useful for killing target cells.

[093] Figure 14: Bioluminescence in MOLM-13 cancer cells in an AML xenograft model; animals treated with CAR-T cells. 1x10⁶ effective anti-E200 P2X7 CAR-T cells or 25x10⁶ non-transduced T cells were administered 3 days after 1x10⁶ MOLM-13 cells. CD33-targeted XmAb1, XmAb2, mAb2 No. 1, or mAb2 No. 2 were injected intraperitoneally at 25 pg / mouse twice weekly (50 pg / mouse per week) into the E200-targeted CAR-T cell group. Bioluminescence imaging was performed twice weekly throughout the treatment. (A) Bioluminescence readings of individual animals during the course of treatment in the group with only anti-E200 P2X7 CAR, with XmAb1, XmAb2, or mAb2 No. 1 targeting CD33. (B) Average animal bioluminescence readings of each group of five treated mice during the course of treatment.(C) The peripheral blood cell subpopulation of mice bearing the MOLM-13 xenograft (and treated as described in Figure 14) was analyzed by flow cytometry when the animal reached the endpoint of the experiment and was euthanized. The blood cell subpopulations were: mouse blood cells (mouse CD45+), MOLM-13 cancer cells (GFP+), injected CAR-T cells (mouse CD45-, human CAR+ CD3+), and human CAR-negative T cells (mouse CD45+, human CAR- CD3+). The percentage of each population relative to the total blood cells was plotted. Animals treated with the asymmetric bridge molecule XmAb2 had better CAR-T cell persistence compared to mice treated with the symmetric molecule mAb2 No. 1. Petition 870250089762, dated 02 / 10 / 2025, pp. 394 / 580 34 / 176

[094] Figure 15: Median fluorescence intensity ratio (MFIR) measured by flow cytometry, after incubation of wild-type and CD33 knockout U937 cells.

[095] Figure 16: Percentage of cell viability of U937 cells after co-incubation with CAR-T BIL-03 cells and various bridging molecules. The results show comparable efficacy of a bispecific CLL1xCD33 crossmab to monospecific CD33 and CLL1 crossmabs mixed 1:1 in U937 WT cells expressing CD33 and CLL1. · = Monospecific CD33 crossmab with E200 tag in a single Fab arm. · = Monospecific CLL1 crossmab with E200 tag in a single Fab arm. ▲ = Bispecific CLL1xCD33 crossmab with E200 tag in the anti-CLL1 Fab arm. ▼ = Monospecific CD33 and CLL1 crossmab mixed in a 1:1 ratio.

[096] Figure 17: Percentage of cell viability of U937 CLL1 KO cells after co-incubation with CAR-T BIL-03 cells and various bridging molecules. · = Monospecific crossmab for CD33 with E200 tag in a single Fab arm. · = Monospecific crossmab for CLL1 with E200 tag in a single Fab arm. ▲ = Bispecific CLL1xCD33 crossmab with E200 tag in the anti-CLL1 Fab arm. ▼ = Monospecific crossmab for CD33 and CLL1 mixed in a 1:1 ratio.

[097] Figure 18: Percentage of cell viability of U937 CD33 KO cells after co-incubation with CAR-T BIL-03 cells and various bridging molecules. · = Monospecific crossmab for CD33 with E200 tag in a single Fab arm. • = Monospecific crossmab for CLL1 with E200 tag in a single Fab arm. ▲ = Bispecific CLL1xCD33 crossmab with E200 tag in the anti-CLL1 Fab arm. ▼ = Monospecific crossmab for CD33 and CLL1 mixed in a 1:1 ratio.

[098] Figure 19: Percentage of cell viability of U937 cells Petition 870250089762, dated 02 / 10 / 2025, pp. 395 / 580 35 / 176 mixed after co-incubation with CAR-T BIL-03 cells and various bridging molecules. U937 WT, CD33KO, and CLL1KO were mixed in a 1:1:1 ratio. · = Monospecific crossmab for CD33 labeled E200 on a single Fab arm. · = Monospecific crossmab for CLL1 labeled E200 on a single Fab arm. ▲ = Bispecific CLL1xCD33 crossmab with E200 label on the anti-CLL1 Fab arm. ▼ = Monospecific crossmab for CD33 and CLL1 mixed in a 1:1 ratio. Sequence Information

[099] Table 1: Sequence information for exemplary heterodimeric molecules (bridge molecules) of the invention comprising E200-based epitope fractions for use with an anti-E200 CAR system.

[100] The expert in the subject will appreciate that the sequence pairing in the table below is: Left-arm sequences = KNOB-side sequences are paired with each other (always a light chain (LC) and a heavy chain (HC)). Right-arm sequences = HOLE-side sequences are paired with each other (always an LC and an HC). HC CROSS-MAB sequences are for pairing only with LC CROSS-MAB sequences. HC DUET-MAB sequences are for pairing only with LC DUET-MAB sequences.

[101] The following superscript number indicates the number of peptide labels on the KNOB side or on the Fab on the left side of the antibody, which theoretically could be 1, 2, 3, 4, 5, 6 or more. The molecule formats XmAb1, XmAb2, XmAb3, XmAb4, XmAb5, XmAb6, XmAbn or DUETMAB1, DUETMAB2, DUETMAB3, DUETMAB4, DUETMAB5, DUETMAB6, DUETMABn provide XmAb and DUETMAB with potency similar to a symmetric mAb in mediating specific effector functions, but having much less undesirable target-independent activation, exhaustion and fratricide than symmetric mAbs. Petition 870250089762, dated 02 / 10 / 2025, pp. 396 / 580 36 / 176 Description Sequence SEQ ID NO: Anti-CD33 Lin HC (left arm) KNOB K147E K213E L234A L235A P329G E356K D399K (pairs with SEQ ID NO: 8, 10, 287, 288) MGWSCIILFLVATATGVHSQVQLVQSGAEVKKPGSS VKVSCKASGYTFTDYNMHWVRQAPGQGLEWIGYI YPYNGGTGYNQKFKSKATITADESTNTAYMELSSLRS EDTAVYYCARGRPAMDYWGQGTLVTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDEKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAK GQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK HHHHHH 1 Anti-CD33 Lin HC (left arm) KNOB + variant Label E200 No26 K147E K213E L234A L235A P329G E356K D399K (pairs with SEQ ID NO: 8, 10,288) MGWSCIILFLVATATGVHSDFPGHNYTTRNILPGLN ITSGSEPKSSDKTHTGSQVQLVQSGAEVKKPGSSVK VSCKASGYTFTDYN MHWVRQAPGQG LEWIGYIYPY NGGTGYNQKFKSKATITADESTNTAYMELSSLRSED TAVYYCARGRPAM DYWGQGTLVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVEDYFPEPVTVSWNSGA LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDEKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQP REPQVYTLPPSRKELTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGKHHH HHH 2 Anti-CD33 Lin HC (left arm) KNOB + Label E200 K147E K213E L234A L235A P329G E356K D399K (pairs with SEQ ID NO: 8, 10,288) MGWSCII LFLVATATGVHSGHNYTTRNILPGLNITS QVQLVQSGAEVKKPGSSVKVSCKASGYTFTDYNMH WVRQAPGQGLEWIGYIYPYNGGTGYNQKFKSKATI TADESTNTAYMELSSLRSEDTAVYYCARGRPAMDY WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAAL GCLVEDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDEKV EPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK VSNKALGAPIEKTISKAKGQPREPQVYTLPPSRKELTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP VLKSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPGKHHHHHH 287 Anti-CD33 Lin HC (right arm) MGWSCIILFLVATATGVHSQVQLVQSGAEVKKPGSS VKVSCKASGYTFTDYNMHWVRQAPGQGLEWIGYI 3, Petition 870250089762, dated 02 / 10 / 2025, pp. 397 / 580 37 / 176 Description Sequence SEQ ID NO: HOLE L234A L235A P329G K329D K409D (pairs with SEQ ID NO: 9) YPYNGGTGYNQKFKSKATITADESTNTAYMELSSLRS EDTAVYYCARGRPAMDYWGQGTLVTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAK GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYDTTPPVLDSDGSFFLYSDLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG KHHHHHH Anti-CD33 Lin HC (right arm) CROSSMAB HOLE Elbow sequence VHCL Q124E +short hinge+ L234A L235A P329G K329D K409D (pairs with SEQ ID NO: 11,12) MGWSCIILFLVATATGVHSQVQLVQSGAEVKKPGSS VKVSCKASGYTFTDYNMHWVRQAPGQGLEWIGYI YPYNGGTGYNQKFKSKATITADESTNTAYMELSSLRS EDTAVYYCARGRPAMDYWGQGTLVTVSSASVAAP SVFIFPPSDEELKSGTASVVCLLNNFYPREAKVQWKV DNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE KHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVD VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTIS KAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYDTTPPVLDSDGSFFLYSD LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGKHHHHHH 4 Anti-CD33 Lin HC (right arm) CROSSMAB HOLE + variant label E200 no26 Elbow sequence VHCL Q124E +short hinge+ L234A L235A P329G K329D K409D (pairs with SEQ ID NO: 11,12) MGWSCIILFLVATATGVHSDFPGHNYTTRNILPGLN ITSGSEPKSSDKTHTGSQVQLVQSGAEVKKPGSSVK VSCKASGYTFTDYN MHWVRQAPGQG LEWIGYIYPY NGGTGYNQKFKSKATITADESTNTAYMELSSLRSED TAVYYCARGRPAM DYWGQGTLVTVSSASVAAPSVF IFPPSDEELKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH KVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPA PEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKA KGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD ​​IAVEWESNGQPENNYDTTPPVLDSDGSFFLYSDLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG KHHHHHH 5 Lin HC (right arm) DUETMAB HOLE F126C C220S L234A L235A P329G K329D K409D MGWSCIILFLVATATGVHSQVQLVQSGAEVKKPGSS VKVSCKASGYTFTDYNMHWVRQAPGQGLEWIGYI YPYNGGTGYNQKFKSKATITADESTNTAYMELSSLRS EDTAVYYCARGRPAMDYWGQGTLVTVSSASTKGPS 6, Petition 870250089762, dated 02 / 10 / 2025, pp. 398 / 580 38 / 176 Description Sequence SEQ ID NO: (pairs with SEQ ID NO: 13, 14) VCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAK GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYDTTPPVLDSDGSFFLYSDLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG KHHHHHH Anti-CD33 Lin HC (right arm) DUETMAB HOLE + variant label E200 no 26 F126C C220S L234A L235A P329G K329D K409D (pairs with SEQ ID NO: 13,14) MGWSCIILFLVATATGVHSDFPGHNYTTRNILPGLN ITSGSEPKSSDKTHTGSQVQLVQSGAEVKKPGSSVK VSCKASGYTFTDYN MHWVRQAPGQG LEWIGYIYPY NGGTGYNQKFKSKATITADESTNTAYMELSSLRSED TAVYYCARGRPAM DYWGQGTLVTVSSASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQP REPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYDTTPPVLDSDGSFFLYSDLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGKHHH HHH 7 Anti-CD33 Lin LC (left arm) KNOB E123KQ124R (pairs with SEQ ID NO: 1, 2,287) METDTLLLWVLLLWVPGSTGDIQMTQSPSSLSASV GDRVTITCRASESVDNYGISFMNWFQQKPGKAPKL LIYAASNQGSGVPSRFSGSGSGTDFTLTISSLQPDDF ATYYCQQSKEVPWTFGQGTKVEIKRTVAAPSVFIFP PSDKRLKSGTASVVCLLNNFYPREAKVQWKVDNAL QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKV YACEVTHQGLSSPVTKSFNRGEC 8 Anti-CD33 Lin LC (right arm) HOLE Q124E (pairs with SEQ ID NO: 3) METDTLLLWVLLLWVPGSTGDIQMTQSPSSLSASV GDRVTITCRASESVDNYGISFMNWFQQKPGKAPKL LIYAASNQGSGVPSRFSGSGSGTDFTLTISSLQPDDF ATYYCQQSKEVPWTFGQGTKVEIKRTVAAPSVFIFP PSDEELKSGTASVVCLLNNFYPREAKVQWKVDNAL QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKV YACEVTHQGLSSPVTKSFNRGEC 9 Lin LC (left arm) + variant label E200 no 26 KNOB E123KQ124R (pairs with SEQ ID NO: 1, 2, 287) M ETDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPG LNITSGSEPKSSDKTHTGSDIQMTQSPSSLSASVG D RVTITCRASESVDNYGISFMNWFQQKPGKAPKLLIY AASNQGSGVPSRFSGSGSGTDFTLTISSLQPDDFATY YCQQSKEVPWTFGQGTKVEIKRTVAAPSVFIFPPSD KRLKSGTASVVCLLNNFYPREAKVQWKVDNALQSG 10, Petition 870250089762, dated 02 / 10 / 2025, pp. 399 / 580 39 / 176 Description Sequence SEQ ID NO: NSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC Lin LC (left arm) + label E200 KNOB E123KQ124R (pairs with SEQ ID NO: 1, 2, 287) M ETDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SDIQMTQSPSSLSASVGDRVTITCRASESVDNYGISF MNWFQQKPGKAPKLLIYAASNQGSGVPSRFSGSGS GTDFTLTISSLQPDDFATYYCQQSKEVPWTFGQGTK VEIKRTVAAPSVFIFPPSDKRLKSGTASVVCLLNNFYP REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGE C 288 Anti-CD33 Lin LC (right arm) CROSSMAB HOLE K147K K213K + elbow sequence VL-CH1 + connecting EPKSC in C-ter (pair with SEQ ID NO: 4, 5) METDTLLLWVLLLWVPGSTGDIQMTQSPSSLSASV GDRVTITCRASESVDNYGISFMNWFQQKPGKAPKL LIYAASNQGSGVPSRFSGSGSGTDFTLTISSLQPDDF ATYYCQQSKEVPWTFGQGTKVEIKSSASTKGPSVFP LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGAL TSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSC 11 Anti-CD33 Lin LC (right arm) CROSSMAB + E200 HOLE K147K K213K label + VL-CH1 elbow sequence + connecting EPKSC to C-ter (pairs with SEQ ID NO: 4,5) METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSDIQMTQSPSSLSASVGDRV TITCRASESVDNYGISFMNWFQQKPGKAPKLLIYAA SNQGSGVPSRFSGSGSGTDFTLTISSLQPDDFATYYC QQSKEVPWTFGQGTKVEIKSSASTKGPSVFPLAPSS KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP SNTKVDKKVEPKSC 12 Anti-CD33 Lin LC (right arm) DUETMAB HOLE S121C Q124E C214S (pairs with SEQ ID NO: 6, 7) METDTLLLWVLLLWVPGSTGDIQMTQSPSSLSASV GDRVTITCRASESVDNYGISFMNWFQQKPGKAPKL LIYAASNQGSGVPSRFSGSGSGTDFTLTISSLQPDDF ATYYCQQSKEVPWTFGQGTKVEIKRTVAAPSVFIFP PCDEELKSGTASVVCLLNNFYPREAKVQWKVDNAL QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKV YACEVTHQGLSSPVTKSFNRGES 13 Anti-CD33 Lin LC (right arm) DUETMAB + Label E200 HOLE S121C Q124E C214S (pairs with SEQ ID NO: 6,7) METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSDIQMTQSPSSLSASVGDRV TITCRASESVDNYGISFMNWFQQKPGKAPKLLIYAA SNQGSGVPSRFSGSGSGTDFTLTISSLQPDDFATYYC QQSKEVPWTFGQGTKVEIKRTVAAPSVFIFPPCDEEL KSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGES 14 Anti-CD19 Tafa HC (left arm) KNOB K147E K213E L234A L235A MGWSCIILFLVATATGVHSEVQLVESGGGLVKPGGS LKLSCAASGYTFTSYVM HWVRQAPGKGLEWIGYIN PYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSE DTAMYYCARGTYYYGTRVFDYWGQGTLVTVSSAST 15, Petition 870250089762, dated 02 / 10 / 2025, pages 400 / 580 40 / 176 Description Sequence SEQ ID NO: P329G E356K D399K (pairs with SEQ ID NO: 22, 24) KGPSVFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLG TQTYICNVNHKPSNTKVDEKVEPKSCDKTHTCPPCP APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKA KGQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSD ​​IAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK HHHHHH Anti-CD19 Tafa HC (left arm) KNOB + Label E200 K147E K213E L234A L235A P329G E356K D399K (pairs with SEQ ID NO: 22, 24) MGWSCIILFLVATATGVHSDFPGHNYTTRNILPGLNI TSGSEPKSSDKTHTGSEVQLVESGGGLVKPGGSLKLS CAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYN DGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTA MYYCARGTYYYGTRVFDYWGQGTLVTVSSASTKGP SVFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDEKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAK GQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK HHHHHH 16 Anti-CD19 Tafa HC (right arm) HOLE L234A L235A P329G K329D K409D (pairs with SEQ ID NO: 23) MGWSCIILFLVATATGVHSEVQLVESGGGLVKPGGS LKLSCAASGYTFTSYVM HWVRQAPGKGLEWIGYIN PYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSE DTAMYYCARGTYYYGTRVFDYWGQGTLVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLG TQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCP APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKA KGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD ​​IAVEWESNGQPENNYDTTPPVLDSDGSFFLYSDLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG KHHHHHH 17 Anti-CD19 Tafa HC (right arm) CROSSMAB HOLE Elbow sequence VHCL Q124E +short hinge+ L234A L235A P329G K329D MGWSCIILFLVATATGVHSEVQLVESGGGLVKPGGS LKLSCAASGYTFTSYVM HWVRQAPGKGLEWIGYIN PYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSE DTAMYYCARGTYYYGTRVFDYWGQGTLVTVSSASV AAPSVFIFPPSDEELKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSTLTLSK 18 Petition 870250089762, dated 02 / 10 / 2025, pp. 401 / 580 41 / 176 Description Sequence SEQ ID NO: K409D (pairs with SEQ ID NO: 25, 26) ADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPI EKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYDTTPPVLDSDGSFF LYSDLTVDKSRWQQG NVFSCSVMH EALHN HYTQK SLSLSPGKHHHHHH Anti-CD19 Tafa HC (right arm) CROSSMAB HOLE + label E200 Elbow sequence VH-CL Q124E +short hinge+ L234A L235A P329G K329D K409D (pairs with SEQ ID NO: 25,26) MGWSCIILFLVATATGVHSDFPGHNYTTRNILPGLNI TSGSEPKSSDKTHTGSEVQLVESGGGLVKPGGSLKLS CAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYN DGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTA MYYCARGTYYYGTRVFDYWGQGTLVTVSSASVAAP SVFIFPPSDEELKSGTASVVCLLNNFYPREAKVQWKV DNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE KHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVD VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTIS KAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYDTTPPVLDSDGSFFLYSD LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGKHHHHHH 19 Anti-CD19 Tafa HC (right arm) DUETMAB HOLE F126C C220S L234A L235A P329G K329D K409D (pairs with SEQ ID NO: 27,28) MGWSCIILFLVATATGVHSEVQLVESGGGLVKPGGS LKLSCAASGYTFTSYVM HWVRQAPGKGLEWIGYIN PYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSE DTAMYYCARGTYYYGTRVFDYWGQGTLVTVSSAST KGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLG TQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCP APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKA KGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD ​​IAVEWESNGQPENNYDTTPPVLDSDGSFFLYSDLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG KHHHHHH 20 Anti-CD19 Tafa HC (right arm) DUETMAB HOLE + label E200 F126C C220S L234A L235A P329G K329D K409D (pairs with SEQ ID NO: 27, 28) MGWSCIILFLVATATGVHSDFPGHNYTTRNILPGLNI TSGSEPKSSDKTHTGSEVQLVESGGGLVKPGGSLKLS CAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYN DGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTA MYYCARGTYYYGTRVFDYWGQGTLVTVSSASTKGP SVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPE 21, Petition 870250089762, dated 02 / 10 / 2025, pp. 402 / 580 42 / 176 Description Sequence SEQ ID NO: AAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAK GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYDTTPPVLDSDGSFFLYSDLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG KHHHHHH Anti-CD19 Tafa LC (left arm) KNOB E123KQ124R (pairs with SEQ ID NO: 15,16) METDTLLLWVLLLWVPGSTGDIVMTQSPATLSLSPG ERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQL LIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFA VYYCMQHLEYPITFGAGTKLEIKRTVAAPSVFIFPPSD KRLKSGTASVVCLLNNFYPREAKVQWKVDNALQSG NSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC 22 Anti-CD19 Tafa LC (braço direito) HOLE Q124E (pareia com SEQ ID NO: 17) METDTLLLWVLLLWVPGSTGDIVMTQSPATLSLSPG ERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQL LIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFA VYYCMQHLEYPITFGAGTKLEIKRTVAAPSVFIFPPSD EELKSGTASVVCLLNNFYPREAKVQWKVDNALQSG NSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC 23 Anti-CD19 Tafa LC (braço esquerdo) + etiqueta E200 KNOB E123KQ124R (pareia com SEQ ID NO: 15,16) METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSDIVMTQSPATLSLSPGERA TLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYR MSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYY CMQHLEYPITFGAGTKLEIKRTVAAPSVFIFPPSDKRL KSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGEC 24 Anti-CD19 Tafa LC (right arm) CROSSMAB HOLE K147K K213K + VL-CH1 elbow sequence + connecting EPKSC at C-ter (pairs with SEQ ID NO: 18,19) METDTLLLWVLLLWVPGSTGDIVMTQSPATLSLSPG ERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQL LIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFA VYYCMQHLEYPITFGAGTKLEIKSSASTKGPSVFPLAP SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC 25 Anti-CD19 Tafa LC (right arm) CROSSMAB + E200 Label HOLE K147K K213K + VL-CH1 elbow sequence + connecting EPKSC in C-ter METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSDIVMTQSPATLSLSPGERA TLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYR MSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYY CMQHLEYPITFGAGTKLEIKSSASTKGPSVFPLAPSSK STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHT FPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPS 26, Petition 870250089762, dated 02 / 10 / 2025, pp. 403 / 580 43 / 176 Description Sequence SEQ ID NO: (pairs with SEQ ID NO: 18, 19) NTKVDKKVEPKSC Anti-CD19 Tafa LC (right arm) DUETMAB HOLE S121C Q124E C214S (pairs with SEQ ID NO: 20, 21) METDTLLLWVLLLWVPGSTGDIVMTQSPATLSLSPG ERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQL LIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFA VYYCMQHLEYPITFGAGTKLEIKRTVAAPSVFIFPPCD EELKSGTASVVCLLNNFYPREAKVQWKVDNALQSG NSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGES 27 Anti-CD19 Tafa LC (right arm) DUETMAB + Label E200 HOLE S121C Q124E C214S (pairs with SEQ ID NO: 20, 21) METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSDIVMTQSPATLSLSPGERA TLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYR MSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYY CMQHLEYPITFGAGTKLEIKRTVAAPSVFIFPPCDEEL KSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGES 28 Heavy chain KNOB CD33_Gemtuzumab MGWSCIILFLVATATGVHSEVQLVQSGAEVKKPGSS VKVSCKASGYTITDSNIHWVRQAPGQSLEWIGYIYP YNGGTDYNQKFKNRATLTVDNPTNTAYMELSSLRS EDTAFYYCVNGNPWLAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT YICVNNHKPSNTKVDEKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPKPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAK GQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK HHHHHH 29 CD33-Gemtuzumab variant etiqueta E200 variant no 26 MGWSCIILFLVATATGVHSDFPGHNYTTRNILPGLN ITSGSEPKSSDKTHTGSEVQLVQSGAEVKKPGSSVK VSCKASGYTITDSNIHWVRQAPGQSLEWIGYIYPYN GGTDYNQKFKN RATLTVDNPTNTAYMELSSLRSEDT AFYYCVNGNPWLAYWGQGTLVTVSSASTKGPSVFP LAPSSKSTSGGTAALGCLVEDYFPEPVTVSWNSGAL TSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDEKVEPKSCDKTHTCPPCPAPEAAG GPSVFLFPKPKDTLMISRTPEVTCVVVDVSHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTRYVVSVLTV LHQDWLNGKEYKCKVSNKALGAPIEKTISKAGQPR EPQVYTLPPSRKELTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRW 321 Petition 870250089762, de 02 / 10 / 2025, pág. 404 / 580 44 / 176 Description Sequência SEQ ID NO: QQGNVFSCSVMHEALHNHYTQKSLSLSPGKHHHH HH CD123_clone_32716 MGWSCIILFLVATATGVHSQIQLVQSGPELKKPGET VKISCKASGYIFTNYGMNWVKQAPGKSFKWMGWI NTYTGESTYSADFKGRFAFSLETSASTAYLHINDLKNE DTATYFCARSGGYDPMDYWGQGTSVTVSSASTKG PSVFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICVNVNHKPSNTKVDEKVEPKSCDKTHTCPPCPAP EAAGGPSVFLFPKPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCCVSNKALGAPIEKTISKAK GQPREVYTLPPSRKELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLKSDGSFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK HHHHHH 30 CD70_Cusatuzumabe MGWSCIILFLVATATGVHSEVQLVESGGGLVQPGG SLRLSCAASGFTFSVYYMNWVRQAPGKGLEWVSDI NNEGGTTYYADSVKGRFTISRDNSKNSLYLQMNSLR AEDTAVYCARDAGYSN HVPIFDSWGQGTLVTVSS ASTKGPSVFPLASSKSTSGGTAALGCLVEDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDEKVEPKSCDKTHTCP PCPAPEAAGGPSVFLFPKPKDTLMISRTPEVTCVVV DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST YRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRKELTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGKHHHHHH 31 LewisY_3S193_MuVH MGWSCIILFLVATATGVHSEVKLVESGGGLVQPGGS LKLSCATSGFTFSDYYMYWVRQTPEKRLEWVAYMS NVGAITDYPDTVKGRFTISRDNAKSTLYLQMSRLRSE DTAMYYCARGTRDGSWFAYWGQGTLVTVSAASTK GPSVFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDEKVEPKSCDKTHTCPPCPA PEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKA KGQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK HHHHHH 32 CD276_hu8H9 MGWSCIILFLVATATGVHSQVQLVQSGAEVVKPGA 33 Petition 870250089762, dated 02 / 10 / 2025, pp. 405 / 580 45 / 176 Descrição Sequência SEQ ID NO: SVKLSCKTSGYTFTNYDINWVRQRPGQGLEWIGWI FPGDDSTQYNEKFKGKATLTTDTSTSTAYMELSSLRS EDTAVYFCARQTTGTWFAYWGQGTLVTVSSASTKG PSVFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICVNHKPSNTKVDEKPKSCDKTCPPCPAP EAAGGPSVFLFPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAK GQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK HHHHHH CD135_4G8 MGWSCIILFLVATATGVHSGHNYTTRNILPGLNITS QVQLQQPGAELVKPGASLKLSCKSSGYTFTSYWMH WVRQRPGHGLEWIGEIDPSDSYKDYNQKFKDKATL TVDRSSNTAYMHLSSLTSDSAVYYCARAITTTPFDF WGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAAL GCLVEDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDEKV EPKSCDKTHTCPPCPEPAEAAGGPSVFLFPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK VSNKALGAPIEKTISKAGQPREPQVYTLPPSRKELTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP VLKSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPGKHHHHHH 34CD19_Tafasitamab MGWSCII LFLVATATGVHSDFPGHNYTTRNILPGLN ITSGSEPKSSDKTHTGSEVQLVESGGGLVKPGGSLKL SCAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYN DGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTA MYYCARGTYYYGTRVFDYWGQGTLVTVSSASTKGP SVFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDEKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAK GQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK HHHHHH 35 CD20_Ofatumumab MGWSCII LFLVATATGVHSDFPGHNYTTRNILPGLN ITSGSEPKSSDKTHTGSEVQLVESGGGLVQPGRSLRL SCAASGFTFNDYAMHWVRQAPGKGLEWVSTISWN 36 Petition 870250089762, dated 02 / 10 / 2025, pp. 406 / 580 46 / 176 Description Sequence SEQ ID NO: SGSIGYADSVKGRFTISRDNAKKSLYLQMNSLRAEDT ALYYCAKDIQYGNYYYGMDVWGQGTTVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDEKVEPKSCDKTHTCPPCPPA PEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSN KALGAPIEKTISKA KGQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSD ​​IAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK HHHHHH CD22_m971-L7 MGWSCIILFLVATATGVHSDFPGHNYTTRNILPGLN ITSGSEPKSSDKTHTGSQVQLQQSGPGMVKPSQTL SLTCAISGDSVSSNSVAWNWIRQSPSRGLEWLGRTY YRSTWYNDYAVSMKSRITINPDTNKNQFSLQLNSVT PE DTAVYYCAREVTG DLEDAFDIWG QGTMVTVSSA STKGPSVFPLAPSSKSTSGGTAALGCLVEDYFPEPVT VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSS LGTQTYICNVNHKPSNTKVDEKVEPKSCDKTHTCPP CPAEAGGPSVFLFPKPKDTLMISRTPEVTCVVVD VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTIS KAKGQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP GKHHHHHH 37CD37_Lilotumab MGWSCII LFLVATATGVHSDFPGHNYTTRNILPGLN ITSGSEPKSSDKTHTGSEIQLQQSG PELVKPGASVKV SCKASGYSFTDYNMYWVKQSHGKSLEWIGYIDPYN GDTTYNQKFKGKATLTDKSSSTAFIHLNSLTSEDSA VYYCARSPYGHYAMDYWGQGTSVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI CNVNHKPSNTKVDEKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQP REPQVYTLPPSRKELTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGKHHH HHH 38 BCMA_11D5 MGWSCII LFLVATATGVHSDFPGHNYTTRNILPGLN ITSGSEPKSSDKTHTGSQIQLVQSGPELKKPGETVKIS CKASGYTFTDYSINWVKRAPGKGLKWMGWINTET 39 Petition 870250089762, dated 02 / 10 / 2025, pp. 407 / 580 47 / 176 Descrição Sequência SEQ ID NO: REPAYAYDFRGRFAFSLETSASTAYLQINNLKYEDTA TYFCALDYSYAMDYWGQGTSVTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVEDYFPEPVTVSWNSGALT SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV NHKPSNTKVDEKVEPKSCDKTHTCPPCPAPEAAGGP SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREP QVYTLPPSRKELTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGKHHHHH H BCMA_CA8_J9M0 MGWSCIILFLVATATGVHSDFPGHNYTTRNILPGLN ITSGSEPKSSDKTHTGSQVQLVQSGAEVKKPGSSVK VSCKGSGYTFTNYWMHWVRQAPGQGLEWIGATY RGHSDTYYNQKFKGRATLTADTSTSTAYMELSSLRSE DTAVYYCTRGAIYDGYDVLDNWGQGTLVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLG TQTYICNVNHKPSNTKVDEKVEPKSCDKTHTCPPCP APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKA KGQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK HHHHHH 40 CD138_nBT062MGWSCII LFLVATATGVHSDFPGHNYTTRNILPGLN ITSGSEPKSSDKTHTGSQVQLQQSGSELM M PGASV KISCKATGYTFSNYWIEWVKQRPGHGLEWIGEILPG TGRTIYNEKFKGKATFTADISSNTVQMQLSSLTSEDS AVYYCARRDYYGNFYYAMDYWGQGTSVTVSSASTK GPSVFPLASSKSTSGGTAALGCLVEDYFPPVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICVNHKPSNTKVDEKVEPKSCDKTHTCPPCPA PEAAGGPSVFLFPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCCVSNKALGAPIEKTISKA KGQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK HHHHHH 41 CD38_Daratumomabe MGWSCII LFLVATATGVHSDFFPGHNYTTRNILPGLN ITSGSEPKSSDKTHTGSEVQLLESSGGGLVQPGGSLRRL SCAVSGFTFNSFAMSWVRQAPGKGLEWVSAISGSG 42 Petition 870250089762, de 02 / 10 / 2025, pág. 408 / 580 48 / 176 Description Sequence SEQ ID NO: GGTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDT AVYFCAKDKILWFGEPVFDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDEKVEPKSCDKTHTCPPPCPA PEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKA KGQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSD ​​IAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK HHHHHH CLL1_1075.7 MGWSCIILFLVATATGVHSDFPGHNYTTRNILPGLN ITSGSEPKSSDKTHTGSDIQLQESGPGLVKPSQSLSLT CSVTGYSITSAYYWNWIRQFPGNKLEWMGYISYDG RNNYNPSLKNRISITRDTSKNQFFLKLNSVTTEDTATY YCAKEGDYDVGNYYAMDYWGQGTSVTVSSASTKG PSVFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVTVPSSSLGTQ TYICVNHKPSNTKVDEKVEPKSCDKTTHTCPPCPAP EAAGGPSVFLFPKPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAK GQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSPGK HHHHHH 43 ROR1_R12MGWSCII LFLVATATGVHSDFPGHNYTTRNILPGLN ITSGSEPKSSDKTHTGSQEQLVESGGRLVTPGGSLTL SCKASGFDFSAYYMSWVRQAPGKGLEWIATIYPSSG KTYYATWVNGRFTISSDNAQNTVDLQMNSLTAADR ATYFCARDSYADDGALFNIWGPGTLVTISSASTKGPS VFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDEKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAK GQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK HHHHHH 44 Light chain KNOB CD33_Gemtuzumab METDTLLLWVLLLWVPGSTGDIQLTQSPSTLSASVG DRVTITCRASESLDNYGIRFLTWFQQKPGKAPKLLM 45 Petition 870250089762, dated 02 / 10 / 2025, pp. 409 / 580 49 / 176 Descrição Sequência SEQ ID NO: YAASNQGSGVPSRFSGSGSGTEFTLTISSLQPDDFAT YYCQQTKEVPWSFGQGTKVEVKRTVAAPSVFIFPPS DRKLKSGTASVVCLLNNFYPREAKVQWKVDNALQS GNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYA CEVTHQGLSSPVTKSFNRGEC CD123_clone_32716 METDTLLLWVLLLWVPGSTGDIVLTQSPASLAVSLG QRATISCRASESVDNYGNTFMHWYQQKPGQPPKLL IYRASNLESGIPARFSGSGSRTDFTLTINPVEADDVAT YYCQQSNEDPPTFGAGTKLELKRTVAAPSVFIFPPSD RKLKSGTASVVCLLNNFYPREAKVQWKVDNALQSG NSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC 46 CD70_Cusatuzumabe METDTLLLWVLLLWVPGSTGQAVVTQEPSLTVSPG GTVTLTCGLKSGSVTSDNFPTWYQQTPGQAPRLLIY NTNTRHSGVPDRFSGSILGNKAALTITGAQADDEAE YFCALFISNPSVEFGGGTQLTVLTVAAPSVFIFPPSDR KLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGN SQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC 47 LewisY_3S193_MuVH METDTLLLWVLLLWVPGSTGDVLMTQTPLSLPVSL GDQASISCRSSQRIVHSNGNTYLEWYLQKPGQSPKL LIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLG VYYCFQGSHVPFTFGSGTKLEIKRTVAAPSVFIFPPSD RKLKSGTASVVCLLNNFYPREAKVQWKVDNALQSG NSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC 48CD276_hu8H9 METDTLLLWVLLLWVPGSTGEIVMTQSPATLSVSPG ERVTLSCRASQSISDYLYWYQQKSHESPRLLIKYASQS ISGIPARFSGSGSGSEFTLTINSVEPEDVGVYYCQNG HSFPLTFGQGTKLELKRTVAAPSVFIFPPSDRKLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGL SSPVTKSFNRGEC 49 CD135_4G8 METDTLLLWVLLLWVPGSTGDIVLTQSPATLSVTPG DSVSLSCRASQSISNNLHWYQQKSHESPRLLIKYASQ SISGIPSRFSGSGSGTDFTLSINSVETEDFGVYFCQQS NTWPYTFGGGTKLEIKRTVAAPSVFIFPPSDRKLKSG TASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG LSSPVTKSFNRGEC 50 CD19_Tafasitamabe METDTLLLWVLLLWVPGSTGDIVMTQSPATLSLSPG ERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQL LIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFA VYYCMQHLEYPITFGAGTKLEIKRTVAAPSVFIFPPSD RKLKSGTASVVCLLNNFYPREAKVQWKVDNALQSG 51 Petição 870250089762, de 02 / 10 / 2025, pág. 410 / 580 50 / 176 Descrição Sequência SEQ ID NO: NSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC CD20_Ofatumumabe METDTLLLWVLLLWVPGSTGEIVLTQSPATLSLSPGE RATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASN RATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR SNWPITFGQGTRLEIKRTVAAPSVFIFPPSDRKLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGL SSPVTKSFNRGEC 52 CD22_m971-L7 METDTLLLWVLLLWVPGSTGDIQMIQSPSSLSASVG DRVTITCRASQTIWSYLNWYRQRPGEAPNLLIYAASS LQSGVPSRFSGRGSGTDFTLTISSLQAEDFATYYCQQ SYSIPQTFGQGTKLEIKRTVAAPSVFIFPPSDRKLKSG TASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG LSSPVTKSFNRGEC 53 CD37_Lilotumabe METDTLLLWVLLLWVPGSTGDIVMTQSHKLLSTSV GDRVSITCKASQDVSTAVDWYQQKPGQSPKLLINW ASTRHTGVPDRFTGSGSGTDYTLTISSMQAEDLALY YCRQHYSTPFTFGSGTKLEIKRTVAAPSVFIFPPSDRK LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNS QESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC 54 BCMA_11D5 METDTLLLWVLLLWVPGSTGDIVLTQSPPSLAMSLG KRATISCRASESVTILGSHLIHWYQQKPGQPPTLLIQL ASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTKLEIKRTVAAPSVFIFPPSDRKL KSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGEC 55 BCMA_CA8_J9M0 METDTLLLWVLLLWVPGSTGDIQMTQSPSSLSASV GDRVTITCSASQDISNYLNWYQQKPGKAPKLLIYYTS NLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQ QYRKLPWTFGQGTKLEIKRTVAAPSVFIFPPSDRKLK SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGEC 56 CD138_nBT062 METDTLLLWVLLLWVPGSTGDIQMTQSTSSLSASLG DRVTISCSASQGINNYLNWYQQKPDGTVELLIYYTST LQSGVPSRFSGSGSGTDYSLTISNLEPEDIGTYYCQQY SKLPRTFGGGTKLEIKRTVAAPSVFIFPPSDRKLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGL SSPVTKSFNRGEC 57 CD38_Daratumomabe METDTLLLWVLLLWVPGSTGEIVLTQSPATLSLSPGE 58 Petição 870250089762, de 02 / 10 / 2025, pág. 411 / 580 51 / 176 Descrição Sequência SEQ ID NO: RATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASN RATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR SNWPPTFGQGTKVEIKRTVAAPSVFIFPPSDRKLKSG TASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG LSSPVTKSFNRGEC CLL1_1075.7 METDTLLLWVLLLWVPGSTGENVLTQSPAIMSASP GEKVTMTCRASSNVISSYVHWYQQRSGASPKLWIY STSNLASGVPARFSGSGSGTSYSLTISSVEAEDAATYY CQQYSGYPLTFGAGTKLELKRAAAAPSVFIFPPSDRK LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNS QESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC 59 ROR1_R12 METDTLLLWVLLLWVPGSTGELVLTQSPSVSAALGS PAKITCTLSSAHKTDTIDWYQQLQGEAPRYLMQVQ SDGSYTKRPGVPDRFSGSSSGADRYLIIPSVQADDEA DYYCGADYIGGYVFGGGTQLTVTGTVAAPSVFIFPPS DRKLKSGTASVVCLLNNFYPREAKVQWKVDNALQS GNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYA CEVTHQGLSSPVTKSFNRGEC 60 Cadeia leve KNOB E200 CD33_Gemtuzumabe METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SDIQLTQSPSTLSASVGDRVTITCRASESLDNYGIRFL TWFQQKPGKAPKLLMYAASNQGSGVPSRFSGSGS GTEFTLTISSLQPDDFATYYCQQTKEVPWSFGQGTK VEVKRTVAAPSVFIFPPSDRKLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL SSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRG EC 61 CD123_clone_32716 METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SDIVLTQSPASLAVSLGQRATISCRASESVDNYGNTF MHWYQQKPGQPPKLLIYRASNLESGIPARFSGSGSR TD FTLTIN PVEADDVATYYCQQSNED PPTFGAGTKL ELKRTVAAPSVFIFPPSDRKLKSGTASVVCLLNNFYPR EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS TLLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 62 CD70_Cusatuzumabe METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SQAVVTQEPSLTVSPGGTVTLTCGLKSGSVTSDNFPT WYQQTPGQAPRLLIYNTNTRHSGVPDRFSGSILGNK AALTITGAQADDEAEYFCALFISNPSVEFGGGTQLTV LTVAAPSVFIFPPSDRKLKSGTASVVCLLNNFYPREAK VQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL SKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 63 LewisY_3S193_MuVH METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SDVLMTQTPLSLPVSLGDQASISCRSSQRIVHSNGNT 64 Petition 870250089762, on 10 / 02 / 2025, page. 412 / 580 52 / 176 Descrição Sequência SEQ ID NO: YLEWYLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSG TDFTLKISRVEAEDLGVYYCFQGSHVPFTFGSGTKLEI KRTVAAPSVFIFPPSDRKLKSGTASVVCLLNNFYPRE AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSST LTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC CD276_hu8H9 METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SEIVMTQSPATLSVSPGERVTLSCRASQSISDYLYWY QQKSHESPRLLIKYASQSISGIPARFSGSGSGSEFTLTI NSVEPEDVGVYYCQNGHSFPLTFGQGTKLELKRTVA APSVFIFPPSDRKLKSGTASVVCLLNNFYPREAKVQW KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC 65 CD135_4G8 METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SDIVLTQSPATLSVTPGDSVSLSCRASQSISNNLHWY QQKSHESPRLLIKYASQSISGIPSRFSGSGSGTDFTLSI NSVETEDFGVYFCQQSNTWPYTFGGGTKLEIKRTVA APSVFIFPPSDRKLKSGTASVVCLLNNFYPREAKVQW KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC 66 CD19_Tafasitamabe METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SDIVMTQSPATLSLSPGERATLSCRSSKSLQNVNGNT YLYWFQQKPGQSPQLLIYRMSNLNSGVPDRFSGSG SGTEFTLTISSLEPEDFAVYYCMQHLEYPITFGAGTKL EIKRTVAAPSVFIFPPSDRKLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 67 CD20_Ofatumumabe METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQ 68 CD22_m971-L7 METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SDIQMIQSPSSLSASVGDRVTITCRASQTIWSYLNWY RQRPGEAPNLLIYAASSLQSGVPSRFSGRGSGTDFTL TISSLQAEDFATYYCQQSYSIPQTFGQGTKLEIKRTVA APSVFIFPPSDRKLKSGTASVVCLLNNFYPREAKVQW KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC 69 ​​CD37_Lilotumabe METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SDIVMTQSHKLLSTSVGDRVSITCKASQDVSTAVDW YQQKPGQSPKLLINWASTRHTGVPDRFTGSGSGTD YTLTISSMQAEDLALYYCRQHYSTPFTFGSGTKLEIKR TVAAPSVFIFPPSDRKLKSGTASVVCLLNNFYPREAK 70 Petition 870250089762, on 10 / 02 / 2025, page. 413 / 580 53 / 176 Descrição Sequência SEQ ID NO: VQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL SKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC BCMA_11D5 METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SDIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIH 71 BCMA_CA8_J9M0 METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SDIQMTQSPSSLSASVGDRVTITCSASQDISNYLNW YQQKPGKAPKLLIYYTSNLHSGVPSRFSGSGSGTDFT LTISSLQPED FATYYCQQYRKLPWTFGQGTKLEIKRT VAAPSVFIFPPSDRKLKSGTASVVCLLNNFYPREAKV QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS KADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 72 CD138_nBT062 METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SDIQMTQSTSSLSASLGDRVTISCSASQGINNYLNW 73 CD38_Daratumomabe METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNITSEIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQ QKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLT ISSLEPEDFAVYYCQQRSNWPPTFGQGTKVEIKRTV AAPSVFIFPPSDRKLKSGTASVVCLLNNFYPREAKVQ WKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 74 CLL1_1075.7 METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SENVLTQSPAIMSASPGEKVTMTCRASSNVISSYVH WYQQRSGASPKLWIYSTSNLASGVPARFSGSGSGTS YSLTISSVEAEDAATYYCQQYSGYPLTFGAGTKLELKR AAAAPSVFIFPPSDRKLKSGTASVVCLLNNFYPREAK VQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL SKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 75 ROR1_R12 METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SELVLTQSPSVSAALGSPAKITCTLSSAHKTDTIDWY QQLQGEAPRYLMQVQSDGSYTKRPGVPDRFSGSSS GADRYLIIPSVQADDEADYYCGADYIGGYVFGGGTQ LTVTGTVAAPSVFIFPPSDRKLKSGTASVVCLLNNFYP REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGE C 76 Petição 870250089762, de 02 / 10 / 2025, pág. 414 / 580 54 / 176 Descrição Sequência SEQ ID NO: Cadeia leve KNOB E-200-NO-26 CD33_Gemtuzumabe METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSDIQLTQSPSTLSASVGD RV TITCRASESLDNYGIRFLTWFQQKPGKAPKLLMYAAS NQGSGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQ QTKEVPWSFGQGTKVEVKRTVAAPSVFIFPPSDRKL KSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGEC 77 CD123_clone_32716 METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSDIVLTQSPASLAVSLGQRA TISCRASESVDNYGNTFMHWYQQKPGQPPKLLIYR ASNLESGIPARFSGSGSRTDFTLTINPVEADDVATYY CQQSNEDPPTFGAGTKLELKRTVAAPSVFIFPPSDRK LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNS QESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC 78 CD70_Cusatuzumabe METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSQAVVTQEPSLTVSPGGTVT LTCGLKSGSVTSDNFPTWYQQTPGQAPRLLIYNTNT RHSGVPDRFSGSILGNKAALTITGAQADDEAEYFCAL FISNPSVEFGGGTQLTVLTVAAPSVFIFPPSDRKLKSG TASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG LSSPVTKSFNRGEC 79 LewisY_3S193_MuVHMETDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSDVLMTQTPLSLPVSLGDQ ASISCRSSQRIVHSNGNTYLEWYLQKPGQSPKLLIYK VSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYY CFQGSHVPFTFGSGTKLEIKRTVAAPSVFIFPPSDRKL KSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGEC 80 CD276_hu8H9 METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSEIVMTQSPATLSVSPGERV TLSCRASQSISDYLYWYQQKSHESPRLLIKYASQSISGI PARFSGSGSGSEFTLTINSVEPEDVGVYYCQNGHSFP LTFGQGTKLELKRTVAAPSVFIFPPSDRKLKSGTASVV CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS KDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT KSFNRGEC 81 CD135_4G8 METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSDIVLTQSPATLSVTPGDSVS LSCRASQSISNNLHWYQQKSHESPRLLIKYASQSISGI PSRFSGSGSGTDFTLSINSVETEDFGVYFCQQSNTW 82 Petição 870250089762, de 02 / 10 / 2025, pág. 415 / 580 55 / 176 Descrição Sequência SEQ ID NO: PYTFGGGTKLEIKRTVAAPSVFIFPPSDRKLKSGTASV VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGEC CD19_Tafasitamabe METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSDIVMTQSPATLSLSPGERA TLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYR MSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYY CMQHLEYPITFGAGTKLEIKRTVAAPSVFIFPPSDRKL KSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGEC 83 CD20_Ofatumumabe METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSEIVLTQSPATLSLSPGERATL SCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATG IPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNW PITFGQGTRLEIKRTVAAPSVFIFPPSDRKLKSGTASV VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGEC 84 CD22_m971-L7 METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSDIQMIQSPSSLSASVGDRV TITCRASQTIWSYLNWYRQRPGEAPNLLIYAASSLQS GVPSRFSGRGSGTDFTLTISSLQAEDFATYYCQQSYSI PQTFGQGTKLEIKRTVAAPSVFIFPPSDRKLKSGTASV VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGEC 85 CD37_Lilotumabe METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSDIVMTQSHKLLSTSVGDRV SITCKASQDVSTAVDWYQQKPGQSPKLLINWASTR HTGVPDRFTGSGSGTDYTLTISSMQAEDLALYYCRQ HYSTPFTFGSGTKLEIKRTVAAPSVFIFPPSDRKLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGL SSPVTKSFNRGEC 86 BCMA_11D5 METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSDIVLTQSPPSLAMSLGKRA TISCRASESVTILGSHLIHWYQQKPGQPPTLLIQLASN VQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCL QSRTIPRTFGGGTKLEIKRTVAAPSVFIFPPSDRKLKS GTASVVCLLNNFYPREAKVQWKVDNALQSGNSQES VTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGEC 87 BCMA_CA8_J9M0 METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL 88 Petição 870250089762, de 02 / 10 / 2025, pág. 416 / 580 56 / 176 Descrição Sequência SEQ ID NO: NITSGSEPKSSDKTHTGSDIQMTQSPSSLSASVGDRV TITCSASQDISNYLNWYQQKPGKAPKLLIYYTSNLHS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYRK LPWTFGQGTKLEIKRTVAAPSVFIFPPSDRKLKSGTA SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTE QDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS SPVTKSFNRGEC CD138_nBT062 METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSDIQMTQSTSSLSASLGDRV TISCSASQGINNYLNWYQQKPDGTVELLIYYTSTLQS GVPSRFSGSGSGTDYSLTISNLEPEDIGTYYCQQYSKL PRTFGGGTKLEIKRTVAAPSVFIFPPSDRKLKSGTASV VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGEC 89 CD38_Daratumomabe METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSEIVLTQSPATLSLSPGERATL SCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATG IPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNW PPTFGQGTKVEIKRTVAAPSVFIFPPSDRKLKSGTASV VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGEC 90 CLL1_1075.7 METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSENVLTQSPAIMSASPGEKV TMTCRASSNVISSYVHWYQQRSGASPKLWIYSTSNL ASGVPARFSGSGSGTSYSLTISSVEAEDAATYYCQQY SGYPLTFGAGTKLELKRAAAAPSVFIFPPSDRKLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGL SSPVTKSFNRGEC 91 ROR1_R12 METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSELVLTQSPSVSAALGSPAKI TCTLSSAHKTDTIDWYQQLQGEAPRYLMQVQSDGS YTKRPGVPDRFSGSSSGADRYLIIPSVQADDEADYYC GADYIGGYVFGGGTQLTVTGTVAAPSVFIFPPSDRKL KSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTH QGLSSPVTKSFNRGEC 92 Cadeia pesada CROSS-MAB HOLE CD33_Gemtuzumabe MGWSCIILFLVATATGVHSEVQLVQSGAEVKKPGSS VKVSCKASGYTITDSNIHWVRQAPGQSLEWIGYIYP YNGGTDYNQKFKNRATLTVDNPTNTAYMELSSLRS EDTAFYYCVNGNPWLAYWGQGTLVTVSSASVAAPS VFIFPPSDEELKSGTASVVCLLNNFYPREAKVQWKV 93. Petição 870250089762, de 02 / 10 / 2025, pág. 417 / 580 57 / 176 Descrição Sequência SEQ ID NO: DNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE KHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVD VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTIS KAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYDTTPPVLDSDGSFFLYSD LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGKHHHHHH CD123_clone_32716 MGWSCIILFLVATATGVHSQIQLVQSGPELKKPGET VKISCKASGYIFTNYGMNWVKQAPGKSFKWMGWI NTYTGESTYSADFKGRFAFSLETSASTAYLHINDLKNE DTATYFCARSGGYDPMDYWGQGTSVTVSSASVAA PSVFIFPPSDEELKSGTASVVCLLNNFYPREAKVQWK VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY EKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVD VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTIS KAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYDTTPPVLDSDGSFFLYSD LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGKHHHHHH 94 CD70_Cusatuzumabe MGWSCIILFLVATATGVHSEVQLVESGGGLVQPGG SLRLSCAASGFTFSVYYMNWVRQAPGKGLEWVSDI NNEGGTTYYADSVKGRFTISRDNSKNSLYLQMNSLR AEDTAVYYCARDAGYSNHVPIFDSWGQGTLVTVSS ASVAAPSVFIFPPSDEELKSGTASVVCLLNNFYPREAK VQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL SKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKT HTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALG APIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYDTTPPVLDSDGS FFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPGKHHHHHH 95 LewisY_3S193_MuVH MGWSCIILFLVATATGVHSEVKLVESGGGLVQPGGS LKLSCATSGFTFSDYYMYWVRQTPEKRLEWVAYMS NVGAITDYPDTVKGRFTISRDNAKSTLYLQMSRLRSE DTAMYYCARGTRDGSWFAYWGQGTLVTVSAASVA APSVFIFPPSDEELKSGTASVVCLLNNFYPREAKVQW KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCP PCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVV 96 Petition 870250089762, dated 02 / 10 / 2025, pp. 418 / 580 58 / 176 Description Sequence SEQ ID NO: DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST YRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKT ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYDTTPPVLDSDGSFFLYS DLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS LSPGKHHHHHH CD276_hu8H9 MGWSCIILFLVATATGVHSQVQLVQSGAEVVKPGA SVKLSCKTSGYTFTNYDINWVRQRPGQGLEWIGWI FPGDDSTQYNEKFKGKATLTTDTSTSTAYMELSSLRS EDTAVYFCARQTTGTWFAYWGQGTLVTVSSASVAA PSVFIFPPSDEELKSGTASVVCLLNNFYPREAKVQWK VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY EKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVD VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTIS KAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYDTTPPVLDSDGSFFLYSD LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGKHHHHHH 97 CD135_4G8 MGWSCIILFLVATATGVHSQVQLQQPGAELVKPGA SLKLSCKSSGYTFTSYWMHWVRQRPGHGLEWIGEI DPSDSYKDYNQKFKDKATLTVDRSSNTAYMHLSSLT SDDSAVYYCARAITTTPFDFWGQGTTLTVSSASVAA PSVFIFPPSDEELKSGTASVVCLLNNFYPREAKVQWK VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADY EKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVD VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTIS KAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYDTTPPVLDSDGSFFLYSD LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGKHHHHHH 98 CD19_Tafasitamab MGWSCIILFLVATATGVHSEVQLVESGGGLVKPGGS LKLSCAASGYTFTSYVM HWVRQAPGKGLEWIGYIN PYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSE DTAMYYCARGTYYYGTRVFDYWGQGTLVTVSSASV AAPSVFIFPPSDEELKSGTASVVCLLNNFYPREAKVQ WKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPI EKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLV 99 Petition 870250089762, dated 02 / 10 / 2025, pp. 419 / 580 59 / 176 Descrição Sequência SEQ ID NO: KGFYPSDIAVEWESNGQPENNYDTTPPVLDSDGSFF LYSDLTVDKSRWQQG NVFSCSVMH EALHN HYTQK SLSLSPGKHHHHHH CD20_Ofatumumabe MGWSCIILFLVATATGVHSEVQLVESGGGLVQPGRS LRLSCAASGFTFNDYAMHWVRQAPGKGLEWVSTIS WNSGSIGYADSVKGRFTISRDNAKKSLYLQMNSLRA EDTALYYCAKDIQYGNYYYGM DVWGQGTTVTVSSA SVAAPSVFIFPPSDEELKSGTASVVCLLNNFYPREAKV QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS KADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGA PIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL 100 CD22_m971-L7 RTYYRSTWYN DYAVSMKSRITINPDTNKNQFSLQLN SVTPEDTAVYYCAREVTGDLEDAFDIWGQGTMVTV SSASVAAPSVFIFPPSDEELKSGTASVVCLLNNFYPRE AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSST LTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP101 CD37_Lilotumab MGWSCIILFLVATATGVHSEIQLQQSGPELVKPGAS VKVSCKASGYSFTDYNMYWVKQSHGKSLEWIGYID PYNGDTTYNQKFKGKATLTVDKSSSTAFIHLNSLTSE DSAVYYCARSPYGHYAMDYWGQGTSVTVSSASVA APSVFIFPPSDEELKSGTASVVCLLNNFYPREAKVQW KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCP PCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVV DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST LSPGKHHHHHH Petition 870250089762, dated 02 / 10 / 2025, pages 420 / 580 60 / 176 Descrição Sequência SEQ ID NO: BCMA_11D5 MGWSCIILFLVATATGVHSQIQLVQSGPELKKPGET VKISCKASGYTFTDYSINWVKRAPGKGLKWMGWIN TETREPAYAYDFRGR FAFSLETSASTAYLQINNLKYED TATYFCALDYSYAMDYWGQGTSVTVSSASVAAPSV FIFPPSDEELKSGTASVVCLLNNFYPREAKVQWKVD NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCP APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKA KGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYDTTPPVLDSDGSFFLYSDLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG KHHHHHH 103 BCMA_CA8_J9M0 MGWSCIILFLVATATGVHSQVQLVQSGAEVKKPGSS VKVSCKGSGYTFTNYWMHWVRQAPGQGLEWIGA TYRGHSDTYYNQKFKGRATLTADTSTSTAYMELSSLR SEDTAVYYCTRGAIYDGYDVLDNWGQGTLVTVSSA SVAAPSVFIFPPSDEELKSGTASVVCLLNNFYPREAKV QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS KADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGA PIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL VKGFYPSDIAVEWESNGQPENNYDTTPPVLDSDGSFFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPGKHHHHHH 104 CD138_nBT062 MGWSCIILFLVATATGVHSQVQLQQSGSELM M PG ASVKISCKATGYTFSNYWIEWVKQRPGHGLEWIGEI LPGTGRTIYNEKFKGKATFTADISSNTVQMQLSSLTS EDSAVYYCARRDYYGN FYYAM DYWGQGTSVTVSS ASVAAPSVFIFPPSDEELKSGTASVVCLLNNFYPREAK VQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL SKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKT HTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALG APIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYDTTPPVLDSDGS FFLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPGKHHHHHH 105 CD38_Daratumomab MGWSCIILFLVATATGVHSEVQLLESGGGLVQPGGS LRLSCAVSGFTFNSFAMSWVRQAPGKGLEWVSAIS GSGGGTYYADSVKGRFTISRDNSKNTLYLQMNSLRA 106 Petition 870250089762, dated 02 / 10 / 2025, pp. 421 / 580 61 / 176 Descrição Sequência SEQ ID NO: EDTAVYFCAKDKILWFGEPVFDYWGQGTLVTVSSAS VAAPSVFIFPPSDEELKSGTASVVCLLNNFYPREAKV QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS KADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTH TCPPCPAPEEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGA PIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL VKGFYPSDIAVEWESNGQPENNYDTTPPVLDSDGSF FLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPGKHHHHHH CLL1_1075.7 MGWSCIILFLVATATGVHSDIQLQESGPGLVKPSQSL SLTCSVTGYSITSAYYWNWIRQFPGNKLEWMGYISY DGRNNYNPSLKNRISITRDTSKNQFFLKLNSVTTEDT ATYYCAKEGDYDVGNYYAMDYWGQGTSVTVSSAS VAAPSVFIFPPSDEELKSGTASVVCLLNNFYPREAKV QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS KADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTH TCPPCPAPEEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGA PIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL VKGFYPSDIAVEWESNGQPENNYDTTPPVLDSDGSF FLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPGKHHHHHH 107 ROR1_R12 MGWSCIILFLVATATGVHSQEQLVESGGRLVTPGGSLTLSCKASGFDFSAYYMSWVRQAPGKGLEWIATIYP SSGKTYYATWV NGRFTISSDNAQNTVD LQM N SLTA ADRATYFCARDSYADDGALFNIWGPGTLVTISSASV AAPSVFIFPPSDEELKSGTASVVCLLNNFYPREAKVQ WKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPI EKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYDTTPPVLDSDGSFF LYSDLTVDKSRWQQG NVFSCSVMH EALHN HYTQK SLSLSPGKHHHHHH 108 Cadeia leve CROSS-MAB HOL E CD33_Gemtuzumabe METDTLLLWVLLLWVPGSTGDIQLTQSPSTLSASVG DRVTITCRASESLDNYGIRFLTWFQQKPGKAPKLLM YAASNQGSGVPSRFSGSGSGTEFTLTISSLQPDDFAT YYCQQTKEVPWSFGQGTKVEVKSSASTKGPSVFPLA PSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS 109 Petição 870250089762, de 02 / 10 / 2025, pág. 422 / 580 62 / 176 Descrição Sequência SEQ ID NO: GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV NHKPSNTKVDKKVEPKSC CD123_clone_32716 METDTLLLWVLLLWVPGSTGDIVLTQSPASLAVSLG QRATISCRASESVDNYGNTFMHWYQQKPGQPPKLL IYRASNLESGIPARFSGSGSRTDFTLTINPVEADDVAT YYCQQSNEDPPTFGAGTKLELKSSASTKGPSVFPLAP SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC 110 CD70_Cusatuzumabe METDTLLLWVLLLWVPGSTGQAVVTQEPSLTVSPG GTVTLTCGLKSGSVTSDNFPTWYQQTPGQAPRLLIY NTNTRHSGVPDRFSGSILGNKAALTITGAQADDEAE YFCALFISNPSVEFGGGTQLTVLSSASTKGPSVFPLAP SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC 111 LewisY_3S193_MuVH METDTLLLWVLLLWVPGSTGDVLMTQTPLSLPVSL GDQASISCRSSQRIVHSNGNTYLEWYLQKPGQSPKL LIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLG VYYCFQGSHVPFTFGSGTKLEIKSSASTKGPSVFPLAP SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC 112 CD276_hu8H9 METDTLLLWVLLLWVPGSTGEIVMTQSPATLSVSPG ERVTLSCRASQSISDYLYWYQQKSHESPRLLIKYASQSISGIPARFSGSGSGSEFTLTINSVEPEDVGVYYCQNG HSFPLTFGQGTKLELKSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSC 113 CD135_4G8 METDTLLLWVLLLWVPGSTGDIVLTQSPATLSVTPG DSVSLSCRASQSISNNLHWYQQKSHESPRLLIKYASQ SISGIPSRFSGSGSGTDFTLSINSVETEDFGVYFCQQS NTWPYTFGGGTKLEIKSSASTKGPSVFPLAPSSKSTS GGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN TKVDKKVEPKSC 114 CD19_Tafasitamabe METDTLLLWVLLLWVPGSTGDIVMTQSPATLSLSPG ERATLSCRSSKLQNVNGNTYLYWFQQKPGQSPQL LIYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFA VYYCMQHLEYPITFGAGTKLEIKSSASTKGPSVFPLAP SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSC 115 CD20_Ofatumumab METDTLLLWVLLLWVPGSTGEIVLTQSPATLSLSPGE 116 Petition 870250089762, de 02 / 10 / 2025, pág. 423 / 580 63 / 176 Description Sequência SEQ ID NO: RATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASN RATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR SNWPITFGQGTRLEIKSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSC CD22_m971-L7 METDTLLLWVLLLWVPGSTGDIQMIQSPSSLSASVG DRVTITCRASQTIWSYLNWYRQRPGEAPNLLIYAASS LQSGVPSRFSGRGSGTDFTLTISSLQAEDFATYYCQQ SYSIPQTFGQGTKLEIKSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSC 117 CD37_Lilotumab METDTLLLWVLLLWVPGSTGDIVMTQSHKLLSTSV GDRVSITCKASQDVSTAVDWYQQKPGQSPKLLINW ASTRHTGVPDRFTGSGSGTDYTLTISSMQAEDLALY YCRQHYSTPFTFGSGTKLEIKSSASTKGPSVFPLAPSS KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP SNTKVDKKVEPKSC 118 BCMA_11D5 METDTLLLWVLLLWVPGSTGDIVLTQSPPSLAMSLG KRATISCRASESVTILGSHLIHWYQQKPGQPPTLLIQL ASNVQTGVPARFSGSGSRTDFTLITDPVEEDDVAVY YCLQSRTIPRTFGGGTKLEIKSSASTKGPSVFPLAPSSK STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHT FPAVLQSSGLYSLSSVVTVPSSLGTQTYICNVNHKPS NTKVDKKVEPKSC119 BCMA_CA8_J9M0 METDTLLLWVLLLWVPGSTGDIQMTQSPSSLSASV GDRVTITCSASQDISNYLNWYQQKPGKAPKLLIYYTS NLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQ QYRKLPWTFGQGTKLEIKSSASTKGPSVFPLAPSSKS TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF PAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPS NTKVDKKVEPKSC 120 CD138_nBT062 METDTLLLWVLLLWVPGSTGDIQMTQSTSSLSASLG DRVTISCSASQGINNYLNWYQQKPDGTVELLIYYTST LQSGVPSRFSGSGSGTDYSLTISNLEPEDIGTYYCQQY SKLPRTFGGGTKLEIKSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSC 121 CD38_Daratumomab METDTLLLWVLLLWVPGSTGEIVLTQSPATLSLSPGE RATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASN RATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR SNWPPTFGQGTKVEIKSSASTKGPSVFPLAPSSKSTS 122 Petition 870250089762, dated 02 / 10 / 2025, pp. 424 / 580 64 / 176 Description Sequence SEQ ID NO: GGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN TKVDKKVEPKSC CLL1_1075.7 METDTLLLWVLLLWVPGSTGENVLTQSPAIMSASP GEKVTMTCRASSNVISSYVHWYQQRSGASPKLWIY STSNLASGVPARFSGSGSGTSYSLTISSVEAEDAATYY CQQYSGYPLTFGAGTKLELKSSASTKGPSVFPLAPSS KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP SNTKVDKKVEPKSC 123 ROR1_R12 METDTLLLWVLLLWVPGSTGELVLTQSPSVSAALGS PAKITCTLSSAHKTDTIDWYQQLQGEAPRYLMQVQ SDGSYTKRPGVPDRFSGSSSGADRYLIIPSVQADDEA DYYCGADYIGGYVFGGGTQLTVTGSSASTKGPSVFP LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGAL TSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSC 124 Exemplary linking domain E200 (P2X7) for inclusion in a CAR (BIL03) EVQLLESGGGLVQPGGSLRLSCAASGFTFRNHDMG WVRQAPGKGLEWVSAISGSGGSTYYANSVKGRFTIS RDNSKNTLYLQMNSLRAEDTAVYYCAEPKPMDTEF DYRSPGTLVTVSS 286 CD276 clone 376.96 CD276 clone 376.96 Cadeia pesada CROSS-MAB KNOB MGWSCIILFLVATATGVHSEVQLVESGGGLVKPGGS LKLSCEASRFTFSSYAMSWVRQTPEKRLEWVAAISG GGRYTYYPDSMKGRFTISRDNAKNFLYLQMSSLRSE DTAMYYCARHYDGYLDYWGQGTTLTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSWNS GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDEKVEPKSCDKTHTCPPCPAPE AAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAK GQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK HHHHHH 299 CD276 clone 376.96 Cadeia pesada CROSS-MAB KNOB_E200 MGWSCIILFLVATATGVHSGHNYTTRNILPGLNITSE VQLVESGGGLVKPGGSLKLSCEASRFTFSSYAMSWV RQTPEKRLEWVAAISGGGRYTYYPDSMKGRFTISRD NAKNFLYLQMSSLRSEDTAMYYCARHYDGYLDYW GQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGC LVEDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDEKVEP KSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMIS RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT 300. Petição 870250089762, de 02 / 10 / 2025, pág. 425 / 580 65 / 176Descrição Sequência SEQ ID NO: KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS NKALGAPIEKTISKAKGQPREPQVYTLPPSRKELTKN QVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPV LKSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPGKHHHHHH CD276 clone 376.96 Cadeia pesada CROSS-MAB KNOB_E200-NO-26 MGWSCIILFLVATATGVHSDFPGHNYTTRNILPGLNI TSGSEPKSSDKTHTGSEVQLVESGGGLVKPGGSLKLS CEASRFTFSSYAMSWVRQTPEKRLEWVAAISGGGR YTYYPDSMKGRFTISRDNAKNFLYLQMSSLRSEDTA MYYCARHYDGYLDYWGQGTTLTVSSASTKGPSVFP LAPSSKSTSGGTAALGCLVEDYFPEPVTVSWNSGAL TSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDEKVEPKSCDKTHTCPPCPAPEAAG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTV LHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPR EPQVYTLPPSRKELTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGKHHHH HH 301 CD276 clone 376.96 Cadeia leve CROSS-MAB KNOB METDTLLLWVLLLWVPGSTGDIVMTQSHKFMSTSI GARVSITCKASQDVRTAVAWYQQKPGQSPKLLIYSA SYRYTGVPDRFTGSGSGTDFTFTISSVQAEDLAVYYC QQHYGTPPWTFGGGTKLEIKRTVAAPSVFIFPPSDR KLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGN SQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC 302 CD276 clone 376.96 Cadeia leve CROSS-MAB KNOB_E200 METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SDIVMTQSHKFMSTSIGARVSITCKASQDVRTAVA WYQQKPGQSPKLLIYSASYRYTGVPDRFTGSGSGTD FTFTISSVQAEDLAVYYCQQHYGTPPWTFGGGTKLEI KRTVAAPSVFIFPPSDRKLKSGTASVVCLLNNFYPRE AKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSST LTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 303 CD276 clone 376.96 Cadeia leve CROSS-MAB KNOB_E200-NO-26 METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSDIVMTQSHKFMSTSIGARV SITCKASQDVRTAVAWYQQKPGQSPKLLIYSASYRYT GVPDRFTGSGSGTDFTFTISSVQAEDLAVYYCQQHY GTPPWTFGGGTKLEIKRTVAAPSVFIFPPSDRKLKSG TASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG LSSPVTKSFNRGEC 304 CD276 clone 376.96 Heavy chain CROSS-MAB HOLE MGWSCIILFLVATATGVHSEVQLVESGGGLVKPGGS LKLSCEASRFTFSSYAMSWVRQTPEKRLEWVAAISG GGRYTYYPDSMKGRFTISRDNAKNFLYLQMSSLRSE 305. Petition 870250089762, dated 02 / 10 / 2025, pp. 426 / 580 66 / 176 Descrição Sequência SEQ ID NO: DTAMYYCARHYDGYLDYWGQGTTLTVSSASVAAPS VFIFPPSDEELKSGTASVVCLLNNFYPREAKVQWKV DNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE KHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVD VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTIS KAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYDTTPPVLDSDGSFFLYSD LTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGKHHHHHH CD276 clone 376.96 Cadeia leve CROSS-MAB HOLE METDTLLLWVLLLWVPGSTGDIVMTQSHKFMSTSI GARVSITCKASQDVRTAVAWYQQKPGQSPKLLIYSA SYRYTGVPDRFTGSGSGTDFTFTISSVQAEDLAVYYC QQHYGTPPWTFGGGTKLEIKSSASTKGPSVFPLAPS SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGV HTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSC 306 CLL1 (CD371) clone 1075.7 CLL1 (CD371) clone 1075.7 Cadeia pesada CROSS-MAB KNOB MGWSCIILFLVATATGVHSDIQLQESGPGLVKPSQSL SLTCSVTGYSITSAYYWNWIRQFPGNKLEWMGYISY DGRNNYNPSLKNRISITRDTSKNQFFLKLNSVTTEDT ATYYCAKEGDYDVGNYYAMDYWGQGTSVTVSSAS TKGPSVFPLAPSSKSTSGGTAALGCLVEDYFPEPVTV SWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDEKVEPKSCDKTHTCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVD VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTIS KAKGQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP GKHHHHHH 307 CLL1 (CD371) clone 1075.7 Cadeia pesada CROSS-MAB KNOB_E200 MGWSCIILFLVATATGVHSGHNYTTRNILPGNLITSD IQLQESGPGLVKPSQSLSLTCSVTGYSITSAYWNWI RQFPGNKLEWMGYISYDGRNNYNPSLKNRISITRDT SKNQFFLKLNSVTTEDTATYYCAKEGDYDVGNYYA MDYWGQGTSSVTVSSASTKGPSVFPLASSKSTSGG TAALGCLVEDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKV DEKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPK DTLMISRTPEVTVCVVDVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE YCKKVSNKALGAPIEKTISKAGQPREPQVYTLPPSR KELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNY 308. Petition 870250089762, de 02 / 10 / 2025, pág. 427 / 580 67 / 176 Descrição Sequência SEQ ID NO: KTTPPVLKSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGKHHHHHH CLL1 (CD371) clone 1075.7 Cadeia pesada CROSS-MAB KNOB_E200-NO-26 MGWSCIILFLVATATGVHSDFPGHNYTTRNILPGLNI TSGSEPKSSDKTHTGSDIQLQESGPGLVKPSQSLSLT CSVTGYSITSAYYWNWIRQFPGNKLEWMGYISYDG RNNYNPSLKNRISITRDTSKNQFFLKLNSVTTEDTATY YCAKEGDYDVGNYYAMDYWGQGTSVTVSSASTKG PSVFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ TYICNVNHKPSNTKVDEKVEPKSCDKTHTCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAK GQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK HHHHHH 309 CLL1 (CD371) clone 1075.7 Cadeia leve CROSS-MAB KNOB METDTLLLWVLLLWVPGSTGENVLTQSPAIMSASP GEKVTMTCRASSNVISSYVHWYQQRSGASPKLWIY STSNLASGVPARFSGSGSGTSYSLTISSVEAEDAATYY CQQYSGYPLTFGAGTKLELKRTVAAPSVFIFPPSDRK LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNS QESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC 310 CLL1 (CD371) clone 1075.7 Cadeia leve CROSS-MAB KNOB_E200 METDTLLLWVLLLWVPGSTGGHNYTTRNILPGLNIT SENVLTQSPAIMSASPGEKVTMTCRASSNVISSYVH WYQQRSGASPKLWIYSTSNLASGVPARFSGSGSGTS YSLTISSVEAEDAATYYCQQYSGYPLTFGAGTKLELKR TVAAPSVFIFPPSDRKLKSGTASVVCLLNNFYPREAK VQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL SKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 311 CLL1 (CD371) clone 1075.7 Cadeia leve CROSS-MAB KNOB_E200-NO-26 METDTLLLWVLLLWVPGSTGDFPGHNYTTRNILPGL NITSGSEPKSSDKTHTGSENVLTQSPAIMSASPGEKV TMTCRASSNVISSYVHWYQQRSGASPKLWIYSTSNL ASGVPARFSGSGSGTSYSLTISSVEAEDAATYYCQQY SGYPLTFGAGTKLELKRTVAAPSVFIFPPSDRKLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGL SSPVTKSFNRGEC 312 CLL1 (CD371) clone 1075.7 Cadeia pesada CROSS-MAB HOLE MGWSCIILFLVATATGVHSDIQLQESGPGLVKPSQSL SLTCSVTGYSITSAYYWNWIRQFPGNKLEWMGYISY DGRNNYNPSLKNRISITRDTSKNQFFLKLNSVTTEDT ATYYCAKEGDYDVGNYYAMDYWGQGTSVTVSSAS VAAPSVFIFPPSDEELKSGTASVVCLLNNFYPREAKV QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS 313. Petição 870250089762, de 02 / 10 / 2025, pág. 428 / 580 68 / 176 Descrição Sequência SEQ ID NO: KADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGA PIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL VKGFYPSDIAVEWESNGQPENNYDTTPPVLDSDGSF FLYSDLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPGKHHHHHH CLL1 (CD371) clone 1075.7 Cadeia leve CROSS-MAB HOLE METDTLLLWVLLLWVPGSTGENVLTQSPAIMSASP GEKVTMTCRASSNVISSYVHWYQQRSGASPKLWIY STSNLASGVPARFSGSGSGTSYSLTISSVEAEDAATYY CQQYSGYPLTFGAGTKLELKSSASTKGPSVFPLAPSS KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP SNTKVDKKVEPKSC 314 Table 2: Exemplary sequences of epitope fractions and exemplary heterodimeric molecules for use with “universal CAR” or alternative CAR systems. Description Sequence SEQ ID NO: Exemplary PNE peptide tag + CD19 Tafasitamab, heavy chain, knob (“left arm of antibody”),presentation N-terminal da etiqueta de peptídeo Sequência de etiqueta PNE NYHLENEVARLKKL 125 CD19_Tafa_HC_KNOB _N-TERMINAL_PNEG4S_LALA_P329G NYHLENEVARLKKL GGGGS EVQLVESGGGLVKPGGSLKLS CAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKY NEKFQGRVTISSSDKSISTAYMELSSLRSEDTAMYYCARGTY YYGTRVFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVEDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDEKVEPK SCDKTHTCPPCPAPEAAGGPSVFLFPKPKPKDTLMISRTPEV TVCVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTIK AKGQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSDIAV EWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGKHHHHHH 126 CD19_Tafa_LC_KNOB_ N-TERMINAL_PNE-G4S NYHLENEVARLKKL GGGGS DIVMTQSPATLSLSPGERATLS CRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIIRMSNLNS GVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITF GAGTKLEIKRTVAAPSVFIFPPSDRKLKSGTASVVCLLNNFY PREAKVQWKVDNALQGSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 127 CD19_Tafa_HC_N- NYHLENEVARLKKLGGGGSEVQLVESGGGLVKPGGSLKLS 128, Petition 870250089762, dated 02 / 10 / 2025, pp. 429 / 580 69 / 176 Descrição Sequência SEQ ID NO: TERMINAL_PNE-G4S CAASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKY NEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTY YYGTRVFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPK SCDKTHTCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVT CVVVGVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYQS TYRVVSVLTVLHQDWLNGKEYKCKVSNKQLPSPIEKTISKA KGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ GNVFSCSVMHEALHNHYTQKSLSLSPGKHHHHHH CD19_Tafa_LC_N- TERMINAL_PNE-G4S NYHLENEVARLKKLGGGGSDIVMTQSPATLSLSPGERATLS CRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNS GVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITF GAGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECHHHHH H 129 CD33_LIN_HC_KNOB_ N-TERMINAL_PNEG4S_LALA_P329G NYHLENEVARLKKL GGGGS QVQLVQSGAEVKKPGSSVKV SCKASGYTFTDYNMHWVRQAPGQGLEWIGYIYPYNGGT GYNQKFKSKATITADESTNTAYMELSSLRSEDTAVYYCARG RPAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDEKVEPKSC DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAK GQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSPGKHHHHHH 130 CD33_LIN_LC_KNOB_ N-TERMINAL_PNE-G4S NYHLENEVARLKKL GGGGS DIQMTQSPSSLSASVGDRVTI TCRASESVDNYGISFMNWFQQKPGKAPKLLIYAASNQGS GVPSRFSGSGSGTDFTLTISSLQPDDFATYYCQQSKEVPWT FGQGTKVEIKRTVAAPSVFIFPPSDRKLKSGTASVVCLLN N F YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 131 CD33_LIN_HC_N- TERMINAL_PNE-G4S NYHLENEVARLKKL GGGGS QVQLVQSGAEVKKPGSSVKV SCKASGYTFTDYNMHWVRQAPGQGLEWIGYIYPYNGGT GYNQKFKSKATITADESTNTAYMELSSLRSEDTAVYYCARG RPAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGT AALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC DKTHTCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCV VVGVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYQSTYRVVSVLTVLHQDWLNGKEYKCKVSNKQLPSPIEKTISKAKG 132 Petition 870250089762, dated 02 / 10 / 2025, pp. 430 / 580 70 / 176 Description Sequence SEQ ID NO: QPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWE SNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPGKHHHHHH CD33_LIN_LC_N- TERMINAL_PNE-G4S NYHLENEVARLKKL GGGGS DIQMTQSPSSLSASVGDRVTI TCRASESVDNYGISFMNWFQQKPGKAPKLLIYAASNQGS GVPSRFSGSGSGTDFTLTISSLQPDDFATYYCQQSKEVPWT FGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNN FYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSST LTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECHHHH HH 133 tag sequence EGFRvIII LEEKKGNYVVTDH 134 tag sequence EGFRvIII LEEKK 280 tag sequence EGFRvIII NYVVTDH 281 CD19_Tafa_HC_KNOB _NTERMINAL_EGFRvIIIG4S_LALA_P329G LEEKKGNYVVTDH GGGGS EVQLVESGGGLVKPGGSLKLSC AASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKY NEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTY YYGTRVFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVEDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDEKVEPK SCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSDIAV EWESNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGKHHHHHH 135 CD19_Tafa_LC_KNOB_ NTERMINAL_EGFRvIIIG4S LEEKKGNYVVTDH GGGGS DIVMTQSPATLSLSPGERATLS CRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNS GVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITF GAGTKLEIKRTVAAPSVFIFPPSDRKLKSGTASVVCLLNNFY PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 136 CD19_Tafa_HC_NTERMINAL_EGFRvIIIG4S LEEKKGNYVVTDH GGGGS EVQLVESGGGLVKPGGSLKLSC AASGYTFTSYVMHWVRQAPGKGLEWIGYINPYNDGTKY NEKFQGRVTISSDKSISTAYMELSSLRSEDTAMYYCARGTY YYGTRVFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPK SCDKTHTCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVT CVVVGVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYQS TYRVVSVLTVLHQDWLNGKEYKCKVSNKQLPSPIEKTISKA KGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ 137 Petição 870250089762, de 02 / 10 / 2025, pág. 431 / 580 71 / 176 Descrição Sequência SEQ ID NO: GNVFSCSVMHEALHNHYTQKSLSLSPGKHHHHHH CD19_Tafa_LC_NTERMINAL_EGFRvIIIG4S LEEKKGNYVVTDH GGGGS DIVMTQSPATLSLSPGERATLS CRSSKSLQNVNGNTYLYWFQQKPGQSPQLLIYRMSNLNS GVPDRFSGSGSGTEFTLTISSLEPEDFAVYYCMQHLEYPITF GAGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECHHHHH H 138 CD33_LIN_HC_KNOB_ NTERMINAL_EGFRvIIIG4S_LALA_P329G LEEKKGNYVVTDH GGGGS QVQLVQSGAEVKKPGSSVKVS CKASGYTFTDYNMHWVRQAPGQGLEWIGYIYPYNGGTG YNQKFKSKATITADESTNTAYMELSSLRSEDTAVYYCARGR PAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVEDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDEKVEPKSCD KTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKG QPREPQVYTLPPSRKELTKNQVSLTCLVKGFYPSDIAVEWE SNGQPENNYKTTPPVLKSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPGKHHHHHH 139 CD33_LIN_LC_KNOB_ NTERMINAL_EGFRvIIIG4S LEEKKGNYVVTDH GGGGS DIQMTQSPSSLSASVGDRVTITCRASESVDNYGISFMNWFQQKPGKAPKLLIYAASNQGSG VPSRFSGSGSGTDFTLTISSLQPDDFATYYCQQSKEVPWTF GQGTKVEIKRTVAAPSVFIFPPSDRKLKSGTASVVCLLNNFY PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 140 CD33_LIN_HC_NTERMINAL_EGFRvIIIG4S LEEKKGNYVVTDH GGGGS QVQLVQSGAEVKKPGSSVKVS CKASGYTFTDYNMHWVRQAPGQGLEWIGYIYPYNGGTG YNQKFKSKATITADESTNTAYMELSSLRSEDTAVYYCARGR PAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD KTHTCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVV VGVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYQSTYR VVSVLTVLHQDWLNGKEYKCKVSNKQLPSPIEKTISKAKG QPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWE SNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPGKHHHHHH 141 CD33_LIN_LC_NTERMINAL_EGFRvIIIG4S LEEKKGNYVVTDH GGGGS DIQMTQSPSSLSASVGDRVTIT CRASESVDNYGISFMNWFQQKPGKAPKLLIYAASNQGSG VPSRFSGSGSGTDFTLTISSLQPDDFATYYCQQSKEVPWTF GQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECHHHHH H 142 Petition 870250089762, dated 02 / 10 / 2025, pp. 432 / 580 72 / 176 Description Sequence SEQ ID NO: CAR directed to EGFRvIII scFv_(HLconfiguration_clone_1 39)_CD8a_CD8TM_CD 28_CD137_CD3zeta EVQVLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQ APGKGLEWVSAISGSGGSTNYADSVKGRFTISRDNSKNTL YLQMN SLRAEDTAVYYCAGSSGWSEYWGQGTLVTVSSG GGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRA SQGIRNNLAWYQQKPGKAPKRLIYAASNLQSGVPSRFTGS GSGTEFTLIVSSLQPEDFATYYCLQHHSYPLTSGGGTKVEIK TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLD FACDIYIWAPLAGTCGVLLLSLVITLYCRSKRSRLLHSDYMN MTPRRPGPTRKHYQPYAPPRDFAAYRSKRGRKKLLYIFKQ PFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAP AYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKP RRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHD GLYQGLSTATKDTYDALHMQALPPR 285 CLDN6 tag sequence WTAHAIIRDFYNPLVAEAQKREL 143 CLDN6 peptide 13mer TAHAIIRDFYNPL 282 CLDN6 peptide 10mer LVAEAQKREL 283 CD19_Tafa_HC_KNOB _N-TERMINAL_CLDN6G4S_LALA_P329G WTAHAIIRDFYNPLVAEAQKREL GGGGS EVQLVESGGGLV KPGGSLKLSCAASGYTFTSYVMHWVRQAPGKGLEWIGYI NPYNDGTKYNEKFQGRVTISSDKSISTAYMELSSLRSEDTA MYYCARGTYYYGTRVFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVEDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDEKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALG APIEKTISKAKGQPREPQVYTLPPSRKELTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKHHH HHH 144 CD19_Tafa_LC_KNOB_ N-TERMINAL_CLDN6G4S WTAHAIIRDFYNPLVAEAQKREL GGGGS DIVMTQSPATLS LSPGERATLSCRSSKSLQNVNGNTYLYWFQQKPGQSPQLL IYRMSNLNSGVPDRFSGSGSGTEFTLTISSLEPEDFAVYYC MQHLEYPITFGAGTKLEIKRTVAAPSVFIFPPSDRKLKSGTA SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSK DSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNR GEC 145 CD33_LIN_HC_KNOB_ N-TERMINAL_CLDN6G4S_LALA_P329G WTAHAIIRDFYNPLVAEAQKREL GGGGS QVQLVQSGAEV KKPGSSVKVSCKASGYTFTDYNMHWVRQAPGQGLEWIG YIYPYNGGTGYNQKFKSKATITADESTNTAYMELSSLRSED TAVYYCARGRPAMDYWGQGTLVTVSSASTKGPSVFPLAP SSKSTSGGTAALGCLVEDYFPEPVTVSWNSGALTSGVHTF PAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKV 148 Petição 870250089762, de 02 / 10 / 2025, pág. 433 / 580 73 / 176 Descrição Sequência SEQ ID NO: DEKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALG APIEKTISKAKGQPREPQVYTLPPSRKELTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLKSDGSFFLYSKLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKHHH HHH CD33_LIN_LC_KNOB_ N-TERMINAL_CLDN6G4S WTAHAIIRDFYNPLVAEAQKREL GGGGS DIQMTQSPSSLS ASVGDRVTITCRASESVDNYGISFMNWFQQKPGKAPKLLI YAASNQGSGVPSRFSGSGSGTDFTLTISSLQPDDFATYYCQ QSKEVPWTFGQGTKVEIKRTVAAPSVFIFPPSDRKLKSGTA SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSK DSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNR GEC 149 CD33_LIN_HC_NTERMINAL_CLDN6G4S WTAHAIIRDFYNPLVAEAQKREL GGGGS QVQLVQSGAEV KKPGSSVKVSCKASGYTFTDYNMHWVRQAPGQGLEWIG YIYPYNGGTGYNQKFKSKATITADESTNTAYMELSSLRSED TAVYYCARGRPAMDYWGQGTLVTVSSASTKGPSVFPLAP SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF PAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKV DKKVEPKSCDKTHTCPPCPAPPVAGPSVFLFPPKPKDTLMI SRTPEVTCVVVGVSHEDPEVKFNWYVDGVEVHNAKTKPR EEQYQSTYRVVSVLTVLHQDWLNGKEYKCKVSNKQLPSPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKHHHH HH 150 CD33_LIN_LC_NTERMINAL_CLDN6G4S WTAHAIIRDFYNPLVAEAQKREL GGGGS DIQMTQSPSSLS ASVGDRVTITCRASESVDNYGISFMNWFQQKPGKAPKLLI YAASNQGSGVPSRFSGSGSGTDFTLTISSLQPDDFATYYCQ QSKEVPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTA SVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSK DSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNR GECHHHHHH 151 CAR direcionado a CLDN6 scFv_(HLconfiguration_clone_A b34)_CD8a_CD8TM_CD2 8_CD137_CD3zeta EVQLLESGGGLVQPGGSMRLSCAASGFTFSNYWMNWV RQAPGKGLEWVAQIRLKSDNYATHYADSVKGRFTISRDDS KNTVYLQMNSLRAEDTGVYYCNDGPPSGYWGQGTLLTV SSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTIT CRISENIYSYLAWYQQKPGKAPKLLIYNAKILVEGVPSRFSG SGSGTDFTLTISSLQPEDFATYYCQHHYTVPWTFGQGTKLE IKTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRG LDFACDIYIWAPLAGTCGVLLLSLVITLYCRSKRSRLLHSDY MNMTPRRPGPTRKHYQPYAPPRDFAAYRSKRGRKKLLYIF KQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSAD APAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGG 284 Petition 870250089762, dated 02 / 10 / 2025, pp. 434 / 580 74 / 176 Description Sequence SEQ ID NO: KPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGH DGLYQGLSTATKDTYDALHMQALPPR Table 3: P2X sequences and epitope fractions thereof. Description Sequence SEQ ID NO: Exemplary sequences of P2X receptor epitope fraction? Human P2X7 receptor MPACCSCSDVFQYETNKVTRIQSMNYGTIKWFFHVIIFSYVCF ALVSDKLYQRKEPVISSVHTKVKGIAEVKEEIVENGVKKLVHSV FDTADYTFPLQGNSFFVMTNFLKTEGQEQRLCPEYPTRRTLCS SDRGCKKGWMDPQSKGIQTGRCVVYEGNQKTCEVSAWCPI EAVEEAPRPALLNSAENFTVLIKNNIDFPGHNYTTRNILPGLNI TCTFHKTQNPQCPIFRLGDIFRETGDNFSDVAIQGGIMGIEIY WDCNLDRWFHHCRPKYSFRRLDDKTTNVSLYPGYNFRYAKY YKENNVEKRTLIKVFGIRFDILVFGTGGKFDIIQLVVYIGSTLSYF GLAAVFIDFLIDTYSSNCCRSHIYPWCKCCQPCVVNEYYYRKKC ESIVEPKPTLKYVSFVDESHIRMVNQQLLGRSLQDVKGQEVPR PAMDFTDLSRLPLALHDTPPIPGQPEEIQLLRKEATPRSRDSPV WCQCGSCLPSQLPESHRCLEELCCRKKPGACITTSELFRKLVLS RHVLQFLLLYQEPLLALDVDSTNSRLRHCAYRCYATWRFGSQ DMADFAILPSCCRWRIRKEFPKSEGQYSGFKSPY 152 E200 exemplary epitope GHNYTTRNILPGLNITC 153 E200 variant epitope peptide (E200') GHNYTTRNILPGLNIT 154 Extended E200 Cys modification to Ser GHNYTTRNILPGLNITS 155 Extended E200 Cys modification to Ser GHNYTTRNILPGLNITSTFHK 156 E200' extended modification Cys to Ser(22 aa) GHNYTTRNILPGLNITSTFHKT 157 Extended E200 Cys modification to Ser GHNYTTRNILPGLNITSTFHKTC 158 Pep16 DFPGHNYTTRNILPGC 159 Pep17 GHNYTTRNILPGLNITSTFHKTS 160 Extended Pep17 (27 aa) GHNYTTRNILPGLNITSTFHKTSGSGK 161 Minimum sequence of E200 peptide NYTTRNILPGL 162 Petition 870250089762, dated 02 / 10 / 2025, pp. 435 / 580 75 / 176 Description Sequence SEQ ID NO: (target epitope) E300 exemplary epitope KYYKENNVEKRTLIK 163 E300 epitope variant peptide (ESQ') KYYKENNVEKRTLIKVF 164 E200 / E300 epitope or exemplary composite GHNYTTRNILPGAGAKYYKENNVEK 165 E200 + G4S ligand GHNYTTRNILPGLNITSGGGGS 166 E200 + 2xG4S ligand GHNYTTRNILPGLNITSGGGGSGGGGS 167 E200 + 3xG4S ligand GHNYTTRNILPGLNITSGGGGSGGGGSGGGGS 168 Extended E200 peptide 17v3 (24 aa) GHNYTTRNILPGLNITSTFHKTGS 169 Peptide E200_extended 17v4 (22 aa) GHNYTTRNILPGLNITSTFHGS 170 E200_extended peptide 17v5 (19 aa) GHNYTTRNILPGLNITSGS 171 E200_extended peptide 17v6 (22 aa) (extended E200 epitope in the N-terminal region) DFPGHNYTTRNILPGLNITSGS 172 E200_extended peptide 17v7 (25 aa) + ligand DFPGHNYTTRNILPGLNITSGGGGS 173 E200_extended peptide 17v8 (30 aa) + ligand DFPGHNYTTRNILPGLNITSGGGGSGGGGS 174 E200_extended peptide 17v9 (35 aa) + ligand DFPGHNYTTRNILPGLNITSGGGGSGGGGSGGGGS 175 Peptide E200_extended 17v10 (30 aa) DFPGHNYTTRNILPGLNITSTFHKTSGSGK 176Extended E200 peptide 17v11(32aa) + DFPGHNYTTRNILPGLNITSTFHKTSGSGKGS ligand 177 Extended E200 peptide DFPGHNYTTRNILPGLNITSTFHKTSGSGKGGGGS 178 Petition 870250089762, dated 02 / 10 / 2025, pp. 436 / 580 76 / 176 Description Sequence SEQ ID NO: 17v12 (35 aa) + ligand E200_extended peptide 17v13 (25 aa) + ligand DFPGHNYTTRNILPGLNITSTFHGGGGS 179 E200_extended peptide 17v14 (22 aa) + ligand GHNYTTRNILPGLNITSTFHGGGGS 180 E200_extended peptide 17v15 (30 aa) + ligand DFPGHNYTTRNILPGLNITSTFHKTGGGGS 181 E200_extended peptide 17v16 (27 aa) + GHNYTTRNILPGLNITSTFHKTGGGGS 182 E200+ hinge IgG (E200 underlined) GHNYTTRNILPGLNITSEP KSSDKTHT 183 E200+ GS ligand_hinge IgG GHNYTTRNILPGLNITSGSEPKSSDKTHT 184 E200+ G4S ligand+hinge IgG GHNYTTRNILPGLNITSGGGGSEPKSSDKTHT 185 E200 extended+_hinge IgG GHNYTTRNILPGLNITSTFHKTSGSGKEPKSSDKTHT 186 E200 extended+ GS ligand+_hinge IgG GHNYTTRNILPGLNITSTFHKTSGSGKGSEPKSSDKTHT 187 E200 extended+ G4S ligand+hinge IgG GHNYTTRNILPGLNITSTFHKTSGSGKGGGGSEPKSSDKTHT 188 E200+hinge IgG+GS ligand GHNYTTRNILPGLNITSEPKSSDKTHTGS 189 E200+GS ligand+IgG hinge+GS ligand GHNYTTRNILPGLNITSGSEPKSSDKTHTGS 190 E200+G4S ligand+hingeIgG+ligand GS GHNYTTRNILPGLNITSGGGGSEPKSSDKTHTGS 191 E200 extended+_dobradiç a IgG+liganteGS GHNYTTRNILPGLNITSTFHKTSGSGKEPKSSDKTHTGS 192 E200 extended + ligand GS+_ GHNYTTRNILPGLNITSTFHKTSGSGKGSEPKSSDKTHTGS 193 Petition 870250089762, of 02 / 10 / 2025, p. 437 / 580 77 / 176 Description Sequence SEQ ID NO: hinge IgG + GS ligand E200 extended + G4S ligand + hinge IgG + GS ligand GHNYTTRNILPGLNITSTFHKTSGSGKGGGGSEPKSSDKTHTG S 194 E200+hinge IgG+G4S ligand GHNYTTRNILPGLNITSEPKSSDKTHTGGGGS 195 E200+GS ligand+hinge IgG+G4S ligand GHNYTTRNILPGLNITSGSEPKSSDKTHTGGGGS 196 E200+G4S ligand+hinge IgG+G4S ligand GHNYTTRNILPGLNITSGGGGSEPKSSDKTHTGGGGS 197 E200 extended+hinge IgG+G4S ligand GHNYTTRNILPGLNITSTFHKTSGSGKEPKSSDKTHTGGGGS 198 E200 extended+ligand GS+IgG hinge+G4S ligand GHNYTTRNILPGLNITSTFHKTSGSGKGSEPKSSDKTHTGGGG S 199 E200 extended+G4S ligand+IgG hinge+G4S ligand GHNYTTRNILPGLNITSTFHKTSGSGKGGGGSEPKSSDKTHTG GGGS 200 E200 extended Nterm +IgG hinge DFPGHNYTTRNILPGLNITSEPKSSDKTHT 201 E200 extended Nterm+GS ligand+IgG hinge DFPGHNYTTRNILPGLNITSGSEPKSSDKTHT 202 E200 extended Nterm +G4S ligand+IgG hinge DFPGHNYTTRNILPGLNITSGGGGSEPKSSDKTHT 203 E200 extended N and C term +IgG hingeDFPGHNYTTRNILPGLNITSTFHKTSGSGKEPKSSDKTHT 204 Extended E200 N and C term + GS ligand + IgG hinge DFPGHNYTTRNILPGLNITSTFHKTSGSGKGSEPKSSDKTHT 205 Extended E200 N and C term + G4S ligand + IgG hinge DFPGHNYTTRNILPGLNITSTFHKTSGSGKGGGGSEPKSSDKT HT 206 Extended E200 N term + IgG hinge + GS ligand DFPGHNYTTRNILPGLNITSEPKSSDKTHTGS 207 Petition 870250089762, dated 02 / 10 / 2025, pp. 438 / 580 78 / 176 Description Sequence SEQ ID NO: E200 extended Nterm+GS ligand+IgG hinge+GS ligand DFPGHNYTTRNILPGLNITSGSEPKSSDKTHTGS 208 E200 extended Nterm+G4S ligand+IgG hinge+GS ligand DFPGHNYTTRNILPGLNITSGGGGSEPKSSDKTHTGS 209 E200 extended Nterm and C term + IgG hinge+GS ligand DFPGHNYTTRNILPGLNITSTFHKTSGSGKEPKSSDKTHTGS 210 E200 extended Nterm and C term+GS ligand + IgG hinge+GS ligand DFPGHNYTTRNILPGLNITSTFHKTSGSGKGSEPKSSDKTHTGS 211 E200 extended Nterm and C term+G4S ligand +IgG hinge+GS ligand DFPGHNYTTRNILPGLNITSTFHKTSGSGKGGGGSEPKSSDKT HTGS 212 Extended E200 Nterm + hinge IgG + G4S ligand DFPGHNYTTRNILPGLNITSEPKSSDKTHTGGGGS 213 Extended E200 Nterm + GS ligand + hinge IgG + G4S ligand DFPGHNYTTRNILPGLNITSGSEPKSSDKTHTGGGGS 214 Extended E200 Nterm + G4S ligand + hinge IgG + G4S ligand DFPGHNYTTRNILPGLNITSGGGGSEPKSSDKTHTGGGGS 215 Extended E200 Nterm and C term + hinge IgG + G4S ligand DFPGHNYTTRNILPGLNITSTFHKTSGSGKEPKSSDKTHTGGGGS GS 216 Extended E200 Nterme C term+GS ligand + IgG hinge+G4S ligand DFPGHNYTTRNILPGLNITSTFHKTSGSGKGGSEPKSSDKTHTG GGGS 217 E200 extended Nterm and C term+G4S ligand DFPGHNYTTRNILPGLNITSTFHKTSGSGKGGGGSEPKSSDKT HTGGGGS 218 Petition 870250089762, dated 02 / 10 / 2025, pp. 439 / 580 79 / 176 Description Sequence SEQ ID NO: +IgG hinge+G4S ligand Extended E200 N-term and C-term for use at the C-terminal end of the light chain LONG SEQUENCE v1 GGGGSEPKSSDKTHTSPPSPAPELLGGGGSDFPGHNYTTRNIL PGLNITS 289 Extended E200 N-term and C-term for use at the C-terminal end of the light chain LONG SEQUENCE v2 GGGGSEPKSSDKTHTSPPSPAPELLGGGGSGHNYTTRNILPGL NITS 290 Extended E200 N-term and C-term for use at the C-terminal end of the light chain LONG SEQUENCE v3 EPKSSDKTHTSPPSPAPELLGGGGSDFPGHNYTTRNILPGLNIT S 291 Extended E200 N-term and C-term for use at the C-terminal end of the light chain LONG SEQUENCE v4 EPKSSDKTHTSPPSPAPELLGGGGSGHNYTTRNILPGLNITS 292 Extended E200 N-term and C-term for use in the C-terminal end of light chain LONG SEQUENCE v5 EPKSSDKTHTSPPSPAPELLDFPGHNYTTRNILPGLNITS 293 Extended E200 N-term and C-term for use in the C-terminal end of light chain LONG SEQUENCE v6 EPKSSDKTHTSPPSPAPELLGHNYTTRNILPGLNITS 294 Extended E200 N-term and C-term forGGGGSEPKSSDKTHTGSDFPGHNYTTRNILPGLNITS 295 Petition 870250089762, dated 02 / 10 / 2025, pp. 440 / 580 80 / 176 Description Sequence SEQ ID NO: use in the C-terminal end of the light chain MEDIUM SEQUENCEv1 Extended E200 Nterm and C-term for use in the C-terminal end of the light chain MEDIUM SEQUENCEv2 EPKSSDKTHTGSDFPGHNYTTRNILPGLNITS 296 Extended E200 Nterm and C-term for use in the C-terminal end of the light chain MEDIUM SEQUENCEv3 GGGGSEPKSSDKTHTGSGHNYTTRNILPGLNITS 297 Extended E200 Nterm and C-term for use in the C-terminal end of the light chain MEDIUM SEQUENCEv4 EPKSSDKTHTGSGHNYTTRNILPGLNITS 298 Table 4: Exemplary sequences of symmetrical molecules (used as controls) Description Sequence SEQ ID NO: CD33_Gem Heavy Chain (In mAb2 no1 paired with CD33_Gem Light Chain_N Term E200_No26 In mAb2 no 2 paired with CD33_Gem Light Chain_C EVQLVQSGAEVKKPGSSVKVSCKASGYTITDSNIHWVRQAPGQSL EWIGYIYPYNGGTDYNQKFKNRATLTVDNPTNTAYMELSSLRSEDT AFYYCVNGNPWLAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVGVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYQSTYRVVSVLTVLHQDWLNGKEYK CKVSNKQLPSPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDK SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 322 Petition 870250089762, dated 02 / 10 / 2025, pp. 441 / 580 81 / 176 Description Sequence SEQ ID NO: Term E200) CD33_Gem Light Chain C Term label E200 DIQLTQSPSTLSASVGDRVTITCRASESLDNYGIRFLTWFQQKPGKA PKLLMYAASNQGSGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQ QTKEVPWSFGQGTKVEVKRTVAAPSVFIFPPSDEQLKSGTASVVCL LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSEPKSSDK THTGSDFPGHNYTTRNILPGLNITS 323 CD33_Gem Light Chain N Term variant Label E200 no 26 DFPGHNYTTRNILPGLNITSGSEPKSSDKTHTGSDIQLTQSPSTLSA SVGDRVTITCRASESLDNYGIRFLTWFQQKPGKAPKLLMYAASNQ GSGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQTKEVPWSFGQ GTKVEVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKV QWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKV YACEVTHQGLSSPVTKSFNRGEC 324 Detailed description of the modalities

[102] Reference will now be made in detail to certain embodiments of the invention. Although the invention is described in conjunction with the embodiments, it will be understood that the intention is not to limit the invention to these embodiments. Rather, the invention is intended to encompass all alternatives, modifications and equivalents that may be included within the scope of the present invention, as defined by the claims.

[103] A person skilled in the art will recognize many methods and materials similar or equivalent to those described in the present invention, which could be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described.

[104] It will be understood that the invention disclosed and defined in this descriptive report extends to all alternative combinations of two or more of the individual features mentioned or evident from the text or drawings. All these different combinations constitute various alternative aspects of the invention.

[105] All patents and publications referred to in this invention are incorporated by reference in their entirety. Petition 870250089762, dated 02 / 10 / 2025, pp. 442 / 580 82 / 176

[106] The present invention is based on the inventors' recognition that undesirable toxicity may arise when using molecules previously described in the art as being useful for bringing CAR-T cells and target cells (such as cancer cells) closer together. Such molecules are typically referred to as bridging molecules or adaptor molecules in the art. More specifically, the inventors recognized that symmetrical bridging molecules with more than one fraction for binding to receptors on immune cells (such as CARs on CAR-T cells) can cause chronic activation, exhaustion, and induction of apoptosis. Without wishing to be limited by theory, the inventors believe that this results from the transactivation of different CAR receptors on individual CAR-T cells, leading to signaling without the presence of a target cell and / or the bridging molecules engaging with two CAR-T cells simultaneously, thus causing target-independent toxicity and fratricide.

[107] The inventors surprisingly discovered that the optimal architecture for a molecule to engage both CAR-T cells and cancer cells is one in which the fractions for binding to CAR-T cells are present only on a single arm of the bridge molecule (e.g., as depicted in Figure 2B) and, preferably, in which the fraction is linked or attached to the Fab portion of the bridge molecule (as opposed to binding to the constant chain of an antibody). In other words, the preferred architecture of a bridge molecule is one that minimizes or prevents the likelihood of the molecule binding to two independent CAR receptors on a single CAR-T cell, and / or to two or more individual CAR-T cells simultaneously.

[108] The inventors believe that bridge molecules with this optimal architecture: Petition 870250089762, dated 02 / 10 / 2025, pp. 443 / 580 83 / 176 1. They exhibit potency similar to bridging molecules based on symmetrical antibodies. 2. They have pharmacokinetics similar to a symmetrical mAb, but 3. They have significantly less unwanted and target-independent activation, exhaustion, and fratricide compared to symmetric mAb-based bridging molecules.

[109] The inventors' findings have implications for a wide range of CAR technologies, including indirect CAR technologies and technologies that repurpose CAR-T cells, as the continuous toxicity of the adapter molecule using symmetrical mAbs compromises the quality of the CAR-T cell product with respect to i) increased undesirable toxicity to the patient, ii) increased toxicity in the CAR-T cell product. Definitions

[110] Unless otherwise defined, the technical and scientific terms used in the present invention have the same meaning as is commonly understood by a person skilled in the art to which this invention pertains.

[111] For the purposes of interpreting this descriptive report, the following definitions will generally apply and, where appropriate, terms used in the singular will also include the plural and vice versa.

[112] P2X7 receptor generally refers to a purinergic receptor formed by three protein subunits or monomers, with at least one of the monomers having an amino acid sequence substantially as shown in SEQ ID NO: 152 below: MPACCSCSDVFQYETN KVTRIQSMNYGTIKWFFHVIIFSYVCFALVSDKLYQRKEPV ISSVHTKVKGIAEVKEEIVENGVKKLVHSVFDTADYTFPLQGNSFFVMTNFLKTEGQEQRL CPEYPTRRTLCSSDRGCKKGWMDPQSKGIQTGRCVVYEGNQKTCEVSAWCPIEAVEEAP RPALLNSAENFTVLIKNNIDFPGHNYTTRNILPGLNITCTFHKTQNPQCPIFRLGDIFRETGD Petition 870250089762, dated 02 / 10 / 2025, pp. 444 / 580 84 / 176 NFSDVAIQGGIMGIEIYWDCNLDRWFHHCRPKYSFRRLDDKTTNVSLYPGYNFRYAKYYK ENNVEKRTLIKVFGIRFDILVFGTGGKFDIIQLVVYIGSTLSYFGLAAVFIDFLIDTYSSNCCRS HIYPWCKCCQPCVVNEYYYRKKCESIVEPKPTLKYVSFVDESHIRMVNQQLLGRSLQDVK GQEVPRPAMDFTDLSRLPLALHDTPPIPGQPEEIQLLRKEATPRSRDSPVWCQCGSCLPS QLPESHRCLEELCCRKKPGACITTSELFRKLVLSRHVLQFLLLYQEPLLALDVDSTNSRLRHC AYRCYATWRFGSQDMADFAILPSCCRWRIRKEFPKSEGQYSGFKSPY Since the P2X7 receptor is formed by three monomers, it is a trimer or trimeric form. The P2X7 receptor encompasses naturally occurring variants of the P2X7 receptor, for example, where the P2X7 monomers are splice variants, allelic variants, SNPs, and isoforms, including naturally occurring truncated or secreted forms of the monomers that form the P2X7 receptor (e.g., a form consisting of the extracellular domain sequence or its truncated form), naturally occurring variant forms (e.g., alternatively spliced ​​forms), and naturally occurring allelic variants. In certain embodiments of the invention, the native sequence P2X7 monomeric polypeptides disclosed in the present invention are mature or full-length native sequence polypeptides comprising the full-length amino acid sequence shown in SEQ ID NO: 152.In certain embodiments, the P2X7 receptor may have an amino acid sequence that is modified; for example, several of the amino acids in the sequence shown in SEQ ID NO: 152 may be substituted, deleted, or a residue may be inserted.

[113] Functional P2X7 receptor generally refers to a form of the P2X7 receptor having three intact binding sites or clefts for ATP binding. When bound to ATP, the functional receptor forms a non-selective sodium / calcium channel that converts into a pore-like structure that allows the entry of calcium ions and molecules up to 900 Da into the cytosol, from which Petition 870250089762, dated 02 / 10 / 2025, pp. 445 / 580 85 / 176 One consequence may be the induction of programmed cell death. In normal homeostasis, the expression of functional P2X7 receptors is generally limited to cells that undergo programmed cell death, such as thymocytes, dendritic cells, lymphocytes, macrophages, and monocytes. There may also be some expression of functional P2X7 receptors in erythrocytes and other cell types.

[114] A dysfunctional P2X7 receptor generally refers to a form of a dysfunctional P2X7 receptor having a conformation distinct from the functional P2X7, whereby the receptor is unable to form an apoptotic pore, but is still able to operate as a non-selective channel by maintaining a single functional ATP binding site located between adjacent monomers. An example arises when one or more of the monomers have a cis isomerization at Pro210 (according to SEQ ID NO: 152). The isomerization can arise from any molecular event that leads to incorrect folding of the monomer, including, for example, mutation of the primary sequence of the monomer or abnormal post-translational processing. A consequence of the isomerization is that the receptor is unable to bind ATP at one, or more particularly two, ATP binding sites on the trimer and, as a consequence, fails to extend the channel opening.Under these circumstances, the receptor cannot form a pore, and this limits the extent to which calcium ions can enter the cytosol. Dysfunctional P2X7 receptors are expressed in a wide range of epithelial and hematopoietic cancers. As used in the present invention, the term dysfunctional P2X7 receptors can be used interchangeably with the term non-functional P2X7 receptors or nfP2X7 receptors.

[115] Cancer-associated P2X7 receptors are generally P2X7 receptors that are found in cancer cells (including pre-cancerous cells) Petition 870250089762, dated 02 / 10 / 2025, pp. 446 / 580 86 / 176 neoplastic, malignant, benign or metastatic), but not in non-cancerous or normal cells.

[116] Epitope E200 generally refers to an epitope having the sequence GHNYTTRNILPGLNITC and variants thereof (e.g., SEQ ID NOs: 153 to 162 and 166 to 218).

[117] Epitope E300 generally refers to an epitope having the sequence KYYKENNVEKRTLIK and variants thereof (SEQ ID NOs: 163 and 164).

[118] A composite epitope generally refers to an epitope that is formed by the juxtaposition of the E200 and E300 epitopes or parts of these epitopes. An example of a composite epitope comprising the E200 and E300 epitopes is GHNYTTRNILPGAGAKYYKENNVEK (SEQ ID NO: 165).

[119] Antibodies or immunoglobulins or Igs are gamma globulin proteins found in the blood or other body fluids of vertebrates that function in the immune system to bind to antigens, identifying and / or neutralizing foreign objects.

[120] Antibodies are generally a heterotetrameric glycoprotein composed of two identical light (L) chains and two identical heavy (H) chains. Each L chain is linked to an H chain by a covalent disulfide bond. The two H chains are linked to each other by one or more disulfide bonds, depending on the isotype of the H chain. Each H and L chain also has regularly spaced intrachain disulfide bridges.

[121] The H and L chains define specific Ig domains. More specifically, each H chain has, at the N-terminal end, a variable domain (VH) followed by three constant domains (CH) for each of the α and γ chains and four CH domains for the μ and ε isotypes. Each L chain has, at the N-terminal end, a variable domain (Vl) followed by a constant domain (CL) at its other end. The Vl is aligned with the Vh and the Cl is Petition 870250089762, dated 02 / 10 / 2025, pp. 447 / 580 87 / 176 aligned with the first constant domain of the heavy chain (CH1).

[122] Antibodies can be assigned to different classes or isotypes. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, with heavy chains designated α, δ, ε, y, and μ, respectively. The y classes are subdivided into subclasses based on relatively minor differences in sequence and function; for example, humans express the following subclasses: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The L chain of any vertebrate species can be assigned to one of two clearly distinct types, called kappa and lambda, based on the amino acid sequences of their constant domains.

[123] The constant domain includes the Fc portion comprising the carboxy-terminal portions of both H chains linked by disulfides. The effector functions of antibodies, such as ADCC, are determined by sequences in the Fc region, which is also the region recognized by Fc receptors (FcR) found on certain cell types.

[124] The pairing of a Vh and a Vl together forms a variable region or variable domain, including the amino-terminal domains of the antibody's heavy or light chain. The variable domain of the heavy chain may be referred to as Vh. The variable domain of the light chain may be referred to as Vl. The V domain contains an antigen-binding site that affects antigen binding and defines the specificity of a particular antibody for its particular antigen. V regions span approximately 110 amino acid residues and consist of relatively invariant stretches called structure regions (FRs) (usually about 4) of 15-30 amino acids, separated by shorter regions of extreme variability called hypervariable regions (usually about 3), each of which is generally 9 to 12 amino acids long. FRs largely adopt a β-sheet configuration and the Petition 870250089762, dated 02 / 10 / 2025, pp. 448 / 580 88 / 176 hypervariable regions form loops that connect and, in some cases, form part of the β-sheet structure.

[125] Hypervariable region refers to regions of an antibody variable domain that are hypervariable in sequence and / or form structurally defined loops. Generally, antibodies comprise six hypervariable regions; three in Vh (H1, H2, H3) and three in Vl (L1, L2, L3).

[126] Structure Residues or FR are those variable domain residues that are not the hypervariable region residues defined in the present invention.

[127] An antigen-binding site generally refers to a molecule that includes at least the hypervariable and structural regions that are necessary to confer antigen-binding function to a V domain. An antigen-binding site may be in the form of an antibody or an antibody fragment (such as an mAb, single-domain mAb (SD)-mAb, dAb, Fab, SD-Fab, Fd, SD-Fv, Fv, F(ab')2 or scFv) in a method described in the present invention.

[128] An intact or complete antibody is one that comprises an antigen-binding site as well as a CL and at least the constant domains of the heavy chain, CH1, CH2 and CH3. Constant domains may be native sequence constant domains (e.g., human native sequence constant domains) or amino acid sequence variant thereof.

[129] Complete antibody fragments including a variable domain include SD-mAb, Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies, single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.

[130] The Fab fragment consists of an entire L chain together Petition 870250089762, dated 02 / 10 / 2025, pp. 449 / 580 89 / 176 with the variable region domain of the H chain (Vh) and the first constant domain of a heavy chain (CH1). Each Fab fragment is monovalent with respect to antigen binding, that is, it has a single antigen-binding site.

[131] A Fab' fragment differs from Fab fragments by having some additional residues at the carboxy-terminal end of the CH1 domain, including one or more cysteines from the antibody hinge region. Fab'-SH is the designation in the present invention for Fab' in which the cysteine ​​residue(s) of the constant domains have a free thiol group.

[132] An F(ab')2 fragment corresponds approximately to two Fab fragments linked by disulfide, with divalent antigen-binding activity, and is also capable of performing antigen crosslinking.

[133] A vF is the smallest antibody fragment that contains a complete antigen recognition and binding site. This fragment consists of a dimer of a variable region domain of the heavy chain and a light chain domain in close, non-covalent association.

[134] In a single-chain Fv species (scFv), a variable domain of the heavy chain and one of the light chain can be covalently linked by a flexible peptide linker, so that the light and heavy chains can associate in a dimeric structure analogous to that of a two-chain Fv species. From the folding of these two domains emanate six hypervariable loops (3 loops from each of the H and L chains) that contribute amino acid residues for antigen binding and confer antigen-binding specificity to the antibody.

[135] Single-chain Fv, also abbreviated as sFv or scFv, are antibody fragments comprising the Vh and Vl antibody domains connected to form a single polypeptide chain. Preferably, the Petition 870250089762, dated 02 / 10 / 2025, pages 450 / 580 The 90 / 176 scFv polypeptide further comprises a polypeptide linker between the Vh and Vl domains that allows scFv to form the desired structure for antigen binding.

[136] A single variable domain is half of an Fv (comprising only three antigen-specific CDRs) that has the ability to recognize and bind to the antigen, although generally with a lower affinity than the whole binding site.

[137] Diabodies refers to antibody fragments with two antigen-binding sites, which fragments comprise a variable heavy chain domain (Vh) connected to a variable light chain domain (Vl) on the same polypeptide chain (Vh-Vl). The small antibody fragments are prepared by constructing sFv fragments (see previous paragraph) with short linkers (about 5-10 residues) between the Vh and Vl domains, so that interchain pairing, but not intrachain pairing, of the V domains is achieved, resulting in a bivalent fragment, i.e., a fragment with two antigen-binding sites.

[138] Diabodies can be bivalent or bispecific. Bispecific diabodies are heterodimers of two sFv crossover fragments in which the Vh and Vl domains of the two antibodies are present on different polypeptide chains. Triabodies and tetrabodies are also commonly known in the art.

[139] An isolated antibody is one that has been identified and separated and / or recovered from a component of its pre-existing environment. Contaminating components are materials that would interfere with the therapeutic uses of the antibody and may include enzymes, hormones, and other protein or non-protein solutes.

[140] A human antibody refers to an antibody that has a Petition 870250089762, dated 02 / 10 / 2025, pp. 451 / 580 91 / 176 amino acid sequence that corresponds to that of an antibody produced by a human being. Human antibodies can be produced using various techniques known in the art, including phage display libraries. Human antibodies can be prepared by administering the antigen to a transgenic animal that has been modified to produce such antibodies in response to an antigenic challenge, but whose endogenous loci have been deactivated.

[141] Humanized forms of non-human antibodies (e.g., from rodents) are chimeric antibodies that contain a minimal sequence derived from the non-human antibody. For the most part, humanized antibodies are human immunoglobulins (receptor antibody) in which residues from a hypervariable region of the receptor are replaced by residues from a hypervariable region of a non-human species (donor antibody), such as a mouse, rat, rabbit, or non-human primate, that possesses the desired antibody specificity, affinity, and capacity. In some cases, residues from the structure region (FR) of the human immunoglobulin are replaced by corresponding non-human residues. In addition, humanized antibodies may comprise residues that are not found in the receptor antibody or the donor antibody. These modifications are made to further refine antibody performance.In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the RFs are those of a human immunoglobulin sequence. The humanized antibody will optionally also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin.

[142] Monoclonal antibody refers to an antibody obtained from a Petition 870250089762, dated 02 / 10 / 2025, pp. 452 / 580 92 / 176 Substantially homogeneous antibody population, meaning that the individual antibodies that make up the population are identical, except for possible naturally occurring mutations that may be present in smaller quantities. Monoclonal antibodies are highly specific, being directed against a single antigenic site or determinant on the antigen. In addition to their specificity, monoclonal antibodies are advantageous because they can be synthesized without contamination by other antibodies. Monoclonal antibodies can be prepared using the hybridoma methodology. Monoclonal antibodies can also be isolated from phage antibody libraries using molecular engineering techniques.

[143] The term P2X7 receptor antibody or an antibody that binds to the P2X7 receptor refers to an antibody that is capable of binding to the P2X7 receptor with sufficient affinity such that the antibody is useful as a diagnostic and / or therapeutic agent in targeting the P2X7 receptor, typically the non-functional P2X7 receptor or a cancer-associated P2X7 receptor. Preferably, the extent of binding of a cancer-associated P2X7 receptor antibody to an unrelated protein is less than 10% of the antibody binding to the P2X7 receptor measured, for example, by a radioimmunoassay (RIA), Enzyme-Linked Immunosorbent Assay (ELISA), Biacore or Flow Cytometry. In certain embodiments, an antibody that binds to a P2X7 receptor has a dissociation constant (Kd) of < 1 μM, < 100 nM, < 10 nM, < 1 nM, or < 0.1 nM.An anti-nfP2X7 receptor antibody is generally one that has some of these serological characteristics and binds to dysfunctional P2X7 receptors but not to functional P2X7 receptors.

[144] A mature affinity antibody is one with one or more alterations in one or more of its hypervariable regions that result in a Petition 870250089762, dated 02 / 10 / 2025, pp. 453 / 580 93 / 176 improvement in antibody affinity for the antigen, compared to a parental antibody that does not have this / these alteration(s). Preferred matured affinity antibodies will have nanomolar or even picomolar affinities for the target antigen. Matured affinity antibodies are produced by procedures known in the art.

[145] A blocking antibody or an antagonist antibody is one that inhibits or reduces the biological activity of the antigen to which it binds. Blocking antibodies or preferred antagonists substantially or completely inhibit the biological activity of the antigen.

[146] An agonist antibody, as used in the present invention, is an antibody that mimics at least one of the functional activities of a polypeptide of interest.

[147] Binding affinity generally refers to the strength of the sum total of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, as used in the present invention, binding affinity refers to intrinsic binding affinity, which reflects a 1:1 interaction between the members of a binding pair (e.g., antibody and antigen). The affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (Kd). Affinity can be measured by common methods known in the art, including those described in the present invention. Low-affinity antibodies generally bind slowly to the antigen and tend to dissociate rapidly, while high-affinity antibodies generally bind more rapidly to the antigen and tend to remain bound for longer.A variety of methods for measuring binding affinity are known in the art, any of which can be used for the purposes of the present invention. Petition 870250089762, dated 02 / 10 / 2025, pp. 454 / 580 94 / 176

[148] As used in the present invention, the term antigen is intended to include substances that bind to or elicit the production of one or more antibodies and may include, but is not limited to, proteins, peptides, polypeptides, oligopeptides, lipids, carbohydrates, and combinations thereof, for example, a glycosylated protein or a glycolipid. The term antigen, as used in the present invention, refers to a molecular entity that can be expressed on a target cell and that can be recognized by the adaptive immune system, including, but not limited to, antibodies or TCRs, or engineered molecules, including, but not limited to, transgenic TCRs, CARs, scFvs or multimers thereof, Fab fragments or multimers thereof, antibodies or multimers thereof, single-chain antibodies or multimers thereof, or any other molecule that can perform high-affinity binding to a structure.

[149] Epitope generally refers to the part of an antigen that is bound by the antigen-binding site of an antibody. An epitope can be linear in the sense that the hypervariable loops of the antibody CDRs that form the antigen-binding site bind to an amino acid sequence as in a primary protein structure. In certain embodiments, the epitope is a conformational epitope, i.e., one in which the hypervariable loops of the CDRs bind to residues as they are presented in the tertiary or quaternary structure of the protein.

[150] The term target cell, as used in the present invention, refers to a cell that expresses a cell surface molecule to which the targeting fraction of the bridge molecule binds. The target cell can be a cancerous cell or any other diseased cell.

[151] The term disorder or condition means an abnormality or functional disturbance in an individual, such as a cancer, a disorder Petition 870250089762, dated 02 / 10 / 2025, pages 455 / 580 95 / 176 autoimmune or an infection caused by a virus, bacteria, parasite, or other organ.

[152] For example, a nucleic acid or peptide naturally present in a living animal is not isolated, but the same nucleic acid or peptide partially or totally separated from the coexisting materials of its natural state is isolated. An isolated nucleic acid or protein may also exist in a non-native environment, such as, for example, in a host cell.

[153] As used in the present invention, the term individual refers to a mammal, such as a mouse, rat, cow, pig, goat, chicken, dog, monkey, or human. Preferably, the individual is a human. The individual may be an individual suffering from a disorder such as cancer (a patient), but the individual may also be a healthy individual.

[154] The term autologous, as used in the present invention, refers to any material derived from the same individual to which it is subsequently reintroduced.

[155] The term allogeneic, as used in the present invention, refers to any material derived from an individual other than the same species as the individual to which the material is reintroduced.

[156] The terms therapeutically effective quantity or therapeutically effective population mean a quantity of, for example, a population of cells that provides a therapeutic benefit in an individual.

[157] The terms binds to, binds specifically to, or specific to with respect to a targeting fraction, as used, for example, in the bridge molecule as disclosed herein, or of a CAR referring to an antigen-binding domain that recognizes and binds to a specific antigen, does not recognize or binds substantially to other molecules in a sample. An antigen-binding domain or targeting fraction that binds Petition 870250089762, dated 02 / 10 / 2025, pp. 456 / 580 96 / 176 specifically bound to an antigen of one species may also bind to that antigen of another species. This cross-reactivity between species is typical of many antibodies and is therefore not contrary to the definition that the antigen-binding domain is specific. An antigen-binding domain that binds specifically to an antigen may also bind to different allelic forms of the antigen (allelic variants, splice variants, isoforms, etc.) or homologous variants of that antigen from the same gene family. This cross-reactivity is typical of many antibodies and is therefore not contrary to the definition that the antigen-binding domain is specific.

[158] The terms manipulated cell and genetically modified cell, as used in the present invention, may be used interchangeably. The terms mean containing and / or expressing a foreign gene or nucleic acid sequence that, in turn, modifies the genotype or phenotype of the cell or its progeny. In particular, the terms refer to the fact that cells, preferably immune cells, can be manipulated by recombinant methods known in the art to stably or transiently express peptides or proteins that are not expressed in these cells in the natural state. For example, immune cells are manipulated to express an artificial construct, such as a chimeric antigen receptor, on their cell surface.For example, CAR sequences can be delivered to cells using an adenoviral vector, based on adeno-associated viruses (AAVs), retrovirals, or lentivirals, or any other pseudotyped variations thereof, or any other gene delivery mechanism, such as electroporation or lipofection with CRISPR / Cas9, transposons (e.g., sleeping-beauty) or their variations. Gene delivery can be in the form of mRNA (transient) or DNA (transient or permanent).

[159] The terms immune cell or immune effector cell refer to Petition 870250089762, dated 02 / 10 / 2025, pp. 457 / 580 97 / 176 a cell that can be part of the immune system and performs a particular effector function, such as alpha-beta T cells, NK cells, NKT cells, B cells, Breg cells, Treg cells, innate lymphoid cells (ILCs), cytokine-induced killer cells (CIKs), lymphokine-activated killer cells (LAKs), gamma-delta T cells, mesenchymal stem cells or mesenchymal stromal cells (MSCs), monocytes or macrophages, or any hematopoietic progenitor cells, such as pluripotent stem cells and subsets of early progenitors that can mature or differentiate into somatic cells. Cells can be naturally occurring or generated by exposure to cytokines, artificial / genetically modified cells (such as iPSCs and other types of artificial cells). The immune cell can be a subset of artificial cells, including induced pluripotent stem cells and cells matured from them.Preferred immune cells are cells with cytotoxic effector function, such as alpha-beta T cells, NK cells, NKT cells, ILC, CIK cells, LAK cells, or gamma-delta T cells. Effector function means a specialized function of a cell; for example, in a T cell, an effector function might be cytolytic activity or helper cell activity, including cytokine secretion.

[160] The term treat (treatment of) a disorder, as used in the present invention, means reducing the frequency or severity of at least one sign or symptom of a disease or disorder experienced by an individual.

[161] The term expression, as used in the present invention, is defined as the transcription and / or translation of a particular nucleotide sequence directed by its promoter in a cell. Exogenous immune cell receptors (e.g., CARs) General Petition 870250089762, dated 02 / 10 / 2025, pages 458 / 580 98 / 176

[162] The present invention finds utility in the context of redirecting immune cells expressing exogenous receptors (such as chimeric antigen receptors, or CARs) toward a target cell. In its simplest form, the invention relates to the use of a bridging molecule, as described in the present invention, to bring a cytotoxic immune cell (such as a T cell and especially a CAR-T cell) closer to a target cell (such as a cancer cell). The bridging molecules of the invention may enable the redirection of a CAR-T cell to a target cell comprising a different antigen than that recognized by the CAR, or may increase the binding affinity of the CAR to the target cell.

[163] Those skilled in the art will appreciate that the exogenous immune cell receptors contemplated for use according to the invention are typically exogenous receptors (i.e., they do not normally occur on the surface of the immune cell or are recombinant / heterologous), which typically comprise an antigen-binding (or recognition) domain and a signaling domain that is stimulated to activate the cytotoxic activity of the cell after binding of the antigen-binding domain. Also contemplated according to the invention is the use of universal CAR systems based on switches or tags, where the CAR does not necessarily comprise an antigen-binding domain, but another means of engaging with a target cell, via an intermediate molecule.

[164] The terms cancer- or tumor-associated CAR targeting, or tumor-specific CAR targeting, refer to the use of a CAR-T cell to bind to a target antigen that is presented on the cell surface of tumor cells, but which is not typically found on the surface of a healthy cell. In other words, normal cells under normal circumstances may be characterized by low expression or absence of the target antigen on the surface of tumor cells. Petition 870250089762, dated 02 / 10 / 2025, pp. 459 / 580 99 / 176 extracellular membrane (and therefore the presence of the antigen on the cell surface cannot be detected). In even more examples, such cells may express the mRNA encoding the antigen at the intracellular level. Since CAR-T cells only recognize antigens expressed on the surface, intracellular expression of the target proteins will not lead to CAR engagement. Alternatively (and particularly in the case of tumor-associated CAR targeting), CAR may be for binding to a target antigen that is overexpressed by cancer cells compared to healthy cells.

[165] An example of an epitope expressed on the surface of cancer cells is the P2X7 receptor. The target epitopes E200 and E300 of the P2X7 receptor are not exposed in the form of the receptor found in healthy tissue and therefore these epitopes can be considered cancer-specific. In other words, the E200 and E300 epitopes are exposed and available for binding only when the P2X7 receptor has an altered non-functional conformation, as occurs in the context of cancer (in this case, the receptor can be referred to as the nfP2X7 receptor). Another example of a cancer-specific target epitope can be derived from the EGFRvIII splicing variant. Yet another example is the CLDN6 antigen, which is mainly restricted to embryonic and fetal life and has very limited expression in healthy cells after the early stages of life, and can be considered highly restricted and relatively overexpressed in cancer.The present invention contemplates binding to any specific tumor or tumor-associated antigen, including P2X7, EGFRvIII and CLDN6, for specific cancer targeting and engagement of CAR-T cells, via bridging molecules described in the present invention, to cancer-associated antigens.

[166] The present invention also contemplates linking a CAR to a tumor-associated antigen. Tumor-associated antigens are known in Petition 870250089762, dated 02 / 10 / 2025, pages 460 / 580 100 / 176 technique (and are typically associated with increased expression in the context of cancer).

[167] In general, a CAR may comprise an extracellular domain (extracellular part) comprising the binding domain (also referred to as antigen-binding domain or antigen recognition domain, or tag-binding domain in the context of a universal CAR), a transmembrane domain and an intracellular signaling domain.

[168] Generally, an antigen recognition domain refers to the region of the CAR that specifically binds to an antigen (and thus is able to target a cell containing the antigen). The CARs of the invention may comprise one or more antigen-binding domains. Generally, the targeting regions in the CAR are extracellular. The antigen-binding domain may comprise an antibody or an antibody-binding fragment thereof. The antigen-binding domain may comprise, for example, full-length heavy chain, Fab fragments, single-chain Fv fragments (scFv), divalent single-chain antibodies, or diabodies. Any molecule that specifically binds to a given antigen, such as antibodies or naturally occurring receptor ligand-binding domains, may be used as an antigen-binding domain. Frequently, the antigen-binding domain is an scFv.Typically, in a scFv, the variable regions of an immunoglobulin heavy and light chain are fused by a flexible linker to form an scFv. Such a linker can be, for example, the (G4 / S1) linker and its variations, but a person skilled in the art will appreciate that various linker sequences and formats can be used.

[169] In some cases, it is beneficial for the antigen-binding domain to be derived from the same species in which the CAR will be used. For example, when planning to use it therapeutically in humans, it may be beneficial that the Petition 870250089762, dated 02 / 10 / 2025, pp. 461 / 580 101 / 176 The antigen-binding domain of the CAR comprises a human or humanized antibody or an antigen-binding fragment thereof. Human or humanized antibodies or antigen-binding fragments thereof can be made by a variety of methods well known in the art. The CAR, as disclosed in the present invention, has an extracellular marker / ligand epitope-binding domain as an antigen-binding domain, enabling it to bind indirectly, via a target cell-binding molecule as disclosed in the present invention, to an antigen expressed on a target cell.

[170] The extracellular domain of a CAR can be linked to the transmembrane domain by a linker. The extracellular domain may also comprise a signal peptide. The extracellular portion of the CAR for use according to the present invention preferably comprises a tumor-associated antigen-binding domain. For example, the antigen may be any of those described in the present invention, including P2X7, EGFRvIII, or CLDN6.

[171] The antigen-binding domain may be a P2X7-binding domain that recognizes the E200 epitope (or E300 or E200-300 composite) as disclosed in the present invention. Specifically, the CAR as disclosed in the present invention has an extracellular nfP2X7 E200-binding domain (or E300 or E200-300 composite) as an antigen-binding domain. Alternatively, the antigen-binding domain may be an EGFRvIII-binding domain that recognizes an epitope resulting from the fusion of the amino acid sequence starting at position 25-29 LEEKK (SEQ ID NO: 280), followed by the insertion of G and the subsequent amino acid sequence 298-304 NYVVTDH (SEQ ID NO: 281), the total epitope being a 13-mer composed of the sequence LEEKKGNYVVTDH (SEQ ID NO: 134). Alternatively, the domain of connection to Petition 870250089762, dated 02 / 10 / 2025, pp. 462 / 580 102 / 176 antigen may be a CLDN6-binding domain that recognizes an epitope in the second extracellular domain of CLDN6 [UniProtKB: P56747 (CLDN6_HUMAN)] via the amino acid sequence of SEQ ID NO: 143.

[172] Typically, the antigen recognition domain includes a binding polypeptide that includes amino acid sequence homology with one or more complementarity-determining regions (CDRs) of an antibody that binds to a tumor-specific or tumor-associated antigen (such as a dysfunctional P2X7 receptor, EGFRvIII, or CLDN6). In any embodiment, the binding polypeptide includes amino acid sequence homology with the CDR1, 2, and 3 domains of the Vh and / or Vl chain of an antibody that binds to a tumor-specific or tumor-associated antigen (such as a dysfunctional P2X7 receptor, EGFRvIII, or CLDN6).

[173] In particularly preferred embodiments of the invention, the antigen recognition domain of the CAR binds to an epitope of the nfP2X7 antigen. In such embodiments, the binding polypeptide comprises the amino acid sequence of the CDRs of the Vh and / or Vl chain of an antibody described in any of: PCT / AU2002 / 000061 or PCT / AU2002 / 001204 (or in any of the corresponding US patents US 7,326,415, US 7,888,473, US 7,531,171, US 8,080,635, US 8,399,617, US 8,709,425, US 9,663,584 or US 10,450,380), PCT / AU2007 / 001540 (or in the corresponding US patent US 8,067,550), PCT / AU2007 / 001541 (or in the corresponding US patent publication US 2010-0036101), PCT / AU2008 / 001364 (or in any of the corresponding US patents US 8,440,186, US 9,181,320, US 9,944,701 or US 10,597,451), PCT / AU2008 / 001365 (or in any of the corresponding US patents US 8,293,491 or US 8,658,385), PCT / AU2009 / 000869 (or in any of the corresponding US patents US 8,597,643, US 9,328,155 or US 10,238,716), PCT / AU2010 / 001070 (or in any of the publications) Petition 870250089762, dated 02 / 10 / 2025, pp. 463 / 580 103 / 176 corresponding WO / 2011 / 020155, US 9,127,059, US 9,688,771, or US 10,053,508), and PCT / AU2010 / 001741 (or in any of the corresponding publications WO 2011 / 075789 or US 8,835,609) the full content of which is incorporated herein by reference. Preferably, the linking polypeptide comprises the amino acid sequence of the CDRs of the VH and / or VL chain of the 2-2-1 antibody described in PCT / AU2010 / 001070 (or in any of the corresponding US patents US 9,127,059, US 9,688,771 or US 10,053,508) or BPM09 described in PCT / AU2007 / 001541 (or in the corresponding US patent publication US 2010-0036101) and produced by the AB253 hybridoma deposited in the European Culture Collection (ECACC) under Accession number 06080101, WO2013185010A1 or WO2019056023.Alternatively, the CAR-binding polypeptide may comprise the amino acid sequences of the sdAbs 2-2-3, 2-472-2 or 2-2-12 antibody CDRs described in WO 2017 / 041143 (also published as US 2019 / 0365805), and WO 2019 / 222796 (corresponding to US application 17 / 057060), incorporated into the present invention by reference.

[174] The CAR-binding polypeptide may comprise the amino acid sequence of the Vh and / or Vl chains of an antibody described in any of: PCT / AU2002 / 000061 or PCT / AU2002 / 001204 (or in any of the corresponding US patents US 7,326,415, US 7,888,473, US 7,531,171, US 8,080,635, US 8,399,617, US 8,709,425, US 9,663,584, or US 10,450,380), PCT / AU2007 / 001540 (or in the corresponding US patent US 8,067,550), PCT / AU2007 / 001541 (or in the corresponding US patent publication US 20100036101), PCT / AU2008 / 001364 (or any of the corresponding US patents US 8,440,186, US 9,181,320, US 9,944,701 or US 10,597,451), PCT / AU2008 / 001365 (or any of the corresponding US patents US 8,293,491 or US 8,658,385), PCT / AU2009 / 000869 (or any of the Petition 870250089762, dated 02 / 10 / 2025, pp. 464 / 580 104 / 176 corresponding US patents US 8,597,643, US 9,328,155 or US 10,238,716), PCT / AU2010 / 001070 (or in any of the corresponding publications WO / 2011 / 020155, US 9,127,059, US 9,688,771, or US 10,053,508) and PCT / AU2010 / 001741 (or in any of the corresponding publications WO 2011 / 075789 or US 8,835,609) the full content of which is incorporated herein by reference. Preferably, the linking polypeptide comprises the amino acid sequence of the Vh and / or Vl chains of antibody 22-1 described in PCT / AU2010 / 001070 (or in any of the corresponding US patents US 9,127,059, US 9,688,771, or US 10,053,508) or BPM09 described in PCT / AU2007 / 001541 (or in the corresponding US patent publication US 2010-0036101) and produced by the AB253 hybridoma deposited in the European Culture Collection (ECACC) under Accession number 06080101, WO2013185010A1 or WO2019056023.Alternatively, the CAR-binding polypeptide may comprise the amino acid sequences of the VH and / or VL chains of the sdAbs 2-2-3, 2-472-2, or 2-212 antibody described in WO 2017 / 041143 (also published as US 2019 / 0365805), and WO 2019 / 222796 (corresponding to US application 17 / 057060), incorporated into the present invention by reference.

[175] The CAR-binding polypeptide may comprise the amino acid sequence of an antibody or fragment thereof described in any of PCT / AU2002 / 000061 or PCT / AU2002 / 001204 (or in any of the corresponding US patents US 7,326,415, US 7,888,473, US 7,531,171, US 8,080,635, US 8,399,617, US 8,709,425, US 9,663,584, or US 10,450,380), PCT / AU2007 / 001540 (or in the corresponding US patent US 8,067,550), PCT / AU2007 / 001541 (or in the corresponding US patent publication US 20100036101), PCT / AU2008 / 001364 (or any of the corresponding US patents US 8,440,186, US 9,181,320, US 9,944,701 or US 10,597,451), Petition 870250089762, dated 02 / 10 / 2025, pp. 465 / 580 105 / 176 PCT / AU2008 / 001365 (or any of the corresponding US patents US 8,293,491 or US 8,658,385), PCT / AU2009 / 000869 (or any of the corresponding US patents 8,597,643, US 9,328,155 or US 10,238,716), PCT / AU2010 / 001070 (or any of the corresponding publications WO / 2011 / 020155, US 9,127,059, US 9,688,771, or US 10,053,508), and PCT / AU2010 / 001741 (or any of the corresponding publications WO 2011 / 075789 or US 8,835,609), the full content of which is incorporated herein by reference. Preferably, the linking polypeptide comprises the sdAb 2-2-1 amino acid sequence described in PCT / AU2010 / 001070 (or in any of the corresponding US patents US 9,127,059, US 9,688,771, or US 10,053).508) or antibody BPM09 described in PCT / AU2007 / 001541 (or in the corresponding US patent publication US 20100036101) and produced by the AB253 hybridoma deposited in the European Culture Collection (ECACC) under Accession number 06080101, WO2013185010A1 or WO2019056023. Alternatively, the linking polypeptide may comprise the sdAbs amino acid sequences 2-2-3, 2-472-2, or 2-2-12 described in WO 2017 / 041143 (also published as US 2019 / 0365805), and WO 2019 / 222796 (corresponding to US application 17 / 057060), incorporated into the present invention by reference. Universal CAR Systems

[176] It will be appreciated that the methods of the present invention also find utility in the context of immune cells comprising so-called universal CARs or switchable CARs. Such CARs are intended for a use similar to that of conventional CARs (e.g., for expression in immune cells and for the killing of target cells), but not antigen-binding domains for direct binding to antigens on target cells. Instead, these exogenous receptors may comprise binding domains for Petition 870250089762, dated 02 / 10 / 2025, pp. 466 / 580 106 / 176 binding to an intermediate (tag or switch) or neoepitope that is attached and thus indirectly allows the targeting (or redirection) of immune cells expressing the receptor to a target cell.

[177] In certain embodiments, the receptor is capable of binding to a neoepitope or exogenous epitope or peptide tag, such as a dye (e.g., fluorescein isothiocyanate, FITC), biotin moiety, AZip tag (or leucine zipper moiety), Blm peptide Latch, SpyTag, the 5B9 tag (i.e., comprising 10 amino acids derived from the human nuclear autoantigen La / SS-B) and the PNE tag (i.e., a 14-amino acid neoepitope peptide derived from a yeast transcription factor) and Domain D-based Universal CAR-T (e.g., ARC-SparX). In the art, such neoepitopes or tags may also be referred to as switch elements in the context of so-called Universal CAR systems. Those skilled in the art will recognize that such neoepitopes are not typically cancer antigen-associated epitopes. Other portions of CARs and Universal CARs

[178] A sequence that encodes a CAR can also encode a signal peptide, which refers to a peptide sequence that directs the transport and location of the protein within a cell, for example, to a specific cell organelle (such as the endoplasmic reticulum) and / or to the cell surface.

[179] Spacer or hinge, as used in the present invention, refers to the hydrophilic region that lies between the antigen-binding domain and the transmembrane domain. The CARs of the invention may comprise an extracellular spacer domain, but it is also possible to omit such a spacer. The spacer may include, for example, Fc fragments of antibodies or fragments thereof, hinge regions of antibodies or fragments of Petition 870250089762, dated 02 / 10 / 2025, pp. 467 / 580 107 / 176 same, CH2 or CH3 regions of antibodies, accessory proteins, artificial spacer sequences, or combinations thereof. A prominent example of a spacer is the CD8alpha hinge or the CD28 hinge.

[180] The transmembrane domain of CAR can be derived from any desired natural or synthetic source for such a domain. When the source is natural, the domain can be derived from any membrane-bound or transmembrane protein. The transmembrane domain can be derived, for example, from CD8alpha or CD28. When the key signaling and antigen recognition modules (domains) are in two (or even more) polypeptides, then CAR can have two (or more) transmembrane domains. The splitting of the key signaling and antigen recognition modules allows for small molecule-dependent, titratable, and reversible control over CAR cell expression (Wu et al, 2015, Science 350: 293-303) due to small molecule-dependent heterodimerization domains in each CAR polypeptide.

[181] The cytoplasmic domain (or intracellular signaling domain) of CAR is responsible for activating at least one of the normal effector functions of the immune cell in which CAR is expressed. Effector function means a specialized function of a cell, for example, in a T cell, an effector function might be cytolytic activity or helper cell activity, including cytokine secretion. The intracellular signaling domain refers to the part of a protein that transduces the effector function signal and directs the cell expressing CAR to perform a specialized function. The intracellular signaling domain may include any complete, mutated, or truncated part of the intracellular signaling domain of a given protein, sufficient to transduce a signal that initiates or blocks the effector functions of the immune cell.

[182] The function of intracellular domains can be pro or anti- Petition 870250089762, dated 02 / 10 / 2025, pages 468 / 580 108 / 176 inflammatory and / or immunomodulatory, or a combination thereof.

[183] ​​Prominent examples of intracellular signaling domains for use in CARs include T cell receptor (TCR) cytoplasmic signaling sequences and coreceptors that initiate signal transduction after antigen receptor engagement.

[184] Generally, T cell activation can be mediated by two distinct classes of cytoplasmic signaling sequences: firstly, those that initiate antigen-dependent primary activation via the TCR (primary cytoplasmic signaling sequences) and secondly, those that act in an antigen-independent manner to provide a secondary or co-stimulatory signal (secondary cytoplasmic signaling sequences, co-stimulatory signaling domain). Therefore, an intracellular signaling domain of a CAR can comprise one or more primary cytoplasmic signaling domains and / or one or more secondary cytoplasmic signaling domains.

[185] Primary cytoplasmic signaling sequences that act in a stimulatory manner may contain ITAMs (immunoreceptor tyrosine-based activation motifs) signaling motifs.

[186] Examples of primary cytoplasmic signaling sequences containing ITAMs frequently used in CARs are those derived from TCR zeta (CD3 zeta), FcR gamma, FcR beta, CD3 gamma, CD3 delta, CD3 epsilon, CD5, CD22, CD79a, CD79b and CD66d. The most prominent is the sequence derived from CD3 zeta.

[187] The cytoplasmic domain of CAR can be designed to comprise the CD3-zeta signaling domain alone or combined with any other desired cytoplasmic domain(s). The cytoplasmic domain of CAR can comprise a portion of the CD3 zeta chain and a Petition 870250089762, dated 02 / 10 / 2025, pp. 469 / 580 109 / 176 Co-stimulatory signaling region. The co-stimulatory signaling region refers to a part of the CAR that comprises the intracellular domain of a co-stimulatory molecule. A co-stimulatory molecule is a cell surface molecule, other than an antigen receptor or its ligands, necessary for an efficient lymphocyte response to an antigen. Examples of a co-stimulatory molecule are CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, and B7-H3.

[188] Cytoplasmic signaling sequences within the cytoplasmic signaling portion of the CAR can be linked to each other with or without a linker, in a random or specified order. A short oligo- or polypeptide linker, preferably between 2 and 10 amino acids in length, can form the linker. A prominent linker is the glycine-serine doublet.

[189] As an example, the cytoplasmic domain may comprise the CD3-zeta signaling domain and the CD28 signaling domain. In another example, the cytoplasmic domain may comprise the CD3-zeta signaling domain and the CD27 signaling domain. In a further example, the cytoplasmic domain may comprise the CD3-zeta signaling domain, the CD28 signaling domain, and the CD27 signaling domain.

[190] As mentioned earlier, both the extracellular portion and the transmembrane domain or cytoplasmic domain of a CAR may also comprise a heterodimerization domain with the aim of splitting the CAR’s key signaling and antigen recognition modules.

[191] Non-limiting examples of CARs that can be used accordingly Petition 870250089762, dated 02 / 10 / 2025, pages 470 / 580 110 / 176 with the present invention are provided in WO 2017 / 041143 (also published as US 2019 / 0365805), and WO 2019 / 222796 (corresponding to US application 17 / 057060), and WO 2022 / 187906, incorporated in the present invention by reference.

[192] A CAR for use in accordance with the present invention, that is, a CAR comprising an nfP2X7 E200 binding domain, may be designed to comprise any portion or part of the aforementioned domains, as described in the present invention, in any order and / or combination that results in a functional CAR.

[193] The CARs as disclosed in the present invention, or polypeptide(s) derived therefrom, recombinant nucleic acid molecule(s) or expression vectors, cells encoding said CARs or cell populations expressing said CARs, may be isolated and / or purified. The term isolated means altered or removed from the natural state. For example, an isolated cell population means an enrichment of such cells and the separation of other cells that are normally associated in their natural occurrence state with said isolated cells. An isolated cell population means a substantially purified cell population that is a more homogeneous population of cells than that found in nature. Preferably, the enriched cell population comprises at least about 90% of the selected cell type.In particular aspects, the cell population comprises at least approximately 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or even 100% of the selected cell type.

[194] The affinity with which the P2X7 receptor-binding domain of CAR binds to the E200 moiety of the bridge molecule can vary, but generally the binding affinity can be in the range of 100 μM, 1 nM, 10 nM or 100 nM, Petition 870250089762, dated 02 / 10 / 2025, pp. 471 / 580 111 / 176 preferably at least about 1 pM or 10 pM, even more preferably at least about 100 pM.

[195] CAR-T cells targeting EGFRvIII can be used to treat solid cancers. EGFRvIII is a frequent splice variant of EGFR that skips exons 2-7. EGFRvIII is relatively tumor-specific and does not occur in healthy cells, as EGFR is tightly regulated in normal cells. EGFRvIII is commonly expressed in glioblastoma, but also in breast cancer and head and neck cancer. The EGFRvIII CAR-T can be targeted to the epitope resulting from the fusion of the amino acid sequence starting at position 25-29 LEEKK (SEQ ID NO: 280), followed by the insertion of G and the subsequent amino acid sequence 298-304 NYVVTDH (SEQ ID NO: 281) the total epitope comprises or consists of the sequence LEEKKGNYVVTDH (SEQ ID NO: 134). The complete EGFR sequence is found in UniProtKB - P00533 (EGFR_HUMAN), and the complete protein has 1210 amino acids in isoform 1. An exemplary anti-EGFRvIII CAR sequence is established in SEQ ID NO: 285.

[196] EGFRvIII-targeted CAR-T cells can be used to treat glioblastoma in a conventional manner to target EGFRvIII in cancer cells, but they can also be redirected to other cancer-associated target antigens via bridging molecules described in the present invention if the EGFRvIII epitope fraction is integrated into the bridging molecule sequence. EGFRvIII CAR-T cells can then be used in the same manner as outlined for the nfP2X7 CAR-targeted approach described in the present invention. The peptide tag can be the 13 mer LEEKKGNYVVTDH peptide or a shortened or extended natural or artificial variant thereof, SEQ ID NO: 134.

[197] The amino acid sequence of EGFRvIII CAR-compatible bridge molecules targeting CD19 and CD33 are described in Table 2 as Petition 870250089762, dated 02 / 10 / 2025, pp. 472 / 580 112 / 176 SEQ ID NO: 135 and 136; 137 and 138; and SEQ ID NOs: 139 and 140; 141 and 142, respectively.

[198] CLDN6-targeted CAR-T cells can be used to treat solid cancers, for example, ovarian cancer. The CLDN6 CAR-T can be targeted to the second extracellular domain of CLDN6 cells [UniProtKB P56747 (CLD6_HUMAN)] directly via amino acid sequence [ECD2, >sp|P56747|138-160 WTAHAIIRDFYNPLVAEAQKREL (SEQ ID NO: 143)], but can also be redirected to other cancer-associated target antigens, for example, CD33 or CD19, via bridging molecules described in the present invention, if the CLDN6 epitope fraction is integrated into the bridging molecule sequence. CLDN6 CAR-T cells can then be used in the same manner as outlined for the nfP2X7 CAR-targeted approach described in the present invention. The peptide tag may be the 23-mer peptide WTAHAIIRDFYNPLVAEAQKREL (SEQ ID NO: 143) or a naturally occurring or artificially shortened or extended variant thereof. An exemplary anti-CLDN CAR sequence is presented in SEQ ID NO: 284.

[199] The amino acid sequence of CAR CLDN6-compatible bridge molecules targeting CD19 and CD33 are described in Table 2 as SEQ ID NO: 144 and 145; and SEQ ID NOs: 148 and 149; 150 and 151, respectively. Bridge molecule (heterodimeric molecule)

[200] It will be appreciated that the bridge molecule of the invention is typically in the form of an antibody (e.g., two Fab arms and optionally one or more Fc regions of an antibody), and in which fragments capable of being bound by an exogenous immune receptor are present in any region of a single Fab arm. Examples of suitable architectures are represented in Figure 3.

[201] Any suitable ligand can be used to link the fractions Petition 870250089762, dated 02 / 10 / 2025, pp. 473 / 580 113 / 176 for binding via the immune cell receptor to the Fab arm. The ligand may comprise a polypeptide, a peptide, or a chemical group.

[202] A linker can be a peptide with a length of up to 20 amino acids. The term linked to or fused to refers to a covalent bond, for example, a peptide bond, formed between two moieties. Consequently, in the context of the present invention, the linker can have a length of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22 amino acids. For example, the bridging molecule provided in the present invention may comprise a linker between each moiety and the Fab arm. Such linkers have the advantage that they can increase the likelihood that the different polypeptides of the fusion protein will fold independently and behave as expected.

[203] Those skilled in the art will be familiar with the design and use of various peptide linkers composed of various amino acids and of various lengths, which would be suitable for use as linkers according to the present invention. The linker may comprise various combinations of repeated amino acid sequences. The linker may be a flexible linker (such as those comprising repeats of glycine and serine residues), a rigid linker (such as those comprising glutamic acid and lysine residues flanking alanine repeats) and / or a cleavable linker (such as sequences that are susceptible to cleavage by protease).

[204] The peptide linker may be one or more Gly-Ser (GS), Gly-Gly-Ser (GGS), Gly-Gly-Gly-Ser (GGGS, SEQ ID NO: 315) or Gly-Gly-GlyGly-Ser (GGGGS, SEQ ID NO: 316) repetitions or variations thereof. In any embodiment, the linker may comprise or consist of the sequence GGGGSGGGGSGGGGS, i.e., (G4S)3 (SEQ ID NO: 146).

[205] In any form, the peptide linker may include the Petition 870250089762, dated 02 / 10 / 2025, pp. 474 / 580 114 / 176 amino acid sequence GGGGGS (a 6-amino acid long linker) or even longer. The linker can be a series of repeating glycine and serine (GS) residues of different lengths, i.e., (GS)n where n is any number from 1 to 15 or more. For example, the linker can be (GS)3 (i.e., GSGSGS, SEQ ID NO: 317) or longer (GS)ii or longer. It will be recognized that n can be any number, including 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or more.

[206] In additional embodiments, the linker may comprise the inclusion of an amino acid that confers rigidity, such as lysine. For example, in certain embodiments, the linker region may also comprise the sequence GSGK (SEQ ID NO: 318).

[207] The peptide linker may consist of a series of Thr-Pro (TP) repeats comprising one or more additional amino acids at the N- and C-terminal ends of the repeat sequence. For example, the linker may comprise or consist of the sequence GTPTPTPTPTGEF (also known as the TP5 linker; SEQ ID NO: 147). In additional aspects, the linker may be a short and / or alpha-helical rigid linker (e.g., A(EAAAK)3A, SEQ ID NO: 319; PAPAP, SEQ ID NO: 320; or a dipeptide such as LE or CC).

[208] In additional embodiments, as an alternative to or in addition to a glycine-serine-based linker region as described above, the targeting moiety may be in the form of a fusion protein, in which the targeting moiety is linked to the epitope moiety of tumor-specific or tumor-associated antigen, preferably to the epitope moiety of the dysfunctional P2X7 receptor, via a hinge region. The linkage between the targeting moiety and the epitope moiety of tumor-specific or tumor-associated antigen may comprise a combination of hinge region and linker regions. Petition 870250089762, dated 02 / 10 / 2025, pp. 475 / 580 115 / 176

[209] Examples of suitable hinge regions include hinge regions derived from immunoglobulins. The hinge region may be derived from an IgG1, IgG2, IgG3, or IgG4, and may comprise one or more amino acid substitutions (e.g., to prevent or reduce the likelihood of disulfide bridge formation). Alternative hinge sequences may be derived from alternative immunoglobulin domains, CD8A, CD8B, CD4 or CD28, TRAC, TRBC, TRGC, TRDC.

[210] Table 5 below provides non-limiting examples of hinge regions suitable for use in joining the targeting fraction and the specific / tumor-associated antigen epitope fraction in the bridging molecules of the invention.

[211] It will be appreciated that the targeting fraction can be joined to the specific / tumor-associated antigen epitope fraction by more than one linker and / or more than one hinge region. For example, the fusion protein may comprise (from the N-terminus to the C-terminus) the specific / tumor-associated antigen epitope fraction, directly conjugated to the targeting fraction. Alternatively, the fusion protein may comprise the specific / tumor-associated antigen epitope fraction, followed by a linker region, and then the targeting fraction. Still further, the fusion protein may comprise the specific / tumor-associated antigen epitope fraction, followed by a linker region, then a hinge region, and then the targeting fraction.In yet another embodiment, the fusion protein may comprise the tumor-specific antigen epitope fraction, followed by a linker region, then a hinge region, an additional linker region, and then the targeting fraction. Table 5: exemplary hinge regions Amino acid sequence SEQ ID NO: Description EPKSSDKTHTSPPSP 219 Sequences derived from IgG1 (this Petition 870250089762, dated 02 / 10 / 2025, pp. 476 / 580 116 / 176 Amino acid sequence SEQ ID NO: Description EPKSSDKTHTSPP 220 is,regiões of IgG1 modifica... EPKSGDKTHTGPPP 229 EPKSGDKTHT 230 DKTHTGPPGP 231 DKTHTGPPP 232 EPKSDKTHTGPPGP 233 EPKSGDKTHTSPPSP 234 EPKSGDKTHTGPPSP 235 EPKSSDKTHTGPPGP 236 EPKSSDKTHTGPPSP 237 EPKSSDKTHTGPPP 238 EPKSGDKTHTSPP 239 EPKSTDKTHTTPPTP 240 EPKSTDKTHTTPP 241 EPKSTDKTHTTPPPP 242 EPKSTDKTHT 243 DKTHTTPPTP 244 DKTHTTPP 245 EPKSDKTHTTPPTP 246 EPKSSDKTHTTPPTP 247 EPKSSDKTHTSPPTP 248 EPKSADKTHTTPPMP 249 EPKSVDKTHTLPPIP 250 EPKSLDKTHTAPPAP 251 EPKSVDKTHTAPP 252 EPKSIDKTHTLPP 253 DKTHTAPPLP 254 DKTHTVPPLP 255 EPKSSDKTHTSP 256 EPKSCDKTHTCPPCP 257 IgG1 do tipo selvagem ERKCCVECPPCP 258 IgG2 ELKTPLGDTTHTXPR 264 IgG3 ELKTPLGDTTHTXPR 264 IgG3 ELKTPLGDTTHTXPR 264 IgG3 ELKTPLGDTTHTXPR Petition 870250089762, on 10 / 02 / 2025, page. 477 / 580 117 / 176 SEQ amino acid sequence ID NO: Description ELKTPLGDTTHT 265 IgG3 mutated EPKSCDTPPPCPRCP 266 IgG3 of the selvagem type EPKSXDTPPPXPRXP 267 IgG3 mutated EPKPRPK3X6StaTPPPXda IgG3 269 ​​IgG3 mutant DTPPPXPRXP 270 IgG3 mutant ESKYGPPCPSCP 271 IgG4 do type selvagem EXKYGPPCPXCP 272 IgG4 mutant EXKYGPPCP 273 IgG4 mutant KYGPPG muSCPKTH4ICXgCP 275 Selvagem-type IgG1 (v2) EPKSCDTPPPCP 276 Selvagem-type IgG3 (v2) ESKYGPPSPSSP 277 Mutated IgG4 ERKSSVESPPSP 278 Mutated IgG2 EPKSSDTPPPSP 279 Mutada IgG Fab regions of the bridge molecule

[212] The Fab regions of the bridge molecule of the invention preferably bind to a cell surface molecule on a target cell. The cell surface molecule may comprise an antigen. The cell surface molecule may be selected from a protein, a lipid fraction, a glycoprotein, a glycolipid, a carbohydrate, a polysaccharide, a nucleic acid, an MHC-bound peptide, or a combination thereof. The cell surface molecule may comprise parts (e.g., envelopes, capsules, cell walls, flagella, fimbriae, and toxins) of bacteria, viruses, and other microorganisms. The cell surface molecule may be expressed by the target cell. The cell surface molecule may not be expressed by the target cell. By way of non-limiting example, the cell surface molecule may be a ligand expressed by a cell other than the target cell and which is bound to the target cell or to a cell surface molecule of the target cell.Furthermore, by way of a non-limiting example, the cell surface molecule may be a toxin, exogenous molecule, or viral protein that is bound to a cell surface or to the cell surface receptor of the target cell. Petition 870250089762, dated 02 / 10 / 2025, pages 478 / 580 118 / 176

[213] In preferred embodiments, the Fab regions of the bridge molecules of the invention are for binding to antigens on the surface of cancer cells.

[214] Bridge molecules can interact with a plurality of target cells. The target cell can be an infected cell. The target cell can be a pathogen-infected cell. The target cell can be a diseased cell. The target cell can be a genetically modified cell. The target cell may not be a host cell. The target cell can come from an invading organism (e.g., yeast, worm, bacteria, fungus). Additionally, bridge molecules that interact with a molecule on a non-cellular target are disclosed in the present invention. The non-cellular target can be a virus or a portion thereof. The non-cellular target can be a fragment of a cell. The non-cellular target can be a component or protein of the extracellular matrix.

[215] The target cell can be derived from a tissue. The tissue can be selected from brain, esophagus, breast, viscera, intestine, colon, lung, glia, ovary, uterus, testes, prostate, gastrointestinal tract, bladder, liver, spleen, thymus, bone, fat, and skin. The target cell can be derived from one or more endocrine glands. Alternatively, or additionally, the target cell can be derived from one or more endocrine glands. The endocrine gland can be a lymphatic gland, pituitary gland, thyroid gland, parathyroid gland, pancreas, gonad, or pineal gland.

[216] The target cell can be selected from a stem cell, a pluripotent cell, a hematopoietic stem cell, or a progenitor cell. The target cell can be a circulating cell. The target cell can be an immune cell.

[217] The target cell may be a cancer stem cell. The target cell may be a cancer cell. The cancer cell may be derived from a Petition 870250089762, dated 02 / 10 / 2025, pages 479 / 580 119 / 176 tissue. The tissue may be selected, by way of non-limiting example, from a brain, an esophagus, a breast, a colon, a lung, glia, an ovary, a uterus, a testicle, a prostate, a gastrointestinal tract, a bladder, a liver, a thyroid, and skin. The cancer cell may be derived from bone. The cancer cell may be derived from blood. The cancer cell may be derived from a B cell, a T cell, a monocyte, a platelet, a leukocyte, a neutrophil, an eosinophil, a basophil, a lymphocyte, a hematopoietic stem cell, or an endothelial cell progenitor. The cancer cell may be derived from a CD19-positive B lymphocyte. The cancer cell may be derived from a stem cell. The cancer cell may be derived from a pluripotent cell. A cancer cell can originate from one or more endocrine glands.An endocrine gland can be a lymphatic gland, pituitary gland, thyroid gland, parathyroid gland, pancreas, gonad, or pineal gland.

[218] The cell surface molecule of the target cell may be a receptor. The receptor may be an extracellular receptor. The receptor may be a cell surface receptor. By way of non-limiting example, the receptor may bind to a hormone, a neurotransmitter, a cytokine, a growth factor, or a cell recognition molecule. The receptor may be a transmembrane receptor. The receptor may be an enzyme-linked receptor. The receptor may be a G protein-coupled receptor (GPCR). The receptor may be a growth factor receptor. By way of non-limiting example, the growth factor receptor may be selected from an epidermal growth factor receptor, a fibroblast growth factor receptor, a platelet-derived growth factor receptor, a nerve growth factor receptor, a transforming growth factor receptor, a growth factor receptor of the growth factor morphogenetic protein Petition 870250089762, dated 02 / 10 / 2025, pp. 480 / 580 120 / 176 bone, a hepatocyte growth factor receptor, a vascular endothelial growth factor receptor, a stem cell factor receptor, an insulin-like growth factor receptor, a somatomedin receptor, an erythropoietin receptor, and homologs and fragments thereof. The receptor may be a hormone receptor. The receptor may be an insulin receptor. By way of non-limiting example, the receptor may be selected from an eicosanoid receptor, a prostaglandin receptor, an estrogen receptor, a follicle-stimulating hormone receptor, a progesterone receptor, a growth hormone receptor, a gonadotropin-releasing hormone receptor, homologs and fragments thereof. The receptor may be an adrenergic receptor. The receptor may be an integrin. The receptor may be an Eph receptor. The receptor may be a luteinizing hormone receptor.The cell surface molecule may be at least 50% homologous to a luteinizing hormone receptor. The receptor may be an immune receptor. By way of non-limiting example, the immune receptor may be selected from a pattern recognition receptor, a Toll-like receptor, a NOD-like receptor, an assassin activator receptor, an assassin inhibitor receptor, an Fc receptor, a B cell receptor, a complement receptor, a chemokine receptor, and a cytokine receptor. By way of non-limiting example, the cytokine receptor may be selected from an interleukin receptor, an interferon receptor, a transforming growth factor receptor, a tumor necrosis factor receptor, a colony-stimulating factor receptor, homologs and fragments thereof. The receptor may be a receptor kinase. The receptor kinase may be a tyrosine kinase receptor. The receptor kinase may be a serine kinase receptor.The receptor kinase may be a threonine receptor. Petition 870250089762, dated 02 / 10 / 2025, pp. 481 / 580 121 / 176 kinase. As a non-limiting example, the receptor kinase may activate a signaling protein selected from a Ras, a Raf, a PI3K, a protein kinase A, a protein kinase B, a protein kinase C, an AKT, an AMPK, a phospholipase, homologs thereof, and fragments thereof. The receptor kinase may activate a MAPK / ERK signaling pathway. The receptor kinase may activate Jak, Stat, or Smad.

[219] The cell surface molecule may be a non-receptor cell surface protein. The cell surface molecule may be a protein of the differentiation cluster. By way of non-limiting example, the cell surface molecule may be selected from CD3, CD4, CD8, CD11a, CD11b, CD13, CD14, CD15, CD16, CD22, CD24, CD25, CD30, CD31, CD33, CD34, CD38, CD45, CD56, CD61, CD91, CD114, CD117, CD182, CD200, fragments thereof and homologs thereof.

[220] The target cell's cell surface molecule may be a molecule that does not comprise a peptide. The cell surface molecule may comprise a lipid. The cell surface molecule may comprise a lipid moiety or a lipid group. The lipid moiety may comprise a sterol. The lipid moiety may comprise a fatty acid. The antigen may comprise a glycolipid. The cell surface molecule may comprise a carbohydrate.

[221] The cell surface molecule of the target cell may be an antigen. The antigen may be at least a portion of a surface antigen or a cell surface marker on a cell. The antigen may be a receptor or a coreceptor on a cell. The antigen may refer to a molecule or molecular fragment that can be bound by a major histocompatibility complex (MHC) and presented to a T cell receptor. The term antigen may also refer to an immunogen. The immunogen Petition 870250089762, dated 02 / 10 / 2025, pages 482 / 580 122 / 176 can elicit an adaptive immune response if injected alone into an individual. The immunogen can induce an immune response on its own. The antigen can be a superantigen, a T-dependent antigen, or a T-dependent antigen. The antigen can be an exogenous antigen. Exogenous antigens are typically antigens that entered the body from the outside, for example, by inhalation, ingestion, or injection. Some antigens may start as exogenous antigens and then become endogenous (e.g., intracellular viruses). The antigen can be an endogenous antigen. An endogenous antigen can be an antigen that was generated within cells as a result of normal cellular metabolism, or because of pathogenic infections (e.g., viral, bacterial, fungal, parasitic). The antigen can be an autoantigen.An autoantigen can be a normal protein or a protein complex (and sometimes DNA or RNA) that is recognized by the immune system of patients suffering from a specific autoimmune disease. These antigens, under normal conditions, should not be targeted by the immune system, but due to genetic and / or environmental factors, normal immune tolerance to such an antigen is not present in these patients. The antigen may be present or overexpressed due to a condition or disease. The condition or disease may be cancer or leukemia. The condition may be an inflammatory disease or condition. The condition may be a metabolic disease. The condition may be a genetic disorder.

[222] The present invention may also find application in the treatment of specific autoimmune diseases associated with the B or T cell lineage, for example, using anti-idiotypic antibodies or fragments thereof or ligands to target the B cell receptor and / or the T cell receptor. Such diseases include myasthenia gravis, systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), multiple sclerosis (MS), hyperacute, acute, chronic rejection. Petition 870250089762, dated 02 / 10 / 2025, pp. 483 / 580 123 / 176 or mixed type solid organ transplant, bone marrow or stem cell transplant rejection, and graft-versus-host disease.

[223] The cell surface molecule of the target cell may be an antigen that has been designated as a tumor antigen. Tumor antigens or neoantigens may be antigens that are presented by MHC I or MHC II molecules on the surface of tumor cells. These antigens may sometimes be presented by tumor cells and never by normal cells. In this case, they are called tumor-specific antigens (TSAs) and generally result from a tumor-specific mutation. More common are antigens that are presented by both tumor cells and normal cells, and are called tumor-associated antigens (TAAs). A TAA-associated antigen is not exclusive to a tumor cell and is instead also expressed in a normal cell under conditions that do not induce a state of immunological tolerance to the antigen. The expression of the antigen in the tumor may occur under conditions that allow the immune system to respond to the antigen.Tumor antigens (TAAs) can be antigens that are expressed on normal cells during fetal development, when the immune system is immature and unable to respond, or they can be antigens that are normally present at extremely low levels on normal cells but are expressed at much higher levels on tumor cells. Cytotoxic T lymphocytes that recognize these antigens may be able to destroy tumor cells before they proliferate or metastasize. Tumor antigens can also be on the surface of the tumor in the form of, for example, a mutated receptor, in which case they can be recognized by B cells.

[224] Non-limiting examples of TSA or TAA antigens include the following: Differentiation antigens such as MART-1 / MelanA (MART-I), gp 100 (Pmel 17), tyrosinase, TRP-1, TRP-2 and specific multilineage antigens of Petition 870250089762, dated 02 / 10 / 2025, pages 484 / 580 124 / 176 tumor antigens such as MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, p15; overexpressed embryonic antigens such as CEA; overexpressed oncogenes and mutated tumor suppressor genes such as p53, Ras, HER-2 / neu; unique tumor antigens resulting from chromosomal translocations such as BCRABL, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR; and viral antigens such as Epstein-Barr virus (EBVA) antigens and human papillomavirus (HPV) E6 and E7 antigens. Other large protein-based antigens include TSP180, MAGE-4, MAGE-5, MAGE-6, RAGE, NY-ESO, p185erbB2, p180erbB-3, c-met, nm-23Hl, PSA, TAG-72, CA 19-9, CA 72-4, CAM 17.1, NuMa, K-ras, beta-Catenin, CDK4, Mum-1, p 15, p 16, 43-9F, 5T4, 791Tgp72, alpha-fetoprotein, beta-HCG, BCA225, BTAA, CA 125, CA 15-3 / CA 27.29\BCAA, CA 195, CA 242, CA-50, CAM43, 20 CD68\Pl, C0-029, FGF-5, G250, Ga733\EpCAM, HTgp-175, M344, MA-50, MG7-Ag, MOV18, NB / 70K, NY-C0-1, RCASl, SDCCAG16, TA-90\Mac-2 binding protein\cyclophilin C-associated protein, TAAL6, TAG72, TLP, and TPS.

[225] The cell surface molecule of the target cell may be an antigen selected from the group consisting of any antigens expressed on the surface. Exemplary target antigens may include, but are not limited to: CD33 (Siglec-3), CD123 (IL3RA), CD135 (FLT-3), CD44 (HCAM), CD44V6, CD47, CD184 (CXCR4), CLEC12A (CLL1), LeY, FRe, MICA / B, CD305 (LAIR-1), CD366 (TIM3), CD96 (TACTILE), CD133, CD56, CD29 (ITGB1), CD44 (HCAM), CD47 (IAP), CD66 (CEA), CD112 (Nectin2), CD117 (c-Kit), CD133, CD146 (MCAM), CD155 (PVR), CD171 (L1CAM), CD200 (OX-2), CD221 (IGF1), CD227 (MUC1), CD243 (MRD1), CD246 (ALK), CD271 (LNGFR), CD19, CD20, GD2, and EGFR. Other target antigens include TCR, sugars, lipids, carbohydrates, or any other molecule expressed on the surface of the target cell.

[226] In preferred embodiments, the target cell is a cancer cell and the cell surface molecule of the cancer cell is an associated antigen. Petition 870250089762, dated 02 / 10 / 2025, pp. 485 / 580 125 / 176 to cancer. The antigen may be a tumor-specific antigen or a tumor-associated antigen. The antigen may be one that is associated with a particular type of cancer. For example, overexpression of the antigen may be associated with a specific cancer or a specific class of cancer. For example, where the cancer is breast cancer, the antigen may be associated with breast cancer but not another form of cancer. Alternatively, the antigen may be associated with a class of cancers, such as solid tumors, but not associated with hematologic (i.e., liquid) tumors, or vice versa. The antigen may be associated with cancers of a particular lineage but not others. For example, the antigen may be associated with sarcomas but not lymphomas or carcinoma. As used in the present invention, the term associated with in relation to cancer will be understood to mean that the expression of the antigen (whether increased or decreased) is considered a marker of cancer.It is appreciated that there may be low levels of expression of an antigen, but this does not equate to the antigen being associated with a particular cancer.

[227] Suitable cancer antigens that can be bound by the Fab regions of the bridge molecule (or indeed by the antigen recognition domain of a CAR) include, but are not limited to, mesothelin (MSLN), prostate-specific membrane antigen (PSMA), prostate stem cell antigen (PCSA), carbonic anhydrase IX (CAIX), carcinoembryonic antigen (CEA), CD5, CD7, CD10, CD19, CD20, CD22, CD30, CD33, CD34, CD38, CD41, CD44, CD49f, CD56, CD74, CD123, CD133, CD138, epithelial glycoprotein (EGP 2), epithelial glycoprotein-40 (EGP-40), epithelial cell adhesion molecule (EpCAM), folate-binding protein (FBP), fetal acetylcholine receptor (AChR), folate receptor-α and β (FRa and β), Ganglioside G2 (GD2), Ganglioside G3 (GD3), Human Epidermal Growth Factor Receptor 2 (HER-2 / ERB2), Petition 870250089762, dated 02 / 10 / 2025, pp. 486 / 580 126 / 176 HER3, Epidermal Growth Factor Receptor vIII (EGFRvIII), ERB3, ERB4, human telomerase reverse transcriptase (hTERT), interleukin-13 receptor alpha-2 subunit (IL-13Ra2), κ light chain, kinase insertion domain receptor (KDR), Lewis A (CA19).9), Lewis Y (LeY), cell adhesion molecule L1 (L1CAM), melanoma-associated antigen 1 (melanoma antigen family A1, MAGE-A1), Mucin 16 (Muc-16), Mucin 1 (Muc-1), NKG2D ligands, testicular cancer antigen NY-ESO-1, oncofetal antigen (h5T4), tumor-associated glycoprotein 72 (TAG-72), vascular endothelial growth factor receptor 2 (VEGF-R2), Wilms tumor protein (WT-1), transmembrane receptor tyrosine kinase type 1 (ROR1), B7-H3 (CD276), B7H6 (Nkp30), chondroitin sulfate proteoglycan-4 (CSPG4), DNAX Accessory Molecule (DNAM-1), Ephrin receptor type A 2 (EpHA2), Fibroblast-associated protein (FAP), Gpl00 / HLA-A2, Glypican 3 (GPC3), HA-1H, HERK-V, IL-1 1Ra, Latent Membrane Protein 1 (LMP1), Neurocellular Adhesion Molecule (NCAM / CD56), and Trail Receptor (TRAIL R). It is understood that these or other cancer antigens can be used for targeting by a bridging molecule in the present invention.

[228] The bridging molecule may be in the form of an immunoglobulin selected from an IgG, an IgA, an IgD, an IgE, an IgM, a fragment thereof, or a modification thereof. The immunoglobulin may be IgG. IgG may be IgG1. IgG may be IgG2. IgG may be IgG3. IgG may be IgG4. IgG may have one or more Fc mutations to modulate the binding of the FcR of endogenous T cells to the bridging molecule. IgG may have one or more Fc mutations to modulate the removal of the Fc binding capacity to the FcR of FcR-positive cells. One or more Fc mutations may remove a glycosylation site. One or more Fc mutations may be selected from E233P, L234V, L235A, delG236, A327G, A330S, P331S, N297Q, and any Petition 870250089762, dated 02 / 10 / 2025, pp. 487 / 580 127 / 176 combination of them. One or more Fc mutations may be in IgG1. One or more Fc mutations in IgG1 may be L234A, L235A, or both. Alternatively, or additionally, one or more Fc mutations in IgG1 may be L234A, L235E, or both. Alternatively, or additionally, one or more Fc mutations in IgG1 may be N297A. Alternatively, or additionally, one or more Fc mutations may be in IgG2. One or more Fc mutations in IgG2 may be V234A, V237A, or both.

[229] The bridging molecule can be a null Fc immunoglobulin or a fragment thereof.

[230] As used in the present invention, the term antibody fragment refers to any form of an antibody other than the full-length form. Antibody fragments in the present invention include antibodies that are smaller components that exist within full-length antibodies, and antibodies that have been manipulated. Antibody fragments include, but are not limited to, Fv, Fc, Fab, and (Fab')2, single-chain Fv (scFv), diabodies, triabodies, tetrabodies, bifunctional hybrid antibodies, CDR1, CDR2, CDR3, combinations of CDRs, variable regions, framework regions, constant regions, heavy chains, light chains, non-antibody alternative scaffold molecules, and bispecific antibodies. Unless specifically noted otherwise, statements and claims using the term antibody or antibodies may specifically include antibody fragment and antibody fragments.

[231] The bridging molecule can be a human, fully human, humanized, engineered human, non-human, and / or chimeric antibody. The non-human antibody can be humanized to reduce immunogenicity to humans while maintaining the specificity and affinity of the parent non-human antibody. Chimeric antibodies may refer to antibodies created Petition 870250089762, dated 02 / 10 / 2025, pages 488 / 580 128 / 176 through the union of two or more antibody genes that originally coded for separate antibodies. A chimeric antibody may comprise at least one amino acid from a first antibody and at least one amino acid from a second antibody, where the first and second antibodies are different. At least a portion of the antibody or antibody fragment may be from a bovine species, a human species, or a murine species. At least a portion of the antibody or antibody fragment may be from a mouse, a goat, a guinea pig, or a rabbit. At least a portion of the antibody or antibody fragment may be from a human. At least a portion of the antibody or antibody fragment may be from a cynomolgus monkey.

[232] The bridge molecule may be based on or derived from an antibody or antibody fragment from a mammal, bird, fish, amphibian, or reptile. Mammals include, but are not limited to, carnivores, rodents, elephants, marsupials, rabbits, bats, primates, seals, anteaters, cetaceans, odd-toed ungulates, and even-toed ungulates. The mammal may be a human, a non-human primate, a mouse, a sheep, a cat, a dog, a cow, a horse, a goat, or a pig.

[233] The bridge molecule can recognize or bind to an antigen selected from, by way of non-limiting example, CD19, Her2, CLL-1, CD33, EGFRvIII, CD20, CD22, BCMA or a fragment thereof. The antigen may comprise a wild-type antigen. The antigen may comprise one or more mutations.

[234] A Fab region of the bridge molecule may be an anti-CD19 antibody or a fragment thereof. The bridge molecule may be an anti-CD22 antibody. A Fab region of the bridge molecule may be an anti-BCMA antibody or a fragment thereof. A Fab region of the bridge molecule may Petition 870250089762, dated 02 / 10 / 2025, pages 489 / 580 129 / 176 being an anti-EGFRvIII antibody or a fragment thereof. A Fab region of the bridge molecule may be an anti-Her2 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD20 antibody or an antibody fragment. The bridge molecule may comprise rituximab. A Fab region of the bridge molecule may comprise an anti-EGFR antibody or an antibody fragment. A Fab region of the bridge molecule may comprise an anti-CEA antibody or an antibody fragment. A Fab region of the bridge molecule may comprise an anti-CLL-1 antibody or an antibody fragment. A Fab region of the bridge molecule may comprise an anti-CD33 antibody or an antibody fragment. A Fab region of the bridge molecule may comprise an anti-EpCAM antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD30 antibody or a fragment thereof.A Fab region of the bridge molecule may comprise an anti-CD79B antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD37 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD38 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD70 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD276 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-GD2 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD371 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD135 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD105 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD123 antibody or a fragment thereof.A Fab region of the molecule. Petition 870250089762, dated 02 / 10 / 2025, pages 490 / 580 A Fab region of the bridge molecule (130 / 176) may comprise an anti-ROR-1 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-PDL1 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-MET-R antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-PDGFRalpha antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-Her3 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-FRalpha antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-GPC3 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-SLAMf7 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-TNFRSF10B antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-GPNMB antibody or a fragment thereof.A Fab region of the bridge molecule may comprise an anti-VEGFR2 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-alpha4beta7 / alphaEbeta7 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CSPG4 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD80 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CCR4 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD115 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-ENOX-2 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD56 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-huVH1-69 antibody or a fragment thereof. A Fab region of the... Petition 870250089762, dated 02 / 10 / 2025, pp. 491 / 580A 131 / 176 bridge molecule may comprise an anti-CD117 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD133 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-MUC1 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-MSLN antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-ROR-2 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-IL13Ra2 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-EPHA2 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-EGFRvIII antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-PSMA antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CEA antibody or a fragment thereof.A Fab region of the bridge molecule may comprise an anti-Lewis Y antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-PSCA antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-MUC1 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD171L1CAM antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-EpCAM antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-ALK antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-IGF-1R CD221 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-Nectin 4 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-FAP antibody or a fragment thereof. A Fab region of the bridge molecule... Petition 870250089762, dated 02 / 10 / 2025, pp. 492 / 580 A Fab region of the 132 / 176 bridge may comprise an anti-AXL antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD138 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CLDN6 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-Her4 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-Claudin 18.2 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-O-acetylated GD2 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-GD3 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-GM2 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-TM4SF1 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD147 antibody or a fragment thereof.A Fab region of the bridge molecule may comprise an anti-CEACAM5 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-VEGFR-1 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-PDPN antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-WT1 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-GPC2 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-FGFR4 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-EphB4 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-STEAP-1 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-STEAP-2 antibody or a fragment thereof. A Fab region of the bridge molecule... Petition 870250089762, dated 02 / 10 / 2025, pp. 493 / 580 A Fab region of the bridge molecule may comprise an anti-MUCl antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-chlorotoxin antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-206 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-ILRAP1 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-MICA antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-MAGE-A1 scTcR antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-MAGE-A1 sTCR antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-MICA antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-TRBC1 / 2 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-B7-H7 antibody or a fragment thereof.A Fab region of the bridge molecule may comprise an anti-CD34 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD7 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-TIM3 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD191 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD66b antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-CD11b antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-EMR2 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-MUC16 antibody or a fragment thereof. A Fab region of the bridge molecule may comprise an anti-NYESO-1 HLA-A2 antibody or a fragment thereof, or a soluble TcR or variant thereof. Petition 870250089762, dated 02 / 10 / 2025, pp. 494 / 580 134 / 176 same. A Fab region of the bridge molecule may comprise an anti-SURVIVIN HLA-A2 antibody or a fragment thereof, or a soluble TcR or variant thereof. A Fab region of the bridge molecule may comprise an anti-CD200 antibody or a fragment thereof.

[235] The bridge molecule may comprise or be derived from any commercially available antibody, such as: trastuzumab (for Her2 binding), alemtuzumab (for CD52 binding), bevacizumab (for VEGF-A binding), brentuximab (for CD30 binding), gemtuzumab (for CD33 binding), ipilimumab (for VTLA-4 binding), ibritumomab (for CD20 binding), panitumumab (for EGFR binding), cetuximab (for EGFR binding), rituximab (for CD20 binding), and fragments thereof.

[236] The bridge molecule may be any one referred to in Table 1 or 2 or an antigen-binding fragment that is at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical thereto.

[237] It will be well within the competence of a person skilled in the art to be able to determine the appropriate design of a bridge molecule as described in the present invention, including the selection of the Fab regions (antigen-binding domains) of the bridge molecule and the fragments to be bound by the exogenous immune cell receptor.

[238] More specifically, the person skilled in the art, having knowledge of the CAR for use in the methods (including redirection as described in the present invention) would be able to select an alternative redirection fraction to which the CAR should be directed, to facilitate treatment. Petition 870250089762, dated 02 / 10 / 2025, pages 495 / 580 135 / 176 of cancer or disease requiring treatment.

[239] As described in the present invention, the Fab region of the bridge molecule can be for binding to any cell surface molecule on a target cell (such as a tumor-associated antigen expressed on a cancer cell). The person skilled in the art can select an appropriate cell surface molecule depending on the disease (e.g., cancer) requiring treatment. For example, if the disease is a Her2+ cancer, then the person skilled in the art can select a Fab that is capable of specifically binding to Her2 on the cancer cell. Similarly, if the disease is a CD19+ cancer, then the person skilled in the art can select a Fab that is capable of specifically binding to CD19 on the cancer cell. The person skilled in the art will be familiar with methods for determining the antigen expression profile of the cancer or condition to be treated in order to identify suitable surface molecules expressed on the target cell.

[240] Having identified a suitable cell surface molecule (target antigen) on the target cell (e.g., CD19, Her2, CLL-1, CD33, EGFRvIII, CD20, CD22, BCMA, or any other tumor-associated antigen described in the present invention or that may be selected for targeting), the person skilled in the art can readily determine the sequence of a Fab-binding domain that can bind to said target antigen, including by reference to commercially available antibodies or published literature describing antigen-binding domains of known antibodies that bind to the antigen. The person skilled in the art can then formulate the structure of a bridge molecule (fusion protein) as described in the present invention, comprising a Fab that binds to the cell surface molecule on a target cell. The composition of the remaining components of the bridge molecule (e.g., the tumor-specific antigen epitope fraction) is described Petition 870250089762, dated 02 / 10 / 2025, pp. 496 / 580 136 / 176 additionally in the present invention.

[241] Finally, a person skilled in the art, having identified a suitable cell surface molecule (target antigen) on the target cell and a tumor-specific antigen epitope fraction, so as to arrive at a bridging molecule of the invention, can readily determine, using routine techniques, whether the bridging molecule: a) binds to the target cell, b) binds to the CAR-T cell, and c) can successfully redirect the CAR-T cell to the target cells to induce cell death. Methods for determining target antigen binding and cytotoxicity are well known in the art. Non-limiting methods and various experimental protocols for determining target cell binding, CAR-T cell binding, and cell death induction are described in detail in the present invention in the Examples. P2X receptor epitope fraction?

[242] The present invention contemplates the use of an antigen epitope fraction in the form of a dysfunctional P2X7 receptor epitope.

[243] A dysfunctional P2X7 receptor epitope fraction can be provided in the form of a dysfunctional P2X7 receptor, or a fragment of a dysfunctional P2X7 receptor, which has at least one of the three ATP binding sites that are formed at the interface between correctly packaged adjacent monomers that are unable to bind to ATP. Such receptors are unable to extend the opening of non-selective calcium channels to apoptotic pores.

[244] A range of peptide fragments of a dysfunctional P2X7 receptor is known and discussed in PCT / AU2002 / 000061 (and in the corresponding publications WO 2002 / 057306 and US 7,326,415, US 7,888,473, US 7,531,171, US 8,080,635, US 8,399,617, US 8,709,425, US 9,663,584 or US 10,450,380), PCT / AU2008 / 001364 (and in the corresponding publications WO 2009 / 033233 and US 8,440,186, US 9,181,320, US 9,944,701 or US 10,597,45) and Petition 870250089762, dated 02 / 10 / 2025, pp. 497 / 580 137 / 176 PCT / AU2009 / 000869 (and in the corresponding publications WO 2010 / 000041 and US 8,597,643, US 9,328,155 or US 10,238,716) whose contents are all fully incorporated. Exemplary peptides within these specifications that include epitopes contemplated for use in this invention are described below. Publication PCT Peptide sequence

[245] WO 2002 / 057306 GHNYTTRNILPGLNITC (SEQ ID NO: 153) (also referred to in the present invention as the epitope E200) WO 2009 / 033233 KYYKENNVEKRTLIK (SEQ ID NO: 163) (also referred to in the present invention as the epitope E300) WO 2010 / 000041 GHNYTTRNILPGAGAKYYKENNVEK (SEQ ID NO: 165) (also referred to in the present invention as the E200 / E300 epitope or composite) In any embodiment, the amino acid sequence of the P2X7 receptor epitope fraction of any bridge molecule described in the present invention comprises or consists of a sequence as set forth in any of the SEQ ID Nos: 155 to 218, or sequences that are at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical thereto. Preferably, the P2X7 receptor epitope fraction comprises at least the sequence with SEQ ID NO: 162.

[246] The P2X7 receptor epitope fragment may have any functional chemical group, such as a carboxyl group, an active ester, an acetamide or maleimide, capable of coupling to a targeting fragment as disclosed in the present invention, for example, an antibody or fragment of Petition 870250089762, dated 02 / 10 / 2025, pp. 498 / 580 138 / 176 even when using NH2 or SH groups for coupling to it.

[247] Several examples of P2X7 receptor epitope fractions are described in the present invention, particularly in Table 3. As is clear from Table 3, the minimum E200 sequence comprises at least the sequence with SEQ ID NO: 162. In preferred embodiments, the minimum E200 sequence comprises at least the sequence with SEQ ID NO: 155 or a variant thereof.

[248] In certain examples, the E200 epitope for inclusion in the bridge molecules of the invention may comprise extensions in the N-terminal and / or C-terminal region. The extensions may be derived from the naturally occurring nfP2X7 receptor sequence and may comprise extensions of at least 1 amino acid residue, at least 2 amino acid residues, at least 3, at least 4, at least 5 or more amino acid residues. For example, the E200 epitope defined in SEQ ID NO: 155 may comprise an N-terminal extension of the DFP sequence, corresponding to the three amino acid residues immediately N-terminal to the E200 sequence in SEQ ID NO: 1. Additionally, the E200 epitope defined in SEQ ID NO: 155 may comprise a C-terminal extension, as represented, for example, in SEQ ID NO: 155 and 156. It will be appreciated that such N- and C-terminal extensions may serve to improve the effectiveness of CAR-T cell binding to the epitope on the bridge molecule.

[249] In any embodiment, the P2X7 receptor epitope fraction is conjugated to the Fab region of the bridge molecule to allow sufficient accessibility for binding by an nfP2X7 receptor CAR. Thus, although the minimal sequence of the dysfunctional P2X7 receptor epitope fraction comprises the minimum E200 sequence at least the SEQ ID NO: 162 sequence, in preferred embodiments, the minimum E200 sequence comprises at least the SEQ ID NO: 154 sequence, or a variant thereof, in addition to Petition 870250089762, dated 02 / 10 / 2025, pp. 499 / 580 139 / 176 one or more linker or hinge regions to allow binding by an nfP2X7 receptor CAR. In certain embodiments, the total length of the dysfunctional P2X7 receptor epitope fraction (including linker and hinge region) is at least 17 amino acids in length, or at least 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 45, 47, 48, 49 or 50 amino acids in length. Preferably, the epitope fraction of the dysfunctional P2X7 receptor is no more than about 100 amino acids in length, more preferably no more than about 99, 98, 97, 96, 95, 94, 93, 92, 91, 90, 89, 88, 87, 86, 85, 84, 83, 82, 81, 80, 79, 78, 77, 76, 75, 74, 73, 72, 71, 70, 69, 68, 67, 66, 65, 64, 63, 62, 61, 60, 59, 58, 57, 56, 55, 54, 53, 52, 51 or 50 amino acids in length.

[250] In the context of a bridge molecule for CD19 targeting, preferably the dysfunctional P2X7 receptor epitope fragment is no more than about 35 amino acids long and at least 20 amino acids long, preferably at least 21, 22, 23, 24, 25 or 26 amino acids long.

[251] In the context of a bridge molecule for targeting CD33, preferably the dysfunctional P2X7 receptor epitope fragment is no more than about 45 amino acids long and at least 20 amino acids long, preferably at least 21, 22, 23, 24, 25 or 26 amino acids long. PNE epitope fraction

[252] In any embodiment, the amino acid sequence of the PNE neoepitope fraction of any bridge molecule described in the present invention comprises or consists of a sequence as set forth in any of the SEQ ID Nos: 125, or sequences at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least Petition 870250089762, dated 02 / 10 / 2025, pp. 500 / 580 140 / 176 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to it. EGFRvIII epitope fraction

[253] In any embodiment, the amino acid sequence of the EGFRvIII epitope portion of any bridge molecule described in the present invention comprises or consists of a sequence as set forth in any of the SEQ ID Nos: 267, or sequences at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical thereto. Preferably, the EGFRvIII epitope portion comprises at least the sequence of SEQ ID NO: 134. CLDN6 epitope fraction

[254] In any embodiment, the amino acid sequence of the CLDN6 epitope fraction of any bridge molecule described in the present invention comprises or consists of a sequence as set forth in any of the SEQ ID Nos: 143, 282 or 283, or sequences at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical thereto. Preferably, the CLDN6 epitope fraction comprises at least the sequence of SEQ ID NO: 143. Exemplary bridge molecules Petition 870250089762, dated 02 / 10 / 2025, pp. 501 / 580 141 / 176

[255] This descriptive report provides several non-limiting examples of tumor-specific antigen epitope fraction (e.g., P2X7, CLDN6 or EGFRIII receptor epitope fractions) / Fab region pairs.

[256] Exemplary bridging molecules of the invention are described in Tables 1 and 2. For the bridging molecules described in Table 1 that include an nfP2X7 epitope fraction, the descriptive report includes the bridging molecules, but with the nfP2X7 epitope fraction replaced by a PNE, EGFRvIII or CLDN6 epitope fraction (in Table 2).

[257] In examples where the bridge molecules comprise a Fab region for binding to CD19, the bridge molecule may comprise or consist of a combination of paired heavy and light variable chains as set forth in Tables 1 or 2 (heavy and light chain, respectively); or sequences that are at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the same.

[258] In any embodiment, the bridge molecule of the invention may comprise the amino acid sequences of SEQ ID NOs: 1, 9 and 10, or of SEQ ID NOs: 2, 8 and 10, or of SEQ ID NOs: 287, 8 and 10, or of SEQ ID NOs: 3 and 9, or of SEQ ID NOs: 4, 11 and 12, or of SEQ ID NOs: 5, 11 and 12, or of SEQ ID NOs: 6, 13 and 14, or of SEQ ID NOs: 7, 13 and 14, or of SEQ ID NOs: 1, 2 and 288, or of SEQ ID NOs 4 and 11 in combination with SEQ ID NOs: 8 and 287, 1 and 288, 2 and 8, 1 and 10 or 287 and 288; or sequences of at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least Petition 870250089762, dated 02 / 10 / 2025, pp. 502 / 580 142 / 176 minus 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to them. Nucleic acids

[259] In a further aspect, the present invention provides a nucleic acid molecule that encodes a bridge molecule of the invention, or part thereof.

[260] The nucleic acid molecule may comprise any polyribonucleotide or polydeoxyribonucleotide, which may be unmodified or modified RNA or DNA. For example, the nucleic acid molecule may include single-stranded and / or double-stranded DNA, DNA that is a mixture of single-stranded and double-stranded regions, single-stranded and double-stranded RNA, and RNA that is a mixture of single-stranded and double-stranded regions, hybrid molecules comprising DNA and RNA that may be single-stranded or, more typically, double-stranded or a mixture of single-stranded and double-stranded regions. In addition, the nucleic acid molecule may comprise triple-stranded regions comprising RNA or DNA or both RNA and DNA. The nucleic acid molecule may also comprise one or more modified bases or DNA or RNA backbones modified for stability or other reasons.A variety of modifications can be made to DNA and RNA; thus, the term nucleic acid molecule encompasses chemically, enzymatically, or metabolically modified forms.

[261] In some embodiments of the second aspect of the invention, the nucleic acid molecule comprises a nucleic acid sequence encoding the amino acid sequence of any of the SEQ ID NOs: 1 to 151. Preferably, the nucleic acid comprises a nucleotide sequence encoding the heavy chain and light chain pairs described above. Petition 870250089762, dated 02 / 10 / 2025, pp. 503 / 580 143 / 176

[262] Additionally, the present invention provides a nucleic acid construct including a nucleic acid molecule encoding a bridge molecule of the invention or part thereof. The nucleic acid construct may further comprise one or more of the following: an origin of replication for one or more hosts; a selection marker gene that is active in one or more hosts; and / or one or more transcriptional control sequences.

[263] As used in the present invention, the term selection marker gene includes any gene that confers a phenotype to a cell in which it is expressed, to facilitate the identification and / or selection of cells that are transfected or transformed with the construct.

[264] Selection marker genes include any nucleotide sequences that, when expressed by a cell transformed with the construct, confer a phenotype to the cell that facilitates the identification and / or selection of these transformed cells. A range of nucleotide sequences encoding suitable selection markers is known in the art (e.g. Mortesen, RM. and Kingston RE. Curr Protoc Mol Biol, 2009; Unit 9.5).Exemplary nucleotide sequences encoding selection markers include: Adenosine deaminase (ADA) gene; Cytosine deaminase (CDA) gene; Dihydrofolate reductase (DHFR) gene; Histidinol dehydrogenase (hisD) gene; Puromycin-N-acetyltransferase (PAC) gene; Thymidine kinase (TK) gene; Xanthine-guanine phosphoribosyltransferase (XGPRT) gene; or antibiotic resistance genes, such as ampicillin resistance genes, puromycin resistance genes, bleomycin resistance genes, hygromycin resistance genes, kanamycin resistance genes, and ampicillin resistance genes; fluorescent reporter genes, such as genes encoding green, red, yellow, or blue fluorescent protein; and reporter genes. Petition 870250089762, dated 02 / 10 / 2025, pp. 504 / 580 144 / 176 based on luminescence, such as the luciferase gene, among others, which allow the optical selection of cells using techniques such as Fluorescence-Activated Cell Sorting (FACS).

[265] In addition, it should be noted that the selection marker gene may be a distinct open reading frame in the construct or may be expressed as a fusion protein with another polypeptide (e.g., CAR).

[266] As set forth above, the nucleic acid construct may also comprise one or more transcriptional control sequences. The term transcriptional control sequence should be understood as including any nucleic acid sequence that affects the transcription of an operationally connected nucleic acid. A transcriptional control sequence may include, for example, a leader, a polyadenylation sequence, a promoter, an enhancer or upstream activation sequence, and a transcription terminator. Typically, a transcriptional control sequence includes at least one promoter. The term “promoter,” as used in the present invention, describes any nucleic acid that confers, activates, or enhances the expression of a nucleic acid in a cell.

[267] In some embodiments, at least one transcriptional control sequence is operationally connected to the nucleic acid molecule of the second aspect of the invention. For the purposes of this descriptive report, a transcriptional control sequence is considered operationally connected to a given nucleic acid molecule when the transcriptional control sequence is capable of promoting, inhibiting, or otherwise modulating the transcription of the nucleic acid molecule. Therefore, in some embodiments, the nucleic acid molecule is under the control of a transcriptional control sequence, such as a constitutive promoter or an inducible promoter.

[268] The nucleic acid construct can be in any form Petition 870250089762, dated 02 / 10 / 2025, pages 505 / 580 145 / 176 suitable, such as in the form of a plasmid, phage, transposon, cosmid, chromosome, vector, etc., that is capable of replication when associated with the appropriate control elements and that can transfer gene sequences, contained in the construct, between cells. Thus, the term includes cloning and expression vehicles, as well as viral vectors. In some embodiments, the nucleic acid construct is a vector. In some embodiments, the vector is a viral vector.

[269] A promoter can regulate the expression of an operationally connected nucleic acid molecule constitutively, or differentially, with respect to the cell, tissue, or organ in which the expression occurs. As such, the promoter may include, for example, a constitutive promoter or an inducible promoter. A constitutive promoter is a promoter that is active under most environmental and physiological conditions. An inducible promoter is a promoter that is active under specific environmental or physiological conditions. The present invention contemplates the use of any promoter that is active in the cell of interest. As such, a wide range of promoters would be readily discerned by one skilled in the art.

[270] Mammalian constitutive promoters may include, but are not limited to, Simian virus 40 (SV40), cytomegalovirus (CMV), P-actin, Ubiquitin C (UBC), elongation factor-1 alpha (EF1A), phosphoglycerate kinase (PGK) and chicken CMV / ε-actin early enhancer (CAGG).

[271] Inducible promoters may include, but are not limited to, chemically inducible promoters and physically inducible promoters. Chemically inducible promoters include promoters whose activity is regulated by chemical compounds such as alcohols, antibiotics, steroids, metal ions, or other compounds. Examples of chemically inducible promoters include: tetracycline-regulated promoters (e.g., see U.S. Patent 5,851,796 and U.S. Patent 5,464,758); promoters responsive to Petition 870250089762, dated 02 / 10 / 2025, pages 506 / 580 146 / 176 steroids, such as glucocorticoid receptor promoters (e.g., see U.S. Patent 5,512,483), ecdysone receptor promoters (e.g., see U.S. Patent 6,379,945) and the like; and metal-responsive promoters, such as metallothionein promoters (e.g., see U.S. 4,940,661, U.S. Patent 4,579,821 and U.S. 4,601,978), among others.

[272] In the context of the present invention, it will be appreciated that it may be desirable, under certain circumstances, that the expression of the bridge molecule be under the control of an inducible promoter. This allows switching on and off the expression of the nucleic acid encoding the bridge molecule.

[273] In certain embodiments, and in the case of an inducible expression construct, an immune cell expressing a CAR can be genetically modified with a) a nucleic acid encoding an antigen-binding receptor and b) an inducible expression construct encoding the bridge molecule. After binding of the dysfunctional P2X7 receptor, the immune cell induces expression of the gene encoding the bridge molecule. In certain embodiments, expression of such a gene facilitates and / or improves cancer treatment.

[274] As mentioned above, control sequences may also include a terminator. The term terminator refers to a DNA sequence at the end of a transcriptional unit that signals the termination of transcription. Terminators are 3'-untranslated DNA sequences that usually contain a polyadenylation signal, which facilitates the addition of polyadenylate sequences to the 3' end of a primary transcript. Like promoter sequences, the terminator can be any terminating sequence that is operational in the cells, tissues, or organs where it is intended to be used. Suitable terminators would be known to a person skilled in the art.

[275] As will be understood, the nucleic acid constructs of Petition 870250089762, dated 02 / 10 / 2025, pp. 507 / 580 147 / 176 The invention may additionally include additional sequences, for example, sequences that allow enhanced expression, cytoplasmic or membrane transport, and localization signals. Specific, non-limiting examples include an Internal Ribosome Entry Site (IRES) or a cleavage site (e.g., P2A, T2A).

[276] The present invention extends to all genetic constructs essentially as described in the present invention. These constructs may additionally include nucleotide sequences intended for the maintenance and / or replication of the genetic construct in eukaryotes and / or for the integration of the genetic construct or part thereof into the genome of a eukaryotic cell.

[277] Methods for the deliberate introduction (transfection / transduction) of exogenous genetic material, such as the nucleic acid construct of the third aspect of the present invention, into eukaryotic cells are known in the art. As will be understood, the most suitable method for introducing the nucleic acid construct into the desired host cell depends on many factors, such as the size of the nucleic acid construct, the type of host cell, the desired efficiency rate of transfection / transduction, and the desired or required final viability of the transfected / transduced cells.Non-limiting examples of such methods include: chemical transfection with chemicals such as cationic polymers, calcium phosphate or structures such as liposomes and dendrimers; non-chemical methods such as electroporation, sonoporation, heat shock or optical transfection; particle-based methods such as gene gun release, magnetofection or impalefection or viral transduction.

[278] The nucleic acid construct will be selected depending on the desired transfection / transduction method. In some modalities of Petition 870250089762, dated 02 / 10 / 2025, pages 508 / 580 148 / 176 third aspect of the invention, the nucleic acid construct is a viral vector, and the method for introducing the nucleic acid construct into a host cell is viral transduction. Methods for using viral transduction to induce the expression of a CAR in a PBMC are known in the art (Parker, LL. et al. Hum Gene Ther. 2000;11: 2377-87) and, more generally, for using retroviral systems for the transduction of mammalian cells (Cepko, C. and Pear, W. Curr Protoc Mol Biol. 2001, unit 9.9). In other embodiments, the nucleic acid construct is a plasmid, a cosmid, an artificial chromosome or similar, and can be transfected into the cell by any suitable method known in the art. Modified cells

[279] As described in the present invention, the invention includes the use of a cell expressing a chimeric antigen receptor comprising a binding domain and a signaling domain. The cell may be a manipulated cell, a genetically modified cell, an immune cell, or an immune effector cell, as described in the present invention. Additionally, the cell may be capable of differentiating into an immune cell. A cell that is capable of differentiating into an immune cell may be a stem cell, a multilineage progenitor cell, or an induced pluripotent stem cell.

[280] In any embodiment, the cell may be a T cell, in which optionally said T cell does not express TcRae, PD1, CD3 or CD96 (for example, by means of knockdown or deletion of one of these genes at the genetic or functional level).

[281] In any modality, the cell may be an immune cell, in which optionally said cell does not express accessory molecules that may be checkpoint, exhaustion or apoptosis-associated signaling receptors, as well as ligands such as PD-1, LAG-3, TIGIT, CTLA4, FAS-L and FAS-R, (for example, by knocking out one of these genes in Petition 870250089762, dated 02 / 10 / 2025, pp. 509 / 580 149 / 176 genetic level or functional level).

[282] In some forms, the genetically modified cell includes two or more different CARs.

[283] In some embodiments of the invention, the genetically modified cell includes a nucleic acid molecule or a nucleic acid construct, which codes for two or more different CARs. In some embodiments of the invention, the genetically modified cell includes two or more nucleic acid molecules, or two or more nucleic acid constructs, each of which codes for a different CAR.

[284] As referred to in the present invention, a genetically modified cell includes any cell comprising a nucleic acid molecule or a nucleic acid construct of unnatural occurrence and / or introduction, encompassed by the present invention. The nucleic acid molecule or the introduced nucleic acid construct may be maintained in the cell as a discrete DNA molecule, or it may be integrated into the cell's genomic DNA.

[285] The genomic DNA of a cell should be understood in its broadest context to include any and all endogenous DNA that makes up the genetic complement of a cell. As such, the genomic DNA of a cell should be understood as including chromosomes, mitochondrial DNA, and the like. Thus, the term genomically integrated encompasses chromosomal integration, mitochondrial DNA integration, and the like. The genomically integrated form of the construct may be the whole or part of the construct. However, in some embodiments, the genomically integrated form of the construct includes at least the nucleic acid molecule of the second aspect of the invention.

[286] As used in the present invention, the term CARs different Petition 870250089762, dated 02 / 10 / 2025, pages 510 / 580 150 / 176 or different chimeric antigen receptors refer to any two or more CARs that have non-identical antigen recognition and / or signaling domains. In one example, different CARs include two CARs with the same antigen recognition domains (e.g., both CARs can recognize a dysfunctional P2X7 receptor) but with different signaling domains, such as one CAR with a signaling domain with a portion of an activating receptor and the other CAR with a signaling domain with a portion of a co-stimulatory receptor. As will be understood, at least one of the two or more CARs within this modality will have an antigen recognition domain that recognizes the dysfunctional P2X7 receptor, and the other CAR(s) can assume any suitable form and can be directed against any suitable antigen.

[287] Therefore, in some embodiments of the invention, the two or more different CARs have different signaling domains and may have identical or different antigen recognition domains. Specifically, the genetically modified cell of the invention may include a first chimeric antigen receptor with a signaling domain that includes a portion derived from an activating receptor and a second chimeric antigen receptor with a signaling domain including a portion derived from a costimulatory receptor.

[288] In some modalities, the activation receptor (from which a portion of the signaling domain is derived) is the CD3 coreceptor complex or is an Fc receptor.

[289] In some modalities, the co-stimulatory receptor (from which a portion of the signaling domain is derived) is selected from the group consisting of CD27, CD28, CD-30, CD40, DAP10, OX40, 4-1BB (CD137) and ICOS.

[290] In some modalities, the co-stimulatory receptor (of which a Petition 870250089762, dated 02 / 10 / 2025, pp. 511 / 580 151 / 176 portion of the signaling domain is derived) is selected from the group consisting of CD28, OX40, or 4-1BB.

[291] In some embodiments, the genetically modified cell is further modified to constitutively express costimulatory receptors.

[292] As described above, a cellular immune response is typically induced only when an activation signal (typically in response to an antigen) and a co-stimulation signal are experienced simultaneously. Therefore, by having a genetically modified cell according to some of the above modalities, which includes two or more CARs that, in combination, provide both an intracellular activation signal and an intracellular co-stimulation signal, it is ensured that a sufficient immune response can be induced in response to recognition by the CAR(s) of its cognate antigen(s). Alternatively, the genetically modified cell may include only one CAR, which has an antigen recognition domain that recognizes a dysfunctional P2X7 receptor, and may constitutively express co-stimulatory receptors, thus increasing the likelihood that co-stimulation will be provided simultaneously when the CAR is activated.Alternatively, the genetically modified cell can be further modified to constitutively express both the costimulatory receptor(s) and its ligand(s). In this way, the cell is continuously experiencing costimulation and only needs the activation of a CAR, with a signaling domain including a portion of an activating receptor, for immune activation of the cell.

[293] Therefore, in some embodiments, the genetically modified cell that expresses CAR is further modified to constitutively express costimulatory receptors. In additional embodiments, the Petition 870250089762, dated 02 / 10 / 2025, pages 512 / 580 152 / 176 genetically modified cells are further modified to express ligands for co-stimulatory receptors, thereby facilitating cell self-stimulation. Examples of CAR-expressing T cells that also express both co-stimulatory receptors and their cognate ligands (so as to induce self-stimulation) are known in the art and include, among others, those disclosed in Stephan MT. et al. Nat Med, 2007; 13: 1440-9.

[294] The potency of a genetically modified cell including a CAR can be increased by further modifying the cell to secrete cytokines, preferably pro-inflammatory or proproliferative cytokines. This cytokine secretion provides both autocrine support for the CAR-expressing cell and alters the local environment around the CAR-expressing cell so that other immune system cells are recruited and activated. Consequently, in some embodiments of the fourth or fifth aspects of the invention, the genetically modified cell is further modified to secrete cytokines. This secretion may be constitutive or may be inducible after recognition by a CAR of its cognate antigen or ligand.

[295] Although one or more cytokines may be selected depending on the desired immune response, preferred cytokines and / or chemokines include IL-2, IL-7, IL-12, IL-15, IL-17, IL-18 and IL-21, CCL19, CCL21 or a combination thereof.

[296] The immune cell of the invention may be any suitable immune cell, or progenitor cell thereof, or may be a homogeneous or heterogeneous cell population. In some embodiments, the cell is a leukocyte, a Peripheral Blood Mononuclear Cell (PBMC), a lymphocyte, a T cell, a CD4+ T cell, a CD8+ T cell, a natural killer cell, a natural killer T cell or a γδ T cell. Petition 870250089762, dated 02 / 10 / 2025, pp. 513 / 580 153 / 176

[297] The immune cell may be a T cell, in which optionally said T cell does not express TcRae, PD1, CD3 or CD96 (for example, by means of knockdown or deletion of one of these genes at the genetic or functional level).

[298] The immune cell may not express accessory molecules that may be checkpoint, exhaustion or apoptosis-associated signaling receptors, as well as ligands such as PD-1, LAG-3, TIGIT, CTLA-4, FAS-L and FASR, (for example, by means of knockout or knockdown of one of these genes at the genetic or functional level). Treatment and administration methods

[299] As further discussed in this document, the present invention finds application in the treatment of a variety of conditions, although preferably in the treatment of cancers.

[300] The present invention also contemplates various scenarios for the use of the two components of the therapeutics described in the present invention.

[301] In one scenario, the individual requiring treatment receives a single composition comprising both CAR-T cells and the bridge molecule.

[302] In additional scenarios, the individual requiring treatment receives a population of CAR-T cells, which cells comprise an expression vector that encodes the bridge molecule. The expression vector can facilitate constitutive or inducible expression of the nucleic acid sequence that encodes the bridge molecule.

[303] Furthermore, the individual requiring treatment may receive CAR-T cells and, at a later date, receive a composition comprising the bridging molecule (e.g., via infusion), or a nucleic acid sequence encoding the bridging molecule. Such a scenario may be appropriate in circumstances where the individual is first treated with the Petition 870250089762, dated 02 / 10 / 2025, pp. 514 / 580 154 / 176 CAR-T cells for the targeted treatment of cancers that are positive for the dysfunctional P2X7 receptor and where subsequent administration of the bridging molecule is intended to redirect CARs to alternative cancer antigens, or to peptides derived from an infectious agent and presented on MHC I or II molecules of the cells.

[304] Thus, a bridging molecule can be administered before, at the same time as, or after the individual receives CART cell treatment.

[305] Where a bridge molecule and CAR-T cells are administered to the individual at the same time, they may be administered by the same route of administration (including in a single composition), or alternatively by different routes of administration. For example, CAR-T cells may be administered by injection into the individual’s bloodstream, while a bridge molecule may be administered by another route of administration, such as intramuscular, intradermal, subcutaneous or intraperitoneal.

[306] A bridge molecule can be produced or expressed within the body by genetically engineered cells that secrete bridge molecules spontaneously or under stimulation via a stimulating agent, for example, a small molecule. Alternatively, cells can continuously secrete bridge molecules and will stop secreting them after the application of a stimulating agent, for example, a small molecule.

[307] It will be clearly understood that, although this descriptive report refers specifically to applications in humans, the invention is also useful for veterinary purposes. Thus, in all respects, the invention is useful for domestic animals such as cattle, sheep, horses and poultry; for companion animals such as cats and dogs; and for zoo animals. Therefore, the general term individual or individual to be / being treated is understood as Petition 870250089762, dated 02 / 10 / 2025, pages 515 / 580 155 / 176 including all animals (such as humans, monkeys, dogs, cats, horses and cows).

[308] The term administered means the administration of a therapeutically effective dose of the aforementioned composition, including the respective cells, to an individual. By therapeutically effective amount is meant a dose that produces the effects for which it is administered. The exact dose will depend on the purpose of the treatment and will be determinable by a person skilled in the art using known techniques. As is known in the art and described above, adjustments may be necessary for systemic versus localized release, age, body weight, general health, sex, diet, time of administration, drug interaction, and the severity of the condition, and will be determinable by routine experimentation by people skilled in the art.

[309] Individuals requiring treatment include those who already have a benign, precancerous, or non-metastatic tumor, as well as those in whom the occurrence or recurrence of cancer must be prevented. Individuals may have metastatic cells, including metastatic cells present in the ascitic fluid and / or lymph nodes.

[310] The goal or outcome of treatment may be to reduce the number of cancer cells; reduce the size of the primary tumor; inhibit (i.e., slow down to some extent and preferably stop) the infiltration of cancer cells into peripheral organs; inhibit (i.e., slow down to some extent and preferably stop) tumor metastasis; inhibit, to some extent, tumor growth; and / or alleviate to some extent one or more of the symptoms associated with the disorder.

[311] The effectiveness of treatment can be measured by assessing the duration of survival, time to disease progression, response rates (RR), duration of response and / or quality of life. Petition 870250089762, dated 02 / 10 / 2025, pp. 516 / 580 156 / 176

[312] The method is particularly useful for extending the time for disease progression.

[313] The method is particularly useful for extending human survival, including overall survival as well as progression-free survival.

[314] The method is particularly useful for providing a complete response to therapy, whereby all signs of cancer in response to treatment have disappeared. This does not always mean that the cancer has been cured.

[315] The method is particularly useful for providing a partial response to therapy, whereby there has been a decrease in the size of one or more tumors or lesions, or in the extent of cancer in the body, in response to treatment.

[316] The goal or outcome of treatment may be any one or more of the following: - to reduce the number of cancer cells; - to reduce the size of the primary tumor; - to inhibit (that is, to delay to some extent and preferably stop) the infiltration of cancer cells into peripheral organs; - to inhibit (that is, to delay to some extent and preferably stop) tumor metastasis; - to inhibit, to some extent, tumor growth; - to alleviate, to some extent, one or more of the symptoms associated with the disorder.

[317] In one modality, individuals requiring treatment include those with a benign, precancerous, or non-metastatic tumor.

[318] In one modality, the cancer is precancerous or preneoplastic.

[319] In one modality, the cancer is a secondary cancer or metastasis. Secondary cancer can be located in any organ or Petition 870250089762, dated 02 / 10 / 2025, pp. 517 / 580 157 / 176 tissue, and particularly in those organs or tissues with relatively higher hemodynamic pressures, such as the lung, liver, kidney, pancreas, intestine, and brain. Secondary cancer may be detected in the ascitic fluid and / or lymph nodes.

[320] In one form, the cancer may be substantially undetectable.

[321] Precancerous or preneoplastic generally refers to a condition or growth that typically precedes or develops into cancer. A precancerous growth may have cells that are characterized by abnormal cell cycle regulation, proliferation, or differentiation, which can be determined by cell cycle markers.

[322] Cancer can be a solid or liquid tumor. In other words, cancer can be a growth in a tissue (carcinoma, sarcoma, adenomas, etc.) or it can be a cancer present in bodily fluids, such as in the blood or bone marrow (e.g., lymphomas and leukemias).

[323] In certain preferred modalities, the cancer requiring treatment may be a cancer characterized by low levels of expression of the dysfunctional P2X7 receptor. Examples of such cancers include Burkitt's lymphoma. However, immunohistochemical analyses of the surface expression of the dysfunctional P2X7 receptor (nfP2X7) in tumor biopsies from patients reveal a range of 1+ to 3+ in the IHC score. Samples with low expression can therefore be found in a wide range of tumor types. Examples are found in solid tumors of various types, including, but not limited to, neuroblastoma, colorectal cancers, lung cancers, kidney cancers, skin cancers, breast cancers, brain cancers, and prostate cancer. Such differences in the level of expression in different tissues may be due to the formation of tumors from Petition 870250089762, dated 02 / 10 / 2025, pages 518 / 580 158 / 176 cells that are in an earlier stage of transformation (tissues with the highest expression of the receptor may be those undergoing the highest rate of proliferation).

[324] Other examples of cancers that can be treated according to the methods of the present invention include blastoma (including medulloblastoma and retinoblastoma), sarcoma (including liposarcoma and synovial cell sarcoma), neuroendocrine tumors (including carcinoid tumors, gastrinoma and islet cell carcinoma), mesothelioma, schwannoma (including acoustic neuroma), meningioma, adenocarcinoma, melanoma, leukemia or lymphoid malignancies, lung cancer, including small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), lung adenocarcinoma and squamous cell carcinoma of the lung, peritoneal cancer, hepatocellular cancer, gastric or stomach cancer, including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer (including metastatic breast cancer), colon cancer, rectal cancer, colorectal cancer,Endometrial or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, liver carcinoma, anal carcinoma, penile carcinoma, testicular cancer, esophageal cancer, biliary tract tumors, as well as head and neck cancer.

[325] In further examples, the treatment methods contemplated within the scope of the present invention include methods for treating or preventing an infectious disease. Thus, the bridging molecules of the invention can be used to redirect CAR-T cells to an additional accessible antigen on the surface, for example, where the antigen is a non-cancer-associated pathogenic antigen presented on an MHC I molecule or Petition 870250089762, dated 02 / 10 / 2025, pp. 519 / 580 159 / 176 MHC II, as described further in the present invention.

[326] An individual requiring treatment for an infectious disease may be at risk or have been diagnosed with the disease. Individuals at risk include those who are immunocompromised. Thus, the methods of the present invention also allow for the prevention of the onset of infectious diseases in individuals receiving therapy (such as for cancer treatment) that renders them immunocompromised and therefore susceptible to infections.

[327] Examples of intracellular pathogens whose peptides are presented on MHC I or MHC II molecules include: viral infections, intracellular bacterial infections, protozoan infections, and intracellular fungal infections.

[328] Examples of viral infections that can be treated using the methods of the present invention include: HIV, hepatitis (e.g., Hepatitis A, B or C), a coronavirus (e.g., SARS-CoV-2), an influenza virus, varicella-zoster virus, mumps virus.

[329] Examples of intracellular bacterial infections that can be treated using the methods of the present invention include: mycobacterial infections (e.g., Mycobacterium tuberculosis), Bartonella henselae, Francisella tularensis, Listeria monocytogenes, Salmonella Typhi, Brucella, Legionella, Nocardia, Neisseria, Rhodococcus, Yersinia, Staphylococcus aureus, Chlamydia, Rickettsia, Coxiella, and Chlamydophila pneumoniae.

[330] Examples of intracellular infections caused by fungal pathogens: Histoplasma capsulatum, Cryptococcus neoformans, and Pneumocystitis jirovecii.

[331] Examples of obligate intracellular protozoan pathogens include: Apicomplexans (Plasmodium spp., Toxoplasma gondii and Cryptosporidium parvum), and Trypanosomatids (Leishmania spp. and Trypanosoma). Petition 870250089762, dated 02 / 10 / 2025, pages 520 / 580 160 / 176 cruzi).

[332] Immune cells that can be targeted to modulate the immune system in the context of cancer and / or autoimmune disease can be B cells (CD19, CD20, CD22), plasma cells (BCMA, CD38, CD138), T cell subsets via (TRBC1 or TRBC2, α4β7 and aEe7, CD7), macrophages and TAMs (CD163 and CD206). In the context of allogeneic stem cell transplantation, immune system-based conditioning can be performed via targeting (CD34, CD117, CD133, CD33 and CD38) especially in the case of non-malignant diseases, for example, thalassemia major or sickle cell anemia and / or in the case of DNA repair defects, such as Fanconi anemia.

[333] Targeting senescent tumor cells via marker (uPAR) will help eliminate resting tumor cells that are likely to expand at later times and promote even faster proliferation of cancer cells, the latter secreting tumor-promoting cytokines and shaping a tumor-suppressing environment that protects new cancerous subclones.

[334] CAR-T cells can be constructed in such a way that they are able to immunosuppress other immune cells, for example, CAR-T TREG cells, or by secreting cytokines (TGFbeta, IL10) and immunosuppressive chemokines by introducing the corresponding inducible expression cassette [NFAT-dependent cytokine secretion] and signaling into the construct.

[335] The bridging molecules of the invention can be formulated for administration to an individual using techniques known to those skilled in the art. The bridging molecule formulations may include pharmaceutically acceptable excipient(s) (carriers or diluents). Examples of commonly used excipients include, without limitation: saline solution, buffered saline solution, dextrose, water for injection, glycerol, ethanol, and combinations thereof. Petition 870250089762, dated 02 / 10 / 2025, pages 521 / 580 161 / 176 of the same, stabilizing agents, solubilizing agents and surfactants, buffers and preservatives, tonicity agents, bulking agents and lubricating agents.

[336] A bridging molecule formulation may include one type of bridging molecule, or more than one type of bridging molecule (i.e., wherein the bridging molecules may have identical or different targeting and / or dysfunctional P2X7 receptor epitope fractions).

[337] Bridge molecules may be administered to an individual using modes and techniques known to those skilled in the art. Exemplary modes include, but are not limited to, intravenous, intraperitoneal, and intratumoral injection. Other modes include, without limitation, intradermal, subcutaneous (sc, sq, sub-Q, Hypo), intramuscular (im), intra-arterial, intramedullary, intracardiac, intra-articular (joint), intrasynovial (joint fluid area), intracranial, intraspinal, and intrathecal (spinal fluids).

[338] Formulations comprising the bridging molecule are administered to an individual in an amount that is effective in treating the specific indication or disorder. In general, formulations comprising at least about 0.01 pg / kg to about 100 mg / kg of body weight of the bridging molecule may be administered to an individual requiring treatment. In most cases, the dosage may be from about 100 pg / kg to about 10 mg / kg of body weight of the bridging molecules daily, taking into account the routes of administration, symptoms, etc. However, the amount of bridging molecules in the formulations administered to an individual may vary within wide limits, depending on the location, source, identity, extent and severity of the disorder, the age and condition of the individual being treated, etc. Ultimately, a physician can determine the appropriate dosages to be used. The bridging molecules may be administered as an infusion. Petition 870250089762, dated 02 / 10 / 2025, pages 522 / 580 162 / 176 continuous or a bolus application.

[339] The time interval between CAR-T cell administration and bridging molecule formulation can vary widely depending on factors including the type of (immune) cells being used, the binding specificity of the CAR, the identity of the targeting fraction and the identity of the target cell, for example, the cancer cell to be treated, the location of the target cell in the individual, the means used to administer the formulations to the individual, and the health, age and weight of the individual to be treated. In fact, the TCBM formulation can be administered before, simultaneously with, or after the genetically engineered (immune) cell formulation.

[340] It will be understood that the invention disclosed and defined in this descriptive report extends to all alternative combinations of two or more of the individual features mentioned or evident from the text or drawings. All these different combinations constitute various alternative aspects of the invention. Examples

[341] Bivalent bridge molecules of various shapes were tested for their ability to stimulate cell death by CAR-T cells. In these examples, the CAR-T cells tested all comprised antigen-binding domains for binding to the E200 epitope of the P2X7 receptor (e.g., comprising an antigen-binding domain as set out in SEQ ID NO: 286, as described in WO 2022 / 187906). Example 1: Doubly tagged bridging molecules can cause unwanted cytotoxicity.

[342] The inventor identified that bridging molecules that have two tags present on each side of a bridging molecule can result in unwanted cytotoxic events. Petition 870250089762, dated 02 / 10 / 2025, pp. 523 / 580 163 / 176

[343] More specifically, a symmetrical antibody architecture of bridge molecules was found to induce significantly higher expression of the activation markers CD25+CD69+ as well as the exhaustion marker PD-1- compared to the bridge molecule formats XmAb1 and XmAb2.

[344] Figure 9A shows the relative change in CD25+CD69+ expression, and Figure 9B shows the relative change in PD-1 expression. This experiment was performed using cytokine-deprived adaptor CAR-T cells for 72h, incubated in a range of bridging molecule concentrations. The reference for calculating the relative change is the cytokine-deprived adaptor CAR-T cell condition without any bridging molecule addition (0 ng / mL). In the other conditions, the various bridging molecule formats, as represented, were added at the indicated concentrations and the fraction positive for CD25 and CD69 (A) or PD-1 (B) was quantified and divided by the condition without bridging molecule (0 ng / mL).

[345] The XmAb1 and XmAb2 bridge molecule formats demonstrated a significantly lower potential to induce unwanted target-independent activation than their symmetrical counterparts. In conclusion, XmAb bridge molecules facilitate enhanced anticancer potency while reducing unwanted activation, exhaustion, and Activation-Induced Cell Death (AICD) induction of CAR-T cells. Example 2: Alternative bridge molecule design

[346] The inventors designed bivalent bridge molecules in antibody format (IgG1). The bridge molecules were generated using the knobs-in-holes (KIH) format in combination with CROSS-mAb or DUET-Mab technologies to control the pairing of heavy and light chains. The bridge molecules also comprised either a single tag for binding. Petition 870250089762, dated 02 / 10 / 2025, pp. 524 / 580 164 / 176 per CAR-T anti-E200 cells or multiple labels.

[347] Figure 3 shows various architectures for various bridge molecules in the form of mono-, double-, triple- or tetra-labeled antibodies.

[348] The inventors determined that the optimal distance between the tags on a single antibody-shaped bridge molecule was about 25 nm. This meant that the location of the tags on the Fab portion of the antibodies was determined to be optimal (e.g., compared to the location of the tag on the Fc portion of the antibody). Furthermore, the inventors determined that the optimal location of the tags on the Fab portion of the antibody was such that multiple tags should all be attached to the same Fab arm of the antibody, so as to prevent cytotoxic events as described in Example 1. Example 3: Cell death by anti-E200 CAR-T cells in the presence of CD33-targeted antibody-containing bridging molecules.

[349] Figure 4 shows the results of cell death experiments of U937 cells (expressing CD33).

[350] In these experiments, the anti-E200 CAR comprised Vh BIL03 2-21 and Vl WTB1 (sequences revealed in WO 2023 / 028653), the hinge region of CD8A, the transmembrane domain of CD28, and the co-stimulatory / signaling domain of CD28, 41BB and CD3zeta.

[351] The results shown in Figure 4 demonstrate that doubly labeled antibody-shaped bridge molecules facilitate greater cell death by anti-E200 CAR-T cells compared to mono-labeled antibody-shaped bridge molecules.

[352] A similar experiment was conducted using the same CAR-T cells and bridging molecules, but with Jeko-1 cells, which do not express CD33. The results (shown in Figure 5) demonstrate that they do not Petition 870250089762, dated 02 / 10 / 2025, pages 525 / 580 165 / 176 there is non-directed cytolysis of CAR-T cells by bridging molecules. Example 4: Cell death by alternative anti-E200 CAR-T cells in the presence of CD33-targeted antibody-containing bridging molecules.

[353] An additional experiment was conducted using an alternative CAR format comprising the single-domain antibody BIL03 (2-2-1) and the hinge region of CD8A, the transmembrane domain of CD28 and the costimulatory / signaling domains of CD28, 41BB (CD137) and CD3zeta.

[354] The results, shown in Figure 6, further demonstrate the usefulness of doubly labeled anti-CD33 antibody-shaped bridge molecules to induce cell death in CD33-expressing cells. Example 5: Consideration of orthogonal label presentation

[355] The inventors further investigated the optimal presentation of the label to maximize the engagement of cancer cells and T cells by the bridge molecules of the invention. Figure 7 shows a schematic representing the optimal (A) and non-optimal (B) presentation of the label.

[356] Figure 8 shows the results of a series of experiments where cell death of U937 or Jeko-1 cells by anti-E200 CAR-T cells was induced by engaging anti-CD33 or anti-CD19 bridging molecules comprising one or two E200 tags.

[357] Green binding domains refer to anti-CD33 binding regions of the bridge molecules. Red binding domains refer to anti-CD19 binding regions of the molecules.

[358] As can be seen in Figure 8, the inventors compared the cell death-increasing capacity of mono- or doubly-labeled monospecific bivalent bridge molecules and mono- or doubly-labeled bispecific bivalent bridge molecules. Petition 870250089762, dated 02 / 10 / 2025, pp. 526 / 580 166 / 176

[359] In the context of U937 cells, there is no cell binding via the red arm of the antibody (since U937 cells do not express CD19). Similarly, in the context of Jeko-1 cells, there is no cell binding via the green arm of the antibody (since Jeko-1 cells do not express CD33).

[360] The results indicate that superior results are obtained by using a bivalent, monospecific IgG with dual tags, where the binding domains of the IgG are for binding to an antigen on the target cancer cell (e.g., CD33) and where the tags are located on the same Fab arm as the IgG.

[361] The results also show that a doubly labeled bispecific bivalent bridge molecule (e.g., as shown in the orange bars) provides greater U937 cell death (e.g., compared to a monolabeled version of the same molecule, as shown in the purple bars), even if there is no binding to the target cell via the anti-CD19 Fab portion of the antibody. This is also shown in the context of Jeko-1 cells, whereby a doubly labeled bispecific bivalent bridge molecule, as shown in the peach bars, provides greater Jeko-1 cell death compared to a monolabeled version of the same molecule, as shown in the pink bars.

[362] These results also demonstrate that, in the context of bispecific bridging molecules, it is preferable that the tags be located in a Fab portion of the antibody that is capable of binding to the target cell. Example 6: Cytolysis of the AML cell line using anti-E200 CAR and bridging molecules comprising the E200 peptide tag.

[363] Cytolysis of the AML U937 cell line by an anti-E200 CAR Petition 870250089762, dated 02 / 10 / 2025, pages 527 / 580 167 / 176 (comprising an antigen-binding domain comprising sdAb BILO3) was measured using a luciferase-based cell death assay at an ET ratio of 0.5:1 (CAR-T to target ratio) in conjunction with and without the represented bridging molecule variants (CD33-targeted, lintuzumab-based) at the indicated concentrations.

[364] The bridge molecules tested were: right side of antibody: HOLE, CROSS-MAb: SEQ ID NO: 4, 11 for all sequences. Left side of antibody: KNOB, CROSS-mAb were: Left side of the antibody: SEQIDNOs: Format XmAb1 No 1 SEQ ID NO: 287.8 Petition 870250089762, dated 02 / 10 / 2025, pages 528 / 580 168 / 176 Left side of the antibody: SEQIDNOs: Format XmAb2No 2 SEQID NO: 287, 10

[365] Control experiments (data shown in Figure 10 Aii, Bii and Cii) make use of a CAR-T cell product that has no ability to bind to the tag.

[366] The results are shown in Figure 10 Ai (cytolysis of the AML U937 cell line at 24 hours) and in Figure lOBi (cytolysis of AML cells at 48 hours) and in Figure lOCi (cytolysis at 72 hours).

[367] This control demonstrates that i) CAR has no impact on the elimination of cancer cells if it cannot bind to the tag sequence present in the bridge molecule and ii) bridge molecules alone (i.e., without a CAR-T capable of recognizing the tag sequence in the bridge molecules) do not impact the growth of cancer cells and are immunologically inert.

[368] In this assay, the inventors demonstrate that the XmAb2 variant was able to eliminate cancer cells with equal efficiency to its more toxic counterparts in a symmetrical mAb format. (Toxicity measured by undesired target-independent activation and exhaustion is illustrated in Figure 9). Petition 870250089762, dated 02 / 10 / 2025, pp. 529 / 580 169 / 176

[369] Figure 11 further shows a comparison of the cytolysis of the AML U937 cell line by an anti-E200 CAR-T cell at 1 ng / mL of the corresponding bridge molecule formates at 24 hours, 48 ​​hours and 72 hours.

[370] Statistical analysis of the various bridging molecules demonstrated a superior function of the symmetrical mAb2, mAb4 and XmAb2 antibodies compared to XmAb1 and Fab1. Thus, XmAb2 has a similar function to the symmetrical mAb2 and mAb4, however, with an enhanced toxicity profile for CAR-T cells (reduced nonspecific activation, AICD) as well as theoretically for the patient, reducing the nonspecific activation of CAR-T cells, unrelated to the target antigen, which are induced via undirected cytokine secretion, which can trigger a cascade of serious life-threatening side effects, such as cytokine release syndrome (CRS), and / or immune effector cell-associated neurotoxicity syndrome (ICANS), and / or macrophage activation syndrome (MAS).

[371] In other words, the superior activity of symmetrically labeled antibodies comes at the price of undesirable target-independent and AICD toxicities. Thus, in the in vitro assay, they may work better, but overall performance is likely lower due to their potential toxicity on the CAR-T cell product and also induce undesirable toxic effects in patients due to non-target-related activation of CAR-T cells. Example 7: Effectiveness of bridging molecules + CAR-T cells in an AML model

[372] In the AML cell line-derived xenograft model (based on MOLM-13), 0.5x106 MOLM-13 cells were injected into the tail vein on day -3. On day 0, treatment was initiated after confirmation of the leukemic graft using bioluminescence imaging. MOLM-13 cells Petition 870250089762, dated 02 / 10 / 2025, pages 530 / 580 170 / 176 were engineered to express eGFP and firefly luciferase as reporter genes for cell line development and use for functional assessment, including BLI (oxidation of D-luciferin potassium after in vivo distribution of the chemical compound). No treatment (tumor-only group), 25 x 10⁶ non-transduced T cells, activated by TransAct and cultured with cytokines (IL₇ and IL₁₅ at 10 ng / mL) and expanded, or 5 x 10⁶ CD33-targeted CAR-T cells (Lintuzumab-based), or 5 x 10⁶ E200-targeted CAR-T cells (BIL₀3 2-2-1-based) were administered intravenously twice weekly. CD33-targeted XmAb₂ (Lintuzumab-based) was injected at 50 pg / mouse (2 x 50 pg / mouse = 100 pg / mouse per week) intraperitoneally into the E200-targeted CAR-T cell group. Bioluminescence was performed twice weekly throughout the treatment period. Overall survival was calculated according to Kaplan-Meier. Figure 12 shows significantly reduced survival in the tumor-only group and the T-cell treatment group compared to the CAR-T CD33 cell group and the E200-targeted CAR treatment plus CD33 XmAb2Lintuzumab. Example 8: Demonstration of the effectiveness of the invention's XmAbs in a universal CAR system (PNE)

[373] The ability of the bridge molecules of the invention to induce cell death was tested in a system using a universal CAR system. The data shown in Figure 13 were generated using cryopreserved CAR-T PNE cells, thawed on the day of the assay and used after a reconstitution time of approximately 6 hours at an effector-to-target ratio of 1:1.

[374] The antibody-based bridging molecules used are Petition 870250089762, dated 02 / 10 / 2025, pp. 531 / 580 171 / 176 indicated at the top of Figure 13 as crossmab XmAb1 (one tag on the N-terminal light chain) and XmAb2 (two tags on the N-terminal light chain and heavy chain). The last antibody or condition on the right of the figure (in brown) did not include a PNE tag, but two E200 tags and was therefore incompatible with CAR PNE. This served as a negative control.

[375] At the indicated concentrations, there was no difference in performance. However, although the cytotoxic potential of XmAbí and XmAb2 at 100 ng / mL is similar to that of symmetrical antibodies (as shown in purple, orange and black in the figure), XmAbí and XmAb2 (red and green in the figure) will induce less undesirable target-independent activation of CAR-T cells (similar to the data shown in Figure 9), leading to continuous activation and exhaustion, as well as fratricide. Example 9: In vivo demonstration of the superiority of the asymmetric bridge molecule design in an AML disease model.

[376] In this series of experiments, 1x10e6 MOLM13 cells were injected into the tail vein of mice on day -3. On day 0, treatment was initiated after confirmation of leukemic grafting using bioluminescence imaging. MOLM-13 cells were primed to express eGFP and firefly luciferase as reporter genes for cell line development and use for functional assessment, including BLI (oxidation of Dluciferin pota...

Claims

CLAIMS 1. A molecule capable of simultaneously binding to a target cell and an immune cell expressing an exogenous cell surface receptor comprising an intracellular signaling domain, the molecule characterized in that it comprises: a heterodimer consisting of a first portion comprising a first Fab arm of an antibody and a second portion comprising a second Fab arm of an antibody, wherein the first and second Fab arms each comprise an antigen-binding domain for binding to a cell surface molecule on a target cell, wherein the first Fab arm comprises at least a portion capable of being bound by the exogenous immune cell receptor,wherein the second Fab arm does not comprise any fragments capable of being bound by the exogenous immune cell receptor and wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains.

2. A molecule capable of simultaneously binding to a target cell and an immune cell expressing an exogenous cell surface receptor comprising an intracellular signaling domain, the molecule characterized in that it comprises: a heterodimer consisting of a first portion comprising a first Fab arm of an antibody and a second portion comprising a second Fab arm of an antibody, wherein the first and second Fab arms each comprise an antigen-binding domain for binding to a cell surface molecule on a target cell, wherein the first Fab arm comprises at least two fragments capable of being bound by the exogenous immune cell receptor, and wherein the second Fab arm does not comprise any fragments capable of being bound by the exogenous immune cell receptor.

3. Molecule, according to claim 2, characterized in that the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains.

4. Two-component therapeutic, characterized in that it comprises: (a) an immune cell or progenitor thereof expressing an exogenous receptor comprising an intracellular signaling domain; and (b) a bridge molecule comprising: a heterodimer consisting of a first portion comprising a first Fab arm of an antibody and a second portion comprising a second Fab arm of an antibody, wherein the first and second Fab arms each comprise an antigen-binding domain for binding to a cell surface molecule on a target cell, wherein the first Fab arm comprises at least a fraction capable of being bound by the exogenous immune cell receptor, wherein the second Fab arm does not comprise any fractions capable of being bound by the exogenous immune cell receptor, and wherein the first and second portions of the heterodimer Petition 870250089762, dated 02 / 10 / 2025, p.539 / 580 3 / 15 comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains.

5. Composition, characterized in that it comprises: (a) an immune cell or progenitor thereof expressing an exogenous receptor comprising and an intracellular signaling domain;and (b) a bridge molecule comprising: a heterodimer consisting of a first portion comprising a first Fab arm and a second portion comprising a second Fab arm, wherein the first and second Fab arms comprise an antigen-binding domain for binding to a cell surface molecule on a target cell, wherein the first Fab arm comprises at least a fraction capable of being bound by the exogenous immune cell receptor, wherein the second Fab arm does not comprise any fractions capable of being bound by the exogenous immune cell receptor, and wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains.

6. Kit, characterized in that it comprises: (a) an immune cell or progenitor thereof expressing an exogenous receptor comprising an intracellular signaling domain; and (b) a bridge molecule comprising: a heterodimer consisting of a first portion comprising a first Fab arm and a second portion Petition 870250089762, dated 02 / 10 / 2025, page.540 / 580 4 / 15 comprising a second Fab arm, wherein the first and second Fab arms comprise an antigen-binding domain for binding to a cell surface molecule on a target cell, wherein the first Fab arm comprises at least a fraction capable of being bound by the exogenous immune cell receptor, wherein the second Fab arm does not comprise any fractions capable of being bound by the exogenous immune cell receptor, and wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains.

7. A molecule according to any one of claims 1 to 3 or a bridge molecule defined in any one of claims 4 to 6, characterized in that the first and second portions comprise an Fc region of an antibody.

8. Molecule according to claim 7, characterized in that the Fc region of an antibody is a CH2 region of an antibody.

9. Molecule according to claim 7, characterized in that the Fc region of an antibody is a CH3 region of an antibody.

10. Molecule according to claim 7, characterized in that the Fc region of an antibody comprises a CH2 region and a CH3 region of an antibody.

11. Molecule, according to any one of claims 7 to 10, characterized in that the antigen-binding domains of the first and second Fab arms of the heterodimeric molecule are identical in amino acid sequence. Petition 870250089762, dated 10 / 02 / 2025, pp. 541 / 580 5 / 15 12. A molecule, according to any one of claims 7 to 10, characterized in that the antigen-binding domains are not identical, but bind to the same epitope of an antigen or bind to different epitopes of the same antigen.

13. Molecule, according to any one of claims 7 to 10, characterized in that the antigen-binding domains of the first and second Fab arms of the heterodimeric molecule are for binding to different antigens.

14. A molecule, according to any one of claims 7 to 13, characterized in that the first and second Fab arms are capable of specifically binding to an antigen on a cancer cell.

15. Molecule, according to any one of claims 7 to 14, characterized in that the first Fab arm of the heterodimeric molecule comprises 3 or 4 or 5.

16. Molecule, according to any one of claims 7 to 15, characterized in that the fractions capable of being bound by the exogenous immune cell receptor are selected from: a peptide, polysaccharide, dye, lipid or small molecule.

17. Molecule, according to any one of claims 7 to 16, characterized in that the fractions capable of being bound by the exogenous immune cell receptor are peptides.

18. Molecule, according to claim 17, characterized in that the fractions capable of being bound by the exogenous immune cell receptor are peptides comprising the amino acid sequence of a neoepitope.

19. Molecule, according to claim 17, characterized in that the fractions capable of being bound by the exogenous immune cell receptor Petition 870250089762, dated 10 / 02 / 2025, page 542 / 580 6 / 15 are peptides comprising the amino acid sequence of a tumor-specific or tumor-associated antigen.

20. Molecule, according to claim 19, characterized in that the antigen is selected from a P2X7, EGFRvIII or CLDN6 antigen and the fractions capable of being bound by the exogenous immune cell receptor are peptides comprising the amino acid sequence of an epitope derived from P2X7, EGFRvIII or CLDN6.

21. Molecule, according to claim 20, characterized in that the fractions capable of being bound by the exogenous immune cell receptor are peptides comprising the amino acid sequence of an epitope derived from the P2X7 receptor.

22. Molecule, according to claim 21, characterized in that the fractions capable of being bound by the exogenous immune cell receptor are peptides comprising the amino acid sequence of an epitope derived from the E200 or E300 epitopes of P2X7.

23. Molecule, according to claim 22, characterized in that the fractions capable of being bound by the exogenous immune cell receptor are peptides comprising or consisting of an amino acid sequence as set forth in any of the SEQ ID NO: 155 to 218.

24. Molecule, according to claim 22, characterized in that the fractions capable of being bound by the exogenous immune cell receptor are peptides comprising or consisting of the amino acid sequence as set forth in SEQ ID NO:

162.

25. Therapeutic, according to claim 4, composition, according to claim 5, kit, according to claim 6 or molecule, according to any of claims 1 to 3 and 7 to 24, characterized in that the exogenous immune cell receptor is a CAR comprising an antigen recognition domain and a signaling domain, wherein the antigen recognition domain is capable of binding to a tumor-specific or tumor-associated antigen.

26. Therapeutic, according to claim 4, composition, according to claim 5, kit, according to claim 6 or molecule, according to any one of claims 1 to 3 and 7 to 24, characterized in that the exogenous immune cell receptor is a CAR comprising an antigen recognition domain for binding to a P2X7 receptor epitope, preferably to a cancer-associated form of the receptor (such as dysfunctional or non-functional forms of the receptor).

27. Therapeutic, composition, kit or molecule, according to claim 26, characterized in that the exogenous receptor binding domain binds to an epitope associated with an adenosine triphosphate (ATP) binding site of the P2X7 receptor.

28. Therapeutic, composition, kit or molecule, according to claim 27, characterized in that the P2X7 epitope comprises or is a derivative of the E200 epitope as defined in the present invention and preferably comprises at least the amino acid sequence as set forth in SEQ ID NO:

162.

29. Therapeutic, composition, kit or molecule, according to claim 26 or 27, characterized in that the antigen recognition domain binds to an epitope that includes proline at amino acid position 210 of the P2X7 receptor.

30. Therapeutic, composition, kit or molecule, according to any of claims 27 to 29, characterized in that the antigen recognition domain binds to an epitope that includes one or more amino acid residues extending from glycine at amino acid position 200 to cysteine ​​at amino acid position 216, inclusive, of the P2X receptor.

31. Therapeutic, composition, kit or molecule, according to any one of claims 27 to 30, characterized in that the antigen recognition domain comprises a binding polypeptide comprising the amino acid sequence of the CDRs of the Vh and / or Vl chains of an antibody, or the amino acid sequence of the Vh and / or Vl chains of an antibody, or the amino acid sequence of an antibody or fragment thereof, wherein the antibody or fragment thereof comprises the amino acid sequences of any antibody described in: PCT / AU2002 / 000061 or PCT / AU2002 / 001204 (or in any of the corresponding US patents US 7,326,415, US 7,888,473, US 7,531,171, US 8,080,635, US 8,399,617, US 8,709,425, US 9,663,584, or US 10,450,380), PCT / AU2007 / 001540 (or in the corresponding US patent US 8,067,550), PCT / AU2007 / 001541 (or in the corresponding US publication US 2010-0036101), PCT / AU2008 / 001364 (or in any of the corresponding US patents US 8,440,186, US 9,181,320,US 9,944,701 or US 10,597,451), PCT / AU2008 / 001365 (or in any of the corresponding US patents US 8,293,491 or US 8,658,385), PCT / AU2009 / 000869 (or in any of the corresponding US patents US 8,597,643, US 9,328,155 or US 10,238,716), PCT / AU2010 / 001070 (or in any of the corresponding publications WO / 2011 / 020155, US 9,127,059, US 9,688,771, or US 10,053,508) and PCT / AU2010 / 001741 (or in any of the corresponding publications WO 2011 / 075789 or US 8,835,609).

32. Therapeutic, composition, kit or molecule, according to any one of claims 27 to 31, characterized in that the antigen recognition domain comprises the CDR amino acid sequences of the 2-2-1 antigen-binding domain described in PCT / AU2010 / 001070 (or in Petition 870250089762, dated 10 / 02 / 2025, pp. 545 / 580 9 / 15, any of the corresponding US patents US 9,127,059, US 9,688,771, or US 10,053,508) or BPM09 described in PCT / AU2007 / 001541 (or in the corresponding US publication US 2010-0036101) and produced by the AB253 hybridoma deposited in the European Collection of Cultures (ECACC) under number Access 06080101.

33. Therapeutic, composition, kit or molecule, according to any one of claims 27 to 32, characterized in that the antigen recognition domain comprises the amino acid sequence of a binding domain as described in WO 2023 / 028653, optionally as defined in SEQ ID NO:

286.

34. Therapeutic, composition, kit or molecule, according to any one of claims 27 to 33, characterized in that the immune cell receptor signaling domain comprises a portion derived from an activating receptor; optionally where the activating receptor is a member of the CD3 coreceptor complex or is an Fc receptor, optionally where the portion derived from the CD3 coreceptor complex is CD3-Z; optionally where the portion derived from the Fc receptor is FcεRI or FcyRI.

35. Therapeutic, composition, kit or molecule, according to any one of claims 27 to 34, characterized in that the signaling domain includes a portion derived from a co-stimulatory receptor; in some embodiments, the signaling domain includes a portion derived from an activating receptor and a portion derived from a co-stimulatory receptor; in some embodiments, the co-stimulatory receptor is selected from the group consisting of CD27, CD28, CD30, CD40, DAP10, OX40, 4-1BB (CD137) and ICOS.

36. Nucleic acid, characterized in that it comprises a sequence of nucleotides that encodes a molecule defined in any one of claims 1 to 3 and 7 to 24.

37. A method of treating a disease or disorder in an individual, the method characterized by the fact that it comprises administering to the individual: (a) an immune cell or progenitor thereof expressing an exogenous receptor comprising an intracellular signaling domain;and (b) a bridge molecule comprising: a heterodimer consisting of a first portion comprising a first Fab arm and a second portion comprising a second Fab arm, wherein the first and second Fab arms comprise an antigen-binding domain for binding to a cell surface molecule on a disease- or disorder-associated target cell, wherein the first Fab arm comprises at least a fraction capable of being bound by the exogenous immune cell receptor, and wherein the second Fab arm does not comprise any fractions capable of being bound by the exogenous immune cell receptor, wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains, thereby treating the disease or disorder in the individual.

38. A method for treating cancer in an individual, the method characterized in that it comprises administering to the individual: (a) an immune cell or progenitor thereof expressing an exogenous receptor comprising an intracellular signaling domain; and (b) a bridging molecule comprising: Petition 870250089762, dated 02 / 10 / 2025, p.547 / 580 11 / 15 a heterodimer consisting of a first portion comprising a first Fab arm and a second portion comprising a second Fab arm, wherein the first and second Fab arms comprise an antigen-binding domain for binding to a cell surface molecule on a tumor cell, wherein the first Fab arm comprises at least two fragments capable of being bound by an exogenous immune cell receptor, and wherein the second Fab arm does not comprise any fragments capable of being bound by an exogenous immune cell receptor, wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains, thereby treating cancer in the individual.

39. Method for killing a target cell, the method characterized in that it comprises exposing or bringing the target cell into contact with: (a) an immune cell or progenitor thereof expressing an exogenous receptor comprising an intracellular signaling domain; and (b) a bridge molecule comprising: a heterodimer consisting of a first portion comprising a first Fab arm and a second portion comprising a second Fab arm, wherein the first and second Fab arms comprise an antigen-binding domain for binding to a cell surface molecule on a tumor cell, wherein the first Fab arm comprises at least a portion Petition 870250089762, dated 02 / 10 / 2025, p.548 / 580 12 / 15 capable of being bound by the exogenous immune cell receptor, and wherein the second portion of the Fab arm does not comprise any fractions capable of being bound by the exogenous immune cell receptor, wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains, thereby killing the target cell.

40. Use, characterized in that it is a heterodimer consisting of a first portion comprising a first Fab arm and a second portion comprising a second Fab arm, wherein the first and second Fab arms comprise an antigen-binding domain for binding to a cell surface molecule on a target cell, wherein the first Fab arm comprises at least a fraction capable of being bound by an exogenous immune cell receptor, wherein the second Fab arm does not comprise any fractions capable of being bound by an exogenous immune cell receptor, and wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains, in the manufacture of a medicament for treating a disorder, wherein preferably the disorder is cancer.

41. Pharmaceutical composition, characterized in that it comprises a heterodimer consisting of a first portion comprising a first Fab arm and a second portion comprising a second Fab arm, wherein the first and second Fab arms comprise an antigen-binding domain for binding to a cell surface molecule on a target cell, wherein the first Fab arm comprises at least a fraction capable of being bound by the exogenous immune cell receptor, wherein the second Fab arm does not comprise any fractions capable of being bound by the exogenous immune cell receptor, and wherein the first and second portions of the heterodimer comprise amino acid sequences to allow heterodimerization via pairing of distinct heavy chains and / or pairing of cognate light chains with the heavy chains, optionally comprising a pharmaceutically acceptable excipient.

42. Pharmaceutical composition, according to claim 41, characterized in that the composition further comprises an immune cell or progenitor thereof expressing an exogenous receptor comprising an intracellular signaling domain and optionally further comprising an antigen recognition domain.

43. Method according to any one of claims 37 to 39, or use according to claim 40, or composition according to claim 41 or 42, characterized in that the first and second portions of the bridge molecule comprise an Fc region of an antibody, such as a CH2 region of an antibody or a CH2 region and a CH3 region of an antibody.

44. Method, use or composition according to claim 43, characterized in that the first and second Fab regions are capable of specifically binding to an antigen on a cancer cell.

45. Method, use or composition, according to claim 43 or 44, characterized in that the fraction capable of being bound by the exogenous immune cell receptor is a peptide.

46. ​​Method, use or composition, according to claim 45, characterized in that the fraction capable of being bound by the exogenous immune cell receptor is a peptide comprising the amino acid sequence of a tumor-specific or tumor-associated antigen.

47. Method, use or composition, according to claim 46, characterized in that the antigen is selected from P2X7, EGFRvIII or CLDN6 and the fraction capable of being bound by the exogenous immune cell receptor is a peptide comprising the amino acid sequence an epitope derived from the antigens P2X7, EGFRvIII or CLDN6.

48. Method, use or composition, according to claim 47, characterized in that the fraction capable of being bound by the exogenous immune cell receptor is a peptide comprising the amino acid sequence an epitope derived from the P2X7 antigen.

49. Method, use or composition according to claim 48, characterized in that the exogenous immune cell receptor-binding fraction is a peptide comprising the amino acid sequence an epitope derived from the E200 or E300 epitopes of P2X?.

50. Method, use or composition, according to claim 49, characterized in that the exogenous immune cell receptor-binding fraction is a peptide comprising or consisting of an amino acid sequence as set forth in any of SEQ ID NO: 155 to 218.

51. Method, use or composition, according to claim 50, Petition 870250089762, dated 10 / 02 / 2025, page 551 / 580 15 / 15 characterized in that the fraction capable of being bound by the exogenous immune cell receptor is a peptide comprising or consisting of the amino acid sequence as set forth in SEQ ID NO:

162.

52. Method, use or composition, according to any one of claims 46 to 51, characterized in that the heterodimeric bridge molecule is bivalent and bispecific.

53. Method, use or composition according to claim 52, characterized in that the epitope fractions located in the molecule are located in a Fab portion that is capable of binding to the target cell.

54. Method, use or composition, according to any one of claims 46 to 51, characterized in that the heterodimeric bridge molecule is bivalent and monospecific for binding to a target cell (such as a cancer cell).

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