TCR mimetic t cell engaging antibodies targeting human leukocyte antigen telomerase reverse transcriptase peptide complex and CD3 and related methods

Abstract

Provided are TCR mimetic T cell engaging antibodies or antigen binding portions thereof that specifically bind antigens relating to TERT pHLA expression, various compositions of such antibodies or antigen binding portions thereof, and methods of their use. The disclosure provides such antibodies, fragments of such antibodies retaining tumor and autoimmune disease associated antigen-binding ability, and pharmaceutical compositions including such antibodies or antigen binding fragments thereof. This disclosure further provides for isolated nucleic acids encoding such antibodies, amino acid sequences of such antibodies, and host cells transformed therewith. Additionally, this disclosure provides for therapeutic methods employing the antibodies and nucleic acids of the disclosure. Finally, the present disclosure provides for recombinant bi specific antibodies capable of specifically binding pHLA-TERT540 and CD3.

Description

Patent ApplicationTCR MIMETIC T CELL ENGAGING ANTIBODIES TARGETING HUMAN LEUKOCYTE ANTIGEN TELOMERASE REVERSE TRANSCRIPTASE PEPTIDE COMPLEX AND CD3 AND RELATED METHODSNUCLEOTIDE AND / OR AMINO ACID SEQUENCE LISTING

[0001] The present application is being filed with a nucleotide and / or amino acid Sequence Listing XML in electronic format. The Sequence Listing XML is provided as a file entitled CROS-007-02WO-Sequence-Listing.xml, created on March 24, 2026 and is 238,632 bytes in size. The Sequence Listing XML is hereby incorporated by reference in its entirety.BACKGROUND

[0002] Telomerase reverse transcriptase (TERT) is part of the telomerase enzyme complex that extends telomeres. It is a self-antigen that is expressed constitutively in a majority of cancers and certain autoimmune diseases, allergies, and diseases exhibiting chronic inflammation.

[0003] TERT expression has been correlated in cancer cells with heightened expression levels measured at every stage of tumor growth and cancer metastases. Further, some cancers are associated with mutations in the TERT promoter region, which may explain the enhanced transcriptional activation of this gene seen in such pathologies.

[0004] The enzyme complex telomerase extends telomeres and comprises two essential components: the telomerase reverse transcriptase (TERT), and an RNA component known as telomerase RNA component (TERC). Other components of the complex include, for example, TCAB1, Dyskerin, Garl, Nhp2, Nop 10, and RHAU. TERT is a limiting component of the telomerase complex, and thus treatments that increase TERT can increase telomerase activity. Telomerase activity is typically measured using the telomeric repeat amplification protocol (TRAP) assay, which quantifies the ability of a cell lysate or other sample to extend a synthetic telomere-like DNA sequence.

[0005] The primary, rate limiting subunit of the human telomerase complex is telomerase reverse transcriptase (TERT or hTERT). Over 85% of human cancers have been associated with aberrant telomerase activity and hTERT expression, in contrast to normal human cells that are unable to indefinitely maintain telomere length. Certain human TERT peptides have also been correlatedPatent Application with cells expressing TERT, including in patients suffering from certain cancers, including breast, colon, lung, melanoma, and prostate cancers. TERT peptides originate from hTERT and form complexes with MHC class I and class II molecules, which are then expressed on the cell surface to elicit an immune response from cytotoxic T lymphocytes (CTLs) and T-cell receptor (TCR) of CD4+helper T (Th) cells.

[0006] High TERT expression is correlated with cancers, and driving tumor growth, proliferating cells and certain inflammatory conditions (e.g., autoimmune diseases). In contrast, normal, healthy tissues generally show little or no expression of TERT (relatively low expression may be seen in normal stem cells and progenitor cells). Consequently, TERT represents a potential target for immunotherapies.BRIEF SUMMARY

[0007] The Summary is provided to introduce a selection of concepts that are further described below in the Detailed Description. This Summary is not intended to identify key or essential features of the claimed subject matter, nor is it intended to be used as an aid in limiting the scope of the claimed subject matter.

[0008] Described herein are antibodies that are capable of specifically binding to TERT pMHC and / or TERT540 pHLA and their incorporation into novel T cell receptor mimetic T cell engagers as described herein. Also described herein are bispecific recombinant antibodies that are capable of specifically binding both TERT540 pHLA and CD3. More preferred aspects of the disclosure include, in certain embodiments, novel T cell receptor (TCR) mimetic (TCRm) T cell engager (TCE) antibodies or fragments thereof that specifically bind to: (i) complexes comprising an HLA class I molecule and a TERT peptide or fragment thereof (e g., TERT540) and / or derivatives of a TERT peptide or fragment; and (ii) at least one immune cell (e.g., via CD3 expressed on the surface of an immune cell). In certain aspects, the distinct complexes comprise HLA class I molecule-TERT540 complexes. As described herein, these TCRm TCE antibodies provide promising therapeutic efficacy.

[0009] The presently disclosed TCRms and antibodies target TERT pHLA and / or TERT540 pMHC. Telomerase reverse transcriptase (TERT) is a self-antigen that is expressed constitutively in cells associated with certain cancers and tumors, as well as cells known to be relevant to the pathogenesis of autoimmune and inflammation disorders such as constitutively activated B cells.Patent Application TERT is part of the telomerase enzyme complex that extends telomeres. The human TERT:540-548 peptide (also referred to herein as “hTERT540”, “hTERT540”, “TERT540”, and “TERT540”), which has the sequence ILAKFLHWL (SEQ ID NO: 15) is an immunodominant HLA-A*02:01-restricted T-cell epitope, correlated with cells expressing TERT, which includes many cancerous cells and those associated with autoimmune and inflammation disorders, such as aberrantly active immune cells (such as B cells). Thus, TERT540 represents a highly specific target for treating diseases associated with TERT expression.

[0010] Tumor cells and certain autoimmune disorder-associated cells often express intracellular antigens, such as TERT540, and may display such antigens on the surface of the cell via Major Histocompatibility Complex (MHC) presentation. MHCs display intracellularly processed protein fragments on the cell surface. In humans, MHC is referred to as Human Leukocyte Antigen (HLA). MHC class I molecules / HLA class I molecules are expressed on the surface of nearly all nucleated cells of humans. MHC class I molecules / HLA class I molecules are dimeric and comprise a transmembrane heavy chain, comprising the peptide antigen-binding cleft, and a smaller extracellular chain, beta2-microglobulin.

[0011] In the canonical pathway, MHC class I molecules / HLA class I molecules present peptides derived from the proteolytic degradation of cellular proteins. Proteins may originate from endogenous de novo biosynthesis or from extra-cellularly sourced proteins through the process of cross-presentation. Canonically, the cleaved peptides are transported into the lumen of the endoplasmic reticulum (ER) by the transporter associated with antigen processing (TAP), where they are bound to the groove of the assembled class I molecule, and the resultant peptide-MHC class I complex is transported to the cell membrane to enable antigen presentation to T lymphocytes. Alternatively, some proteins contain leader or signal sequences that insert directly into the ER, where they are cleaved by signal peptidases resident in the ER and can then bind to MHC class I molecules in a TAP-independent manner for presentation on the cell surface.

[0012] MHC class I / HLA class I genes are highly polymorphic across humans, comprising multiple common alleles for each individual gene. Each MHC allele-peptide complex comprising a specific HLA subtype and a specific peptide fragment presents a protein structure on the cell surface that can be targeted by an immunotherapeutic antigen-binding protein. However, the polymorphic nature of MHC allele / peptide complexes limits the number of possible recipients of therapeutics designed to bind specific MHC allele-peptide complexes.Patent Application

[0013] The TCR mimetic antibodies of the disclosure include those referred to herein as TCRm, TCR mimetic TCEs, in the TERT 1+1 TCRm TCE, TERT 2+1 TCRm TCE formats. TCRm TCEs are characterized in that, in addition to targeting autoimmune disorder-associated and tumor-associated peptides / antigens (TAAs) presented by the MHC, they also have one or more binding domains that target an immune cell associated protein, e.g., an immune cell receptor (e.g., T cell receptor) protein, an immune cell activator, an immune cell engager, and the like, as described in detail herein and / or known in the art. The terms “(1+1)” and “(2+1)” when used in the name of a particular anti-TERT540 TCRm, e.g., as found in the specific TCRms such as “TERT (1+1)” and “TERT(2+1)”, in particular, refer to specific TCRm formats. In the TERT 1+1 TCRm TCE format, the TCRm has a binder targeting a peptide-HLA complex, in this case a TERT pHLA complex (e.g., a TERT540 pHLA complex) and a second binder that targets an immune cell (e.g., via an anti-CD3 binder). In the TERT 2+1 TCRm TCE format, the first and third binders target a peptide-HLA complex, in this case a TERT pHLA complex (e.g., a TERT540 pHLA complex), while a second binder binds to an immune cell (e.g., via an anti-CD3 binder). In the TERT 1+1 TCRm TCE format, there is only one binder that binds to an HLA-TAA-peptide or HLA-autoimmune disorder-associated peptide complex, while a second binder binds to an immune cell (e.g., via an anti-CD3 binder). In the TERT 2+1 TCRm TCE format, the first and third binders bind to an HLA-TAA-peptide or HLA-autoimmune disorder-associated peptide complex, while a second binder binds to an immune cell (e.g., via an anti-CD3 binder).

[0014] Surprisingly, the present inventors have found that the presently disclosed TCRm TCEs have: (i) pHLA complex binding with sufficiently high affinity and avidity; (ii) showed sub-nM and selective activation of T cells only in the presence of target peptide antigen; (iii) provided a clear cytotoxicity window; (iv) specific cytotoxicity in in vitro assays; (v) exhibited robust in vivo efficacy in heme and solid tumor CDX models in mice; (vi) triggered T-cell proliferation in human HLA-A*02:01 PBMCs in vitro, (vii) demonstrated a positive safety profile by not inducing unwanted IFNy in the absence of the TERT540 pHLA complex; (viii) showing no alloreactivity; (ix) no demonstrable interactions with predicted cross-reactive peptides.

[0015] In certain preferred aspects, the present disclosure includes methods, compositions, formulations, and various techniques for producing TCR-mimetic (TCRm) bispecific T Cell Engager (TCE) antibodies (Ab) that bind to the TERTS4O / HLA-A2*O2: O1 pMHC complex with high affinity. The TCRm-based molecules of the disclosure, including the specific TCRm-basedPatent Application TCEs provided herein, induce potent T cell mediated killing of cancer cells and other cells expressing TERT, including cells known to be relevant to the pathogenesis of autoimmune disorders such as constitutively activated B cells, with presumed varying levels of target expression, at sub-nanomolar EC50levels.

[0016] In certain aspects, the present disclosure is based, in part, on the recognition of TERT having a forward feedback regulation with the NF-KB signaling pathway. Within the loop, chronic inflammation leads to telomere / telomerase dysfunction, while telomere / telomerase dysfunction leads to inflammation. Activation of NF-KB has been shown to up-regulate expression of TERT, while expression of TERT is significantly down-regulated upon contact with an NF-KB inhibitor. Thus, TERT expression and telomerase activity are linked to NF-KB signaling. Moreover, other components of the NF-KB signaling pathway, such as interleukin (IL)-6 and tumor necrosis factor (TNF)-a have been shown to upregulate TERT transcription and telomerase activity. There also appears to be a link to NF-KB signaling and the p38 MAPK signaling pathway, mitochondrial damage, and / or reactive oxygen species (ROS) which further link to the TERT / telomerase-inflammation feedback loop. Through this pathway, TERT expression is linked to certain inflammatory conditions, particularly chronic autoimmune conditions.

[0017] In certain aspects, the TCRm-based TCEs (e.g., TCRm TCEs) of the disclosure comprise at least: (i) a first binding domain that binds to an HLA class I molecule-TERT peptide complex; (ii) a second binding domain that binds to an immune cell associated protein (e.g., an immune cell engager such as an anti-CD3 binding domain). In preferred aspects, the second binding domain binds to CD3 on an immune cell.

[0018] In certain aspects, the TCRm-based TCEs (e.g., TCRm TCEs) of the disclosure comprise at least: (i) a first and third binding domains that bind to an HLA class I molecule-TERT peptide complex; (ii) a second binding domain that binds to an immune cell associated protein (e.g., an immune cell engager such as an anti-CD3 binding domain). In preferred aspects, the second binding domain binds to CD3 on an immune cell. In certain aspects, the HLA class I molecule-TERT peptide complex is an HLA class I molecule-TERT54o complex. In certain aspects, the HLA class I molecule-TERT complex includes a TERT and / or a TERT540 peptide that has one or more amino acid modifications relative to the wildtype TERT or TERT540 peptide.

[0019] In further aspects, the disclosed TCR mimetic antibody comprises a third binding domain in addition to the first and second binding domains, wherein the third binding domain binds anPatent Application HLA-TERT peptide complex and, in particular aspects, binds an HLA-TERT540 peptide complex. In alternative aspects, the third binding domain binds to an immune cell receptor. In certain aspects, the immune cell receptor bound by the third binding domain is a different immune cell receptor than bound by the second binding domain. In certain aspects, the second antigen binding domain binds to CD3 or CD28 and the third antigen binding domain binds to CD3 or CD28.

[0020] Described herein are also antibodies, antigen binding regions, and fragments thereof, that may be incorporated into a TCRm-based molecule as described herein, that are capable of specifically binding to TERT540 pHLA. Such antibodies can be monoclonal, synthetic / recombinant, or other type of antibody. Also described herein are bispecific recombinant antibodies, antigen binding regions, and fragments thereof that are capable of specifically binding both TERT540 and CD3 and incorporated into a TCRm-based molecule as described herein.

[0021] Isolated antibodies or antibody fragments as described herein can be a human antibody or humanized antibody or human antibody fragment or humanized antibody fragment.

[0022] The disclosure also provides a TCRm, antibody, or antigen binding portion thereof that specifically binds to a TERT antigen, wherein the TCRm, antibody, or antigen binding portion thereof comprises a heavy chain variable region comprising an amino acid sequence that is at least 90% identical to any one of SEQ ID NOS: 1 and 200-227 and a light chain variable region comprising an amino acid sequence that is at least 90% identical to any one of SEQ ID NOS: 2 and 228-255.

[0023] The disclosure also provides a TCRm, antibody, or antigen binding portion thereof that specifically binds to a TERT antigen, wherein the TCRm, antibody, or antigen binding portion thereof comprises a heavy chain variable region comprising an amino acid sequence that is at least 95% identical to any one of SEQ ID NOS: 1 and 200-227 and a light chain variable region comprising an amino acid sequence that is at least 95% identical to any one of SEQ ID NOS: 2 and 228-255.

[0024] The disclosure also provides a TCRm, antibody, or antigen binding portion thereof that specifically binds to a TERT antigen, wherein the TCRm, antibody, or antigen binding portion thereof comprises a heavy chain variable region comprising an amino acid sequence that is at least 97% identical to any one of SEQ ID NOS: 1 and 200-227 and a light chain variable region comprising an amino acid sequence that is at least 97% identical to any one of SEQ ID NOS: 2 and 228-255.Patent Application

[0025] The disclosure also provides a TCRm, antibody, or antigen binding portion thereof that specifically binds to a TERT antigen, wherein the TCRm, antibody, or antigen binding portion thereof comprises a heavy chain variable region comprising an amino acid sequence that is at least 98% identical to any one of SEQ ID NOS: 1 and 200-227 and a light chain variable region comprising an amino acid sequence that is at least 98% identical to any one of SEQ ID NOS: 2 and 228-255.

[0026] The disclosure also provides a TCRm, antibody, or antigen binding portion thereof that specifically binds to a TERT antigen, wherein the TCRm, antibody, or antigen binding portion thereof comprises a heavy chain variable region comprising an amino acid sequence that is at least 99% identical to any one of SEQ ID NOS: 1 and 200-227 and a light chain variable region comprising an amino acid sequence that is at least 99% identical to any one of SEQ ID NOS: 2 and 228-255.

[0027] The disclosure also provides a TCRm, antibody, or antigen binding portion thereof that specifically binds to a TERT antigen, wherein the TCRm, antibody, or antigen binding portion thereof comprises a heavy chain variable region comprising an amino acid sequence as set forth in any one of SEQ ID NOS: 1 and 200-227 and a light chain variable region as set forth in any one of SEQ ID NOS: 2 and 228-255.

[0028] In additional disclosed aspects, the heavy chain variable region comprises an HC CDR1 having an amino acid sequence as set forth in any one of SEQ ID NO: 3 and SEQ ID NOS: 58–72, an HC CDR2 having an amino acid sequence as set forth in any one of SEQ ID NO: 4 and SEQ ID NOS: 73–90, and an HC CDR3 having an amino acid sequence as set forth in any one of SEQ ID NO: 5 and SEQ ID NOS: 91–118, and wherein the light chain variable region comprises an LC CDR1 having an amino acid sequence as set forth in any one of SEQ ID NO: 6 and SEQ ID NOS: 119–143, an LC CDR2 having an amino acid sequence as set forth in any one of SEQ ID NO: 7 and SEQ ID NOS: 144–171, and an LC CDR3 having an amino acid sequence as set forth in any one of SEQ ID NO: 8 and SEQ ID NOS: 172–199. In these aspects, the disclosed TCR mimetic antibodies encompass any antibody or fragment thereof that incorporates the recited CDR combinations irrespective of framework selection, humanization state, or overall antibody format, provided that binding to the HLA-TERT540 peptide complex is retained.

[0029] The disclosure further encompasses TCR mimetic antibodies and fragments thereof that are assembled from multiple polypeptide chains. In certain aspects, the TCR mimetic antibodyPatent Application comprises a first polypeptide chain, a second polypeptide chain, and a third polypeptide chain, wherein the first polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 12, the second polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 13, and the third polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 14. In alternative aspects, the first polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 56, the second polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 57, and the third polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 14.

[0030] In such bispecific and / or multivalent configurations, the TCR mimetic antibody may comprise a first polypeptide and a second polypeptide, wherein the first polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 10 and the second polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 11. In alternative bispecific configurations, the first polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 54 and the second polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 55. In still further aspects, the TCR mimetic antibody comprises a first polypeptide chain, a second polypeptide chain, and a third polypeptide chain, wherein the first polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 10, the second polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 258 and the third polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 14.Patent Application

[0031] The disclosure also encompasses single-chain or alternative compact formats, wherein the TCR mimetic antibody or fragment thereof comprises a polypeptide having an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 9, thereby capturing all claimed levels of sequence identity for such embodiments. In additional aspects involving third binding domains, the TCR mimetic antibody comprises a first polypeptide and a second polypeptide, wherein the first polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 256 and the second polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 257.

[0032] Isolated antibodies or antibody fragments as described herein can be a human antibody or humanized antibody or human antibody fragment or humanized antibody fragment.

[0033] In certain aspects, the TCRm-based antibodies, isolated antibodies, or antigen binding portions thereof comprise one or more of a monovalent scFv (single chain fragment variable) antibody, divalent scFv, Fab fragment, F(ab’)2 fragment, F(ab’)3 fragment, Fv fragment, or single chain antibody.

[0034] In certain aspects, the TCRm-based TCE comprises an Fc domain. In certain aspects, the Fc domain comprises one or more amino acid substitutions.

[0035] In preferred aspects, the TCRm-based TCE are in a 1+1 TCRm TCE format, or a 2+1 TCRm TCE format as disclosed herein. In certain aspects, the TCRm is selected from the exemplary TCRms provided as TERT(2+1)’, TERT(1+1), TERT(1+1)’, TERT(2+1), TERT(2+1)”, TERT(2+1)*, TERT-a, TERT-b, TERT-c, TERT-d, TERT-e, TERT-f, TERT-g, TERT-h, TERT-I, TERT-j, TERT-k, TERT-1, TERT-m, TERT-n, TERT-o, TERT-p, TERT-q, TERT-r, TERT-s, TERT-t, TERT-u, TERT-v, TERT-w, TERT-x, TERT-y, TERT-z, TERT-aa, TERT-ab, and TERT-ac set forth in Tables la, lb, and Id and Table 2.

[0036] In certain aspects, the TCRm can be or comprise a chimeric antibody, bispecific antibody, trispecific or other multi-specific antibody, or BiTE. In certain aspects, the TCRm can be an IgG antibody or a recombinant IgG antibody or antibody fragment.

[0037] In certain aspects, the TCRm-based antibodies or antibody fragments thereof can exhibit increased binding affinity for TERT540 presented by HLA-A*02:01 compared to sample from aPatent Application healthy subject or subject not having a condition associated with TERT expression, such as an autoimmune disease or cancer.

[0038] In certain aspects, antibodies, including TCRms, as described herein can be conjugated or fused to an imaging agent, a cytotoxic agent, a metal, or a radioactive moiety. In certain aspects, the imaging agent can be a fluorophore. In certain aspects, the radioactive moiety can comprise at least one ofZr-89, Cu-64, F-18, Y-90, Lu-177, At-211, Ac-225, or Pb-212. In certain aspects, the antibody can be an immune conjugate or a radio-immune conjugate. In certain aspects, the antibody is an antibody-drug conjugate.

[0039] In certain aspects, described herein are hybridoma or engineered cells comprising a nucleic acid encoding any of the TCRm-based antibodies or antibody fragments thereof. In certain aspects, described herein is a hybridoma or engineered cell comprising a nucleic acid encoding any amino acid sequence, antibody or antibody fragment thereof, as described herein.

[0040] In certain aspects, also described herein are pharmaceutical preparations. In some aspects, pharmaceutical preparations according to the present disclosure comprise (a) a pharmaceutically acceptable carrier; and (b) a TCRm-based antibody or antigen binding portion thereof of any embodiment as described herein. In some aspects, pharmaceutical preparations according to the present disclosure comprise (a) a pharmaceutically acceptable carrier; and (b) a TCRm-based antibody or antigen binding portion thereof of any embodiment as described herein, wherein said TCRm antibody or antigen binding portion thereof is in a 1+1 TCRm TCE format. In some aspects, pharmaceutical preparations according to the present disclosure comprise (a) a pharmaceutically acceptable carrier; and (b) a TCRm-based antibody or antigen binding portion thereof of any embodiment as described herein, wherein said TCRm antibody or antigen binding portion thereof is in a 2+1 TCRm TCE format.

[0041] In certain aspects, also described herein are methods for treating a subject having a TERT expression-associated disease or disorder, e.g., an autoimmune disease or cancer. Methods according to the present disclosure can comprise administering to a subject in need thereof a therapeutically effective amount of any pharmaceutical preparation or any TCRm-based antibody or any TCRm antibody fragment as described herein..

[0042] Also described herein are methods of making TCRm-based antibodies or antibody fragments thereof. Methods as described herein can comprise, for example, culturing a hybridoma or engineered cell as described herein under conditions that allow expression of the TCRm-basedPatent Application antibody or a fragment thereof and, optionally, isolating the TCRm-based antibody or fragment from the culture.

[0043] In certain aspects, the TCR mimetic-based antibody comprises first and third antigen binding domains as a bispecific antibody in a heterodimer format, wherein the first and third antigen binding domains independently bind to an HLA class I molecule TERT / TERT540peptide complex. In certain aspects, the TCRm-based antibody comprises a second binding domain that comprises a single-chain variable fragment (scFv) or Fab that binds to CD3. In preferred aspects, the first and / or third binding domains comprise an scFv that binds to the plurality of HLA class I molecule TERT / TERT540peptide antigens. In preferred aspects, the first and third binding domains comprise an scFv that binds to the plurality of HLA class I TERT / TERT540peptide antigens.

[0044] In certain TCRm-based antibodies (i.e., TCRm-based T-cell engagers), the mimetic comprises: a first polypeptide comprising the first binding domain and the second polypeptide comprising the second binding domain. In certain aspects, the first polypeptide includes scFv of the first domain. In certain aspects, the second polypeptide includes scFv of the second domain. In certain aspects, the first and second polypeptide each comprise a hinge-CH2-CH3 and form a heterodimer.

[0045] In certain TCRm-based antibodies (i.e., TCRm-based T-cell engagers), the mimetic comprises: a first polypeptide comprising the first binding domain; a second polypeptide and a third polypeptide comprising the second binding domain. In certain aspects, the third polypeptide is a light chain. In certain aspects, the first polypeptide includes scFv of the first domain. In certain aspects, the second polypeptide and third polypeptide include Fab of the second domain. In certain aspects, the first and second polypeptide each comprise a hinge-CH2-CH3 and form a heterodimer.

[0046] In certain TCRm-based antibodies (i.e., TCRm-based T-cell engagers), the mimetic comprises: a first polypeptide comprising the first binding domain; and a second polypeptide comprising the second and third binding domains. In certain aspects, the first polypeptide includes the scFv of the first domain. In certain aspects, the second polypeptide includes the scFvs of the second and third binding domains in a tandem format. In certain aspects, the first and second polypeptide each comprise a hinge-CH2-CH3 and form a heterodimer.

[0047] In certain TCRm-based antibodies (i.e., TCRm-based T-cell engagers), the mimetic comprises: a first polypeptide comprising the first and third binding domain; and a second polypeptide comprising the second binding domains. In certain aspects, the first polypeptidePatent Application includes the scFvs of the first and third binding domains in a tandem format. In certain aspects, the second polypeptide includes scFv of the second domain. In certain aspects, the first and second polypeptide each comprise a hinge-CH2-CH3 and form a heterodimer.

[0048] In certain TCRm-based antibodies (i.e., TCRm-based T-cell engagers), the mimetic comprises: a first polypeptide comprising the first binding domain; a second polypeptide and a third polypeptide comprising the second and third binding domains. In certain aspects, the first polypeptide includes the scFv of the first binding domain. In certain aspects, the second and the third polypeptide include the Fab of the second binding domain and the scFv of third binding domain in a tandem format. In certain aspects, the first and second polypeptide each comprise a hinge-CH2-CH3 and form a heterodimer. In certain aspects, the third polypeptide is a light chain.

[0049] In certain TCRm-based antibodies (i.e., TCRm-based T-cell engagers), the Fc region comprises a C-terminal lysine. In certain aspects, the Fc region comprises a C-terminus without a C-terminal lysine.

[0050] In some embodiments, the TCRm-based antibody is or comprises a single domain antibody, a human single domain antibody, or a humanized single domain antibody. In some embodiments, the TCRm-based antibody is or comprises a murine antibody, a chimeric antibody, a camelid antibody, a humanized antibody, or a human antibody. In some embodiments, the antibody is part of a multispecific antibody or a multifunctional antibody.

[0051] In certain aspects, TCRm-based antibodies of the disclosure comprise one or more Fc silencing mutations, e.g., L234A / L235A / K322A. For example, in certain aspects, the one or more silencing mutations are L234A and L235A. In certain aspects, a TCR mimetic-based antibody or fragment thereof of the disclosure comprises a C-terminal lysine. Alternatively, a TCR mimeticbased antibody or fragment thereof of the disclosure does not comprise a C-terminal lysine. TCR mimetic-based antibodies and other anti-TERT antibodies of the disclosure, including their constituent domains, parts, and peptides, which are exemplified by amino acid sequence herein contemplate variations that do and do not include such a C-terminal lysine.

[0052] In some embodiments, selective binding of a TCR mimetic antibody disclosed herein to the complex comprising the HLA class I molecule and the peptide induces a cellular response; optionally wherein the cellular response is selected from: an immune response, TERT expressing cell (e.g., a cancer cell or autoimmune disorder-associated cell) death, virally-infected cell death, antibody dependent cytotoxicity, immune cell proliferation, and cytokine release. In somePatent Application embodiments, the immune response comprises activation of T cells. In some embodiments, the T cell is a CD3+ T cell. In some embodiments, the immune response comprises activation of cytotoxic T cells (CTLs). In some embodiments, the TCR mimetic antibody binds a peptide-HLA class I molecule complex on a target TERT expressing cell (e.g., a cancer cell or autoimmune disease associated cell). In some embodiments, the TCR mimetic antibody binds an HLA class I molecule-peptide complex on a virally infected cell.

[0053] Described herein, in certain embodiments, are nucleic acids encoding the TCR mimetic antibody or fragment thereof described herein, or a therapeutic fusion polypeptide described herein.

[0054] In some embodiments, the TCR mimetic-based antibody is a bispecific antibody in a heterodimeric format. In some embodiments, the TCRm-based antibody comprises a heterodimeric hinge-CH2-CH3. In some embodiments, the TCR mimetic antibody is a bispecific antibody comprising a homodimeric or heterodimeric format. In some embodiments, the TCRm-based antibody comprises at least one antibody or fragment thereof that binds an immune cell.

[0055] In certain preferred aspects, a TCR mimetic-based antibody or fragment thereof comprises one or more silencing Fc mutations, e.g., L234A / L235A / K322A.

[0056] Described herein, in certain embodiments, are pharmaceutical compositions comprising the TCR mimetic-based antibody or fragment thereof described herein, the therapeutic fusion polypeptide described herein, the nucleic acid described herein, or the cell engager described herein, and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical compositions comprise a plurality of T Cell Receptor (TCR) mimetic-based antibodies or fragments thereof described herein, wherein each member of such plurality has a first binding affinity for a first complex comprising a first peptide-HLA class I molecule and a second binding affinity for a second complex comprising a second, distinct peptide-HLA class I molecule, wherein the first complex and the second complex comprise the same peptide. In some embodiments, the plurality of TCR mimetic antibodies or fragments thereof are operably connected in a single polypeptide or a polypeptide complex. In some embodiments, the pharmaceutical compositions comprise an amalgam of polypeptides each comprising the TCR mimetic antibodies or fragments thereof. In some embodiments, the first binding affinity is at least 10% to 1000% greater than the second binding affinity.Patent Application

[0057] Described herein, in certain embodiments, are methods of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the pharmaceutical composition described herein. Described herein, in certain embodiments, are methods of treating an autoimmune disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the pharmaceutical composition described herein.

[0058] Described herein, in certain embodiments, are methods of inducing T cell dependent cellular cytotoxicity (TDCC) or Redirected T cell cytotoxicity (RTCC) in a subject in need thereof, comprising: administering to the subject an effective amount of the pharmaceutical composition described herein. In some embodiments, the method further comprises administering to the subject a checkpoint inhibitor prior to, subsequent to, or simultaneously with administration of the TCR mimetic antibody or fragment thereof.

[0059] Described herein, in certain embodiments, are pharmaceutical compositions comprising in one or more polypeptide chains or polypeptide complexes a plurality of T cell receptor (TCR) mimetic-based antibodies or fragment thereof, each member of the plurality specifically binds a plurality of complexes comprising an HLA class I molecule and a TERT or TERT540peptide. In some embodiments, each member of such plurality has a first binding affinity for a first peptide-HLA class I molecule and a second binding affinity for a second, distinct peptide-HLA class I molecule.BRIEF DESCRIPTION OF THE DRAWINGS

[0060] The present application includes the following figures. The figures are intended to illustrate certain embodiments and / or features of the compositions and methods, and to supplement any description(s) of the compositions and methods. The figures do not limit the scope of the compositions and methods, unless the written description expressly indicates that such is the case.

[0061] FIG. 1 provides a schematic of an exemplary TCRm TCE of the disclosure in a TERT 1+1 TCRm TCE format, with one TERT pHLA antigen binding domain and optionally one or more Fc domain modifications and / or with or without a C-terminal lysine.

[0062] FIG. 2 provides a schematic of an exemplary TCRm TCE of the disclosure in a TERT 2+1 TCRm TCE format, with two TERT pHLA antigen binding domains and optionally one or more Fc domain modifications and / or with or without a C-terminal lysine.Patent Application

[0063] FIG. 3 provides schematics of exemplary TCRm TCEs of the disclosure with one TERT pHLA antigen binding domain and optionally one or more Fc domain modifications.

[0064] FIGS. 4A-4B provide schematics of exemplary TCRm TCEs of the disclosure with two TERT pHLA antigen binding domains and optionally one or more Fc domain modifications.

[0065] FIGS. 5A-5D show that the TERT TCRms of the disclosure have sub-nM affinity for the TERT540-pHLA complex.

[0066] FIGS. 6A-6C show potent and specific activity of TERT TCRms in vitro against multiple cancer cell lines (myeloid and solid tumor) with presumed varying levels of target expression, highlighting the therapeutic potential of TERT TCRms in oncology.

[0067] FIGS. 7A-7B show that TERT TCRms induce potent and specific T-cell activation in vitro.

[0068] FIGS. 8A-8B show activity of TERT TCRms against NSCLC cells in a spheroid model of lung cancer.

[0069] FIGS. 9A-9B show that the pharmacokinetic (PK) profile of TERT TCRms in human FcRn mice is comparable to conventional IgG-like antibodies.

[0070] FIGS. 10A-10B show in vivo efficacy of TERT TCRms in a mouse model of disseminated AML (U937-A2).

[0071] FIGS. 11A-11C show that TERT TCRms are highly specific for the TERT540 peptide as presented in a pHLA complex based on multi-positional X-scan analysis.

[0072] FIG. 12 shows that TERT TCRms are highly specific for TERT540 as presented in a pHLA complex when compared to peptides computationally predicted to pose the highest cross-reactive risk, based on an in vitro cytotoxicity assay against peptide-loaded T2 cells.

[0073] FIGS. 13A-13B show that TERT TCRms do not induce IFNγ production when applied to a broad panel of normal HLA-A*02:01 positive normal primary cells derived from multiple vital organs and tissues.

[0074] FIGS. 14A-14B show that TERT TCRms are not allo-reactive when tested against B-LCL lines expressing multiple HLA-A haplotypes.

[0075] FIGS. 15A-15B show cytokine release results (TNF, IFNγ, IL-6, and IL-10) for TERT TCRms incubated with HLA-A*02:01 positive PBMCs in the presence and absence of TERT-expressing tumor cells (THP-1).

[0076] FIGS. 16A-16B show that TERT TCRms trigger depletion of CD3-negative populations only in HLA-A*02:01 positive PBMCs, likely reflecting pHLA recognition.Patent Application

[0077] FIG. 17 shows that TERT TCRms induce dose-dependent T-cell proliferation in vitro only in HLA-A*02:01 positive PBMCs, as expected.

[0078] FIGS. 18A-18B show that, in in vitro experiments with HLA-A*02:01 positive PBMCs, TERT TCRms primarily directly target human monocytes and B-cells.

[0079] FIG. 19 shows that TERT TCRms are moderately active in a Colony -Forming Unit (CFU) assay, indicative of targeting of bone marrow progenitor cells.

[0080] FIG. 20 shows that TERT TCRms are expected to possess a wide therapeutic index (~12 or greater) based on their relative in vitro potency against target-positive cancer cell lines and colony-forming units.

[0081] FIGS. 21A-21B show that a single administration of TERT TCRms is well tolerated in human CD34+ mice (“huCD34+” mice) as assessed by body weight.

[0082] FIGS. 22A-22B show transient induction of serum cytokines in huCD34+ mice with TERT(2+1), but not with TERT(1+1).

[0083] FIGS. 23A-23B show broad leukopenia in huCD34+ mice with TERT TCRms, consistent with the previously detected activity against bone marrow progenitor cells.

[0084] FIGS. 24A-24B show that TERT TCRms have no meaningful or sustained effects on red blood cells in huCD34+ mice.

[0085] FIGS. 25A-25B show dose-dependent decreases in human T-cells, B-cells, and monocytes with TERT TCRms in the peripheral blood of huCD34+ mice, consistent with CBC results.

[0086] FIGS. 26A-26B provide a conversion of huCD34+ mouse CBC data to clinical grading of cytopenia, which shows transient significant leukopenia and neutropenia (grades 3-4) with TERT TCRms.

[0087] FIG. 27 summarizes known on-target effects of targeting TERT, for example in TERT KO mice, mice treated with TERT -targeted TCR-T cells, and in patients treated with the TERT inhibitor imetelstat.

[0088] FIGS. 28-29 show that both TERT TCRms met all baseline and stress developability criteria for a therapeutic targeting the pHLA-TERT540peptide complex as a T cell engager.

[0089] FIGS. 30A-30B detail an experimental design and the primary AML samples used in an ex vivo assay to assess TERT(2+1) TCRm at clinically-relevantE: T ratios against primary AML cells.

[0090] FIGS. 31A-31F provide results from the ex vivo assay against primary AML cells. As shown, treatment with the TERT(2+1) TCRm leads to robust T cell activation, T cell proliferation,Patent Application cytotoxicity against primary AML blasts, and cytotoxicity against AML leukemic stem cells (LSC) in this study.

[0091] FIGS. 32A-32C outline an in vivo assay and provide results show that the TERT(2+1) TCRm of the disclosure exhibits robust in vivo efficacy in a cell-derived xenograft (CDX) model of disseminated AML (U937-A2) in mice, as demonstrated by inhibition of tumor growth and by survival benefit. Efficacy is observed with 0.03 mg / kg and 0.1 mg / kg TERT(2+1) TCRm; higher efficacy is observed in the 0.03 mg / kg cohort.

[0092] FIGS. 33A-33E set forth an in vivo assay and provide data to assess repeated dosing of TERT(2+1) TCRm in non-tumor-bearing huCD34 mice. As shown, repeated administration of TERT(2+1) is well tolerated in these mice.

[0093] FIG. 34 shows flow cytometry data indicating that, on average, cancer cell lines that are sensitive to TERT TCRms of the disclosure in vitro (referred to as “responders”) exhibit higher expression of HLA-A*02 compared to cell lines that are not sensitive (referred to as “nonresponders”).

[0094] FIGS. 35A-35B outline an in vivo study and provide results show that the TERT(2+1) TCRm of the disclosure exhibits robust in vivo efficacy in a CDX model of NSCLC (COR-L23-A2) in mice, as demonstrated by marked inhibition of tumor growth and tumor regression compared to isotype control.

[0095] FIGS. 36A-36B outline an in vivo study and provide results show that the TERT(2+1) TCRm of the disclosure exhibits in vivo efficacy in an additional CDX model of NSCLC (NCI-H1703) in mice, as demonstrated by inhibition of tumor growth compared to isotype control.

[0096] FIGS. 37A-37B show results from a Membrane Proteome Array (MPA) study to evaluate potential off-target binding of the TCRm articles of the disclosure.

[0097] FIGS. 38A-38B provide cytokine release data in PBMCs, in the presence and absence of target-positive cancer cells, using either a TERT TCRm or a comparator CD 123 TCE.

[0098] FIG. 39 provides data from a TDCC assay performed by IncuCyte showing that different formats of the TERT TCRms of the disclosure induce potent cytotoxicity against the targetpositive NSCLC cell line NCI-H1755.

[0099] FIGS. 40A-40F show the results of a comparability study performed with TERT(2+1) and TERT(2+1)’ TCRms demonstrating that they have equivalent biological properties as assessed by target binding, specificity, and functional activity.Patent Application

[0100] FIG. 41 shows the mean serum concentration profiles of a TERT TCRm of the disclosure in male cynomolgus monkeys following a single intravenous bolus administration of the TCRm at 0.01, 0.1 and 1 mg / kg.

[0101] FIG. 42 shows the mean serum concentration profiles of a TERT TCRm of the disclosure in male cynomolgus monkeys following a single subcutaneous administration of the TCRm at 0.01, 0.1 and 1 mg / kg.

[0102] FIG. 43 shows anti-drug antibody (ADA) results from the PK study in cynomolgus monkeys using a TERT TCRm of the disclosure, including IV and SC administration groups.

[0103] FIG. 44 shows that administration of TERT TCRms in huCD34+ mice leads to decreased levels of circulating human IgG (hlgG), which suggests the ability of TERT TCRms to decrease circulating autoantibody levels produced by plasma cells in the context of autoimmune disease.

[0104] FIG. 45 provides an overview of an in vivo study with a mouse model of Systemic Lupus Erythematosus (SLE) that could be carried out to evaluate the therapeutic potential of TERT TCRms in autoimmune disease.

[0105] FIGS. 46A-46B show that a TERT TCRm of the disclosure depletes B cells and monocytes (both of which are relevant to SLE pathophysiology) from PBMC samples obtained from SLE patients, while a CD19-targeting TCE included as comparator depletes B cells but not monocytes.

[0106] FIG. 47 shows that a TERT TCRm of the disclosure leads to decreased STING agonist-induced IFNa secretion from PBMCs obtained from SLE and RA patients, which represents a functional consequence of monocyte depletion, while a CD19-targeting TCE does not.

[0107] FIGS. 48A-48C show that, in a mouse model of SLE, treatment with TERT(2+1)’ TCRm leads to depletion of B cells and monocytes, decreased autoantibody production (i.e. serological improvement of SLE), and relief of splenomegaly (i.e. improvement of SLE-related organ dysfunction), highlighting the therapeutic potential of the TERT TCRms in autoimmune disease in general and SLE in particular.DETAILED DESCRIPTION

[0108] The following description recites various aspects and embodiments of the present compositions and methods. No particular embodiment is intended to define the scope of the compositions and methods. Rather, the embodiments merely provide non-limiting examples of various compositions and methods that are at least included within the scope of the disclosedPatent Application compositions and methods. The description is to be read from the perspective of one of ordinary skill in the art; therefore, information well known to the skilled artisan is not necessarily included.

[0109] Described herein are antibodies that are capable of specifically binding to the complexes that comprise HLA class I molecule and TERT-derived peptide TERT540 and their incorporation into novel T cell receptor mimetic T cell engagers as described herein. Also described herein are bispecific recombinant antibodies that are capable of specifically binding both TERT-pHLA and CD3. In certain aspects, the present disclosure provides novel T cell receptor (TCR) mimetic (TCRm) T cell engager (TCE) antibodies or fragments thereof that specifically bind to: (i) complexes comprising an HLA class I molecule and a TERT peptide or fragment thereof (e.g., TERT540) and / or derivatives of a TERT peptide or fragment; and (ii) at least one immune cell (e.g., via CD3 expressed on the surface of an immune cell). In certain aspects, the distinct complexes comprise HLA class I molecule-TERT54o complexes. As described herein, these TCRm TCE antibodies provide promising therapeutic efficacy.

[0110] Most normal cells exhibit a low, base-level of telomerase activity. Evidence indicates that TERT is a limiting component of telomerase and its expression is highly regulated, and its expression is needed to reactivate telomerase activity in telomerase negative cells. The majority of somatic cells lack telomerase activity. Telomerase activity tends to be restricted to actively proliferating cells, e.g., stem cells, germ cells, and tumor cells. TERT expression and telomerase activity are required to maintain telomere length and maintain genome integrity, which may promote cell immortalization and / or malignant transformation. Telomerase reactivation of telomerase activity-negative cells has been correlated with numerous pathologies and pathogenic processes. TERT reactivation is largely associated with cancers, particularly carcinomas. However, surprisingly, TERT reactivation is associated with inflammatory disorders, chronic infection, liver and kidney diseases, and several autoimmune diseases.

[0111] Inflammation, especially chronic inflammation, has been shown to affect and even alter the human immune response. Inflammation involves inflammatory cytokines such as tumor necrosis factor-alpha (TNF-a), interleukin (IL)-6, IL-8, IL-ip, IL-12, IL-23, interferon families, and others. A major role of these pro-inflammatory cytokines is identifying and communicating the presence of an infection or injury to surrounding tissues. In contrast, anti-inflammatory cytokines, e.g., IL-4, IL-10, IL-13, IL-11, IL-1, and IL- Ira, inhibit inflammatory signals. A number of inflammatory factors, such as IL-2 and IL-6, are known to promote expression of telomerase inPatent Application T cells, and subsequently maintain this increased expression. The nuclear factor-kappaB (NF-KB) signaling pathway is a critical pathway in regulating the inflammatory response. NF-KB signals a number of downstream genes and causes the expression of inflammatory factors, including IL-6, IL-8, and TNF-a. Overexpression of these inflammatory factors may cause immune dysfunctions.

[0112] The present disclosure is based, in part, on the recognition of TERT having a forward feedback regulation with the NF-KB signaling pathway. Within the loop, chronic inflammation leads to telomere / telomerase dysfunction, while telomere / telomerase dysfunction leads to inflammation. Activation of NF-KB has been shown to up-regulate expression of TERT, while expression of TERT is significantly down -regulated upon contact with an NF-KB inhibitor. Thus, TERT expression and telomerase activity is linked to NF-KB signaling. Moreover, other components of the NF-KB signaling pathway, such as interleukin (IL)-6 and tumor necrosis factor (TNF)-a, have been shown to upregulate TERT transcription and telomerase activity. There also appears to be a link to NF-KB signaling and the p38 MAPK signaling pathway, mitochondrial damage, and / or reactive oxygen species (ROS) which further link to the TERT / telomerase-inflammation feedback loop. Through this pathway, TERT expression is linked to certain inflammatory conditions, particularly chronic autoimmune conditions.

[0113] For example, the transcriptional regulation of NF-KB by TERT is, in part, responsible for the expression of genes that cause metabolic syndrome— a cluster of conditions that increase the risk of developing chronic diseases, such as heart disease, stroke, and type 2 diabetes. Chronic inflammation in COPD has been associated with increased expression of feedback loop-relevant inflammatory factors such as TNF-a, IL-ip, IL-6, GM-CSF, and IL-8, which are all highly expressed in COPD patients. In some cases, the hTERT gene, which encodes for TERT, has been found to be mutated in patients with severe COPD, which downregulated telomerase activity. In alcoholic liver disease, TERT has been shown to up-regulate the expression levels of TNF-a and IL-ip through the NF-KB signaling pathway. TNF-a and IL- 1 P, in turn, up-regulate the expression of TERT. In Rheumatoid arthritis (RA), synovial infiltrating lymphocytes and peripheral blood leukocytes (PBLs) have been shown to exhibit increased telomerase activity in patients. There has also been a documented association of disease severity progressing with increasing telomerase activity and an actual decrease in telomere length in cells of patients with systemic lupus, particularly in B lymphocytes. In patients with allergic rhinitis (AR), the expression of IL- 10 inPatent Application dendritic cells was negatively correlated with the levels of TERT expression. While the levels of TERT mRNA in the dendritic cells were associated with the allergic response in patients with AR.

[0114] In fact, chronic inflammatory diseases such as chronic obstructive pulmonary disease, diabetes, atherosclerosis, allergies, rheumatoid arthritis, inflammatory bowel disease (IBD), alcoholic liver disease, and chronic renal diseases have all been associated with telomere / telomerase dysfunction and the TERT- NF-KB signaling feedback loop.

[0115] Accordingly, the present disclosure provides methods and compositions for treating diseases or disorders associated with TERT expression, such as certain cancers and autoimmune and chronic inflammatory conditions, wherein said methods may alleviate TERT / telomerase dysfunction and / or inflammation, thereby slowing or stopping the feedback loop.

[0116] Articles “a” and “an” are used herein to refer to one or to more than one (i.e., at least one) of the grammatical object of the article. By way of example, “an element” means at least one element and can include more than one element.

[0117] The use herein of the terms “including,” “comprising,” or “having,” and variations thereof, is meant to encompass the elements listed thereafter and equivalents thereof as well as additional elements. Embodiments recited as “including,” “comprising,” or “having” certain elements are also contemplated as “consisting essentially of’ and “consisting of those certain elements.” As used herein, “and / or” refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations where interpreted in the alternative (“or”).

[0118] As used herein, the transitional phrase “consisting essentially of’ (and grammatical variants) is to be interpreted as encompassing the recited materials or steps “and those that do not materially affect the basic and novel character! stic(s)” of the present disclosure or features of the claims. See, for example, In re Herz, 537 F.2d 549, 551-52, 190 U. S. P. Q. 461, 463 (CCPA 1976) (emphasis in the original); see also MPEP §2111.03. Thus, the term “consisting essentially of’ as used herein should not be interpreted as equivalent to “comprising.”

[0119] Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. For example, if a concentration range is stated as 1% to 50%, it is intended that values such as 2% to 40%, 10% to 30%, or 1% to 3%, etc., are expressly enumerated in thisPatent Application specification. These are only examples of what is specifically intended, and all possible combinations of numerical values between and including the lowest value and the highest value enumerated are to be considered to be expressly stated in this disclosure.

[0120] The terms “about” and “approximately” as used herein shall generally mean an acceptable degree of error for the quantity measured given the nature or precision of the measurements. Exemplary degrees of error are within 20% (%); preferably, within 10%; and more preferably, within 5% of a given value or range of values. Any reference to “about X” or “approximately X” specifically indicates at least the values X, 0.95X, 0.96X, 0.97X, 0.98X, 0.99X, 1.01X, 1.02X, 1.03X, 1.04X, and 1.05X. Thus, expressions “about X” or “approximately X” are intended to teach and provide written support for a claim limitation of, for example, “0.98X.” Numerical quantities given herein are approximate unless stated otherwise, meaning that the term “about” or “approximately” can be inferred when not expressly stated. When “about” is applied to the beginning of a numerical range, it applies to both ends of the range.

[0121] The terms “polypeptide” and “peptide” are used interchangeably herein to refer to a polymer of amino acid residues in a single chain. The terms apply to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers. Amino acid polymers may comprise entirely L-amino acids, entirely D-amino acids, or a mixture of L- and D-amino acids. The term “protein” as used herein refers to either a polypeptide or a dimer (i.e., two) or multimer (i.e., three or more) of single chain polypeptides. The single chain polypeptides of a protein may be joined by a covalent bond, e.g., a disulfide bond, or non-covalent interactions. The terms “portion” and “fragment” are used interchangeably herein to refer to parts of a polypeptide, nucleic acid, or other molecular construct.

[0122] The amino acids in the polypeptides described herein can be any of the 20 naturally occurring amino acids, D-stereoisomers of the naturally occurring amino acids, unnatural amino acids and chemically modified amino acids. Unnatural amino acids (that is, those that are not naturally found in proteins) are also known in the art, as set forth in, for example, Zhang et al. “Protein engineering with unnatural amino acids,” Curr. Opin. Struct. Biol. 23(4): 581-87 (2013); Xie et al. “Adding amino acids to the genetic repertoire,” Curr. Opin. Chem. Biol. 9(6): 548-54 (2005); and all references cited therein. Beta and gamma amino acids are known in the art and are also contemplated herein as unnatural amino acids.Patent Application

[0123] As used herein, a chemically modified amino acid refers to an amino acid whose side chain has been chemically modified. For example, a side chain can be modified to comprise a signaling moiety, such as a fluorophore or a radiolabel. A side chain can also be modified to comprise a new functional group, such as a thiol, carboxylic acid, or amino group. Post-translationally modified amino acids are also included in the definition of chemically modified amino acids.

[0124] The term “identity” “similarity” or “substantial identity,” as used in the context of a polynucleotide or polypeptide sequence described herein, refers to a sequence that has at least 60% sequence identity / similarity to a reference sequence. Alternatively, percent similarity / identity can be any integer from 60% to 100%. Exemplary embodiments include at least: 60%, 65%, 70%, 75%, 80%, 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, similar as compared to a reference sequence using the programs described herein; preferably BLAST using standard parameters, as described below. One of skill will recognize that these values can be appropriately adjusted to determine corresponding identity of proteins encoded by two nucleotide sequences by taking into account codon degeneracy, amino acid similarity, reading frame positioning and the like.

[0125] For sequence comparison, typically one sequence acts as a reference sequence to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. Default program parameters can be used, or alternative parameters can be designated. The sequence comparison algorithm then calculates the percent sequence identities for the test sequences relative to the reference sequence, based on the program parameters.

[0126] A “comparison window,” as used herein, includes reference to a segment of any one of a number of contiguous positions selected from the group consisting of from 20 to 600, usually about 50 to about 200, more usually about 100 to about 150 in which a sequence may be compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned. Methods of alignment of sequences for comparison are well-known in the art. Optimal alignment of sequences for comparison may be conducted by the local homology algorithm of Smith & Waterman Add. APL. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch J. Mol. Biol. 48:443 (1970), by the search for similarityPatent Application method of Pearson & Lipman Proc. Natl. Acad. Sci. (U. S. A.) 85: 2444 (1988), by computerized implementations of these algorithms (e.g., BLAST), or by manual alignment and visual inspection.

[0127] Algorithms that are suitable for determining percent sequence identity / sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al. (1990) J. Mol. Biol. 215: 403-10 and Altschul et al. (1977) Nucleic Acids Res. 25: 3389-402, respectively. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (NCBI) web site. The algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul et al. (1977)). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are then extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for mismatching residues; always <0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a word size (W) of 28, an expectation (E) of 10, M=l, N=-2, and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a word size (W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89:10915 (1989)).

[0128] The BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin & Altschul, Proc. Nat'l. Acad. Sci. USA 90:5873-5787 (1993)). One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by which a match between two nucleotide or amino acid sequences would occur by chance. For example, a nucleic acid is considered similar to a reference sequence if the smallest sum probability in a comparison of the test nucleic acid to thePatent Application reference nucleic acid is less than about 0.01, more preferably less than about 10-5, and most preferably less than about 10-20.

[0129] The term “antigen binding protein” or “ABP” is used herein in its broadest sense and includes certain types of molecules such as TCR mimetic-based antibodies comprising one or more antigen-binding domains that specifically bind to an antigen or epitope.

[0130] The term “peptide-Human Leukocyte Antigen (HLA)-class I molecule complex”, “peptide-HLA class I molecule”, “HLA class I molecule peptide complex”, and “peptide and HLA class I molecule complex”, and similarly described terms as used herein, refer to the complex of both the HLA class I molecule bound to the presented peptide antigen.

[0131] The term “HLA-TERT540 peptide antigen” or “HLA class I molecule-TERTso complex” or “TERT pHLA” and such similar terms as used herein refers to the peptide-HLA class I molecule complex comprising both the HLA class I molecule bound to a TERT540 peptide described herein. The term “HLA- TERT peptide antigen” or “HLA class I molecule-TERT complex” and such similar terms as used herein refers to the peptide-HLA class I molecule complex comprising both the HLA class I molecule bound to a TERT peptide, which may be TERT540, described herein.

[0132] The term “TCR mimetic antibody”, “TCRm-based antibody”, “TCRm-based molecule”, “T Cell Receptor mimetic antibody”, “TCRm”, “TCRm antibody”, “TCR mimetic T cell engager (TCE)” or “TCR mimetic binding protein”, “1+1 TCRm TCE”, “2+1 TCRm TCE”, “TERT 1+1 TCRm TCE”, “TERT 2+1 TCRm TCE”, and the like, as used herein, refers to an ABP that binds at least one peptide MHC-Class I / pHLA molecule complex on a target cell and moiety expressed or presented on an immune cell. Certain exemplary TCRms of the disclosure are set forth in Table 2.

[0133] The term “T cell engager” or “TCE”, as used herein, refers to a molecule, preferably a bispecific or other multi-specific antibody, that binds an immune cell and at least one peptide MHC-Class I / pHLA molecule complex on a different cell. A TCRm-based antibody, as disclosed herein is a TCE that binds to a peptide MHC-Class 1 / pHLA molecule complex on a target cell and that includes an immune cell engager or other binding protein (e.g., an immune cell activator or co-stimulatory molecule) that binds to an immune cell (e.g., a T cell).

[0134] A “HLA- TERT TCR mimetic antibody”, “Anti-HLA-TERT TCR mimetic antibody”, “TERT TCRm”, “TERT 1+1 TCRm TCE”, “TERT 2+1 TCRm TCE”, “anti-TERT TCRm”, “HLA- TERT peptide ABP,” “anti-HLA-TERT peptide ABP,” or “HLA-TERT peptide-specificPatent Application ABP” is an ABP, as provided herein, which specifically binds to the peptide-HLA class T molecule complex bound to a TERT peptide, which includes variant, truncated, or otherwise engineered TERT peptides, such as the TERT540 peptide. An “HLA-TERT540 TCR mimetic antibody”, “TERT TCRm”, “TERT 1+1 TCRm TCE”, “TERT 2+1 TCRm TCE”, “anti-TERT TCRm”, “Anti-HLA-TERT540 TCR mimetic antibody”, “HLA-TERT540 peptide ABP,” “anti-HLA-TERTs4o peptide ABP,” or “HLA-TERT540 peptide-specific ABP” is an ABP, as provided herein, which specifically binds to the peptide-HLA class I molecule complex bound to a TERT540 peptide.

[0135] As used herein, “variable region” refers to a variable sequence that arises from a recombination event, for example, an antibody heavy-chain variable region includes recombined V, D, and J gene segments, and an antibody light-chain variable region includes recombined V and J gene segments. The antibody variable regions contribute to antigen specificity and, together with the corresponding partner chain, form the antigen-binding site.

[0136] The term “antigen-binding domain” means the portion of an ABP that is capable of specifically binding to an antigen or epitope. An antigen-binding domain can include heavy chain and light chain variable regions that include CDRs, e.g., HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3.

[0137] The amino acid sequence boundaries of an antibody, e.g., a TCRm antibody, CDR can be determined by one of skill in the art using any of a number of known numbering schemes, including but not limited to the Kabat numbering system, as described by Kabat et al. in Sequences of Proteins of Immunological Interest, U. S. Department of Health and Human Services, NIH Publication No. 91-3242 (1991), which is incorporated herein by reference. The Kabat system defines CDRs based on sequence variability observed across antibody variable regions and assigns specific residue ranges for CDR1, CDR2, and CDR3 within both heavy and light chains. This scheme is widely used in antibody engineering and characterization to identify and manipulate antigen-binding regions.

[0138] As used herein, the term autoimmune disease refers to a pathological condition arising from an aberrant or inappropriate immune response directed against one or more self-antigens, selftissues, or endogenous cellular components of a subject. In an autoimmune disease, mechanisms of immune tolerance that normally prevent immune recognition of self-structures are impaired or lost, resulting in activation of immune effector pathways that target the host’s own cells, tissues, or organs. Such immune responses may involve autoreactive T lymphocytes, autoreactive BPatent Application lymphocytes, the production of autoantibodies, immune complex formation, complement activation, inflammatory cytokine signaling, or other immune-mediated processes that lead to inflammation, cellular injury, or destruction of normal tissue. Autoimmune diseases may be organspecific, in which the immune response is directed primarily against antigens expressed in a particular tissue or organ, or systemic, in which multiple tissues or organ systems are affected. In various contexts described in patent literature, autoimmune diseases encompass disorders in which immune recognition of self results in chronic inflammation, tissue degeneration, impaired physiological function, or progressive organ damage, including diseases mediated by autoantibodies, autoreactive T cells, or other immune effector mechanisms that recognize endogenous proteins, peptides, nucleic acids, lipids, carbohydrates, or other host-derived molecules.

[0139] “Affinity” refers to the strength of the summed total of non-covalent interactions between a single binding site of a molecule (e.g., a T cell engager protein) and its binding partner (e.g., an antigen or epitope). Unless indicated otherwise, as used herein, “affinity” refers to intrinsic binding affinity, which reflects a 1:1 interaction between members of a binding pair (e.g., T cell engager protein and antigen or epitope). The affinity of a molecule X for its partner Y can be represented by the dissociation equilibrium constant (KD). The kinetic components that contribute to the dissociation equilibrium constant are described in more detail below. Affinity can be measured by common methods known in the art, including those described herein, such as surface plasmon resonance (SPR) technology (e.g., BIACORE®) or biolayer interferometry (e.g., FORTEBIO®).

[0140] With regard to the binding of a TCR mimetic antibody, T cell engager protein or ABP to a target molecule, the terms “bind,” “specific binding,” “specifically binds to,” “specific for,” “selectively binds,” and “selective for” a particular antigen (e.g., a polypeptide target) or an epitope on a particular antigen mean binding that is measurably different from a non-specific or non-selective interaction (e.g., with a non-target molecule). Specific binding can be measured, for example, by measuring binding to a target molecule and comparing it to binding to a non-target molecule. Specific binding can also be determined by competition with a control molecule that mimics the epitope recognized on the target molecule. In that case, specific binding is indicated if the binding of the ABP to the target molecule is competitively inhibited by the control molecule.

[0141] The term “kd” (sec-1), as used herein, refers to the dissociation rate constant of a particular ABP - antigen interaction. This value is also referred to as the koffvalue.Patent Application

[0142] The term “ka” (M'Nsec-1), as used herein, refers to the association rate constant of a particular ABP -antigen interaction. This value is also referred to as the konvalue.

[0143] The term “KD” (M), as used herein, refers to the dissociation equilibrium constant of a particular ABP -antigen interaction. KD = kd / ka. In some embodiments, the affinity of an ABP is described in terms of the KD for an interaction between such ABP and its antigen. For clarity, as known in the art, a smaller KD value indicates a higher affinity interaction, while a larger KD value indicates a lower affinity interaction.

[0144] The term “KA” (M'1), as used herein, refers to the association equilibrium constant of a particular ABP-antigen interaction. KA = ka / kd.

[0145] An “immunoconjugate” is an ABP conjugated to one or more heterologous molecule(s), such as a therapeutic (cytokine, for example).

[0146] When used herein in the context of two or more ABPs, the term “competes with” or “crosscompetes with” indicates that the two or more ABPs compete for binding to an antigen (e g., HLA-TERT peptide). In some embodiments, one ABP reduces binding of another ABP to its antigen by at least 25%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, or at least 95%. A skilled artisan can select the concentrations of the ABPs used in competition assays based on the affinities of the ABPs for HLA-TERT peptide and the valency of the ABPs. A skilled artisan can utilize any suitable assay to determine if ABPs compete with each other. Suitable assays are described, for example, in Cox et al., “Immunoassay Methods,” in Assay Guidance Manual [Internet], Updated December 24, 2014 (www.ncbi.nlm.nih.gov / books / NBK92434 / ; accessed September 29, 2015); Silman et al., Cytometry, 2001, 44:30-37; andFinco et al., J. Pharm. Biomed. Anal., 2011, 54:351-358; each of which is incorporated by reference in its entirety.

[0147] The term “epitope” means a portion of an antigen that specifically binds to an ABP.Epitopes frequently consist of surface-accessible amino acid residues and / or sugar side chains and may have specific three-dimensional structural characteristics, as well as specific charge characteristics. Conformational and non-conformational epitopes are distinguished in that the binding to the former but not the latter may be lost in the presence of denaturing solvents. An epitope may comprise amino acid residues that are directly involved in the binding, and other amino acid residues, which are not directly involved in the binding. The epitope to which an ABP binds can be determined using known techniques for epitope determination such as, for example, testing for ABP binding to HLA-TERT540 peptides in the context of an HLA Class I molecule.Patent Application

[0148] The term “vector,” as used herein, refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes the vector as a self-replicating nucleic acid structure as well as the vector incorporated into the genome of a host cell into which it has been introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operatively linked. Such vectors are referred to herein as “expression vectors.”

[0149] The terms “host cell,” “host cell line,” and “host cell culture” are used interchangeably and refer to cells into which an exogenous nucleic acid has been introduced, and the progeny of such cells. Host cells include “transformants” (or “transformed cells”) and “transfectants” (or “transfected cells”), which each include the primary transformed or transfected cell and progeny derived therefrom. Such progeny may not be completely identical in nucleic acid content to a parent cell, and may contain mutations.

[0150] The term “treating” (and variations thereof such as “treat” or “treatment”) refers to clinical intervention in an attempt to alter the natural course of a disease or condition in a subject in need thereof. Treatment can be performed both for prophylaxis and during the course of a clinical pathology. Desirable effects of treatment include preventing occurrence or recurrence of disease, alleviation of symptoms, diminishment of any direct or indirect pathological consequences of the disease, preventing metastasis, decreasing the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis.

[0151] As used herein, the term “therapeutically effective amount” or “effective amount” refers to an amount of a TCRm-based antibody or pharmaceutical composition provided herein that, when administered to a subject, is effective to treat a disease or disorder.

[0152] As used herein, the term “subject” means a mammalian subject. Exemplary subjects include humans, monkeys, dogs, cats, mice, rats, cows, horses, camels, goats, rabbits, and sheep. In certain embodiments, the subject is a human. In some embodiments the subject has a disease or condition that can be treated with a TCRm-based antibody provided herein. In some aspects, the disease or condition is a cancer. In some aspects, the disease or condition is a viral infection or associated with a viral infection. In some cases, the disease or condition is an autoimmune disease or other inflammatory condition associated with TERT / telom erase and / or the TERT feedback loop.

[0153] The term “tumor” refers to all neoplastic cell growth and proliferation, whether malignant or benign, and all pre-cancerous and cancerous cells and tissues. The terms “cancer,” “cancerous,”Patent Application “cell proliferative disorder,” “proliferative disorder” and “tumor” are not mutually exclusive as referred to herein. The terms “cell proliferative disorder” and “proliferative disorder” refer to disorders that are associated with some degree of abnormal cell proliferation. In some embodiments, the cell proliferative disorder is a cancer. In some aspects, the tumor is a solid tumor. In some aspects, the tumor is a hematologic malignancy.

[0154] The term “pharmaceutical composition” refers to a preparation which is in such form as to permit the biological activity of an active ingredient contained therein to be effective in treating a subject, and which contains no additional components which are unacceptably toxic to the subject in the amounts provided in the pharmaceutical composition.

[0155] The terms “modulate” and “modulation” refer to reducing or inhibiting or, alternatively, activating or increasing, a recited variable.

[0156] The terms “increase” and “activate” refer to an increase of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold, or greater in a recited variable.

[0157] The terms “reduce” and “inhibit” refer to a decrease of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold, or greater in a recited variable.

[0158] The term “agonize” refers to the activation of receptor signaling to induce a biological response associated with activation of the receptor. An “agonist” is an entity that binds to and agonizes a receptor.

[0159] The term “antagonize” refers to the inhibition of receptor signaling to inhibit a biological response associated with activation of the receptor. An “antagonist” is an entity that binds to and antagonizes a receptor.

[0160] As used herein the term “antigen” is a substance that induces an immune response. An antigen can be a neoantigen. An antigen can be a “shared antigen” that is an antigen found among a specific population, e.g., a specific population of cancer or autoimmune disease patients. Antigens can include HLA-TERT540 peptide antigens.

[0161] As used herein the term “neoantigen” is an antigen that has at least one alteration that makes it distinct from the corresponding wild-type antigen, e.g., via mutation in a tumor / autoimmune cell or post-translational modification specific to a tumor / autoimmune cell. In some embodiments, the alteration occurs in tumor or cancer cells. In some embodiments, thePatent Application alteration does not occur in a non-tumor or a non-cancer cell. In some embodiments, the alteration occurs in cells associated with an autoimmune disease or disorder. In some embodiments, the alteration is absent from normal tissue. A neoantigen can include a polypeptide sequence or a nucleotide sequence. Neoantigens can include HLA-TERT540 peptide neoantigens.

[0162] As used herein the term “tumor antigen” is an antigen present in a subject’s tumor cells or tumor subset of cells or cancer cells or tissue(s) but not in the subject’s corresponding normal but not present, expressed or present at an amount (higher or lower) than present in the subject’s corresponding normal cells or subset of cells or tissue(s), or derived from a polypeptide known to or have been found to have altered expression in a cancer cell, tumor cell or cancerous tissue in comparison to a normal / healthy cell or tissue.

[0163] As used herein the terms “autoimmune-associated antigen”, “autoimmune antigen” is an antigen present in a subject’s cells or subset of cells or tissue(s) but not present, expressed or present at an amount (higher or lower) than present in the subject’s corresponding normal cells or subset of cells or tissue(s), or derived from a polypeptide known to or have been found to have altered expression in a cell or tissue in comparison to a normal / healthy cell or tissue, which is associated with an autoimmune disease or disorder, and / or chronic inflammation, and / or an allergy, and / or an allergic reaction.

[0164] As used herein the term “candidate antigen” is a mutation or other aberration giving rise to a sequence that may represent an antigen.

[0165] As used herein the term “coding region” is the portion(s) of a gene that encode protein.

[0166] As used herein the term “coding mutation” is a mutation occurring in a coding region.

[0167] As used herein the term “ORF” means open reading frame.

[0168] As used herein the term “NEO-ORF” is a tumor-specific ORF arising from a mutation or other aberration such as splicing.

[0169] As used herein the term “missense mutation” is a mutation causing a substitution from one amino acid to another.

[0170] As used herein the term “nonsense mutation” is a mutation causing a substitution from an amino acid to a stop codon or causing removal of a canonical start codon.

[0171] As used herein the term “frameshift mutation” is a mutation causing a change in the frame of the protein.

[0172] As used herein the term “indel” is an insertion or deletion of one or more nucleic acids.Patent Application

[0173] As used herein the term “non-stop or read-through” is a mutation causing the removal of the natural stop codon.

[0174] In an aspect, provided herein are ABPs and TCR mimetic antibodies (the TCRm-based TCE antibodies disclosed herein) or fragments thereof that bind HLA-TERT, preferably HLA-TERT540, peptide antigens disclosed herein. In certain aspects, the TCRms bind to a plurality of HLA-TERT540 peptide antigen complexes, wherein each of the plurality of HLA-TERT540 peptide antigen complexes comprises a distinct HLA subtype. In preferred aspects, an ABP or TCR mimetic-based antibody disclosed herein specifically binds to an HLA-TERT peptide comprising TERT540 peptide complexed with an HLA class I molecule.

[0175] In certain embodiments, the HLA-TERT or HLA-TERT540 peptide is located in the peptide binding groove of an α1 / α2 domain of the HLA class I molecule heavy chain.

[0176] In certain aspects, described herein are TCR mimetic-based antibodies or fragments thereof that specifically bind a plurality of complexes comprising an HLA class I molecule and a TERT540 peptide.

[0177] In certain embodiments, the ABP or TCR mimetic-based antibody does not have a binding affinity to (i) the HLA class I molecule alone; or (ii) the TERT or TERT540 peptide alone. Thus, in some embodiments, the ABP or TCR mimetic-based antibody does not bind to the HLA in the absence of the TERT540 peptide. In some embodiments, the ABP or TCR mimetic-based antibody does not bind HLA-TERT540 peptide in the absence of the HLA. In some embodiments, the ABP or TCR mimetic-based antibody binds a tumor cell or autoimmune disease / disorder associated cell presenting the HLA-TERT540 peptide complex, optionally wherein the HLA-TERT540 peptide is a tumor antigen characterizing the cancer or an autoimmune disease associated antigen characterizing the autoimmune disease. In some aspects, the ABP or TCR mimetic-based antibody binds a complex comprising HLA and TERT540 peptide when naturally presented on a cell such as a tumor cell or autoimmune disorder associated cell.

[0178] The HLA-TERT540 peptide antigen can be expressed on the surface of any suitable target cell including a tumor cell or autoimmune disease / disorder associated cell. In some embodiments, the ABP or TCR mimetic-based antibody specifically binds a complex comprising HLA and a TERT540 peptide, e.g., derived from a tumor or autoimmune disease / disorder associated cell.

[0179] An ABP or TCR mimetic-based antibody can bind to each portion of an HLA-TERT540 peptide complex (i.e., HLA and peptide representing each portion of the complex), which whenPatent Application bound together form a novel target and protein surface for interaction with and binding by the TCR mimetic-based antibody, distinct from a surface presented by the peptide alone or an HLA subtype alone. Generally, the novel target and protein surface formed by binding of HLA to peptide does not exist in the absence of each portion of the HLA-TERT540 peptide complex. In some embodiments, the ABP or TCR mimetic-based antibody binds to the HLA-TERT540 peptide antigen through at least one contact point with an HLA class I molecule and through at least one contact point with the HLA-TERT540 peptide.

[0180] In some aspects, the affinity of an HLA-TERT540 peptide TCR mimetic-based antibody for a non-target molecule is less than about 70% (e.g., 60%, 50%, 40%, 30%, 20%, 10%, or less) 50% of the affinity for HLA-TERT540 peptide. In some aspects, the affinity of an HLA-TERT540 peptide ABP for a non-target molecule is less than about 40% of the affinity for HLA-TERT540 peptide. In some aspects, the affinity of an HLA-TERT540 peptide ABP for a non-target molecule is less than about 30% of the affinity for HLA-TERT540 peptide. In some aspects, the affinity of an HLA-TERT540 peptide ABP for a non-target molecule is less than about 20% of the affinity for HLA-TERT540 peptide. In some aspects, the affinity of an HLA-TERT540 peptide ABP for a non-target molecule is less than about 10% of the affinity for HLA-TERT540 peptide. In some aspects, the affinity of an HLA-TERT540 peptide ABP for a non-target molecule is less than about 1% of the affinity for HLA-TERT540 peptide. In some aspects, the affinity of an HLA-TERT540 peptide ABP for a non-target molecule is less than about 0.1% of the affinity for HLA-TERT540 peptide. In preferred aspects, the non-target molecule is TERT and / or pHLA-TERT, which does not include TERT540 or pHLA-TERT540.

[0181] In some embodiments, the HLA class I molecule is HLA-A*02:01.

[0182] In certain embodiments, the ABP or TCR mimetic-based antibody binds HLA-TERT540 peptide antigen with a higher affinity than its affinity to another HLA- TERT peptide. In certain embodiments, the ABP or TCR mimetic-based antibody binds HLA-A*O2: O1-TERT54O peptide antigen with a higher affinity than its affinity to an HLA-TERT peptide.

[0183] In certain embodiments, the ABP or TCR mimetic-based antibody binds HLA-A*02:01-TERT540 peptide antigen with a higher affinity than its affinity to a TERT peptide.

[0184] In certain embodiments, the ABP or TCR mimetic-based antibody binds HLA-TERT540 peptide antigen with a higher affinity than its affinity to a TERT540 peptide. In certainPatent Application embodiments, the ABP or TCR mimetic-based antibody binds HLA-A*02:01-TERT540peptide antigen with a higher affinity than its affinity to a TERT540peptide.

[0185] In certain embodiments, the ABP or TCR mimetic-based antibody binds HLA-TERT540 peptide antigen with a higher affinity than its affinity to HLA-A*02:01 without a complexed peptide (e.g., TERT540).

[0186] In some embodiments, the higher affinity is at least 1.5-fold, at least 2-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 50-fold, at least 100-fold, at least 200-fold, at least 500-fold, at least 1000-fold, at least 10,000-fold, at least 100,000-fold, or at least 1 X 106-fold. In some embodiments, the dissociation constant (KD) of the higher affinity interaction ranges from at least 10'4to 10'5Molar (M), 10'5to 10'6Molar (M), 10'6to 10'7Molar (M), at least 10'7to 10'8Molar (M), at least 10-8to 10-9Molar (M), at least 10-9to 10-10Molar (M), at least 10-10to 10-11Molar (M), at least 10'11to 10'12Molar (M), or at least ICT12to 10’13Molar (M). In certain embodiments, the ABP or TCR mimetic-based antibody binds HLA-TERT540 peptide antigen of 2, 3, 4, 5, 6, 7, 8, 9, 10 or more distinct HLA class I molecules, where the peptide of the peptide-HLA class I complex shares at least 6 identical amino acids. In some embodiments, the HLA class I molecule is HLA-A*02:01. In certain embodiments, the ABP or TCR mimetic-based antibody binds HLA-TERT540 peptide antigen of 2, 3, 4, 5, 6, 7, 8, 9, 10 or more distinct HLA class I molecules, having a dissociation constant (KD) of at least 10'4to 10'5Molar (M), 10'5to 10'6Molar (M), 10'6to 10'7Molar (M), at least 10'7to 10'8Molar (M), at least 10'8to 10'9Molar (M), at least 10’9to IO'10Molar (M), at least IO'10to 10'11Molar (M), at least 10'11to 10'12Molar (M), or at least 10'12to 10’13Molar (M). In certain embodiments, the ABP or TCR mimetic-based antibody binds HLA-TERT540 of each distinct HLA class I molecule with a KD of at least 10'4to 1013Molar (M), wherein the binding of each distinct HLA class I molecule to the ABP or TCR mimetic-based antibody has a distinct K.

[0187] Affinity differences can be determined by any means known in the art. In some embodiments, such affinity differences are assessed by MSD-ECL, SPR, BLI, or flow cytometry.

[0188] In some embodiments, the ABP or TCR mimetic-based antibody is an ABP or TCR mimetic-based antibody that competes with an illustrative ABP or TCR mimetic-based antibody provided herein. In some aspects, the ABP or TCR mimetic-based antibody that competes with the illustrative ABP or TCR mimetic-based antibody provided herein binds the same epitope as an illustrative ABP or TCR mimetic-based antibody provided herein.Patent Application

[0189] In some embodiments, the ABPs or TCR mimetic-based antibodies described herein are referred to herein as “variants.” In some embodiments, a variant is derived from any of the sequences provided herein, wherein one or more conservative amino acid substitutions are made. Conservative amino acid substitutions are described herein. In preferred embodiments, the nonconservative amino acid substitution does not interfere with or inhibit the biological activity of the functional variant. In yet more preferred embodiments, the non-conservative amino acid substitution enhances the biological activity of the functional variant, such that the biological activity of the functional variant is increased as compared to the parent ABP or TCR mimeticbased antibody.

[0190] The ABP or TCR mimetic-based antibody can be isolated and purified by any suitable method known in the art.

[0191] The ABP or TCR mimetic-based antibody can be a multi-specific antibody. The ABP or TCR mimetic-based antibody can bind to 2, 3, 4, 5, 6, 7, 8, 9, 10 or more distinct HLA-TERT540 peptide antigens. In certain embodiments, the ABP or TCR mimetic-based antibody binds 1, 2, 3, 4, 5 or more antigens that are not peptide-HLA class I molecule complex antigens. In certain embodiments, ABP or TCR mimetic-based antibody can bind to 2, 3, 4, 5, 6, 7, 8, 9, 10 or more distinct HLA-TERT540 peptide antigens and one additional antigen that is not an HLA-peptide complex antigen. In certain embodiments, TCR mimetic-based antibody can bind to 2, 3, 4, 5, 6, 7, 8, 9, 10 or more distinct HLA-TERT540 peptide antigens and one additional antigen that is not an HLA-peptide complex antigen on an immune cell (e.g., CD3). The immune cell can be a T cell (e.g., a CD8+ T cell or a cytotoxic T cell). In certain embodiments, the TCR mimetic-based antibody binds an HLA class I TERT540 peptide complex on a cancer cell or autoimmune disease / disorder associated cell. In certain embodiments, the TCR mimetic-based antibody is a multifunctional antibody.

[0192] In certain embodiments, the ABP or TCR mimetic-based antibody further comprises a conjugated therapeutic moiety. In certain embodiments, the therapeutic moiety is known in the art for use in cancer treatment and / or inducing cancer cell death. In certain embodiments, the therapeutic moiety is known in the art for use in autoimmune disease treatment and / or inducing autoimmune disease cell death.

[0193] In certain embodiments, the selective binding of the TCR mimetic-based antibody to the complex comprising the HLA class I molecule and the TERT540 peptide induces an immunePatent Application response in a cell. In certain embodiments, the immune response comprises activation of T cells. The T cell can be a CD8+ T cell, cytotoxic T cell (CTL).

[0194] In certain embodiments, the ABP or TCR mimetic-based antibody or fragment is a murine antibody, a chimeric antibody, a camelid antibody, a humanized antibody, or a human antibody.

[0195] In some embodiments, the ABP or TCR mimetic-based antibody comprises a monovalent antibody fragment comprising a single target molecule binding arm and an Fc region. In some embodiments, the ABP or TCR mimetic-based antibody comprises a fragment antigen-binding region. In some embodiments, the ABP or TCR mimetic-based antibody comprises a single chain antibody or other antibody derivative retaining the antigen specificity and the lower hinge region or a variant thereof. In some embodiments, the ABP or TCR mimetic-based antibody comprises a single chain variable fragment antibody or derivative thereof. In some embodiments, the ABP or TCR mimetic-based antibody comprises a single chain variable fragment antibody in tandem format. In certain embodiments, the ABP or TCR mimetic-based antibody comprises a single domain antibody. In certain embodiments, the ABP or TCR mimetic antibody comprises a humanized single domain antibody.

[0196] In certain embodiments, the TCR mimetic-based antibody comprises: i) a bispecific antibody, and optionally ii) a dimerized hinge-CH2-CH3; wherein the bispecific antibody comprises: i) at least one antibody fragment that selectively binds HLA-TERT540 peptide antigen; and ii) at least one antibody fragment that binds CD3.

[0197] In certain embodiments, the TCR mimetic-based T cell engager antibody is in a heterodimer format. In certain embodiments, the antibody is in a heterodimer format, and comprises: (i) an antibody fragment that binds HLA-TERT540 peptide antigen, (ii) an antibody fragment that binds CD3; and (iii) a heterodimeric hinge-CH2-CH3. In certain embodiments, the antibody comprises (i) two antibody fragments that bind HLA-TERT540 peptide antigen; (ii) an antibody fragment that binds CD3, and (iii) a heterodimeric hinge-CH2-CH3. In certain embodiments, the antibody is in a homodimer format.

[0198] In certain embodiments, the antibody comprises (i) two antibody fragments that bind HLA-TERT540 peptide antigen, (ii) two antibody fragments that bind CD3, and (iii) a homodimeric hinge-CH2-CH3. In certain embodiments, the antibody is in a single chain format. In certain embodiments, the TCR mimetic T cell engager protein does not comprise a dimerized hinge-CH2-CH3. In certain embodiments, the TCR mimetic T cell engager protein comprises an antibodyPatent Application fragment that binds CD3, an antibody fragment that binds HLA-TERT540 peptide antigen, and a 6XHis-tag.

[0199] In certain embodiments, a TCRm antibody of the disclosure comprises a first polypeptide that comprises a first antigen binding domain that binds to HLA-TERT540 peptide antigen and a second polypeptide that comprises a second antigen binding domain that binds to CD3. In certain aspects, the first antigen binding domain comprises a first scFv that binds to HLA-TERT540 peptide antigen and the second antigen binding domain comprises a second scFv that binds to CD3. In certain embodiments, the first polypeptide comprises a first hinge-CH2-CH3 and the second polypeptide comprises a second hinge-CH2-CH3; and wherein the first and second hinge-CH2-CH3 form a heterodimer.

[0200] In certain embodiments, a TCRm antibody of the disclosure comprises a first polypeptide that comprises a first antigen binding domain that binds to HLA-TERT540 peptide antigen, a second and a third polypeptide that comprises a second antigen binding domain that binds to CD3. In certain aspects, the first antigen binding domain comprises a first scFv that binds to HLA-TERT540 peptide antigen and the second antigen binding domain comprises a second Fab that binds to CD3. In certain embodiments, the first polypeptide comprises a first hinge-CH2-CH3 and the second polypeptide comprises a second hinge-CH2-CH3; and wherein the first and second hinge-CH2-CH3 form a heterodimer. In certain embodiments, the third polypeptide is a light chain.

[0201] In certain embodiments, a TCRm antibody of the disclosure comprises a first polypeptide that comprises a first antigen binding domain that binds to HLA-TERT540 peptide antigen, a second polypeptide that comprises a second antigen binding domain that binds to CD3 and a third antigen binding domain that binds to HLA-TERT540 peptide antigen. In certain aspects, the first antigen binding domain comprises a first scFv that binds to HLA-TERT540 peptide antigen, the second antigen binding domain comprises a second scFv that binds to CD3, and the third antigen binding domain comprises a third scFv that binds to HLA-TERT540 peptide antigen. In certain embodiments, the first polypeptide comprises a first hinge-CH2-CH3 and the second polypeptide comprises a second hinge-CH2-CH3; and wherein the first and second hinge-CH2-CH3 form a heterodimer.

[0202] In certain embodiments, a TCRm antibody of the disclosure comprises a first polypeptide that comprises a first and a third antigen binding domains that bind to HLA-TERT540 peptide antigen, and a second polypeptide that comprises a second antigen binding domain that binds toPatent Application CD3. In certain aspects, the first and the third antigen binding domains comprise a first scFv and a third scFv that binds to HLA-TERT540 peptide antigen, the second antigen binding domain comprises a second scFv that binds to CD3. In certain embodiments, the first polypeptide comprises a first hinge-CH2-CH3 and the second polypeptide comprises a second hinge-CH2-CH3; and wherein the first and second hinge-CH2-CH3 form a heterodimer.

[0203] In certain embodiments, a TCRm antibody of the disclosure comprises a first polypeptide that comprises a first antigen binding domain that binds to HLA-TERT540 peptide antigen, a second polypeptide and a third polypeptide that comprise a second antigen binding domain that binds to CD3 and a third antigen binding domain that binds to HLA-TERT540 peptide antigen. In certain aspects, the first antigen binding domain comprises a first scFv that binds to HLA-TERT540 peptide antigen, the second antigen binding domain comprises a second Fab that binds to CD3, and the third antigen binding domain comprises a third scFv that binds to HLA-TERT540 peptide antigen. In certain embodiments, the first polypeptide comprises a first hinge-CH2-CH3 and the second polypeptide comprises a second hinge-CH2-CH3; and wherein the first and second hinge-CH2-CH3 form a heterodimer. In certain embodiments, the third polypeptide is a light chain.

[0204] In certain aspects, the TCR mimetic-based T cell engager comprises an Fc domain with one or more Fc silencing mutations or substitutions, such as the “LALA”, “LALAGA”, “LALAKA” and “LALAPA” Fc substitutions.

[0205] In certain alternative embodiments, the second polypeptide does not comprise an antibody fragment that binds CD3 or an antibody fragment that binds HLA-TERT540 peptide antigen. In certain embodiments, the second polypeptide further comprises a third scFv comprising a second antibody fragment that binds HLA-TERT540 peptide antigen. In certain embodiments, the TCR mimetic T cell engager protein comprises two second polypeptides, wherein each second polypeptide comprises an scFv that binds CD3 and an scFv that binds HLA-TERT540 peptide antigen. In certain embodiments, the antibody fragment that binds CD3 comprises a Fab.

[0206] In certain embodiments, the TCR mimetic-based antibody comprises i) a first polypeptide comprising an scFv that binds HLA-TERT540 peptide antigen and ii) a second polypeptide and a third polypeptide comprising a Fab that binds to an immune cell (e.g., via CD3). In certain embodiments, the TCR mimetic T cell engager protein comprises a second polypeptide and a third polypeptide, wherein the second polypeptide and the third polypeptide comprise a Fab that binds to an immune cell (e g., via CD3) and an scFv that binds HLA-TERT540 peptide antigen.Patent Application

[0207] In certain embodiments, the antibody fragment that binds CD3 binds human CD3. In certain embodiments, the antibody fragment that binds CD3 binds CD3 with an affinity of a KD of less than 10 nM as determined by ELISA, SPR, BLI, flow cytometer or equivalent analysis. In certain embodiments, the antibody fragment that binds CD3 binds CD3 with an affinity of a KD of less than 50 nM as determined by ELISA, SPR, BLI, flow cytometer or equivalent analysis. In certain embodiments, the antibody fragment that binds CD3 with an affinity of a KD of greater than 50 nM as determined by ELISA, SPR, BLI, flow cytometer or equivalent analysis.

[0208] In certain embodiments, the Fc region (CH2-CH3) is silenced. In certain aspects, the Fc region comprises a C-terminal lysine. In certain aspects, the Fc region comprises a C-terminus without a C-terminal lysine.

[0209] In certain embodiments, the TCR mimetic-based antibody comprises a first polypeptide and a second polypeptide; wherein the first polypeptide and the second polypeptide form a dimer through at least one disulfide bond between a first hinge-CH2-CH3 of the first polypeptide and a second hinge-CH2-CH3 of the second polypeptide. In certain embodiments, the first hinge-CH2-CH3 and / or second hinge-CH2-CH3 comprises one or more amino acid substitutions that increase dimerization of the first polypeptide with the second polypeptide. In certain embodiments, the first polypeptide and second polypeptide form a homodimer through at least one disulfide bond between the first hinge-CH2-CH3 and the second hinge-CH2-CH3. In certain embodiments, the first polypeptide and second polypeptide form a heterodimer through at least one disulfide bond between the first hinge-CH2-CH3 and the second hinge-CH2-CH3. In certain embodiments, the immune cell binding domain (e.g., via CD3) comprises a Fab, which comprises a disulfide bond between the VH and the VL. In certain embodiments, the hinge region comprises a C220S mutation.

[0210] In certain embodiments, the antibody fragment that selectively binds the HLA-TERT540 peptide antigen comprises a first and third single chain variable fragment (scFv).

[0211] A composition comprising TCR mimetic-based antibodies described herein may be a pharmaceutical composition. Such a composition may comprise multiple TCR mimetic-based antibodies. Exemplary pharmaceutical compositions are described herein. The composition may be capable of eliciting an immune response. The composition may comprise an adjuvant. Suitable adjuvants include, but are not limited to 1018 ISS, alum, aluminium salts, Amplivax, AS15, BCG, CP-870,893, CpG7909, CyaA, dSLIM, GM-CSF, IC30, IC31, Imiquimod, ImuFact IMP321, ISPatent Application Patch, ISS, ISCOMATRIX, Juvlmmune, LipoVac, MF59, monophosphoryl lipid A, Montanide IMS 1312, Montanide ISA 206, Montanide ISA 50V, Montanide ISA-51, OK-432, OM-174, OM-197-MP-EC, ONTAK, PepTel vector system, PLG microparticles, resiquimod, SRL172, Virosomes and other Virus-like particles, YF-17D, VEGF trap, R848, beta-glucan, Pam3Cys, Aquila's QS21 stimulon (Aquila Biotech, Worcester, Mass., USA) which is derived from saponin, mycobacterial extracts and synthetic bacterial cell wall mimics, and other proprietary adjuvants such as Ribi's Detox. Quil or Superfos. Adjuvants such as incomplete Freund's or GM-CSF are useful. Several immunological adjuvants (e.g., MF59) specific for dendritic cells and their preparation have been described previously (Dupuis M, et al., Cell Immunol. 1998; 186( 1): 18-27; Allison A C; Dev Biol Stand. 1998; 92:3-11). In certain aspects, alternatively or additionally, cytokines are used. Several cytokines have been directly linked to influencing dendritic cell migration to lymphoid tissues (e.g., TNF-alpha), accelerating the maturation of dendritic cells into efficient antigen-presenting cells for T-lymphocytes (e.g., GM-CSF, IL-1 and IL-4) (U. S. Pat. No.5,849,589, specifically incorporated herein by reference in its entirety) and acting as immunoadjuvants (e.g., IL-12) (Gabrilovich D I, et al., J Immunother Emphasis Tumor Immunol.1996 (6):414-418). HLA surface expression and processing of intracellular proteins into peptides to present on HLA can also be enhanced by interferon-gamma (IFN-y). See, e.g., York IA, Goldberg AL, Mo XY, Rock KL. Proteolysis and class I major histocompatibility complex antigen presentation. Immunol Rev. 1999;172:49-66; and Rock KL, Goldberg AL. Degradation of cell proteins and the generation of MHC class I-presented peptides. Ann Rev Immunol. 1999;17: 12.739-779, which are incorporated herein by reference in their entirety.

[0212] Also provided herein are host cells comprising an ABP or TCR mimetic-based antibody or fragment thereof disclosed herein. In some embodiments, the host cell comprises a polynucleotide encoding an ABP or TCR mimetic-based antibody or fragment thereof. In some embodiments, the polynucleotide is heterologous to the host cell. In some embodiments, the host cell does not comprise endogenous MHC. In some embodiments, the host cell comprises an exogenous HLA class I molecule. In some embodiments, the host cell is a cultured cell from a tumor cell line. In some embodiments, the tumor cell line expresses an HLA subtype as defined by the HLA-TERT540 peptide antigen.

[0213] Also provided herein are cell culture systems comprising a host cell disclosed herein and a cell culture medium. In some embodiments, the host cell expresses the HLA class I subtype asPatent Application defined by the HLA-TERT540 peptide antigen and the cell culture medium comprises the HLA-TERT540 peptide as defined by the HLA-TERT540 peptide antigen.

[0214] An overview of the present disclosure is to identify TCR like antibodies binding to MHC-1 restricted tumor / autoimmune disease specific peptide TERT540 derived from TERT and engineered into a TCR mimetic (TCRm) T-cell engager (TCE), as defined herein.

[0215] Telomerase reverse transcriptase (TERT) is a self-antigen that is expressed constitutively in cells associated with certain cancers, tumors, or autoimmune diseases. TERT is part of the telomerase enzyme complex that extends telomeres. In addition to TERT, the complex includes an RNA component known as telomerase RNA component (TERC).

[0216] Telomerase synthesizes telomeric DNA, thereby compensating for its loss with each cell division. Nevertheless, telomeres shorten with successive cell divisions. Thus, telomere length reveals the replicative history of a cell / cellular population.

[0217] Maintaining constant telomere length is associated with chromosomal stability, and attenuates cellular aging. Unfortunately, telomerase activation is also correlated with malignant cell transformation. Though telomerase is, itself, not tumorigenic, its expression has an oncogenic role in facilitating proliferation of pre-cancerous cells, which may then allow proliferation of autoreactive or cancerous cells. Oncogenesis may occur in high-TERT expressing cell lines, by allowing pre-cancerous cells to proliferate continuously and become immortal. Thus, high telomerase activity is found in several forms of cancer and tumors. In contrast, normal tissues display little or no telomerase activity (relatively low activity may be seen in normal stem cells and progenitor cells).

[0218] Unsurprising given the importance of telomere length and maintenance in limiting cellular senescence, apoptosis and resulting cellular dysfunction, genetic mutations of TERT are associated with a range of pathologies. For example, TERT expression has been linked to inheritable disorders associated with inadequate telomere maintenance, such as forms of idiopathic pulmonary fibrosis, dyskeratosis congenita, and aplastic anemia. Further, telomerase / TERT driven premature cellular senescence / apoptosis (e.g., due to short telomeres) is associated with an increased risk of developing certain cancers and certain autoimmune conditions. Abundant correlative data has linked short telomeres (e.g., as a result of a TERT disorder) with increased risk of developing cancer, such as in aplastic anemia, in which evidence suggests that critically short telomeres may cause chromosomal instability, which increases the risk of malignant transformation.Patent Application

[0219] Studies have also shown that TERT expression can be elevated in immune cells like dendritic cells in autoimmune patients, potentially promoting their activation and contributing to inflammation. Associations between elevated TERT expression and several autoimmune diseases have been observed, including rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), and systemic sclerosis (SSc).

[0220] Human cells with little or no telomerase activity transfected with vectors encoding human TERT (hTERT) were found to subsequently express telomerase, display elongated telomeres, proliferatively expand, and display reduced senescence compared non-transfected cells. Normal cells possess a limited ability to divide and replicate, which represents a portion of the biological processes that inhibit cancer development. The end-point of cellular replication is senescence. Senescent cells often exhibit an altered pattern of gene expression, altered morphology, and reduced or abrogated ability to perform their normal functions. Thus, telomere shortening directly is a cause of cellular senescence in addition to its association with cancers and autoimmune diseases. Consequently, an immunotherapy targeting cells with high TERT expression must do so in a way that is targeted, to prevent accelerated senescence in healthy cells and the subsequent development of new pathologies associated with telomere shortening.

[0221] The human TERT:s4o-548 peptide (also referred to herein as “hTERT540” or “TERT540”), which has the sequence ILAKFLHWL (SEQ ID NO: 15) is an immunodominant HLA-A*02:01-restricted T-cell epitope, correlated with cells expressing TERT, which includes many cancerous cells and cells that express chronic inflammatory signals and aberrantly active immune cells (such as B cells) associated with autoimmune diseases and disorders. Thus, TERT540 represents a highly specific target for treating diseases associated with TERT expression.

[0222] According to the present disclosure, cells associated with certain cancers that express antigens contemplated by the present disclosure (TERT540, for example) include, for example, Hepatocellular Carcinoma (HCC), melanoma, lung cancer, colon cancer, pancreatic cancer, NonSmall Cell Lung Cancer (NSCLC), breast cancer, colorectal cancer, head and neck cancer, prostate cancer, kidney cancer, cholangiocarcinoma, and other advanced solid tumors, other than NSCLC, chemo-resistant advanced solid tumors, renal cancer, Acute Myeloid Leukemia (AML). Compositions and methods according to the present disclosure can treat or otherwise be useful for treating cancers and tumors and autoimmune diseases, allergies, and conditions associated with chronic inflammation, in particular those characterized by high expression of TERT540 (highPatent Application expression compared to a WT or non-pathogenic subject). Compositions and methods according to the present disclosure can treat or otherwise be useful for treating a cancer selected from pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, lung cancer, colon cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease, non-Hodgkin's lymphoma, cancer of the esophagus, cancer of the gastroesophageal junction, cancer of the small intestine, ampullary cancer, hepatocellular carcinoma, cholangiocarcinoma, intrahepatic cholangiocarcinoma, gallbladder cancer, biliary tract cancer, mesothelioma, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, chronic or acute leukemias including acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, solid tumors of childhood, lymphocytic lymphoma, myelodysplastic syndromes, myeloproliferative neoplasms, multiple myeloma, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, neoplasm of the central nervous system (CNS), glioma, high-grade glioma, glioblastoma, astrocytoma, oligodendroglioma, ependymoma, medulloblastoma, neuroblastoma, primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma, environmentally induced cancers including those induced by asbestos, and combinations of said cancers.

[0223] Compositions, pharmaceutical compositions, and methods according to the present disclosure can be utilized, for example, to treat a cancer. It is recognized that TERT540 pHLA may be an antigen for one of several types of cancer, including Hepatocellular carcinoma (HCC), Melanoma, Lung cancer, Pancreatic cancer, Non-small cell lung cancer (NSCLC), Breast cancer, colorectal cancer, head and neck cancers, prostate cancer, renal cancers, cholangiocarcinoma, certain advanced solid tumors other than NSCLC, certain chemo-resistant advanced solid tumors, and acute myeloid leukemia, the various compositions and methods provided herein would be useful with respect to other cancers as well.

[0224] Compositions and methods according to the present disclosure can treat or otherwise be useful for treating autoimmune diseases, such as rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), and systemic sclerosis (SSc), metabolic syndrome, chronic obstructivePatent Application pulmonary disease, diabetes, atherosclerosis, allergies, rheumatoid arthritis, inflammatory bowel disease (IBD), alcoholic liver disease, and chronic renal diseases, all of which may be associated with cells expressing high TERT / TERT540 levels. Such cells are also contemplated by the present disclosure as cells expressing antigens of interest.

[0225] The present disclosure provides compositions and methods for treating a subject having or suspected of having a TERT -expression related disorder, an animal model, an in vitro tissue culture model, and the like. TCRm-based antibodies or antigen binding portions thereof of the disclosure that specifically or selectively bind antigen targets related to TERT expression. In certain aspects, antibodies as described herein are human antibodies. In embodiments of the present disclosure, the provided TCRm-based antibodies and fragments thereof that specifically bind to TERT540 pHLA as a tumor antigen. In further embodiments of the present disclosure, novel TCRm-based antibodies (or antibody fragments) specifically bind to TERT540-PHLA as a tumor antigen, and can specifically bind to an antigen that can engage T-cells, for example CD3 expressed on the surface of a T-cell. In the context of this disclosure, TERT540 pHLA may be a tumor antigen or antigen associated with an autoimmune disease, allergy, or chronic inflammation.

[0226] An antibody, as used herein, can refer to an intact antibody (e.g., an intact immunoglobulin), a portion of a TCRm-based antibody, and antibody fragment, for example, an antigen binding fragment, or a bispecific antibody, including as part of a TCRm-based antibody. Antigen binding fragments can comprise at least one antigen binding domain. One example of an antigen binding domain is an antigen binding domain formed by a VH-VL dimer. A TCRm-based antibody, or antigen binding fragment can be described by the antigen to which they specifically bind. In some embodiments, the antigen binding fragments provided herein can comprise any of the antigen binding portions (also referred to as antigen binding domains) described below.

[0227] The VH and VL regions can be further subdivided into regions of hypervariability (hypervariable regions (HVRs), also called complementarity determining regions (CDRs)) interspersed with regions that are more conserved. The more conserved regions are called framework regions (FRs). Each VH and VL generally comprises three CDRs and four FRs, arranged in the following order (from N-terminus to C-terminus): FR1 - CDR1 - FR2 - CDR2 - FR3 - CDR3 - FR4. The CDRs are involved in antigen binding and confer antigen specificity and binding affinity to the antibody. (See Kabat et al. (1991) Sequences of Proteins of Immunological Interest 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD.) CDR sequences onPatent Application the heavy chain (VH) may be designated as CDRH1, 2, 3, while CDR sequences on the light chain (Vv) may be designated as CDRL1, 2, 3.

[0228] In each case, where a specific amino acid sequence is recited, embodiments comprising a sequence having at least 90% (e.g. 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) similarity / identity to the recited sequence are also provided. In some cases, where a specific amino acid sequence is recited, embodiments comprising a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) similarity / identity to the recited sequence are also provided. In each case, where a specific amino acid sequence is recited, embodiments consisting essentially of a sequence having at least 90% (e.g. 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) similarity / identity to the recited sequence are also provided. In some cases, where a specific amino acid sequence is recited, embodiments consisting essentially of a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) similarity / identity to the recited sequence are also provided. In each case, where a specific amino acid sequence is recited, embodiments consisting of a sequence having at least 90% (e.g. 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) similarity / identity to the recited sequence are also provided. In some cases, where a specific amino acid sequence is recited, embodiments consisting of a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) similarity / identity to the recited sequence are also provided.

[0229] The disclosure also provides a TCRm, antibody, or antigen binding portion thereof that specifically binds to a TERT antigen, wherein the TCRm, antibody, or antigen binding portion thereof comprises a heavy chain variable region comprising an amino acid sequence that is at least 90% identical / similar (for example, at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% similar / identical) to any one of SEQ ID NOS: 1 and 200-227 and a light chain variable region comprising an amino acid sequence that is at least 90% similar / identical (for example, at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% similar / identical) to any one of SEQ ID NO: 2 and 228-255. Sequence details can be found in the Examples below.

[0230] The disclosure also provides a TCRm, antibody, or antigen binding portion thereof that specifically binds to a TERT antigen, wherein the TCRm, antibody, or antigen binding portion thereof comprises a heavy chain variable region comprising an amino acid sequence that is at least 90% identical to any one of SEQ ID NOS: 1 and 200-227 and a light chain variable regionPatent Application comprising an amino acid sequence that is at least 90% identical to any one of SEQ ID NOS: 2 and 228-255.

[0231] The disclosure also provides a TCRm, antibody, or antigen binding portion thereof that specifically binds to a TERT antigen, wherein the TCRm, antibody, or antigen binding portion thereof comprises a heavy chain variable region comprising an amino acid sequence that is at least 95% identical to any one of SEQ ID NOS: 1 and 200-227 and a light chain variable region comprising an amino acid sequence that is at least 95% identical to any one of SEQ ID NOS: 2 and 228-255.

[0232] The disclosure also provides a TCRm, antibody, or antigen binding portion thereof that specifically binds to a TERT antigen, wherein the TCRm, antibody, or antigen binding portion thereof comprises a heavy chain variable region comprising an amino acid sequence that is at least 97% identical to any one of SEQ ID NOS: 1 and 200-227 and a light chain variable region comprising an amino acid sequence that is at least 97% identical to any one of SEQ ID NOS: 2 and 228-255.

[0233] The disclosure also provides a TCRm, antibody, or antigen binding portion thereof that specifically binds to a TERT antigen, wherein the TCRm, antibody, or antigen binding portion thereof comprises a heavy chain variable region comprising an amino acid sequence that is at least 98% identical to any one of SEQ ID NOS: 1 and 200-227 and a light chain variable region comprising an amino acid sequence that is at least 98% identical to any one of SEQ ID NOS: 2 and 228-255.

[0234] The disclosure also provides a TCRm, antibody, or antigen binding portion thereof that specifically binds to a TERT antigen, wherein the TCRm, antibody, or antigen binding portion thereof comprises a heavy chain variable region comprising an amino acid sequence that is at least 99% identical to any one of SEQ ID NOS: 1 and 200-227 and a light chain variable region comprising an amino acid sequence that is at least 99% identical to any one of SEQ ID NOS: 2 and 228-255.

[0235] The disclosure also provides a TCRm, antibody, or antigen binding portion thereof that specifically binds to a TERT antigen, wherein the TCRm, antibody, or antigen binding portion thereof comprises a heavy chain variable region comprising an amino acid sequence as set forth in any one of SEQ ID NOS: 1 and 200-227 and a light chain variable region as set forth in any one of SEQ ID NOS: 2 and 228-255.Patent Application

[0236] In additional disclosed aspects, the heavy chain variable region comprises an HC CDR1 having an amino acid sequence as set forth in any one of SEQ ID NO: 3 and SEQ ID NOS: 58–72, an HC CDR2 having an amino acid sequence as set forth in any one of SEQ ID NO: 4 and SEQ ID NOS: 73–90, and an HC CDR3 having an amino acid sequence as set forth in any one of SEQ ID NO: 5 and SEQ ID NOS: 91–118, and wherein the light chain variable region comprises an LC CDR1 having an amino acid sequence as set forth in any one of SEQ ID NO: 6 and SEQ ID NOS: 119–143, an LC CDR2 having an amino acid sequence as set forth in any one of SEQ ID NO: 7 and SEQ ID NOS: 144–171, and an LC CDR3 having an amino acid sequence as set forth in any one of SEQ ID NO: 8 and SEQ ID NOS: 172–199. In these aspects, the disclosed TCR mimetic antibodies encompass any antibody or fragment thereof that incorporates the recited CDR combinations irrespective of framework selection, humanization state, or overall antibody format, provided that binding to the HLA-TERT540 peptide complex is retained.

[0237] In certain aspects, an anti-TERT TCRm of the disclosure comprises a heavy chain variable region and a light chain variable region, wherein:the heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 3, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 4, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 5, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 6, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 7, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 8;the heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 58, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 73, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 91, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 119, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 144, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 172; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 59, an HC CDR2 sequence comprising an amino acid sequence ofPatent Application SEQ ID NO: 74, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 92, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 120, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 145, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 173; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 58, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 75, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 93, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 121, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 146, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 174; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 58, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 75, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 94, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 122, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 147, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 175; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 58, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 76, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 95, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 123, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 148, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 176; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 59, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 75, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 96, and the light chain variable region comprises an LC CDR1 sequence comprising anPatent Application amino acid sequence of SEQ ID NO: 124, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 149, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 177; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 60, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 77, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 97, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 125, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 150, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 178; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 61, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 75, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 98, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 126, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 151, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 179; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 62, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 78, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 99, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 127, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 152, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 180; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 63, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 79, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 100, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 128, an LC CDR2 sequence comprising an amino acidPatent Application sequence of SEQ ID NO: 153, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 181; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 64, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 80, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 101, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 129, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 154, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 182; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 63, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 78, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 102, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 130, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 155, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 183; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 63, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 81, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 103, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 131, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 156, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 184; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 58, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 82, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 104, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 132, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 157, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 185; orPatent Application the heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 65, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 83, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 105, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 133, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 158, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 186; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 3, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 4, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 106, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 134, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 159, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 187; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 59, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 75, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 107, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 135, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 160, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 188; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 59, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 75, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 108, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 136, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 161, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 189; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 59, an HC CDR2 sequence comprising an amino acid sequence ofPatent Application SEQ ID NO: 75, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 109, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 137, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 162, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 190; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 58, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 82, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 110, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 138, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 163, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 191; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 66, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 84, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 111, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 134, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 164, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 192; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 58, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 74, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 112, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 139, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 165, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 193; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 67, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 85, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 113, and the light chain variable region comprises an LC CDR1 sequence comprising anPatent Application amino acid sequence of SEQ ID NO: 134, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 166, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 194; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 68, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 86, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 114, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 140, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 167, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 195; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 69, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 87, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 115, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 141, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 168, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 196; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 70, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 88, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 116, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 142, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 169, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 197; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 71, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 89, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 117, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 142, an LC CDR2 sequence comprising an amino acidPatent Application sequence of SEQ ID NO: 170, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 198; orthe heavy chain variable region comprises an HC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 72, an HC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 90, and an HC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 118, and the light chain variable region comprises an LC CDR1 sequence comprising an amino acid sequence of SEQ ID NO: 143, an LC CDR2 sequence comprising an amino acid sequence of SEQ ID NO: 171, and an LC CDR3 sequence comprising an amino acid sequence of SEQ ID NO: 199.

[0238] In further aspects, the disclosed TCR mimetic antibody comprises a third binding domain in addition to the first and second binding domains. In certain aspects, the third binding domain binds an HLA-TERT peptide complex and, in particular aspects, binds an HLA-TERT540peptide complex. In certain aspects, the third binding domain binds to an immune cell receptor. In certain aspects, the immune cell receptor bound by the third binding domain is a different immune cell receptor than bound by the second binding domain. In certain aspects, the second binding domain binds to CD3 or CD28 and the third binding domain binds to CD3 or CD28. In certain aspects, a binding domain that binds to CD3 comprises an amino acid sequence having at least 90%, 95%, 98%, 99% or 100% similarity to the amino acid sequence of SEQ ID NO: 259.

[0239] The disclosure further encompasses TCR mimetic antibodies and fragments thereof that are assembled from multiple polypeptide chains. In certain aspects, the TCR mimetic antibody comprises a first polypeptide chain, a second polypeptide chain, and a third polypeptide chain, wherein the first polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 12, the second polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 13, and the third polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 14. In alternative aspects, the first polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 56, the second polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, orPatent Application identical to SEQ ID NO: 57, and the third polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 14.

[0240] In further aspects, the disclosed TCR mimetic antibody comprises a third binding domain in addition to the first and second binding domains, wherein the third binding domain binds an HLA-TERT peptide complex and, in particular aspects, binds an HLA-TERT540 peptide complex. In such bispecific or multivalent configurations, the TCR mimetic antibody may comprise a first polypeptide and a second polypeptide, wherein the first polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 10 and the second polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 11. In alternative bispecific configurations, the first polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 54 and the second polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 55. In still further aspects, the TCR mimetic antibody comprises a first polypeptide chain, a second polypeptide chain, and a third polypeptide chain, wherein the first polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 10, the second polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 258, and the third polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 14.

[0241] The disclosure also encompasses single-chain or alternative compact formats, wherein the TCR mimetic antibody or fragment thereof comprises a polypeptide having an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 9, thereby capturing all claimed levels of sequence identity for such embodiments. In additional aspects involving third binding domains, the TCR mimetic antibody comprises a first polypeptide and a second polypeptide, wherein the first polypeptide chain comprises an amino acid sequence that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 256 and the second polypeptide chain comprises an amino acid sequencePatent Application that is at least 90% identical, at least 95% identical, at least 98% identical, or identical to SEQ ID NO: 257.

[0242] In certain aspects, the TCRm is selected from the specific TCRms identified as TERT(2+1)’, TERT(1+1), TERT(1+1)’, TERT(2+1), TERT(2+1)”, TERT(2+1)*, -, TERT-a, TERT-b, TERT-c, TERT-d, TERT-e, TERT-f, TERT-g, TERT-h, TERT-I, TERT-j, TERT-k, TERT-1, TERT-m, TERT-n, TERT-o, TERT-p, TERT-q, TERT-r, TERT-s, TERT-t, TERT-u, TERT-v, TERT-w, TERT-x, TERT-y, TERT-z, TERT-aa, TERT-ab, and TERT-ac in Tables la, lb and Id and Table 2.

[0243] Across all disclosed aspects, the immune cell receptor bound by the second binding domain is CD3, thereby enabling recruitment and activation of T cells. In certain aspects, the disclosed TCR mimetic antibody or fragment thereof does not exhibit substantial binding affinity to the HLA class I molecule in the absence of the TERT540 peptide and does not exhibit substantial binding affinity to the TERT540 peptide in the absence of the HLA class I molecule, such that binding specificity is directed to the intact HLA-TERT540 peptide complex. In further aspects, the TERT540 peptide comprises the amino acid sequence ILAKFLHWL (SEQ ID NO: 15).

[0244] In preferred aspects, the antigen is a telomerase reverse transcriptase (TERT), including human telomerase reverse transcriptase (hTERT), derived immunogenic peptide. In certain aspects, the TERT immunogenic peptide is selected from the TERT immunogenic peptides of Table 1c and / or having an amino acid sequence as set forth in any one of SEQ ID NOS: 15-53.

[0245] In preferred aspects, the antigen is TERT540, a peptide derived from telomerase reverse transcriptase (TERT), and presented by HLA class I, specifically HLA-A*02:01. Provided herein are such TCRms and antigen binding portions thereof that specifically bind to antigens that are or are related to an autoimmune disease antigen or tumor associated antigen as well as another antigen, for example, CD3. Such TCRms also comprise an antibody or antigen binding portion thereof that specifically binds to a non-tumor / non-autoimmune associated antigen, such as CD3.

[0246] The amino acid residue sequences provided herein are set forth in single-letter amino acid code which can be used interchangeably with three-letter amino acid code. An amino acid refers to any monomer unit that can be incorporated into a peptide, polypeptide, or protein. The twenty natural or genetically encoded alpha-amino acids are as follows: alanine (Ala or A), arginine (Arg or R), asparagine (Asn or N), aspartic acid (Asp or D), cysteine (Cys or C), glutamine (Gin or Q), glutamic acid (Glu or E), glycine (Gly or G), histidine (His or H), isoleucine (He or I), leucine (LeuPatent Application or L), lysine (Lys or K), methionine (Met or M), phenylalanine (Phe or F), proline (Pro or P), serine (Ser or S), threonine (Thr or T), tryptophan (Trp or W), tyrosine (Tyr or Y), and valine (Vai or V). The structures of these twenty natural amino acids are shown in, e.g., Stryer et al., Biochemistry, 5thed., Freeman and Company (2002). The term amino acid also includes unnatural amino acids, modified amino acids (e.g., having modified side chains and / or backbones), and amino acid analogs.

[0247] As with all peptides, polypeptides, and proteins, including fragments thereof, it is understood that additional modifications in the amino acid sequence of the autoimmune / tumor associated antigen-specific antibodies or antigen binding fragments thereof described herein, for example, in the heavy chain variable region and / or light chain variable region, can occur that do not alter the nature or function of the antibodies or antigen binding fragments thereof. Such modifications include conservative amino acids substitutions, such that each recited sequence optionally contains one or more conservative amino acid substitutions. The list provided below identifies examples of groups that contain amino acids that are conservative substitutions for one another; these groups are exemplary as other conservative substitutions are known to those of skill in the art.1) Alanine (A), Glycine (G);2) Aspartic acid (D), Glutamic acid (E);3) Asparagine (N), Glutamine (Q);4) Arginine (R), Lysine (K);5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V);6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W);7) Serine (S), Threonine (T); and8) Cysteine (C), Methionine (M).

[0248] By way of example, when an aspartic acid at a specific residue is mentioned, also contemplated is a conservative substitution at the residue, for example, glutamic acid. Nonconservative substitutions, for example, substituting a proline with glycine, are also contemplated.

[0249] In some instances, the affinity of tumor associated antigen-specific antibodies or antigen binding fragments thereof may be optimized through mutations to increase or decrease affinity as desired based on one or more of the known characteristics of the binding interaction with the cognate antigen, the structure of either or both of the antibodies or fragments thereof, or thePatent Application antigen. In some instances, the mutations permit facile elution of purified antibodies or fragments thereof under desirable elution conditions during isolation and purification.

[0250] Methods of generating and screening for antibodies and antigen binding fragments thereof as provided in this disclosure are described in the Examples and are well-known in the art. Methods of further modifying antibodies for enhanced properties (e.g., enhanced affinity, chimerization, humanization) as well as generating antigen binding fragments, as described herein, are also well-known in the art.

[0251] The present disclosure also encompasses TCRms, antibodies or fragments thereof that bind to the same epitope of an antigen (e.g., HLA-TERT540peptide antigen) as the antibodies disclosed herein. Such antibodies can be identified using routine techniques known in the art, including, for example, competitive binding assays.

[0252] The present disclosure also encompasses bi-specific and other multi-specific antibodies or fragments thereof that bind to the same antigen epitopes as the antibodies disclosed herein, as well as other antigens. Such antibodies can be identified using routine techniques known in the art, including, for example, competitive binding assays.

[0253] The term epitope, as used herein, means a component of an antigen capable of specific binding to an antibody or antigen binding fragment thereof. Such components optionally comprise one or more contiguous amino acid residues and / or one or more non-contiguous amino acid residues. Epitopes frequently consist of surface-accessible amino acid residues and / or sugar side chains and can have specific three-dimensional structural characteristics, as well as specific charge characteristics. Conformational and non-conformational epitopes are distinguished in that the binding to the former but not the latter is lost in the presence of denaturing solvents. An epitope can comprise amino acid residues that are directly involved in the binding, and other amino acid residues, which are not directly involved in the binding. The epitope to which an antigen binding protein binds can be determined using known techniques for epitope determination such as, for example, testing for antigen binding protein binding to antigen variants with different point mutations.

[0254] The present disclosure also provides chimeric antibodies. The term chimeric antibody refers to an antibody in which a component of the heavy and / or light chain is derived from a particular source or species, while the remainder of the heavy and / or light chain is derived from a different source or species.Patent Application

[0255] A human antibody is one that possesses an amino acid sequence corresponding to that of an antibody produced by a human or a human cell, or derived from a non-human source that utilizes a human antibody repertoire or human antibody-encoding sequences (e.g., obtained from human sources, genetically modified non-human sources or designed de novo). Human antibodies specifically exclude humanized antibodies.

[0256] In some embodiments, a TCRm, antibody, or antigen binding fragment thereof provided herein can include a heavy (H) chain variable domain sequence (abbreviated herein as VH), and a light (L) chain variable domain sequence (abbreviated herein as VL). In some embodiments, an antibody molecule comprises or consists of a heavy chain and a light chain (referred to as a half antibody). In another example, a TCRm, antibody, or antigen binding fragment thereof includes two heavy (H) chain variable domain sequences and two light (L) chain variable domain sequence, thereby forming two antigen binding sites, such as Fab, Fab', F(ab')2, Fc, Fd, Fd', Fv, single chain antibodies (scFv, for example), single variable domain antibodies, diabodies (Dab) (bivalent and bispecific), and chimeric (e.g., humanized) antibodies, which may be produced by the modification of whole antibodies or synthesized de novo using recombinant DNA technologies. These functional antibody fragments retain the ability to selectively bind with their respective antigen. A TCRm, antibody, or antigen binding fragment thereof can be from any class of antibodies including, but not limited to, IgG, IgA, IgM, IgD, and IgE, and from any subclass (e.g., IgGl, IgG2, IgG3, and IgG4) of antibodies. The preparation of antibody molecules can be monoclonal or polyclonal. An antibody molecule can also be a human, humanized, CDR-grafted, or an in vitro generated antibody. A TCRm, antibody, or antigen binding fragment thereof can have a heavy chain constant region chosen from, e.g., IgGl, IgG2, IgG3, or IgG4. A TCRm, antibody, or antigen binding fragment thereof can also have a light chain chosen from either kappa or lambda light chains.

[0257] As used herein, the term monoclonal antibody refers to an antibody from a population of substantially homogeneous antibodies. A population of substantially homogeneous antibodies comprises antibodies that are the same or substantially similar and that bind the same epitope(s), except for variants that can normally arise during production of the monoclonal antibody. Such variants are generally present in only minor amounts. A monoclonal antibody is typically obtained by a process that includes the selection of a single antibody from a plurality of antibodies. For example, the selection process can be the selection of a unique clone from a plurality of clones,Patent Application such as a pool of yeast clones, phage clones, bacterial clones, mammalian cell clones, hybridoma clones, or other recombinant DNA clones. The selected antibody can be further altered, for example, to improve affinity for the target, for example, by affinity maturation, to humanize the antibody, to improve its production in cell culture, and / or to reduce its immunogenicity in a subject.

[0258] Antigen binding fragments, e.g., of a TCRm or of an antibody molecule are well known in the art, and include, for example, (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CHI domains; (ii) a F(ab')2fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CHI domains; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a diabody (dAb) fragment, which consists of a VH domain; (vi) a camelid or camelized variable domain; (vii) a single chain Fv (scFv) (see e.g., Bird et al. (1988) Science 242:423-426; Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883); (viii) a single domain antibody. These antibody fragments are obtained using conventional techniques known to those skilled in the art, and the fragments are screened for utility in the same manner as are intact antibodies.

[0259] In certain embodiments, a TCRm, antibody, or antigen binding fragment thereof, and compositions comprising TCRms as provided herein are distinguishable from naturally occurring antibodies and compositions in one or more respects. Such distinguishable antibodies and compositions may be referred to as “synthetic,” or may be identified by the proviso that the antibody or composition “is not naturally occurring” or affirmatively as “non-naturally occurring.” As used herein the terms “corresponding antibody,” and “corresponding to” describes the relationship between (1) an antibody characterized by six specific CDR sequences of the antibodies described in the Examples below and (2) a synthetic antibody comprising the same six CDR sequences. Synthetic antibodies of this disclosure may differ in structure from naturally occurring antibodies with the same CDRs. That is, synthetic antibodies identified by specified CDRs may be structurally different from antibodies comprising the specified CDRs that are described in the Examples below. Possible differences for synthetic antibodies include variable region sequences that differ corresponding naturally occurring antibodies, different light chain sequences (i.e. lambda type instead of kappa type or vice versa), different isotypes, different allotypes, and different constant domain variants. These differences are discussed in more detail below. In some embodiments, the synthetic antibody is an engineered polypeptide, also referred to as aPatent Application recombinant polypeptide, that is made using conventional protein and antibody engineering molecular biology, chemical, and biochemical methods as described below, including, but not limited to, those described in the Examples below.

[0260] In one approach, a TCRm, antibody, or antigen binding fragment as provided in this disclosure comprises one or more CDRs of an antibody / molecule described in Tables la, Table lb, Table Id, Table le, and Table 2.Patent Application Table la: Embodiments of Heavy-chain CDRs (Kabat annotation) according to the present disclosureHeavy chainSEQ SEQ SEQAntibodyID VHCDR1 ID VHCDR2 ID VHCDR3 ID NO: NO: NO:TERT-a 3 GYDMH 4 AISSRGGTIDYADSVEG 5 ELVGIGFDY GGYYYYGMD TERT-b 58 SDYWS 73 YIDYTGSTDYNPSLKS 91V59 DSSDTNYGM TERT-c SSYWS 74 YID YTGTTD YNP SLK S 92DV58 GGYGFDYYY TERT-d SDYWS 75 YVDYTGSTDYNPSLKS 93GMDV58 GGGDYYYYG TERT-e SDYWS 75 YVDYTGSTDYNPSLKS 94MDV58 GGDYDFWTY TERT-f SDYWS 76 YIDYTGNADYNPSLKS 95GMDV59 LPDISGWPFD TERT-g SSYWS 75 YVDYTGSTDYNPSLKS 96S TERT-h 60 KYSMH 77 AISGSGAYIDYADSVEG 97 DGISEGFDM DSSYTGHGLD TERT-i 61 SHYWS 75 YVDYTGSTDYNPSLKS 98V WIDPTTGTTNYAQKLQ GDDYYYYAM TERT-j 62 SYGIS 78 99G DV WIDPTTGDTNYAQKLQ VVGDYGGNY TERT-k 63 SYDTS 79 100G YYYMDV VMNPTTGDTIYAQKFQ TERT-1 64 NYWIH 80 101 EYEYGDYWI G WIDPTTGTTNYAQKLQ VMGYTGDYV TERT-m 63 SYDIS 78 102G GYYYAMDV WIDPTTGQTNYAQKLQ EIIAAAGTGD TERT-n 63 SYDIS 81 103G AFDI GGGNDFWSG TERT-o 58 SDYWS 82 YMDYTGSTDYNPSLKS 104SRGYFDL EGGYSGYEFD TERT-p 65 SSGIH 83 GINPSAGSTIYAQKFQG 105Y TERT-q 3 GYDMH 4 AISSRGGTIDYADSVEG 106 ESAGIGFDY GIGGSSWYFG TERT-r 59 SSYWS 75 YVDYTGSTDYNPSLKS 107LELMGDAFDIPatent Application VSYNWNVQE TERT-s 59 SSYWS 75 YVDYTGSTDYNPSLKS 108GAFDI GMGYFSVSH TERT-t 59 SSYWS 75 YVDYTGSTDYNPSLKS 109MDV GGSGWYGAF TERT-u 58 SDYWS 82 YMDYTGSTDYNPSLKS 110DI TERT-v 66 SYGMH 84 AISGSGDATDYADSVEG 111 EVVGVGFQH GAPGILYGMD TERT-w 58 SDYWS 74 YIDYTGTTDYNPSLKS 112V TERT-x 67 NAYMH 85 AISPSGGSTDYADSVEG 113 VDYDAFDI GEHSSGWYA TERT-y 68 EHDMH 86 AISSGSRYIDYADSVEG 114DAFDI TERT-z 69 AYDMH 87 AISGSSTTKDYADSVEG 115 DAVAGTLDY AIWSSSSSTTY TERT-aa 70 SYAMS 88 SISPEGSEYYPDSVKG 116YFDY VGRQWLNWY TERT-ab 71 MYTMS 89 SINDAGSSTYYPDSVKG 117FDL TVAGTADAF TERT-ac 72 NDWMS 90 SILGSGDYTYYPDSVKG 118DITable lb: Embodiments of Light-chain CDRs (Kabat annotation) according to the present disclosureLight chainAntibody SEQ VLCDR1 SEQ VLCDR2 SEQ VLCDR3 ID ID ID ID NO: NO: NO:TERT-a 6 RASQVVSNWLA 7 GATTLQS 8 QQYSEFPLT 119 144 172TERT-b RASETVSTNLA DASHRAP QHIHTWPIT 120 145 173TERT-c RASQSVTSSSLA GATTRAA QQYTTSPRT 121 146 174 HQYHETPL TERT-d RASQSLAGNLA DASNRATT122 147 175 QQYSVLPW TERT-e RASQPVSSNLA GASSRAST123 148 176 QQYNYFPP TERT-f RASQNIYSNLA DASTRADT124 149 177TERT-g RASQSVSNLA AASNRAP QNYNIAPYTPatent Application 125 150 178 QQYDNVPY TERT-h RASQDVSNWLA GASRLQST126 151 179TERT-i RASQGVARNLA ETSKRAT QQYHSIPQT 127 152 180 SSYAGSTW TERT-j TGTNIGRGSVS GDNIRPSG128 153 181 SSFTNRDTL TERT-k TGTTIGSTSVS GTSNRLPV129 154 182 QQYDRAPR TERT-1 RASRDIRNYLA KTSTLAST130 155 183 SSYAGTYT TERT-m TGTALPEQYVS ATDQRPSV131 156 184 GTWDASLG TERT-n TGTNLGNRYVS DTTNRHPAWV132 157 185 QQTYVTPR TERT-o RASQSIGSNLA GESNRAAA133 158 186 QQYVNVPL TERT-p RASQGISNWLA GASNLEST134 159 187TERT-q RASQDVSNWLA AATALQS QQYSEFPLT 135 160 188TERT-r RASQNIGHSLA GASYRAP QQYGSLPPT 136 161 189TERT-s RASQGISSNLA DASIRAP QQYSTSPYS 137 162 190TERT-t RASHSISDNLA DASERAP QQYVSRPST 138 163 191TERT-u RASQNIGSNLA DASTRAA QQYSTYPFT 134 164 192 QQVDTVPF TERT-v RASQDVSNWLA AATNLQTT139 165 193 QQYVTSPY TERT-w RASQSVSTKLA DTSNRATS134 166 194TERT-x RASQDVSNWLA GASNLQS VQSYSTPLT 140 167 195TERT-y RASQDIRTYLA GASTSQS HQSSSYPFT 141 168 196 QQGYSTPQ TERT-z RASQDISNWLA GASNLEAA142 RSSRSLVHGSGD 169 197TERT-aa DAHNRAT MQSFQFPLT NYLH142 RSSRSLVHGSGD 170 198 MQGSQWPH TERT-ab DTSNRAPNYLH T143 RSSQTLLHSSGY 171 199 KQALQFPW TERT-ac DASVRAPNYLH TPatent ApplicationTable lc: Embodiments of TERTPeptide Sequence TERT HLA SEQ ID NO position subtype(s)presentingpeptideMPRAPRCRA 1-9 HLA-B7 SEQ ID NO 16 RLGPQGWR 30-37 HLA-A2 SEQ ID NO 17 RLGPQGWRV 30-38 HLA-A2 SEQ ID NO 18 APSFRQVSCL 68-77 HLA-B7 SEQ ID NO 19 APSFRQVSCLKELVA 68-82 HLA-DR SEQ ID NO 20 AYQVCGPPL 167-175 HLA-A24 SEQ ID NO 21 RPAEEATSL 277-285 HLA-B7 SEQ ID NO 22 VYAETKHFL 324-332 HLA-A24 SEQ ID NO 23 YLEPACAKY 325-333 HLA-A1 SEQ ID NO 24 RPSFLLSSL 342-350 HLA-B7 SEQ ID NO 25 RPSLTGARRL 351-360 HLA-B7 SEQ ID NO 26 YWQMRPLFLELLGNH 386-400 HLA-DP SEQ ID NO 27 DPRRLVQLL 444-452 HLA-B7 SEQ ID NO 28 VYGFVRACL 461-469 HLA-A24 SEQ ID NO 29 FVRACLRRL 464-472 HLA-B7 SEQ ID NO 30 ILAKFLHWL 540-548 HLA-A2 SEQ ID NO 15 LAKFLHWLMSVYWE 541-555 HLA-DP SEQ ID NO 31 LLRSFFYN 555-563 HLA-A2 SEQ ID NO 32 RLFFYRKSV 572-580 HLA-A2 SEQ ID NO 33 YLFFYRKSV 572-580 HLA-A2 SEQ ID NO 34 LFFYRKSVWSKLQSI 573-584 HLA-DP SEQ ID NO 35 EARPALLTSRLRFIPK 611-626 HLA- SEQ ID NO 36DR, DQ, DP RPALLTSRLRFIPKP 613-627 HLA-DP SEQ ID NO 37 DYWGARTF 637-645 HLA-A24 SEQ ID NO 38 ALFSVLNYERARRPGLLGASVLGLDDIHRA 660-689 HLA- SEQ ID NO 39A2, DRSVLNYERARRPGLLG 663-677 HLA-DR SEQ ID NO 40 RPGLLGASVLGLDDI 672-686 HLA- SEQ ID NO 41DR1,7,15 PGLLGASVLGLDD1H 673-687 HLA- SEQ ID NO 42A2, DRGLLGASVLGL 674-683 HLA-A2 SEQ ID NO 43LLGASVLGL 675-683 HLA-A2 SEQ ID NO 44Patent Application LTDLQPYMRQFVAHL 766-780 HLA- SEQ ID NO 45DR1,7,15CYGDMENKL 845-853 HLA-A24 SEQ ID NO 46 RLVDDFLLV 865-873 HLA-A2 SEQ ID NO 47 KLFGVLRLK 973-981 HLA- SEQ ID NO 48A2. A3DLQVNSLQTV 988-997 HLA-A2 SEQ ID NO 49 YLQVNSLQTV 988-997 HLA-A2 SEQ ID NO 50 TYVPLLGSL 1088-1096 HLA-A24 SEQ ID NO 51 LPGTTLTAL 1107-1115 HLA-B7 SEQ ID NO 52LPSDFKTIL 1123-1131 HLA-B7 SEQ ID NO 53Table Id - Amino Acid VH / VL sequences of exemplary TERT TCRms of the disclosureFor SEQ ID NOs.: 1, 2 and 200-255: CDRs 1-3 are in bold and FRs 1-4 are underlined.> TERT-a VH amino acid sequence (SEQ ID NO:1) EVOLVESGGGLVOPGRSLRLSCAASGFTFSGYDMHWVRQAPGKGLEWVSAISSRGG TIDYADSVEGRFTISRDNARNSLYLQMNSLRAEDTAVYYCARELVGIGFDYWGOGT LVTVSS> TERT-b VH amino acid sequence (SEQ ID N0:200) OVOLQESGPGLVKPSOTLSLTCTVSGASITSDYWSWIROPPGKGLEWIGYIDYTGSTD YNPSLKSRVTMSVDTSKNQFSLKVNSVTAADTAVYYCARGGYYYYGMDVWGOGT TVTVS S> TERT-c VH amino acid sequence (SEQ ID NO:201) QVQLQESGPGLVKPSOTLSLTCTVSGASITSSYWSWIROPPGKGLEWIGYIDYTGTTD YNPSLKSRVTMSVDTSKNQFSLKVNSVTAADTAVYYCARDSSDTNYGMDVWGOGT TVTVS S> TERT-d VH amino acid sequence (SEQ ID NO:202) OVOLQESGPGLVKPSOTLSLTCTVSGASVSSDYWSWIROPPGKGLEWIGYVDYTGST DYNPSLKSRVTMSVDTSKNQFSLKVNSVTAADTAVYYCARGGYGFDYYYGMDVW GQGTTVTVSS> TERT-e VH amino acid sequence (SEQ ID NO:203) OVOLQESGPGLVKPSOTLSLTCTVSGASITSDYWSWIROPPGKGLEWIGYVDYTGSTD YNPSLKSRVTMSVDTSKNQFSLKVNSVTAADTAVYYCARGGGDYYYYGMDVWGOGTTVTVSSPatent Application> TERT-f VH amino acid sequence (SEQ ID NO:204) OVOLQESGPGLVKPSOTLSLTCTVSGASISSDYWSWIRQPPGKGLEWIGYIDYTGNAD YNPSLKSRVTMSVDTSKNOFSLKVNSVTAADTAVYYCARGGDYDFWTYGMDVWG QGTTVTVSS> TERT-g VH amino acid sequence (SEQ ID NO:205) OVOLQESGPGLVKPSOTLSLTCTVSGASITSSYWSWIROPPGKGLEWIGYVDYTGSTD YNPSLKSRVTMSVDTSKNQFSLKVNSVTAADTAVYYCARLPDISGWPFDSWGOGTL VTVSS> TERT-h VH amino acid sequence (SEQ ID NO:206) EVOLVESGGGLVQPGRSLRLSCAASGFTFSKYSMHWVROAPGKGLEWVSAISGSGA YIDYADSVEGRFTISRDNAKNSLYLOMNSLRAEDTAVYYCARDGISEGFDMWGOGT MVTVSS> TERT-i VH amino acid sequence (SEQ ID NO:207) OVOLQESGPGLVKPSOTLSLTCTVSGESVTSHYWSWIROPPGKGLEWIGYVDYTGST DYNPSLKSRVTMSVDTSKNQFSLKVNSVTAADTAVYYCARDSSYTGHGLDVWGOG TMVTVSS> TERT-j VH amino acid sequence (SEQ ID NO:208) EVOLVOSGAEVKKPGASVKVSCKASDYTFASYGISWVRQAPGOGLEWMGWIDPTT GTTNYAOKLQGRGTMTTDPSTSTAYMELRSLRSDDTAVYYCARGDDYYYYAMDV WGQGTTVTVSS> TERT-k VH amino acid sequence (SEQ ID NO:209) EVOLVOSGAEVKKPGASVKVSCKASEYTFSSYDISWVRQAPGOGLEWMGWIDPTTG DTNYAOKLQGRGTMTTDPSTSTAYMELRSLRSDDTAVYYCARVVGDYGGNYYYY MDVWGKGTTVTVSS> TERT-1 VH amino acid sequence (SEQ ID NO:210) OVOLVOSGAEVKKPGASVKVSCKVSGYTFANYWIHWVROAPGKGLEWMGVMNPT TGDTIYAQKFQGRVTMTEDTSTDTAYMELSSLKSEDTAVYYCAIEYEYGDYWIWG QGTMVTVSS> TERT-m VH amino acid sequence (SEQ ID NO:211) EVOLVOSGAEVKKPGASVKVSCKASRYTFSSYDISWVRQAPGOGLEWMGWIDPTTG TTNYAOKLOGRGTMTTDPSTSTAYMELRSLRSDDTAVYYCARVMGYTGDYVGYYYAMDVWGQGTTVTVSSPatent Application> TERT-n VH amino acid sequence (SEQ ID NO:212) EVOLVOSGAEVKKPGASVKVSCKASDYTFASYDISWVRQAPGOGLEWMGWIDPTT GOTNYAQKLQGRGTMTTDPSTSTAYMELRSLRSDDTAVYYCAREHAAAGTGDAF DIWGQGTMVTVSS> TERT-o VH amino acid sequence (SEQ ID NO:213) OVOLQESGPGLVKPSOTLSLTCTVSGASISSDYWSWIROPPGKGLEWIGYMDYTGST DYNPSLKSRVTMSVDTSKNQFSLKVNSVTAADTAVYYCARGGGNDFWSGSRGYFD LWGRGTLVTVSS> TERT-p VH amino acid sequence (SEQ ID NO:214) OVOLVOSGAEVKKPGASVKVSCKVSGFTLTSSGIHWVRQAPGKGLEWMGGINPSAG STIYAOKFOGRVTMTEDTSTDTAYMELSSLKSEDTAVYYCAREGGYSGYEFDYWG QGTLVTVSS> TERT-q VH amino acid sequence (SEQ ID NO:215) EVOLVESGGGLVQPGRSLRLSCAASGFTFSGYDMHWVROAPGKGLEWVSAISSRGG TIDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARESAGIGFDYWGOGT LVTVSS> TERT-r VH amino acid sequence (SEQ ID NO:216) OVOLQESGPGLVKPSOTLSLTCTVSGVSISSSYWSWIROPPGKGLEWIGYVDYTGSTD YNPSLKSRVTMSVDTSKNQFSLKVNSVTAADTAVYYCARGIGGSSWYFGLELMGD AFDIWGQGTMVT VS S> TERT-s VH amino acid sequence (SEQ ID NO:217) QVOLQESGPGLVKPSOTLSLTCTVSGASITSSYWSWIROPPGKGLEWIGYVDYTGSTD YNPSLKSRVTMSVDTSKNQFSLKVNSVTAADTAVYYCARVSYNWNVQEGAFDIWG QGTMVTVSS> TERT-t VH amino acid sequence (SEQ ID NO:218) QVQLQESGPGLVKPSQTLSLTCTVSGVSIASSYWSWIROPPGKGLEWIGYVDYTGSTD YNPSLKSRVTMSVDTSKNOFSLKVNSVTAADTAVYYCARGMGYFSVSHMDVWGQ GTTVTVSS> TERT-u VH amino acid sequence (SEQ ID NO:219) OVOLQESGPGLVKPSOTLSLTCTVSGASISSDYWSWIROPPGKGLEWIGYMDYTGST DYNPSLKSRVTMSVDTSKNOFSLKVNSVTAADTAVYYCARGGSGWYGAFDIWGOGTRVTVSSPatent Application> TERT-v VH amino acid sequence (SEQ ID NO:220) EVOLVESGGGLVOPGRSLRLSCAASGVSFSSYGMHWVRQAPGKGLEWVSAISGSGD ATDYADSVEGRFTISRDNAKNSLYLOMNSLRAEDTAVYYCAREVVGVGFOHWGQG TLVTVSS> TERT-w VH amino acid sequence (SEQ ID NO:221) OVOLQESGPGLVKPSOTLSLTCTVSGASITSDYWSWIROPPGKGLEWIGYIDYTGTTD YNPSLKSRVTMSVDTSKNOFSLKVNSVTAADTAVYYCARGAPGILYGMDVWGOGT TVTVS S> TERT-x VH amino acid sequence (SEQ ID NO:222) EVOLVESGGGLVQPGRSLRLSCAASGFTFRNAYMHWVROAPGKGLEWVSAISPSGG STDYADSVEGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARVDYDAFDIWGOGTM VTVSS> TERT-y VH amino acid sequence (SEQ ID NO:223) EVOLVESGGGLVQPGRSLRLSCAASGFTFSEHDMHWVROAPGKGLEWVSAISSGSR YIDYADSVEGRFTISRDNAKNSLYLOMNSLRAEDTAVYYCARGEHSSGWYADAFDI WGOGTMVTVSS> TERT-z VH amino acid sequence (SEQ ID NO:224) EVOLVESGGGLVQPGRSLRLSCAASGFSFSAYDMHWVROAPGKGLEWVSAISGSST TKDYADSVEGRFTISRDNAKNSLYLOMNSLRAEDTAVYYCARDAVAGTLDYWGQG TLVTVSS> TERT-aa VH amino acid sequence (SEQ ID NO:225) EVOLVESGGGLVQPGGSLRLSCAASGFTFGSYAMSWVROAPGKGLELVASISPEGSE YYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAKAIWSSSSSTTYYFDYW GOGTL VTVSS> TERT-ab VH amino acid sequence (SEQ ID NO:226) EVOLVESGGGLVQPGGSLRLSCAASGFTFSMYTMSWVROAPGKGLELVASINDAGS STYYPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARVGRQWLNWYFDL WGQGTL VTVSS> TERT-ac VH amino acid sequence (SEQ ID NO:227) EVOLVESGGGLVOPGGSLRLSCAASGFTFGNDWMSWVRQAPGKGLELVASILGSGD YTYYPDSVKGRFTISRDNAKNSLYLOMNSLRAEDTAVYYCARTVAGTADAFDIWGQGTL VTVSSPatent Application> TERT-a VL amino acid sequence (SEQ ID NO:2) DIQMTOSPSSLSASVGDRVTITCRASOVVSNWLAWYOQKPGKAPKLLIYGATTLOSG VPSRFSGSGLGTDFTLTISSLQPEDVATYYCOOYSEFPLTFGGGTKVEIK> TERT-b VL amino acid sequence (SEQ ID NO:228)EIVMTQ SPATL SL SPGERATL SCRASET VSTNLAW YQQKPGQ APRLLIYDASHRAPGI PARFSGSGSGTDFTLTISSLEPEDFAVYYCOHIHTWPITFGGGTKVEIK> TERT-c VL amino acid sequence (SEQ ID NO:229) EIVMTOSPATLSLSPGERATLSCRASQSVTSSSLAWYOQKPGOAPRLLIYGATTRAAG IPARFSGSGSGTDFTLTISSLEPEDFAVYYCOQYTTSPRTFGGGTKVEIK> TERT-d VL amino acid sequence (SEQ ID NO:230) E1VMTQSPATLSLSPGERATLSCRASOSLAGNLAWYQQKPGQAPRLL1YDASNRATG1 PARFSGSGSGTDFTLTISSLEPEDFAVYYCHQYHETPLTFGGGTKVEIK> TERT-e VL amino acid sequence (SEQ ID NO:231) EIVMTOSPATLSLSPGERATLSCRASQPVSSNLAWYOQKPGOAPRLLIYGASSRASGIP ARFSGSGSGTDFTLTISSLEPEDFAVYYCQOYSVLPWTFGGGTKVEIK> TERT-f VL amino acid sequence (SEQ ID NO:232)EIVMTQ SPATL SL SPGERATL SCRASONIYSNLAW YQQKPGQ APRLLIYDASTRADGI PARFSGSGSGTDFTLTISSLEPEDFAVYYCOOYNYFPPTFGGGTKVEIK> TERT-g VL amino acid sequence (SEQ ID NO:233) EIVMTOSPATLSLSPGERATLSCRASOSVSNLAWYOQKPGQAPRLLIYAASNRAPGIP ARFSGSGSGTDFTLTISSLEPEDFAVYYCQNYNIAPYTFGGGTKVEIK> TERT-h VL amino acid sequence (SEQ ID NO:234) DIOMTOSPSSLSASVGDRVTITCRASODVSNWLAWYOQKPGKAPKLLIYGASRLQSG VPSRFSGSGSGTDFTLTISSLQPEDVATYYCOOYDNVPYTFGGGTKVEIK> TERT-i VL amino acid sequence (SEQ ID NO:235) EIVMTOSPATLSLSPGERATLSCRASOGVARNLAWYOOKPGOAPRLLIYETSKRATG IPARFSGSGSGTDFTLTISSLEPEDFAVYYCOOYHSIPOTFGGGTKVEIK> TERT-j VL amino acid sequence (SEQ ID NO:236)ESALTOPASVSGSPGOSITISCTGTNIGRGSVSWYOQHPGKAPKLMIYGDNIRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYAGSTWGFGGGTKLTVLPatent Application> TERT-k VL amino acid sequence (SEQ ID NO:237) ESALTQPASVSGSPGOSITISCTGTTIGSTSVSWYOOHPGKAPKLMIYGTSNRLPGVS NRFSGSKSGNTASLTISGLOAEDEADYYCSSFTNRDTLVFGGGTKLTVL> TERT-1 VL amino acid sequence (SEQ ID NO:238) DIQMTQSPSSVSASVGDRVTITCRASRDIRNYLAWYQQKPGKAPKLLIYKTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDFANYYCQQYDRAPRTFGGGTKVEIK> TERT-m VL amino acid sequence (SEQ ID NO:239) ESALTQPASVSGSPGQSITISCTGTALPEQYVSWYQQHPGKAPKLMIYATDQRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYAGTYTVFGGGTKLTVL> TERT-n VL amino acid sequence (SEQ ID NO:240) ESALTQPASVSGSPGQSTITISCTGTNLGNRYVSWYQQHPGKAPKLMIYDTTNRHPGVSNRFSGSKSGNTASLTISGLQAEDEADYYCGTWDASLGAWVFGGGTKLTVL> TERT-o VL amino acid sequence (SEQ ID NO:241) EIVMTQSPATLSLSPGERATLSCRASQSIGSNLAWYQQKPGQAPRLLIYGESNRAAGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQTYVTPRAFGGGTKVEIK> TERT-p VL amino acid sequence (SEQ ID NO:242) DIQMTQSPSSVSASVGDRVTITCRASQGISNWLAWYQQKPGKAPKLLIYGASNLESG VPSRFSGSGSGTDFTLTISSLQPEDFANYYCQQYVNVPLTFGGGTKVEIK> TERT-q VL amino acid sequence (SEQ ID NO:243) DIQMTQSPSSLSASVGDRVTITCRASQDVSNWLAWYQQKPGKAPKLLIYAATALQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQQYSEFPLTFGGGTKVEIK> TERT-r VL amino acid sequence (SEQ ID NO:244)EIVMTQ SPATL SL SPGERATL SCRASQNIGHSLAWYQQKPGQ APRLLIYGASYRAPGI PARFSGSGSGTDFTLTISSLEPEDFAVYYCQQYGSLPPTFGGGTKVEIK> TERT-s VL amino acid sequence (SEQ ID NO:245)EIVMTQ SPATLSL SPGERATL SCRASQGISSNLAW YQQKPGQ APRLLIYDASIRAPGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQYSTSPYSFGGGTKVEIK> TERT-t VL amino acid sequence (SEQ ID NO:246)EIVMTQ SPATL SL SPGERATL SCRASHSISDNLAW YQQKPGQ APRLLIYDASERAPGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQYVSRPSTFGGGTKVEIKPatent Application> TERT-u VL amino acid sequence (SEQ ID NO:247)EIVMTQ SPATL SL SPGERATL SCRASQNIGSNLAW YQQKPGQ APRLLIYDASTRAAGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQYSTYPFTFGGGTKVEIK> TERT-v VL amino acid sequence (SEQ ID NO:248) DIQMTQSPSSLSASVGDRVTITCRASQDVSNWLAWYQQKPGKAPKLLIYAATNLQTGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCQQVDTVPFTFGGGTKVEIK> TERT-w VL amino acid sequence (SEQ ID NO:249)EIVMTQ SPATL SL SPGERATL SCRASQS VSTKLAWYQQKPGQ APRLLIYDTSNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQYVTSPYSFGGGTKVEIK> TERT-x VL amino acid sequence (SEQ ID NO:250) DIQMTQSPSSLSASVGDRVTITCRASQDVSNWLAWYQQKPGKAPKLLIYGASNLQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCVQSYSTPLTFGGGTKVEIK> TERT-y VL amino acid sequence (SEQ ID NO:251) DIQMTQSPSSLSASVGDRVTITCRASQDIRTYLAWYQQKPGKAPKLLIYGASTSQSGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCHQSSSYPFTFGGGTKVEIK> TERT-z VL amino acid sequence (SEQ ID NO:252)DIQMTQ SP S SL S AS VGDRVTITCRASQDISNWLAWYQQKPGKAPKLLIYGASNLEAG VPSRFSGSGSGTDFTLTISSLQPEDVATYYC QQGYSTPQAFGGGTKVEIK> TERT-aa VL amino acid sequence (SEQ ID NO:253) DIVMTQSPLSLPVTPGEPASISCRSSRSLVHGSGDNYLHWYLQKPGQSPQLLIYDAHNRATGIPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQSFQFPLTFGGGTKVEIK> TERT-ab VL amino acid sequence (SEQ ID NO:254) DIVMTQSPLSLPVTPGEPASISCRSSRSLVHGSGDNYLHWYLQKPGQSPQLLIYDTSNRAPGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQGSQWPHTFGGGTKVEIK> TERT-ac VL amino acid sequence (SEQ ID NO:255)DIVMTQSPLSLPVTPGEPASISCRSSQTLLHSSGYNYLHWYLQKPGQSPQLLIYDASVRAPGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCKQALQFPWTFGGGTKVEIKPatent Application Table le- Additional Amino Acid sequences of exemplary TERT TCRms of the disclosure (Amino Acid Sequences of Linkers Bold and Underlined)> TERT(2+1)7 TERT(1+1) / TERT(2+1) / TERT(2+1)’7 TERT(2+1)* anti-TERT scFv (VL-linker-VH) Sequences (SEQ ID NO:9)DIQMTQ SP S SL S AS VGDRVTITCRASQ VVSNWL AWYQQKPGKAPKLLIYGATTLQ SG VPSRFSGSGLGTDFTLTISSLOPEDVATYYCQQYSEFPLTFGGGTKVEIKGGGGSGGG GSGGGGSGGGGSEVOLVESGGGLVQPGRSLRLSCAASGFTFSGYDMHWVRQAPGK GLEWVSAISSRGGTIDYADSVEGRFTISRDNARNSLYLQMNSLRAEDTAVYYCARELV GIGFDYWGQGTLVTVSS> TERT(2+1)7 TERT(2+1)* Chain-1 anti-TERT scFv-linker-silent FC IgGl (SEQ ID NO:10)DIQMTQ SP S SL SAS VGDRVTITCRASQ VVSNWL AWYQQKPGKAPKLLIYGATTLQ SG VPSRFSGSGLGTDFTLTISSLQPEDVATYYCOOYSEFPLTFGGGTKVEIKGGGGSGGG GSGGGGSGGGGSEVQLVESGGGLVQPGRSLRLSCAASGFTFSGYDMHWVRQAPGKGLEWVSAISSRGGTIDYADSVEGRFTISRDNARNSLYLQMNSLRAEDTAVYYCARELVGIGFDYWGQGTLVTVSSGSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPGK> TERT(2+1)’ Chain-2 anti-TERT scFv- linker- anti-CD3 scFv-linker-silent FC IgGl(SEQ ID NO:11)DIQMTQ SP S SL SAS VGDRVTITCRASQ VVSNWL AWYQQKPGKAPKLLIYGATTLQ SG VPSRFSGSGLGTDFTLT1SSLOPEDVATYYCOOYSEFPLTFGGGTKVEIKGGGGSGGG GSGGGGSGGGGSEVQLVESGGGLVQPGRSLRLSCAASGFTFSGYDMHWVRQAPGK GLEWVSAISSRGGTIDYADSVEGRFTISRDNARNSLYLQMNSLRAEDTAVYYCARELV GIGFDYWGQGTLVTVSSGGGGSGGGGSAIQMTQSPSSLSASVGDRVTITCRASQDIRN YLNWYQQKPGKAPKLLIYYTSRLESGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQ QGNTLPWTFGQGTKVEIKGGGGSGGGGSGGGGSGGGGSGGGSEVQLVESGGGLV QPGGSLRLSCAASGYSFTGYTMNWVRQAPGKGLEWVALINPYKGVSTYNQKFKDRF TISVDKSKNTAYLQMNSLRAEDTAVYYCARSGYYGDSDWYFDVWGQGTLVTVSSGS EPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALP APIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PGK> TERT(1+1)’ Chain-1 anti-TERT scFv-linker-silent FC IgGl (SEQ ID NO: 12) DIQMTQ SP S SL SAS VGDRVTITCRASQ VVSNWL AW YQQKPGKAPKLLIYGATTLQ SG VPSRFSGSGLGTDFTLTISSLOPEDVATYYCOOYSEFPLTFGGGTKVEIKGGGGSGGG GSGGGGSGGGGSEVQLVESGGGLVQPGRSLRLSCAASGFTFSGYDMHWVRQAPGKGLEWVSAISSRGGTIDYADSVEGRFTISRDNARNSLYLQMNSLRAEDTAVYYCARELVPatent Application GIGFDYWGQGTLVTVSSGSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPGK> TERT(1+1)’ Chain-2 anti-CD3 VH CHl-hinge_silent FC IgGl (SEQ ID NO: 13) EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYTMNWVRQAPGKGLEWVALINPYKG VSTYNQKFKDRFTISVDKSKNTAYLQMNSLRAEDTAVYYCARSGYYGDSDWYFDV WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVD GVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTIS KAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK> TERT(1+1) / TERT(1+1)’ / TERT(2+1)* Chain-3 anti-CD3 VL CK (SEQ ID NO:14) AIQMTQ SP S SL S AS VGDRVTITCRASQDIRNYLNWYQQKPGK APKLLIYYT SRLESGV PSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPWTFGQGTKVEIKRTVAAPSVFI FPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYS LSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC> TERT(2+1) Chain-1 anti- TERT scFv- linker- anti-CD3 scFv-linker-silent FC IgGl(no terminal K) (SEQ ID NO:54)DIQMTQ SP S SL S AS VGDRVTITCRASQ VVSNWL AWYQQKPGKAPKLLIYGATTLQ SG VPSRFSGSGLGTDFTLTISSLOPEDVATYYCOQYSEFPLTFGGGTKVEIKGGGGSGGG GSGGGGSGGGGSEVQLVESGGGLVQPGRSLRLSCAASGFTFSGYDMHWVRQAPGKGLEWVSAISSRGGTIDYADSVEGRFTISRDNARNSLYLQMNSLRAEDTAVYYCARELVGIGFDYWGQGTLVTVSSGSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPG> TERT(2+1) Chain-2 anti-TERT scFv- linker- anti-CD3 scFv-linker-silent FC IgGl(no terminal K) (SEQ ID NO:55)DIQMTQ SP S SL SAS VGDRVTITCRASQ VVSNWL AWYQQKPGKAPKLLIYGATTLQ SG VPSRFSGSGLGTDFTLTISSLOPEDVATYYCQQYSEFPLTFGGGTKVEIKGGGGSGGG GSGGGGSGGGGSEVQLVESGGGLVQPGRSLRLSCAASGFTFSGYDMHWVRQAPGKGLEWVSAISSRGGTIDYADSVEGRFTISRDNARNSLYLQMNSLRAEDTAVYYCARELVGIGFDYWGQGTLVTVSSGGGGSGGGGSAIQMTQSPSSLSASVGDRVTITCRASQDIRN YLNWYQQKPGKAPKLLIYYTSRLESGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQ QGNTLPWTFGQGTKVEIKGGGGSGGGGSGGGGSGGGGSGGGSEVQLVESGGGLV QPGGSLRLSCAASGYSFTGYTMNWVRQAPGKGLEWVALINPYKGVSTYNQKFKDRFTISVDKSKNTAYLQMNSLRAEDTAVYYCARSGYYGDSDWYFDVWGQGTLVTVSSGSPatent Application EPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALP APIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLS PG> TERT(1+1) Chain- l anti-TERT scFv-linker-silent FC IgGl(no terminal K) (SEQ ID NO:56)DIQMTQ SP S SL S AS VGDRVTITCRASQ VVSNWL AW YQQKPGKAPKLLIYGATTLQ SG VPSRFSGSGLGTDFTLTISSLOPEDVATYYCOQYSEFPLTFGGGTKVEIKGGGGSGGG GSGGGGSGGGGSEVQLVESGGGLVQPGRSLRLSCAASGFTFSGYDMHWVRQAPGK GLEWVSAISSRGGTIDYADSVEGRFTISRDNARNSLYLQMNSLRAEDTAVYYCARELV GIGFDYWGQGTLVTVSSGSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPG> TERT(1+1) Chain-2 anti-CD3 VH CHl-hinge_silent FC IgGl(no terminal K) (SEQ ID NO:57) EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYTMNWVRQAPGKGLEWVALINPYKG VSTYNQKFKDRFTISVDKSKNTAYLQMNSLRAEDTAVYYCARSGYYGDSDWYFDV WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVD GVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTIS KAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTT PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG> TERT(2+1)” Chain-1 anti- TERT scFv- linker- anti- TERT scFv-linker-silent FC IgGl (SEQ ID NO:256)DIQMTQ SP S SL SAS VGDRVTITCRASQ VVSNWL AWYQQKPGKAPKLLIYGATTLQ SG VPSRFSGSGLGTDFTLTISSLOPEDVATYYCOOYSEFPLTFGGGTKVEIKGGGGSGGG GSGGGGSGGGGSEVQLVESGGGLVQPGRSLRLSCAASGFTFSGYDMHWVRQAPGK GLEWVSAISSRGGTIDYADSVEGRFTISRDNARNSLYLQMNSLRAEDTAVYYCARELV GIGFDYWGQGTLVTVSSGGGGSGGGGSDIOMTQSPSSLSASVGDRVTITCRASQVVS NWLAWYQQKPGKAPKLLIYGATTLQSGVPSRFSGSGLGTDFTLTISSLQPEDVATYYC QQYSEFPLTFGGGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGR SLRLSCAASGFTFSGYDMHWVRQAPGKGLEWVSAISSRGGTIDYADSVEGRFTISRDN ARNSLYLQMNSLRAEDTAVYYCARELVGIGFDYWGQGTLVTVSSGSEPKSSDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKG QPREPQ VCTLPP SRDELTKNQ VSL SC A VKGF YP SDIAVEWESNGQPENNYKTTPP VLD SDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKPatent Application > TERT(2+1)” Chain-2 anti-CD3 scFv-linker-silent FC IgGl (SEQ ID NO:257) AIQMTQ SP S SL S AS VGDRVTITCRASQDIRNYLNW YQQKPGKAPKLLIYYT SRLESGV PSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPWTFGQGTKVEIKGGGGSGGG GSGGGGSGGGGSGGGSEVQLVESGGGLVQPGGSLRLSCAASGYSFTGYTMNWVRO APGKGLEWVALINPYKGVSTYNQKFKDRFTISVDKSKNTAYLQMNSLRAEDTAVYY CARSGYYGDSDWYFDVWGQGTLVTVSSGSEPKSSDKTHTCPPCPAPEAAGGPSVFLF PPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDE LTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDK SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK> TERT(2+1)* Chain-2 anti-TERT scFv- linker anti-CD3 VH CHl-hinge_silent FC IgGl (SEQ ID NO:258)DIQMTQ SP S SL S AS VGDRVTITCRASQ VVSNWL AW YQQKPGK APKLLIYGATTLQ SG VPSRFSGSGLGTDFTLTISSLOPEDVATYYCOQYSEFPLTFGGGTKVEIKGGGGSGGG GSGGGGSGGGGSEVQLVESGGGLVQPGRSLRLSCAASGFTFSGYDMHWVRQAPGK GLEWVSAISSRGGTIDYADSVEGRFTISRDNARNSLYLQMNSLRAEDTAVYYCARELV GIGFDYWGQGTLVTVSSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGYSF TGYTMNWVRQAPGKGLEWVALINPYKGVSTYNQKFKDRFTISVDKSKNTAYLQMNS LRAEDTAVYYCARSGYYGDSDWYFDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSG GTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPA VLQS SGL YSLS S VVTVPS S SLGT QTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMI SRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGK> TERT(2+1) / TERT(2+1)7 TERT(2+1)” anti-CD3_scFv sequence (SEQ ID NO: 259) AIQMTQ SP S SL S AS VGDRVTITCRASQDIRNYLNWYQQKPGKAPKLLIYYT SRLESGV PSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPWTFGOGTKVEIKGGGGSGGG GSGGGGSGGGGSGGGSEVQLVESGGGLVQPGGSLRLSCAASGYSFTGYTMNWVRQ APGKGLEWVALINPYKGVSTYNQKFKDRFTISVDKSKNTAYLQMNSLRAEDTAVYYCARSGYYGDSDWYFDVWGQGTLVTVSSTable 2 Exemplary anti-TERT TCRms of the disclosureMolecule ID Chain -1 Chain -2 Chain-3 TERT(2+1)’ Seq. ID 10 Seq. ID 11TERT(1+1) Seq. ID 56 Seq. ID 57 Seq. ID 14 TERT(1+1)’ Seq. ID 12 Seq. ID 13 Seq. ID 14 TERT(2+1) Seq. ID 54 Seq. ID 55TERT(2+1)” Seq. ID 256 Seq. ID 257Patent Application TERT(2+1)* Seq. ID 10 Seq. ID 258 Seq. ID 14Patent Application

[0261] In some embodiments, the TCRm antibody comprises a heavy chain variable region sequence and a light chain variable region sequence that are derived from a semi -synthetic human antibody library

[0262] In some embodiments, the TCRm antibody comprises a heavy chain variable region sequence and a light chain variable region sequence that are derived from an immunoglobulin-producing human B cell, and further comprises a heavy chain constant region having an IgG isotype (e.g., IgGl), an IgA isotype (e.g., IgAl), an IgM isotype, an IgD isotype, or that is derived from an IgG, IgA, IgM, or IgD isotype e.g., is a modified IgGl constant region). It will be appreciated by a person of ordinary skill in the art that the different heavy chain isotypes (IgA, IgD, IgE, IgG, and IgM) have different effector functions that are mediated by the heavy chain constant region, and that for certain uses it may be desirable to have an antibody that has the effector function of a particular isotype (e.g., IgG).

[0263] In some embodiments, the TCRm antibody comprises a native (i.e., wild-type) human IgG, IgA, IgM, or IgD constant region. In some embodiments, the antibody comprises a native human IgGl constant region, a native human IgG2 constant region, a native human IgG3 constant region, a native human IgG4 constant region, a native human IgAl constant region, a native human IgA2 constant region, a native human IgM constant region, or a native human IgD constant region. In some embodiments, the antibody comprises a heavy chain constant region that comprises one or more modifications. It will be appreciated by a person of ordinary skill in the art that modifications such as amino acid substitutions can be made at one or more residues within the heavy chain constant region that modulate effector function. In some embodiments, the modification reduces effector function, e.g., results in a reduced ability to induce certain biological functions upon binding to an Fc receptor expressed on an effector cell that mediates the effector function. In some embodiments, the modification (e.g., amino acid substitution) prevents ex vivo Fab arm exchange, which can introduce undesirable effects and reduce the therapeutic efficacy of the antibody. See, e.g., Silva et al., J Biol Chem, 2015, 280:5462-5469.

[0264] In some embodiments, the TCRm antibody comprises a native (i.e., wild-type) human IgM constant region, human IgD constant region, human IgG constant region that is derived from IgGl, IgG2, IgG3, or IgG4, or human IgA constant region that is derived from IgAl or IgA2 and comprises one or more modifications that modulate effector function. In some embodiments the antibody comprises a human IgM constant region, human IgD constant region, human IgGPatent Application constant region that is derived from IgG1, IgG2, IgG3, or IgG4, or human IgA constant region that is derived from IgAl or IgA2. In some embodiments, the antibody comprises a native (i.e., wildtype) human IgM constant region, human IgD constant region, human IgG constant region that is derived from IgGl, IgG2, IgG3, or IgG4, or human IgA constant region that is derived from IgAl or IgA2 and comprises one, two, three, four, five, six, seven, eight, nine, ten or more modifications (e.g., amino acid substitutions). In some embodiments the constant regions includes variations (e.g., one, two, three, four, five, six, seven, eight, nine, ten or more amino acid substitutions) that reduce effector function.

[0265] Synthetic TCRm antibodies of this disclosure may comprise variations in heavy chain constant regions to change the properties of the synthetic antibody relative to the corresponding naturally occurring antibody. Exemplary changes include mutations to modulate antibody effector function (e.g, complement-based effector function or FcyR-based effector function), alter half-life, modulate co-engagement of antigen and FcyRs, introduce or remove glycosylation motifs (glyco-engineering). See Fonseca et al., 2018, “Boosting half-life and effector functions of therapeutic antibodies by Fc-engineering: An interaction-function review” Int J Biol Macromol.19:306-311; Wang et al., 2018, “IgG Fc engineering to modulate antibody effector functions” Protein Cell 2018, 9(1):63- 73; Schlothauer, 2016, “Novel human IgGl and IgG4 Fc-engineered antibodies with completely abolished immune effector functions,” Protein Engineering, Design and Selection 29(10):457-466; Tam et al., 2017, “Functional, Biophysical, and Structural Characterization of Human IgGl and IgG4 Fc Variants with Ablated Immune Functionality” Antibodies 6, 12, each incorporated herein by reference for all purposes.

[0266] In some embodiments, the heavy chain variable region and / or the light chain variable region of the TCRm antibody has an identical sequence to the heavy chain variable region and / or the light chain variable region encoded by the immunoglobulin producing single B cell from the human subject having a cancer or autoimmune disease. In some embodiments, the heavy chain variable region and / or the light chain variable region of the TCRm antibody comprises one or more modifications, e.g., amino acid substitutions, deletions, or insertions.

[0267] The heavy chain variable region sequence and / or light chain variable region sequence of a TCRm antibody described herein can be engineered to comprise one or more variations in the heavy chain variable region sequence and / or light chain variable region sequence. In some embodiments, the engineered variation(s) improves the binding affinity of the TCRm antibody forPatent Application a target antigen and / or immune cell. In some embodiments, the engineered variation(s) improves the cross-reactivity of the TCRm antibody for an antigen.

[0268] In some embodiments, the engineered variation is a variation in one or more CDRs, e.g., an amino acid substitution in a heavy chain CDR and / or a light chain CDR as described herein. In some embodiments, the engineered variation is a variation in one or more framework regions, e.g., an amino acid substitution in both heavy chain and light chain framework regions or in only a heavy chain framework region or only a light chain framework region. In some embodiments, the engineered variation is a reversion of a region of the heavy chain and / or light chain sequence to the inferred naive sequence. Methods for determining an inferred naive immunoglobulin sequence are described in the art. See, e.g., Magnani et al., PLoS Negl Trop Dis, 2017, 11:e0005655, doi: 10.1371 / journal. pntd.0005655

[0269] In some embodiments, affinity maturation is used to engineer further mutations that enhance the binding affinity of the antibody for an antigen or enhance the cross-reactivity of the antibody for a second antigen. Methods for performing affinity maturation are known in the art. See, e.g., Renaut et al., Methods Mol Biol, 2012, 907:451-461.

[0270] TCRm antibody molecules can also be or comprise single domain antibodies. Single domain antibodies can include antibodies whose complementary determining regions are part of a single domain polypeptide. Examples include, but are not limited to, heavy chain antibodies, antibodies naturally devoid of light chains, single domain antibodies derived from conventional 4-chain antibodies, engineered antibodies and single domain scaffolds other than those derived from antibodies. Single domain antibodies may be any of the art, or any future single domain antibodies. Single domain antibodies may be derived from any species including, but not limited to mouse, rat, guinea, pig, human, camel, llama, fish, shark, goat, rabbit, and bovine. Single domain antibodies are described, for example, in International Application Publication No. WO 94 / 04678. For clarity reasons, this variable domain derived from a heavy chain antibody naturally devoid of light chain is known herein as a VHH or nanobody to distinguish it from the conventional VH of four chain immunoglobulins. Such a VHH molecule can be derived from antibodies raised in Camelidae species (e.g., camel, llama, dromedary, alpaca and guanaco) or other species besides Camelidae.

[0271] In some embodiments, an antigen binding fragment can also be or can also comprise, e.g., a non-antibody, scaffold protein. These proteins are generally obtained through combinatorialPatent Application chemistry-based adaptation of preexisting antigen-binding proteins. For example, the binding site of human transferrin for human transferrin receptor can be diversified using the system described herein to create a diverse library of transferrin variants, some of which have acquired affinity for different antigens. See, e.g., Ali et al. (1999) J. Biol. Chem. 274:24066-24073. The portion of human transferrin not involved with binding the receptor remains unchanged and serves as a scaffold, like framework regions of antibodies, to present the variant binding sites. The libraries are then screened, as an antibody library is screened, and in accordance with the methods described herein, against a target antigen of interest to identify those variants having optimal selectivity and affinity for the target antigen. See, e.g., Hey et al. (2005) TRENDS Biotechnol 23(10):514-522.

[0272] Synthetic TCRm antibodies of this disclosure may differ from naturally occurring compositions in at least one or more of the following respects: (i) composition comprises antibodies that are purified, i.e., separated from tissue or cellular material with which they are associated in the human body, and optionally in an manufactured excipient or medium; and / or (ii) antibody compositions according to the present disclosure contain a single species of antibody (are monoclonal) such that all antibodies in the composition have the same structure and specificity.

[0273] The antibodies and antigen binding fragments thereof disclosed herein may be produced by recombinant expression in a human or non-human cell. Synthetic antibody-producing cells include non-human cells expressing heavy chains, light chains, or both heavy and light chains; human cells that are not immune cells; heavy chains, light chains, or both heavy and light chains; and human B cells that produce heavy chains or light chains, but not both heavy and light chains. Synthetic antibodies of this disclosure may be heterologously expressed, in vitro or ex vivo, in cells other than human B cells, such as non-human cells and human cells other than B cells, optionally other than immune cells, and optionally in cells other than cells in a B cell lineage.

[0274] The antigen-specific antibodies or antigen binding fragments thereof disclosed herein (e.g., the TCRm antibodies disclosed herein) can be produced using a variety of techniques known in the art of molecular biology and protein chemistry. For example, a nucleic acid encoding the antibody or antigen binding fragment thereof can be inserted into an expression vector that contains transcriptional and translational regulatory sequences, which include, e.g., promoter sequences, ribosomal binding sites, transcriptional start and stop sequences, translational start and stop sequences, transcription terminator signals, polyadenylation signals, and enhancer or activator sequences. The regulatory sequences include a promoter and transcriptional start and stopPatent Application sequences. In addition, the expression vector can include more than one replication system, such that it can be maintained in two different organisms, for example, in mammalian or insect cells for expression and in a prokaryotic host for cloning and amplification.

[0275] Several possible vector systems are available for the expression of cloned heavy chain and light chain polypeptides from nucleic acids in mammalian cells. One class of vectors relies upon the integration of the desired gene sequences into the host cell genome. Cells that have stably integrated DNA can be selected by simultaneously introducing drug resistance genes such as E. coli (Mulligan and Berg (1981) Proc Natl Acad Sci USA 78:2072) or Tn5 neo (Southern and Berg (1982) Mol Appl Genet 1:327). The selectable marker gene can be either linked to the DNA gene sequences to be expressed or introduced into the same cell by co-transfection (Wigler et al. (1979) Cell 16:77). A second class of vectors utilizes DNA elements that confer autonomously replicating capabilities to an extrachromosomal plasmid. These vectors can be derived from animal viruses, such as bovine papillomavirus (Sarver et al. (1982) Proc Natl Acad Sci USA, 79:7147), CMV, polyoma virus (Deans et al. (1984) Proc Natl Acad Sci USA 81:1292), or SV40 virus (Lusky and Botchan (1981) Nature 293:79).

[0276] The expression vectors can be introduced into cells in a manner suitable for subsequent expression of the nucleic acid. The method of introduction is largely dictated by the targeted cell type, discussed below. Exemplary methods include CaPO4precipitation, liposome fusion, cationic liposomes, electroporation, nucleoporation, viral infection, dextran-mediated transfection, polybrene-mediated transfection, protoplast fusion, and direct microinjection.

[0277] Appropriate host cells for the expression of antibodies or antigen binding fragments thereof include yeast, bacteria, insect, plant, and mammalian cells. Of particular interest are bacteria such as E. coli, fungi such as Saccharomyces cerevisiae and Pichia pastoris, insect cells such as SF9, mammalian cell lines (e.g., human cell lines, CHO cell), as well as primary cell lines.

[0278] In some embodiments, an antibody or fragment thereof can be expressed in, and purified from, transgenic animals (e.g., transgenic mammals). For example, an antibody can be produced in transgenic non-human mammals (e.g., rodents) and isolated from milk as described in, e.g., Houdebine (2002) Curr Opin Biotechnol 13(6):625-629; van Kuik-Romeijn et al. (2000) Transgenic Res 9(2): 155-159; and Pollock et al. (1999) J Immunol Methods 231(1 -2): 147-157.

[0279] The antibodies and fragments thereof can be produced from the cells by culturing a host cell transformed with the expression vector containing nucleic acid encoding the antibodies orPatent Application fragments, under conditions, and for an amount of time, sufficient to allow expression of the proteins. Such conditions for protein expression vary with the choice of the expression vector and the host cell and are easily ascertained by one skilled in the art through routine experimentation. For example, antibodies expressed in E. coli can be refolded from inclusion bodies (see, e.g., Hou et al. (1998) Cytokine 10:319-30). Bacterial expression systems and methods for their use are known in the art (see Ausubel et al. (1988) Current Protocols in Molecular Biology, Wiley & Sons; and Green and Sambrook (2012) Molecular Cloning— A Laboratory Manual, 4th Ed., Cold Spring Harbor Laboratory Press, New York (2001)). The choice of codons, suitable expression vectors and suitable host cells vary depending on a number of factors and may be easily optimized as needed. An antibody (or fragment thereof) described herein can be expressed in mammalian cells or in other expression systems including but not limited to yeast, baculovirus, and in vitro expression systems (see, e.g., Kaszubska et al. (2000) Protein Expression and Purification 18:213-220).

[0280] In vitro methods are also suitable for preparing monovalent antibodies, or fragments thereof. Digestion of antibodies to produce fragments thereof, particularly, Fab fragments, can be accomplished using routine techniques known in the art. For instance, digestion can be performed using papain. Examples of papain digestion are described in International Application Publication No. WO 94 / 29348, U. S. Patent No. 4,342,566, and Harlow and Lane, Antibodies, A Laboratory Manual, Cold Spring Harbor Publications, New York, (1988). Papain digestion of antibodies typically produces two identical antigen binding fragments, called Fab fragments, each with a single antigen binding site, and a residual Fc fragment. Pepsin treatment yields a fragment, called the F(ab’)2 fragment that has two antigen combining sites and is still capable of cross-linking antigen.

[0281] The Fab fragments produced in antibody digestion can also contain the constant domains of the light chain and the first constant domain of the heavy chain. Fab’ fragments differ from Fab fragments by the addition of a few residues at the carboxy terminus of the heavy chain domain including one or more cysteines from the antibody hinge region. The F(ab’)2 fragment is a bivalent fragment comprising two Fab’ fragments linked by a disulfide bridge at the hinge region. Fab’-SH is the designation herein for Fab’ in which the cysteine residue(s) of the constant domains bear a free thiol group.Patent Application

[0282] One method of producing proteins comprising the provided antibodies or fragments is to link two or more peptides or polypeptides together by protein chemistry techniques (or recombinant DNA techniques). For example, peptides or polypeptides can be chemically synthesized using currently available laboratory equipment using either Fmoc (9-fluorenylmethyl-oxycarbonyl) or Boc (tert-butyloxycarbonoyl) chemistry (Applied Biosystems, Inc.; Foster City, CA). Those of skill in the art readily appreciate that a peptide or polypeptide corresponding to the antibody provided herein, for example, can be synthesized by standard chemical reactions. For example, a peptide or polypeptide can be synthesized and not cleaved from its synthesis resin whereas the other fragment of an antibody can be synthesized and subsequently cleaved from the resin, thereby exposing a terminal group that is functionally blocked on the other fragment. By peptide condensation reactions, these two fragments can be covalently joined via a peptide bond at their carboxyl and amino termini, respectively, to form an antibody, or fragment thereof. (Grant GA (1992) Synthetic Peptides: A User Guide. W. H. Freeman and Co., N. Y. (1992); Bodansky M and Trost B., Ed. (1993) Principles of Peptide Synthesis. Springer Verlag Inc., NY). Alternatively, the peptide or polypeptide can by independently synthesized ex vivo. Once isolated, these independent peptides or polypeptides may be linked to form an antibody or fragment thereof via similar peptide condensation reactions.

[0283] For example, enzymatic ligation of cloned or synthetic peptide segments can allow relatively short peptide fragments to be joined to produce larger peptide fragments, polypeptides, or whole protein domains (Abrahmsen et al., Biochemistry, 30:4151 (1991)). Alternatively, native chemical ligation of synthetic peptides can be utilized to synthetically construct large peptides or polypeptides from shorter peptide fragments. This method consists of a two-step chemical reaction (Dawson et al., Science, 266:776779 (1994)). The first step is the chemoselective reaction of an unprotected synthetic peptide a thioester with another unprotected peptide segment containing an amino terminal Cys residue to give a thioester linked intermediate as the initial covalent product. Without a change in the reaction conditions, this intermediate undergoes spontaneous, rapid intramolecular reaction to form a native peptide bond at the ligation site. Application of this native chemical ligation method to the total synthesis of a protein molecule is illustrated by the preparation of human interleukin 8 (IL-8) (Baggiolini et al., FEBS Lett. 307:97-101 (1992); Clark et al., J. Biol. Chem. 269:16075 (1994); Clark et al., Biochemistry 30:3128 (1991); Rajarathnam et al., Biochemistry 33:6623-30 (1994)).Patent Application

[0284] Alternatively, unprotected peptide segments can be chemically linked where the bond formed between the peptide segments as a result of the chemical ligation is an unnatural (nonpeptide) bond (Schnolzer et al., Science 256:221 (1992)). This technique has been used to synthesize analogs of protein domains as well as large amounts of relatively pure proteins with full biological activity (deLisle et al., Techniques in Protein Chemistry IV. Academic Press, New York, pp. 257-267 (1992)).

[0285] Recombinant techniques can also be used to modify antibodies or antigen binding fragments thereof. For example, amino acids found to not contribute to either the activity or the binding specificity or affinity of the antibody can be deleted without a loss in the respective activity. Insertions, deletions, substitutions, or other selected modifications of particular regions or specific amino acids residues can also be made (and are contemplated by the present disclosure), provided the activity of the fragment is not significantly altered or impaired compared to the nonmodified antibody, or antigen binding fragment thereof can be made. Such methods are readily apparent to a skilled practitioner in the art and can include site specific mutagenesis of the nucleic acid encoding the antibody or fragment thereof. (Zoller et al., Nucl. Acids Res. 10:6487-500 (1982)).

[0286] Following expression, the antibodies and fragments thereof can be isolated. An antibody or fragment thereof can be isolated or purified in a variety of ways known in the art depending on what other components are present in the sample. Standard purification methods include electrophoretic, molecular, immunological, and chromatographic techniques, including ion exchange, hydrophobic, affinity, and reverse-phase HPLC chromatography. For example, an antibody can be purified using a standard anti-antibody column (e.g., a protein-A or protein-G column). Ultrafiltration and diafiltration techniques, in conjunction with protein concentration, are also useful. See, e.g., Scopes (1994) Protein Purification, 3rdedition, Springer-Verlag, New York City, New York. The degree of purification necessary varies depending on the desired use. In some instances, no purification of the expressed antibody or fragments thereof is necessary.

[0287] Methods for determining the yield or purity of a purified antibody or fragment thereof are known in the art and include, e.g., Bradford assay, UV spectroscopy, Biuret protein assay, Lowry protein assay, amido black protein assay, high pressure liquid chromatography (HPLC), mass spectrometry (MS), and gel electrophoretic methods (e.g., using a protein stain such as Coomassie Blue or colloidal silver stain).Patent Application

[0288] Any of the TCRm antibodies or antigen binding fragments thereof described herein can be modified. The modifications can be covalent or non-covalent modifications, and can include one or more amino acid substitutions that change the properties of the antigen-specific antibodies or antigen binding fragments thereof. Such modifications can be introduced into the TCRm antibodies or antigen binding fragments by, e.g., reacting targeted amino acid residues of the polypeptide with an organic derivatizing agent that is capable of reacting with selected side chains or terminal residues, or base-pair mutations in a nucleotide sequence encoding the antigen-specific antibodies or antigen binding fragments thereof. Suitable sites for modification can be chosen using any of a variety of criteria including, e.g., structural analysis or amino acid sequence analysis of the TCRm antibodies or fragments. In some instances, the TCRm or the antigen binding fragments may be labeled by a variety of means for use in pharmaceutical applications.

[0289] In some embodiments, the TCRm antibodies or antigen binding fragments thereof described herein may have a modification comprising one or more amino acid substitutions that provide reduced hydrophobicity and reduce the potential for aggregation, thereby improving the binding or therapeutic capacity of a TCRm antibody or an antigen-binding fragment thereof. Such amino acid substitutions can be introduced by changing one or more nucleotides in the polynucleotide encoding the TCRm antibody or antigen-binding fragment such that the triplet codon for the amino acid residue position where the modification is to be introduced is replaced with the triplet codon encoding the amino acid substitution. In some embodiments, the modification may comprise a single amino acid substitution. In some embodiments, the modification may comprise multiple amino acid substitutions. In some embodiments, the modification may be a substitution of 1 amino acid, 2 amino acids, 3 amino acids, 4 amino acids, or 5 amino acids. In some embodiments, the amino acid substitution may be located in a CDR.

[0290] In some embodiments, the amino acid mutations may include aspartic acid to serine modification. In some embodiments, the modification may include an alanine to glycine. One skilled in the art may understand that any modification to the antibody sequence that reduced the hydrophobicity of an amino acid sequence may fall under the scope of such modifications. Described above modifications are provided herein not to limit the scope of the invention but merely provided as examples of modifying an amino acid sequence for altering the hydrophobicity.

[0291] In some embodiments, TERT540 pHLA antibodies of the disclosure may comprise two peptide chains, the sequences of which are provided in Table le. Exemplary CDR sequences ofPatent Application TERT540 pHLA binders antibodies of the disclosure are provided in Tables la and lb. The variable heavy chain (VH) amino acid sequences of certain exemplary TERT540 pHLA antibodies of the disclosure are provided in Table Id, while exemplary variable light chain (LC) amino acid sequences are provided in Table Id. The sequences of certain TERT peptides are provided in Table 1c.

[0292] As provided in Table le, exemplary TERT540 pHLA binders and TERT TCRm antibodies of the disclosure may include one or more linker sequences. As provided in Tables le, such linker sequences may be incorporated within the polypeptide sequence of an antigen binder (e.g., a TERT540 pHLA binder), for example linking the VL and VH (VL-Linker-VH or VH-Linker-VL) of an antigen binder, such as a single chain variable fragment antigen binder. Alternatively or additionally, one or more linker sequences may be used to link different antigen binding domains of a TERT540 pHLA antibody of the disclosure. For example, a linker sequence may be used to link a TERT540 pHLA antigen binding domain to an immune cell binding domain of a TERT TCRm antibody. The linker sequences provided in Table le are exemplary, and other linkers or the use of no linkers is contemplated by the present disclosure.

[0293] In certain aspects, antigen binding domains described herein may be linked to each other in any orientation of N- to C-terminus.

[0294] In certain aspects, a linker comprises a short oligo- or polypeptide linker, for example, between 2 and 40 amino acids (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids) in length may form the linkage between the domains. The linker is a peptide of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more than 30 amino acid residues. Non-limiting examples of amino acids found in linkers include Gly, Ser, Glu, Gin, Ala, Leu, Iso, Lys, Arg, Pro, and the like.

[0295] The linker may be [(Gly)nlSer]n2, where nl and n2 may be any number (e.g. nl and n2 may independently be 1, 2, 4, 5, 6, 7, 8, 9, 10 or more than 10). The linker may be flexible polypeptide linker that is a Gly / Ser linker and comprises the amino acid sequence (Gly-Gly-Ser) n, (Gly-Gly-Gly-Ser) n, or (Gly4-Ser)n which can be repeated n times, where n is a positive integer equal to or greater than 1. For example, n=l, n=2, n=3, n=4, n=5, n=6, n=7, n=8, n=9 and n=10. The linker may include multiple repeats of (Gly Gly Ser), (Gly Ser) or (Gly Gly Gly Ser). Also included within the scope of the invention are linkers described in WO2012 / 138475 (incorporated herein by reference). In some embodiments, the flexible polypeptide linkers include, but are notPatent Application limited to, GGS, GGGGS, (GGGGS)2, (GGGGS)3, (GGGGS)4, (GGGGS)3GG. Tn some embodiments, the linkers include multiple repeats of (GGS), (GS) or (GGGS).

[0296] The linker sequence may comprise a long linker (LL) sequence. The long linker sequence may comprise GGGGS, repeated four times. The long linker sequence may comprise GGGGS, repeated three times. The linker sequence may comprise a short linker (SL) sequence. The short linker sequence may comprise GGGGS. A glycine-serine doublet can be used as a suitable linker. Alternatively, antigen binding domains / peptides are fused directly to each other via peptide bonds without use of a linker. In some embodiments, the TCRm antibodies or antigen binding fragments thereof can be conjugated to a heterologous moiety. The heterologous moiety can be, e.g., a heterologous polypeptide, a therapeutic agent (e.g., a toxin or a drug), or a detectable label such as, but not limited to, a radioactive label, an enzymatic label, a fluorescent label, a heavy metal label, a luminescent label, or an affinity tag such as biotin or streptavidin. Suitable heterologous polypeptides include, e.g., an antigenic tag (e.g., FLAG, polyhistidine, hemagglutinin, glutathione-S-transferase (GST), or maltose-binding protein (MBP)) for use in purifying the antibodies or fragments. Suitable radioactive labels include, e.g.,32P,33P,14C,125I,131I,35S, and3H. Suitable fluorescent labels include, without limitation, fluorescein, fluorescein isothiocyanate (FITC), green fluorescent protein (GFP), DyLight™ 488, phycoerythrin (PE), propidium iodide (PI), PerCP, PE-Alexa Fluor® 700, Cy5, allophycocyanin, and Cy7. Luminescent labels include, e.g., any of a variety of luminescent lanthanide (e.g., europium or terbium) chelates. For example, suitable europium chelates include the europium chelate of diethylene triamine pentaacetic acid (DTP A) or tetraazacyclododecane- 1,4, 7, 10-tetraacetic acid (DOTA). Enzymatic labels include, e.g., alkaline phosphatase, chloramphenicol acetyl transferase (CAT), luciferase, and horseradish peroxidase. Another labeling technique which may result in greater sensitivity consists of coupling the antibodies to low molecular weight haptens. These haptens can then be specifically altered by means of a second reaction. For example, it is common to use haptens such as biotin, which reacts with avidin, or di nitrophenol, pyridoxal, or fluorescein, which can react with specific anti-hapten antibodies.

[0297] Two proteins (e.g., an antibody and a heterologous moiety) can be cross-linked using any of a number of known chemical cross linkers. Examples of such cross linkers are those that link two amino acid residues via a linkage that includes a “hindered” disulfide bond. In these linkages, a disulfide bond within the cross-linking unit is protected (by hindering groups on either side ofPatent Application the disulfide bond) from reduction by the action, for example, of reduced glutathione or the enzyme disulfide reductase. One suitable reagent, 4-succinimidyloxycarbonyl-a-methyl-a(2-pyridyldithio) toluene (SMPT), forms such a linkage between two proteins utilizing a terminal lysine on one of the proteins and a terminal cysteine on the other. Heterobifunctional reagents that cross-link by a different coupling moiety on each protein can also be used. Other useful cross-linkers include, without limitation, reagents which link two amino groups (e.g., N-5-azido-2-nitrobenzoyloxysuccinimide), two sulfhydryl groups (e.g., 1,4-bis-maleimidobutane), an amino group and a sulfhydryl group (e.g., m-maleimidobenzoyl-N-hydroxy succinimide ester), an amino group and a carboxyl group (e.g., 4-[p-azidosalicylamido]butylamine), and an amino group and a guanidinium group that is present in the side chain of arginine (e.g., p-azidophenyl glyoxal monohydrate).

[0298] In some embodiments, a radioactive label can be directly conjugated to the amino acid backbone of the TCRm antibody. Alternatively, the radioactive label can be included as part of a larger molecule (e.g.,125I in meta-[125I]iodophenyl-N-hydroxysuccinimide ([125I]mIPNHS), which binds to free amino groups to form meta-iodophenyl (mIP) derivatives of relevant proteins (see, e.g., Rogers et al. (1997) J Nucl Med 38:1221-1229) or chelate (e.g., to DOTA or DTP A), which is in turn bound to the protein backbone. Methods of conjugating the radioactive labels or larger molecules / chelates containing them to the antibodies or antigen binding fragments described herein are known in the art. Such methods involve incubating the proteins with the radioactive label under conditions (e.g., pH, salt concentration, and / or temperature) that facilitate binding of the radioactive label or chelate to the protein (see, e.g., U. S. Patent No. 6,001,329).

[0299] Methods for conjugating a fluorescent label (sometimes referred to as a fluorophore) to a protein (e.g., an antibody) are known in the art of protein chemistry. For example, fluorophores can be conjugated to free amino groups (e.g., of lysines) or sulfhydryl groups (e.g., cysteines) of proteins using succinimidyl (NHS) ester or tetrafluorophenyl (TFP) ester moieties attached to the fluorophores. In some embodiments, the fluorophores can be conjugated to a heterobifunctional cross-linker moiety such as sulfo-SMCC. Suitable conjugation methods involve incubating an antibody protein or fragment thereof with the fluorophore under conditions that facilitate binding of the fluorophore to the protein. See, e.g., Welch and Redvanly (2003) Handbook of Radiopharmaceuticals: Radiochemistry and Applications, John Wiley and Sons.Patent Application

[0300] In some embodiments, the TCRm antibodies or fragments can be modified, e.g., with a moiety that improves the stabilization and / or retention of the antibodies in circulation, e.g., in blood, serum, or other tissues. For example, the antibody or fragment can be PEGylated as described in, e.g., Lee et al. (1999) Bioconjug Chem 10(6): 973-8; Kinstler et al. (2002) Advanced Drug Deliveries Reviews 54:477-485; and Roberts et al. (2002) Advanced Drug Delivery Review s 54:459-476, or HESylated (Fresenius Kabi, Germany) (see, e.g., Pavisic et al. (2010) Int J Pharm 387(1-2):110-119). The stabilization moiety can improve the stability, or retention of, the antibody (or fragment) by at least 1.5 (e.g., at least 2, 5, 10, 15, 20, 25, 30, 40, or 50 or more) fold.

[0301] In some embodiments, the TCRm antibodies or antigen-binding fragments thereof described herein can be glycosylated. In some embodiments, an antibody or antigen-binding fragment thereof described herein can be subjected to enzymatic or chemical treatment, or produced from a cell, such that the antibody or fragment has reduced or absent glycosylation. Methods for producing antibodies with reduced glycosylation are known in the art and described in, e.g., U. S. Patent No. 6,933,368; Wright et al. (1991) EMBO J 10(10):2717-2723; and Co et al. (1993) Mol Immunol 30:1361.

[0302] The antibodies and antigen binding fragments thereof and molecules comprising such antibodies and antigen binding fragments thereof discussed above (e.g., Abs, bi-specific Abs,) may be produced by recombinant expression in a human or non-human cell. Synthetic antibodyproducing cells include non-human cells expressing heavy chains, light chains, or both heavy and light chains; human cells that are not immune cells expressing heavy chains, light chains, or both heavy and light chains; and human B cells that produce heavy chains or light chains, but not both heavy and light chains. Synthetic antibodies of this disclosure may be heterologously expressed, in vitro or ex vivo, in cells other than human B cells, such as non-human cells and human cells other than B cells, optionally other than immune cells, and optionally in cells other than cells in a B cell lineage.

[0303] The antibodies and antigen binding fragments thereof and molecules comprising them described herein can be produced using a variety of techniques known in the art of molecular biology and protein chemistry. For example, a nucleic acid encoding the antibody or antigen binding fragment thereof can be inserted into an expression vector that contains transcriptional and translational regulatory sequences, which include, e.g., promoter sequences, ribosomal binding sites, transcriptional start and stop sequences, translational start and stop sequences, transcriptionPatent Application terminator signals, polyadenylation signals, and enhancer or activator sequences. The regulatory sequences include a promoter and transcriptional start and stop sequences. In addition, the expression vector can include more than one replication system, such that it can be maintained in two different organisms, for example, in mammalian or insect cells for expression and in a prokaryotic host for cloning and amplification.

[0304] Compositions comprising a TCRm antibody or antigen binding fragment thereof of the present disclosure and a pharmaceutically acceptable carrier are also provided. The compositions may further comprise a diluent, solubilizer, emulsifier, preservative, and / or adjuvant to be used with the methods disclosed herein. Such compositions can be used in a subject with a malignancy, cancer, autoimmune, or other TERT expression-associated disorder that would benefit from any of the antigen-specific antibodies or antigen binding fragments thereof described herein.

[0305] In certain embodiments, acceptable formulation materials preferably are nontoxic to recipients at the dosages and concentrations employed. In certain embodiments, the formulation material(s) are for s.c. and / or I. V. administration. In certain embodiments, the pharmaceutical composition can contain formulation materials for modifying, maintaining or preserving, for example, the pH, osmolality, viscosity, clarity, color, isotonicity, odor, sterility, stability, rate of dissolution or release, adsorption or penetration of the composition. In certain embodiments, suitable formulation materials include, but are not limited to, amino acids (such as glycine, glutamine, asparagine, arginine or lysine); antimicrobials; antioxidants (such as ascorbic acid, sodium sulfite or sodium hydrogen- sulfite); buffers (such as borate, bicarbonate, Tris-HCl, citrates, phosphates or other organic acids); bulking agents (such as mannitol or glycine); chelating agents (such as ethylenediamine tetraacetic acid (EDTA)); complexing agents (such as caffeine, polyvinylpyrrolidone, beta-cyclodextrin or hydroxypropyl-beta- cyclodextrin); fillers; monosaccharides, disaccharides, and other carbohydrates (such as glucose, mannose or dextrins); proteins (such as serum albumin, gelatin or immunoglobulins); coloring, flavoring and diluting agents; emulsifying agents; hydrophilic polymers (such as polyvinylpyrrolidone); low molecular weight polypeptides; salt-forming counterions (such as sodium); preservatives (such as benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid or hydrogen peroxide); solvents (such as glycerin, propylene glycol or polyethylene glycol); sugar alcohols (such as mannitol or sorbitol); suspending agents; surfactants or wetting agents (such as pluronics, PEG, sorbitan esters, polysorbates such asPatent Application polysorbate 20, polysorbate 80, triton, tromethamine, lecithin, cholesterol, tyloxapal); stability enhancing agents (such as sucrose or sorbitol); tonicity enhancing agents (such as alkali metal halides, preferably sodium or potassium chloride, mannitol sorbitol); delivery vehicles; diluents; excipients and / or pharmaceutical adjuvants. (Allen (2012) Remington - The Science and Practice of Pharmacy, 22d Edition, Lloyd V, Allen, ed., The Pharmaceutical Press). In certain embodiments, the optimal pharmaceutical composition is determined by one skilled in the art depending upon, for example, the intended route of administration, delivery format and desired dosage. See, for example, Allen (2012) Remington - The Science and Practice of Pharmacy, 22d Edition, Lloyd V, Allen, ed., The Pharmaceutical Press. In certain embodiments, such compositions may influence the physical state, stability, rate of ex vivo release and / or rate of ex vivo clearance of the tumor / autoimmune associated antigen-specific antibody or antigen binding fragment thereof.

[0306] In certain embodiments, the primary vehicle or carrier in a pharmaceutical composition can be either aqueous or non-aqueous in nature. For example, in certain embodiments, a suitable vehicle or carrier can be water for injection, physiological saline solution or artificial cerebrospinal fluid, possibly supplemented with other materials common in compositions for parenteral administration. In certain embodiments, the saline comprises isotonic phosphate-buffered saline. In certain embodiments, neutral buffered saline or saline mixed with serum albumin are further exemplary vehicles. In certain embodiments, pharmaceutical compositions comprise a pH controlling buffer such phosphate-buffered saline or acetate-buffered saline. In certain embodiments, a composition comprising a tumor / autoimmune associated antigen-specific antibody or antigen binding fragment thereof disclosed herein can be prepared for storage by mixing the selected composition having the desired degree of purity with optional formulation agents (see Allen (2012) Remington - The Science and Practice of Pharmacy, 22d Edition, Lloyd V, Allen, ed., The Pharmaceutical Press) in the form of a lyophilized cake or an aqueous solution. Further, in certain embodiments, a composition comprising a tumor / autoimmune associated antigen-specific antibody or antigen binding fragment thereof disclosed herein can be formulated as a lyophilizate using appropriate excipients. In some instances, appropriate excipients may include a cryo-preservative, a bulking agent, a surfactant, or a combination of any thereof. Exemplary excipients include one or more of a polyol, a disaccharide, or a polysaccharide, such as, for example, mannitol, sorbitol, sucrose, trehalose, and dextran 40. In some instances, the cryo-Patent Application preservative may be sucrose or trehalose. In some instances, the bulking agent may be glycine or mannitol. In one example, the surfactant may be a polysorbate such as, for example, polysorbate-20 or polysorbate-80.

[0307] In certain embodiments, the pharmaceutical composition can be selected for parenteral delivery. In certain embodiments, the compositions can be selected for inhalation or for delivery through the digestive tract, such as orally. The preparation of such pharmaceutically acceptable compositions is within the ability of one skilled in the art.

[0308] In certain embodiments, the formulation components are present in concentrations that are acceptable to the site of administration. In certain embodiments, buffers are used to maintain the composition at physiological pH or at a slightly lower pH, typically within a pH range of from about 5 to about 8. For example, the pH may be 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8. 6.9, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, or 8.5. In some instances, the pH of the pharmaceutical composition may be in the range of 6.6-8.5 such as, for example, 7.0-8.5, 6.6-7.2, 6.8-7.2, 6.8-7.4, 7.2-7.8, 7.0-7.5, 7.5-8.0, 7.2-8.2, 7.6-8.5, or 7.8-8.3. In some instances, the pH of the pharmaceutical composition may be in the range of 5.5-7.5 such as, for example, 5.5-5.8, 5.5-6.0, 5.7-6.2, 5.8-6.5, 6.0-6.5, 6.2-6.8, 6.5-7.0, 6.8-7.2, or 6.8-7.5. In some instances, the pH of the pharmaceutical composition may be in the range of 4.0-5.5 such as, for example, 4.0-4.3, 4.0-4.5, 4.2-4.8, 4.5-4.8, 4.5-5.0, 4.8-5.2, or 5.0-5.5. In an embodiment, the pH is 7.2.

[0309] In certain embodiments when parenteral administration is contemplated, a therapeutic composition can be in the form of a pyrogen-free, parenterally acceptable aqueous solution comprising a tumor / autoimmune associated antigen-specific antibody or antigen binding fragment thereof in a pharmaceutically acceptable vehicle. In certain embodiments, a vehicle for parenteral injection is sterile distilled water in which a tumor / autoimmune associated antigen-specific antibody or antigen binding fragment thereof is formulated as a sterile, isotonic solution and properly preserved. In certain embodiments, the preparation can involve the formulation of the desired molecule with an agent, such as injectable microspheres, bio-erodible particles, polymeric compounds (such as polylactic acid or polyglycolic acid), beads or liposomes, that can provide for the controlled or sustained release of the product which can then be delivered via a depot injection. In certain embodiments, hyaluronic acid can also be used, and can have the effect of promotingPatent Application sustained duration in the circulation. In certain embodiments, implantable drug delivery devices can be used to introduce the desired molecule.

[0310] In certain embodiments, a pharmaceutical composition can be formulated for inhalation. In certain embodiments, an antigen-specific antibody or antigen binding fragment thereof can be formulated as a dry powder for inhalation. In certain embodiments, an inhalation solution comprising a tumor / autoimmune associated antigen-specific antibody or antigen binding fragment thereof can be formulated with a propellant for aerosol delivery. In certain embodiments, solutions can be nebulized. Pulmonary administration is further described in International Application Publication No. WO / 1994 / 020069, which describes pulmonary delivery of chemically modified proteins.

[0311] In certain embodiments, it is contemplated that formulations can be administered orally. In certain embodiments, an antigen-specific antibody or antigen binding fragment thereof that is administered in this fashion can be formulated with or without carriers customarily used in compounding solid dosage forms, such as tablets and capsules. In certain embodiments, a capsule can be designed to release the active portion of the formulation at the point in the gastrointestinal tract when bioavailability is maximized, and pre-systemic degradation is minimized. In certain embodiments, at least one additional agent can be included to facilitate absorption of a tumor / autoimmune associated antigen-specific antibody or antigen binding fragment thereof. In certain embodiments, diluents, flavorings, low melting point waxes, vegetable oils, lubricants, suspending agents, tablet disintegrating agents, and binders can also be employed.

[0312] In certain embodiments, a pharmaceutical composition can involve an effective quantity of an antigen-specific antibody or antigen binding fragment thereof in a mixture with non-toxic excipients suitable for the manufacture of tablets. In certain embodiments, by dissolving the tablets in sterile water or other appropriate vehicle, solutions can be prepared in unit-dose form. In certain embodiments, suitable excipients include, but are not limited to, inert diluents, such as calcium carbonate, sodium carbonate or bicarbonate, lactose, or calcium phosphate; or binding agents, such as starch, gelatin, or acacia; or lubricating agents such as magnesium stearate, stearic acid, or talc.

[0313] Additional pharmaceutical compositions can be selected by one skilled in the art, including formulations involving an antibody or antigen binding fragment thereof in sustained- or controlled-delivery formulations. In certain embodiments, techniques for formulating a variety of other sustained- or controlled-delivery means, such as liposome carriers, bio-erodible microparticles orPatent Application porous beads and depot injections, are also known to those skilled in the art. See for example, International Application Publication No. WO / 1993 / 015722, which describes the controlled release of porous polymeric microparticles for the delivery of pharmaceutical compositions. In certain embodiments, sustained-release preparations can include semipermeable polymer matrices in the form of shaped articles, e.g., films, or microcapsules. Sustained release matrices can include polyesters, hydrogels, polylactides (see, e.g., U. S. Patent No. 3,773,919; U. S. Patent No. 5, 594,091; U. S. Patent No. 8,383,153; U. S. Patent No. 4,767,628; International Application Publication No. WO1998043615, Calo, E. et al. (2015) Eur. Polymer 765:252-267 and European Patent No. EP 058,481), including, for example, chemically synthesized polymers, starch based polymers, and polyhydroxyalkanoates (PHAs), copolymers of L-glutamic acid and gamma ethyl-L-glutamate (Sidman et al. (1993) Biopolymers 22:547-556), poly (2-hydroxyethyl-methacrylate) (Langer et al. (1981) J Biomed Mater Res. 15: 167-277; and Langer (1982) Chem Tech 12:98-105), ethylene vinyl acetate (Hsu and Langer (1985) J Biomed Materials Res 19(4):445-460) or poly-D(-)-3-hydroxybutyric acid (European Patent No. EP0133988). In certain embodiments, sustained release compositions can also include liposomes, which can be prepared by any of several methods known in the art. (See, e.g., Eppstein et al. (1985) Proc. Natl. Acad. Sci. USA 82:3688-3692; European Patent No. EP 036,676; and U. S. Patent Nos. 4,619,794 and 4,615,885).

[0314] The pharmaceutical composition to be used for ex vivo administration typically is sterile. In certain embodiments, sterilization is accomplished by filtration through sterile filtration membranes. In certain embodiments, where the composition is lyophilized, sterilization using this method can be conducted either prior to or following lyophilization and reconstitution. In certain embodiments, the composition for parenteral administration can be stored in lyophilized form or in a solution. In certain embodiments, parenteral compositions generally are placed into a container having a sterile access port, for example, an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle.

[0315] In certain embodiments, once the pharmaceutical composition has been formulated, it can be stored in sterile vials as a solution, suspension, gel, emulsion, solid, or as a dehydrated or lyophilized powder. In certain embodiments, such formulations can be stored either in a ready-to-use form or in a form (e.g., lyophilized) that is reconstituted prior to administration.

[0316] In certain embodiments, kits are provided for producing a single-dose administration unit. In certain embodiments, the kit can contain both a first container having a dried protein and aPatent Application second container having an aqueous formulation. In certain embodiments, kits containing single and multi-chambered pre-filled syringes are included.

[0317] In certain embodiments, the effective amount of a pharmaceutical composition comprising a antibody or antigen binding fragment thereof to be employed therapeutically depends, for example, upon the therapeutic context and objectives. One skilled in the art will appreciate that the appropriate dosage levels for treatment, according to certain embodiments, vary depending, in part, upon the molecule delivered, the indication for which a antibody or antigen binding fragment thereof is being used, the route of administration, and the size (body weight, body surface or organ size) and / or condition (the age and general health) of the patient. The clinician can titer the dosage and modify the route of administration to obtain the optimal therapeutic effect.

[0318] The clinician also selects the frequency of dosing, taking into account the pharmacokinetic parameters of the antibody or antigen binding fragment thereof in the formulation used. In certain embodiments, a clinician administers the composition until a dosage is reached that achieves the desired effect. In certain embodiments, the composition can therefore be administered as a single dose or as two or more doses (which may or may not contain the same amount of the desired molecule) over time, or as a continuous infusion via, for example, an implantation device or catheter. Further refinement of the appropriate dosage is routinely made by those of ordinary skill in the art and is within the ambit of tasks routinely performed by them. In certain embodiments, appropriate dosages can be ascertained through use of appropriate dose-response data.

[0319] In certain embodiments, the route of administration of the pharmaceutical composition is in accord with known methods, e.g., orally, through injection by intravenous, intraperitoneal, intracerebral (intra-parenchymal), intracerebral, intraventricular, intramuscular, subcutaneously, intra-ocular, intraarterial, intraportal, or intralesional routes; by sustained release systems or by implantation devices. In certain embodiments, the compositions can be administered by bolus injection or continuously by infusion, or by implantation device. In certain embodiments, individual elements of a combination therapy may be administered by different routes.

[0320] In certain embodiments, the composition can be administered locally, e.g., during surgery or topically. Optionally local administration is via implantation of a membrane, sponge, or another appropriate material onto which the desired molecule has been absorbed or encapsulated. In certain embodiments, where an implantation device is used, the device can be implanted into any suitablePatent Application tissue or organ, and delivery of the desired molecule can be via diffusion, timed-release bolus, or continuous administration.

[0321] In certain embodiments, it can be desirable to use a pharmaceutical composition comprising an antigen-specific antibody or antigen binding fragment thereof in an ex vivo manner. In such instances, cells that have been removed from a subject may be exposed to a pharmaceutical composition comprising a tumor / autoimmune associated antigen-specific antibody or antigen binding fragment thereof after which the cells are subsequently implanted back into the subject.

[0322] In certain embodiments, a tumor / autoimmune associated antigen-specific antibody or antigen binding fragment thereof can be delivered by implanting certain cells that have been genetically engineered, using methods such as those described herein, to express and secrete the polypeptides. In certain embodiments, such cells can be animal or human cells, and can be autologous, heterologous, or xenogeneic. In certain embodiments, the cells can be immortalized. In certain embodiments, in order to decrease the chance of an immunological response, the cells can be encapsulated to avoid infiltration of surrounding tissues. In certain embodiments, the encapsulation materials are typically biocompatible, semi-permeable polymeric enclosures or membranes that allow the release of the protein product(s) but prevent the destruction of the cells by a subject’s immune system or by other detrimental factors from the surrounding tissues.

[0323] As described herein, the present disclosure provides a method of treating a subject with a cancer or an autoimmune disorder, allergy, or other disorder related to chronic inflammation, comprising administering to the subject a therapeutically effective amount of a TERT540TCRm antibody or antigen binding fragment thereof of the present disclosure.

[0324] The compositions described herein are useful in, inter alia, methods for treating disorders correlated with TERT expression inasubject. As used herein, theterm subject means a mammalian subject. Exemplary subjects include, but are not limited to humans, monkeys, dogs, cats, mice, rats, cows, horses, camels, goats and sheep. In some embodiments, the subject is a human. In some embodiments, the subject has or is suspected to have cancer. In some embodiments, the subject is diagnosed with a cancer or an autoimmune disorder, allergy, and / or condition associated with chronic inflammation. In some embodiments, the subject is a human that is suspected of having a cancer or an autoimmune disorder, allergy, and / or condition associated with chronic inflammation correlated with elevated TERT expression.Patent Application

[0325] As used herein, administer or administration refers to the act of injecting or otherwise physically delivering a substance as it exists outside the body (e.g., a TERT pHLA-specific antibody or antigen binding fragment provided herein or a construct encoding same) into a patient, such as by mucosal, intradermal, intravenous, intramuscular, subcutaneous delivery and / or any other method of physical delivery described herein or known in the art. When a disease, or a symptom thereof, is being treated, administration of the substance typically occurs after the onset of the disease or symptoms thereof. When a disease, or symptoms thereof, are being prevented, administration of the substance typically occurs before the onset of the disease or symptoms thereof.

[0326] The compositions can be administered to a subject, e.g., a human subject, using a variety of methods that depend, in part, on the route of administration. The route can be, e.g., intravenous injection or infusion (IV), subcutaneous injection (SC), intraperitoneal (IP) injection, intramuscular injection (IM), intradermal injection (ID), subcutaneous, transdermal, intracavity, oral, intracranial injection, or intrathecal injection (IT). The injection can be in a bolus or a continuous infusion. Techniques for preparing injectate or infusate delivery systems containing antibodies are well known to those of skill in the art. Generally, such systems should utilize components which will not significantly impair the biological properties of the antibodies, such as the paratope binding capacity (see, for example, Remington's Pharmaceutical Sciences, 8th edition, 1990, Mack Publishing). Those of skill in the art can readily determine the various parameters and conditions for producing antibody injectates or infusates without resort to undue experimentation.

[0327] Administration can be achieved by, e.g., topical administration, local infusion, injection, or by means of an implant. The implant can be of a porous, non-porous, or gelatinous material, including membranes, such as sialastic membranes, or fibers. The implant can be configured for sustained or periodic release of the composition to the subject. See, e.g., U. S. Patent Application Publication No. 20080241223; U. S. Patent Nos. 5,501,856; 5,164,188; 4,863,457; and 3,710,795. The composition can be delivered to the subject by way of an implantable device based on, e.g., diffusive, erodible, or convective systems, e.g., osmotic pumps, biodegradable implants, electrodiffusion systems, electroosmosis systems, vapor pressure pumps, electrolytic pumps, effervescent pumps, piezoelectric pumps, erosion-based systems, or electromechanical systems.Patent Application In some embodiments, an antibody or antigen binding fragment of the present disclosure is therapeutically delivered to a subject by way of local administration.

[0328] Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic / aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer’s dextrose, dextrose and sodium chloride, lactated Ringer’s or fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer’s dextrose), and the like. Preservatives and other additives may also be present such as, for example, antimicrobials, antioxidants, chelating agents, and the like.

[0329] Treating or treatment of any disease or disorder refers to ameliorating a disease or disorder that exists in a subject or a symptom thereof. The term ameliorating refers to any therapeutically beneficial result in the treatment of a disease state, e.g., a cancer or an autoimmune disorder, allergy, and / or condition associated with chronic inflammation, lessening in the severity or progression, promoting remission or durations of remission, or curing thereof. Thus, treating or treatment includes ameliorating at least one physical parameter or symptom. Treating or treatment includes modulating the disease or disorder, either physically (e.g., stabilization of a discernible symptom) or physiologically (e.g., stabilization of a physical parameter) or both. Treating or treatment includes delaying or preventing metastasis. Thus, in the disclosed methods, treatment can refer to a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% reduction in the severity of an established disease or condition or symptom of the disease or condition. For example, a method for treating a cancer or an autoimmune disorder, allergy, and / or condition associated with chronic inflammation in a subject by administering an antibody as described in this disclosure is considered to be a treatment if there is a 10% reduction in one or more symptoms of the cancer or autoimmune disorder, allergy, and / or condition associated with chronic inflammation in a subject as compared to a control. Thus, the reduction can be a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any percent reduction in between 10% and 100% as compared to native or control levels. It is understood that treatment does not necessarily refer to a cure or complete ablation of the disease, condition, or symptoms of the disease or condition.Patent Application

[0330] The principal symptoms can include (without intending to be limiting) bone pain, nausea, constipation, loss of appetite, mental fogginess or confusion, fatigue, frequent infections, weight loss, weakness or numbness in the legs, excessive thirst, easily fractured or broken bones, anemia, leukopenia, thrombocytopenia, excessive urination, hypercalcemia, spinal cord compression, kidney dysfunction, hyperviscosity, and the like.

[0331] As used herein, the term “therapeutically effective amount” or effective amount refers to an amount of an antigen-specific antibody or antigen binding fragment thereof that, when administered to a subject, is effective to treat a disease or disorder such that the symptoms of the disease are ameliorated, or the likelihood of the disease developing or progressing is decreased. A therapeutically effective amount is not, however, a dosage so large as to cause adverse side effects, such as hyperviscosity syndromes, pulmonary edema, congestive heart failure, and the like. A suitable dose of an antibody or fragment thereof described herein, which dose is capable of treating a cancer or an autoimmune disorder, allergy, and / or condition associated with chronic inflammation in a subject, can depend on a variety of factors including the particular construct used and whether it is used concomitantly with other therapeutic agents. For example, a different dose of a whole cancer- or autoimmune-disorder-, allergy-, and / or condition associated with chronic inflammation- antigen-specific antibody may be required to treat a subject with a cancer or an autoimmune disorder, allergy, and / or condition associated with chronic inflammation as compared to the dose of a fragment of the antigen-specific antibody (e.g., Fab’ antibody fragment) required to treat the same subject. Other factors affecting the dose administered to the subject include, e g., the type or extent of the cancer or an autoimmune disorder, allergy, and / or condition associated with chronic inflammation. For example, a subject that has had a previous cancer (or other TERT-associated disease) may require administration of a different dosage of cancer antigen-specific antibody or antigen binding fragment thereof than a subject who has not previously had the cancer. Generally, a therapeutically effective amount may vary with the subject’s age, condition, and sex, as well as the extent of the disease in the subject and can be determined by one of skill in the art. Other factors can include, e.g., other medical disorders concurrently or previously affecting the subject, the general health of the subject, the genetic disposition of the subject, diet, time of administration, rate of excretion, drug combination, and any other additional therapeutics that are administered to the subject. It should also be understood that a specific dosage and treatment regimen for any particular subject also depends upon thePatent Application judgment of the treating medical practitioner (e.g., doctor or nurse). A therapeutically effective amount is also one in which any toxic or detrimental effects of the composition are outweighed by the therapeutically beneficial effects. The dosage of the therapeutically effective amount may be adjusted by the individual physician or veterinarian in the event of any complication. In some instances, a therapeutically effective amount may vary from about 0.001 mg / kg to about 50 mg / kg, preferably from about 0.001 mg / kg to about 20 mg / kg, most preferably from about 0.002 mg / kg to about 2 mg / kg, in one or more dose administrations daily, at least once a day, once every two days, once every three days, once every four days, once every five days, once every six days, once a week, bi-weekly, or monthly. In some instances, the cancer antigen-specific antibody or antigen binding fragment thereof is administered for 2 to 5 or more consecutive days, weeks or months in order to avoid “rebound” of a disease or other pathology being treated from occurring.

[0332] A pharmaceutical composition can include a therapeutically effective amount of a cancer or autoimmune disorder, allergy, and / or condition associated with chronic inflammation antigenspecific antibody or antigen binding fragment thereof described herein. Such effective amounts can be readily determined by one of ordinary skill in the art as described above. Considerations include the effect of the administered cancer or autoimmune disorder, allergy, and / or condition associated with chronic inflammation antigen-specific antibody or antigen binding fragment thereof, or the combinatorial effect of the antigen-specific antibody or antigen binding fragment thereof with one or more additional active agents, if more than one agent is used in or with the pharmaceutical composition. In certain aspects, the doses can be about 1, about 0.5, about 0.1, about 0.05, or about 0.01, or about 0.005, or about 0.0001 mg / kg, or any intervening dose between about 0.001 mg / kg and 1 mg / kg.

[0333] Suitable human doses of any of the cancer or autoimmune disorder, allergy, and / or condition associated with chronic inflammation antigen-specific antibody or antigen binding fragment thereof described herein can further be evaluated in, e.g., Phase I dose escalation studies. See, e.g., van Gurp et al. (2008) Am J Transplantation 8(8): 1711-1718; Hanouska et al. (2007) Clin Cancer Res 13(2, part 1): 523 -531; and Hetherington et al. (2006) Antimicrobial Agents and Chemotherapy 50(10): 3499-3500.

[0334] Toxicity and therapeutic efficacy of such cancer or autoimmune disorder, allergy, and / or condition associated with chronic inflammation antigen-specific antibodies or antigen binding fragments thereof can be determined by known pharmaceutical procedures in cell cultures orPatent Application experimental animals (e.g., animal models of any of the cancers described herein). These procedures can be used, e.g., for determining the LD₅₀ (the dose lethal to 50% of the population) and the ED₅₀ (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index, and it can be expressed as the ratio LD₅₀ / ED₅₀. An antigen-specific antibody or antigen binding fragment thereof that exhibits a high therapeutic index is preferred. While constructs that exhibit toxic side effects may be used, care should be taken to design a delivery system that targets such constructs to the site of affected tissue and to minimize potential damage to normal cells and, thereby, reduce side effects.

[0335] The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage generally within a range of circulating concentrations of the antigen-specific antibody or antigen binding fragment that include the ED50 with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. The therapeutically effective dose can be estimated initially from cell culture assays. A dose can be formulated in animal models to achieve a circulating plasma concentration range that includes the EC50(i.e., the concentration of the construct - e.g., antibody - which achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by high performance liquid chromatography. In some embodiments, e.g., where local administration is desired, cell culture or animal models can be used to determine a dose required to achieve a therapeutically effective concentration within the local site.

[0336] In some embodiments, an antibody or antigen binding fragment thereof described herein can be administered to a subject as a monotherapy. Alternatively, the antibody or antigen binding fragment thereof can be administered in conjunction with other therapies for a cancer or an autoimmune disorder, allergy, and / or condition associated with chronic inflammation (combination therapy). For example, the composition can be administered to a subject at the same time, prior to, or after, a second therapy. In some embodiments, the antibody or antigen binding fragment thereof and the one or more additional active agents are administered at the same time. Optionally, the antibody or antigen binding fragment thereof is administered first in time and the one or more additional active agents are administered second in time. In some embodiments, the one or more additional active agents are administered first in time and the antibody or antigenPatent Application binding fragment thereof is administered second in time. Optionally, the antibody or antigen binding fragment thereof and the one or more additional agents are administered simultaneously in the same or different routes. For example, a composition comprising the antibody or antigen binding fragment thereof optionally contains one or more additional agents.

[0337] An antibody or antigen binding fragment thereof described herein can replace or augment a previously or currently administered therapy. For example, upon treating with an antibody or antigen binding fragment thereof, administration of the one or more additional active agents can cease or diminish, e.g., be administered at lower levels or dosages. In some embodiments, administration of the previous therapy can be maintained. In some embodiments, a previous therapy is maintained until the level of the antibody or antigen binding fragment thereof reaches a level sufficient to provide a therapeutic effect.

[0338] Monitoring a subject (e g., a human patient) for an improvement, as defined herein, means evaluating the subject for a change in a disease parameter, e.g., a reduction in one or more symptoms of cancer or an autoimmune disorder, allergy, and / or condition associated with chronic inflammation exhibited by the subject. In some embodiments, the evaluation is performed at least one (1) hour, e.g., at least 2, 4, 6, 8, 12, 24, or 48 hours, or at least 1 day, 2 days, 4 days, 10 days, 13 days, 20 days or more, or at least 1 week, 2 weeks, 4 weeks, 10 weeks, 13 weeks, 20 weeks or more, after an administration. The subject can be evaluated in one or more of the following periods: prior to beginning of treatment; during the treatment; or after one or more elements of the treatment have been administered. Evaluation can include evaluating the need for further treatment, e.g., evaluating whether a dosage, frequency of administration, or duration of treatment should be altered. It can also include evaluating the need to add or drop a selected therapeutic modality, e.g., adding or dropping any of the treatments described herein.

[0339] In some instances, the antibody or antigen binding fragment thereof can be administered via virus-like particles. Virus-like particles (VLPs) comprise viral protein(s) derived from the structural proteins of a virus. Methods for making and using virus like particles are described in, for example, Garcea and Gissmann, Current Opinion in Biotechnology 15:513-7 (2004).

[0340] In some instances, the antibody or antigen binding fragment thereof can be administered by subviral dense bodies (DBs). DBs transport proteins into target cells by membrane fusion. Methods for making and using DBs are described in, for example, Pepperl-Klindworth et al., Gene Therapy 10:278-84 (2003).Patent Application

[0341] In some instances, the antibody or antigen binding fragment thereof can be administered by tegument aggregates. Methods for making and using tegument aggregates are described in International Publication No. WO 2006 / 110728.

[0342] In another aspect, provided is a method of treating a subject with cancer or autoimmune disorder, the method comprising administering to the patient immune cells that express a CAR comprising an antibody or antigen binding fragment thereof as described herein.

[0343] In another aspect, provided is a method of treating a subject with cancer or autoimmune disorder, the method comprising administering to the patient cells that have been genetically engineered, using methods such as those described herein, to express and secrete an antibody or antigen binding fragment thereof as described in this disclosure.

[0344] In another aspect, provided is a method of treating a subject with a cancer or an autoimmune disorder, allergy, and / or condition associated with chronic inflammation, the method comprising administering to the patient a vector comprising a nucleic acid sequence encoding an antibody or antigen binding fragment thereof as described in this disclosure.

[0345] There are a number of compositions and methods which can be used to deliver the nucleic acid molecules and / or polypeptides to cells, either in vitro or ex vivo via, for example, expression vectors. These methods and compositions can largely be broken down into two classes: viral based delivery systems and non-viral based delivery systems. Such methods are well known in the art and readily adaptable for use with the compositions and methods described herein.

[0346] As used herein, plasmid or viral vectors are agents that transport the disclosed nucleic acids into the cell without undesired degradation and include a promoter yielding expression of the nucleic acid molecule and / or adapter polypeptide in the cells into which it is delivered. Viral vectors are, for example, Adenovirus, Adeno-associated virus, herpes virus, Vaccinia virus, Polio virus, Sindbis, and other RNA viruses, including these viruses with the HIV backbone. Also preferred are any viral families which share the properties of these viruses which make them suitable for use as vectors. Retroviral vectors, in general are described by Coffin et al., Retroviruses, Cold Spring Harbor Laboratory Press (1997), which is incorporated by reference herein for the vectors and methods of making them. The construction of replication-defective adenoviruses has been described (Berkner et al., J. Virology 61:1213-20 (1987); Massie et al., Mol. Cell. Biol. 6:2872-83 (1986); Haj-Ahmad et al., J. Virology 57:267-74 (1986); Davidson et al., J. Virology 61:1226-39 (1987); Zhang et al., BioTechniques 15:868-72 (1993)). The benefit and thePatent Application use of these viruses as vectors is that they are limited in the extent to which they can spread to other cell types, since they can replicate within an initial infected cell, but are unable to form new infections viral particles. Recombinant adenoviruses have been shown to achieve high efficiency after direct, ex vivo delivery to airway epithelium, hepatocytes, vascular endothelium, CNS parenchyma, and a number of other tissue sites. Other useful systems include, for example, replicating and host-restricted non-replicating vaccinia virus vectors. In some instances, the nucleic acid molecules encoding the antibody or antigen binding fragment thereof can be delivered via virus-like particles.

[0347] Non-viral based delivery methods, can include expression vectors comprising nucleic acid molecules and nucleic acid sequences encoding the adapter polypeptides, wherein the nucleic acids are operably linked to an expression control sequence. Suitable vector backbones include, for example, those routinely used in the art such as plasmids, artificial chromosomes, BACs, YACs, or PACs. Numerous vectors and expression systems are commercially available from such corporations as Novagen (Madison, WI), Clonetech (Pal Alto, CA), Stratagene (La Jolla, CA), and Invitrogen / Life Technologies (Carlsbad, CA). Vectors typically contain one or more regulatory regions. Regulatory regions include, without limitation, promoter sequences, enhancer sequences, response elements, protein recognition sites, inducible elements, protein binding sequences, 5’ and 3’ untranslated regions (UTRs), transcriptional start sites, termination sequences, polyadenylation sequences, and introns.

[0348] Preferred promoters controlling transcription from vectors in mammalian host cells may be obtained from various sources, for example, the genomes of viruses such as polyoma, Simian Virus 40 (SV40), adenovirus, retroviruses, hepatitis B virus, and most preferably cytomegalovirus (CMV), or from heterologous mammalian promoters (e.g., β-actin promoter or EF1α promoter), or from hybrid or chimeric promoters (e.g., CMV promoter fused to the β-actin promoter). Of course, promoters from the host cell or related species are also useful herein.

[0349] Enhancer generally refers to a sequence of DNA that functions at no fixed distance from the transcription start site and can be either 5’ or 3’ to the transcription unit. Furthermore, enhancers can be within an intron as well as within the coding sequence itself. They are usually between 10 and 300 bp in length, and they function in cis. Enhancers usually function to increase transcription from nearby promoters. Enhancers can also contain response elements that mediate the regulation of transcription. While many enhancer sequences are known from mammalianPatent Application genes (globin, elastase, albumin, fetoprotein, and insulin), typically one will use an enhancer from a eukaryotic cell virus for general expression. Preferred examples are the SV40 enhancer on the late side of the replication origin, the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers.

[0350] The promoter and / or the enhancer can be inducible (e.g., chemically or physically regulated). A chemically regulated promoter and / or enhancer can, for example, be regulated by the presence of alcohol, tetracycline, a steroid, or a metal. A physically regulated promoter and / or enhancer can, for example, be regulated by environmental factors, such as temperature and light. Optionally, the promoter and / or enhancer region can act as a constitutive promoter and / or enhancer to maximize the expression of the region of the transcription unit to be transcribed. In certain vectors, the promoter and / or enhancer region can be active in a cell type specific manner. Optionally, in certain vectors, the promoter and / or enhancer region can be active in all eukaryotic cells, independent of cell type. Preferred promoters of this type are the CMV promoter, the SV40 promoter, the beta-actin promoter, the EFla promoter, and the retroviral long terminal repeat (LTR).

[0351] The vectors also can include, for example, origins of replication and / or markers. A marker gene can confer a selectable phenotype, e.g., antibiotic resistance, on a cell. The marker product is used to determine if the vector has been delivered to the cell and once delivered is being expressed. Examples of selectable markers for mammalian cells are dihydrofolate reductase (DHFR), thymidine kinase, neomycin, neomycin analog G418, hygromycin, puromycin, and blasticidin. When such selectable markers are successfully transferred into a mammalian host cell, the transformed mammalian host cell can survive if placed under selective pressure. Examples of other markers include, for example, the E. coli lacZ gene, green fluorescent protein (GFP), and luciferase. In addition, an expression vector can include a tag sequence designed to facilitate manipulation or detection (e.g., purification or localization) of the expressed polypeptide. Tag sequences, such as GFP, glutathione S-transferase (GST), polyhistidine, c-myc, hemagglutinin, or FLAG™ tag (Kodak; New Haven, CT) sequences typically are expressed as a fusion with the encoded polypeptide. Such tags can be inserted anywhere within the polypeptide including at either the carboxyl or amino terminus.

[0352] In certain embodiments, the effective amount of a pharmaceutical composition comprising an antibody or antigen binding fragment thereof to be employed therapeutically depends, forPatent Application example, upon the therapeutic context and objectives. One skilled in the art will appreciate that the appropriate dosage levels for treatment, according to certain embodiments, vary depending, in part, upon the molecule delivered, the indication for which a antibody or antigen binding fragment thereof is being used, the route of administration, and the size (body weight, body surface or organ size) and / or condition (the age and general health) of the patient. The clinician can titer the dosage and modify the route of administration to obtain the optimal therapeutic effect.

[0353] The clinician also selects the frequency of dosing, taking into account the pharmacokinetic parameters of the antibody or antigen binding fragment thereof in the formulation used. Such pharmacokinetic parameters are well known in the art, i.e., the rate of absorption, bioavailability, metabolism, clearance, and the like (see, e.g., Hidalgo-Aragones (1996) J. Steroid Biochem. Mol. Biol. 58:611-617; Groning (1996) Pharmazie 51:337-341; Fotherby (1996) Contraception 54:59-69; Johnson (1995) J. Pharm. Sci. 84:1144-1146; Rohatagi (1995) Pharmazie 50:610-613; Brophy (1983) Eur. J. Clin. Pharmacol. 24:103-108; the latest Remington's, supra). In certain embodiments, a clinician administers the composition until a dosage is reached that achieves the desired effect. In certain embodiments, the composition can therefore be administered as a single dose or as two or more doses (which may or may not contain the same amount of the desired molecule) over time, or as a continuous infusion via, for example, an implantation device or catheter. Further refinement of the appropriate dosage is routinely made by those of ordinary skill in the art and is within the ambit of tasks routinely performed by them. In certain embodiments, appropriate dosages can be ascertained through use of appropriate dose-response data.

[0354] In some cases, the dosage (of the active component) ranges from about 0.0001 to 100 mg / kg, and more usually 0.0002 to 20 mg / kg, of the patient’s body weight. For example, dosages can be 0.0003 mg / kg body weight, 0.0001 mg / kg body weight, 0.0003 mg / kg body weight, 0.0005 mg / kg body weight, 0.001 mg / kg body weight or within the range of 0.0001-20 mg / kg. In certain examples, the antibody or antigen binding fragment thereof can be administered at a dose of 0.0001 mg / kg, 0.0002 mg / kg, 0.0003 mg / kg, 0.0004 mg / kg, or 0.0005 mg / kg once every other day, week, or month at least four times. An exemplary treatment regime may include administration once per day, once per week, twice a week, once every two weeks, once every three weeks, once every four weeks, once a month, once every 3 months, or once every three to 6 months. In some cases, the treatment comprises administering antibody or antigen binding fragment thereof according to one of the aforementioned dosing regimens for a first period and another of the aforementioned dosingPatent Application regimens for a second period. In some cases, the treatment discontinues for a period of time before the same or a different dosing regimen resumes. For example, a patient may be on an antibody or antigen binding fragment thereof -specific antibody dosing regimen for two weeks, off for a week, on for another two weeks, and so on. Dosage regimens for antibodies or antigen binding fragments thereof of this disclosure include 0.0001 mg / kg body weight, 0.0003 mg / kg body weight, 0.0002 mg / kg body weight, 0.0004 mg / kg body weight, or 0.001 mg / kg via intravenous administration, with the antibodies or antigen binding fragments thereof being given using one of the following dosing schedules: (i) every four weeks for six dosages, then every three months; (ii) every three weeks; (iii) 3 mg / kg body weight once followed by up to 0.001 mg / kg body weight every three weeks.

[0355] In certain embodiments, the route of administration of the pharmaceutical composition is in accord with known methods, e.g., orally, through injection by intravenous, intraperitoneal, intracerebral (intra-parenchymal), intracerebral, intraventricular, intramuscular, subcutaneously, intra-ocular, intraarterial, intraportal, or intralesional routes; by sustained release systems or by implantation devices. In certain embodiments, the compositions can be administered by bolus injection or continuously by infusion, or by implantation device. In certain embodiments, individual elements of a combination therapy may be administered by different routes.

[0356] In certain embodiments, the composition can be administered locally, e.g., during surgery or topically. Optionally local administration is via implantation of a membrane, sponge, or another appropriate material onto which the desired molecule has been absorbed or encapsulated. In certain embodiments, where an implantation device is used, the device can be implanted into any suitable tissue or organ, and delivery of the desired molecule can be via diffusion, timed-release bolus, or continuous administration.

[0357] In certain embodiments, it can be desirable to use a pharmaceutical composition comprising an antibody or antigen binding fragment thereof in an ex vivo manner. In such instances, cells that have been removed from a subject may be exposed to a pharmaceutical composition comprising an antibody or antigen binding fragment thereof after which the cells are subsequently implanted back into the subject.

[0358] In some instances, the provided methods may include administering to the subject an antibody or antigen binding fragment thereof that is conjugated to a therapeutic agent. ThePatent Application therapeutic agent may be at least one of a cytotoxic agent, a chemotherapeutic agent, or an immunosuppressive agent. Such therapeutic agents are described below.

[0359] In some instances, the provided methods may include administering an antibody or antigen binding fragment thereof and a second form of therapy to the subject. The second form of therapy may include a cytotoxic agent, a chemotherapeutic agent, an immune-activating agent (including immune checkpoint inhibitors), or radiation therapy. In some embodiments, the second form of therapy is an antibody (e.g., a monoclonal antibody). Monoclonal antibodies which may be administered as a second form of therapy include, but are not limited to, rituximab (e.g., for treatment of B-cell lymphomas), trastuzumab (e.g., for treatment of breast cancer), and cetuximab (e.g., for treatment of lung cancer).

[0360] In some embodiments, the antibody or antigen binding fragment thereof is conjugated to a moiety that specifically binds to an immune cell. In some embodiments, provided is a bispecific antibody comprising an antibody or antigen binding fragment thereof as described herein and an antibody or antigen binding fragment thereof that specifically binds to an immune cell. In some embodiments, the bispecific antibody comprises an antibody or antigen binding fragment thereof and an antibody moiety that specifically binds to T cells. Such a molecule is referred to as a bispecific T cell engager and may induce T cell-mediated cytotoxicity of hematological malignancy antigen-expressing cancer cells (see, e.g., Zhou et al., 2021, Biomarker Research 9:38).

[0361] In certain aspects, antibody or antigen binding fragment thereof according to the present disclosure can be administered as a co-therapy with other therapeutic agents. Other examples of therapeutic agents include chemotherapeutic agents, radiotherapeutic agents, and immunotherapeutic agents, as well as combinations thereof. In this way, the antibody or peptide complex delivered to the subject can be multifunctional, in that it exerts one therapeutic effect by binding to the hematological malignancy antigen protein and a second therapeutic effect by delivering a supplemental therapeutic agent.

[0362] The therapeutic agent can act extracellularly, for example by initiating or affecting an immune response, or it can act intracellularly, either directly by translocating through the cell membrane or indirectly by, for example, affecting transmembrane cell signaling. The therapeutic agent is optionally cleavable from the antibody or antigen binding fragment thereof. Cleavage can be autolytic, accomplished by proteolysis, or affected by contacting the cell with a cleavage agent.Patent Application

[0363] In some instances, the therapeutic agent is a cytotoxic agent. A cytotoxin or cytotoxic agent includes any agent that is detrimental to cells. Examples of toxins or toxin moieties include diphtheria, ricin, streptavidin, and modifications thereof. Additional examples include paclitaxel, cisplatin, carboplatin, cytochalasin B, gramicidin D, ethidium bromide, emetine, etoposide, tenoposide, colchicin, dihydroxy anthracin di one, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, puromycin and analogs or homologs thereof. Therapeutic agents include, but are not limited to, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil, decarbazine), alkylating agents (e.g., mechlorethamine, thiotepa, chlorambucil, melphalan, carmustine (BSNU) and lomustine (CCNU), cyclothosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, and cis-dichlorodiamine platinum (II) (DDP) cisplatin), anthracyclines (e. g., daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)), and anti -mitotic agents (e.g., vincristine and vinblastine).

[0364] As referred to herein, a chemotherapeutic agent is a chemical compound useful in the treatment of cancer. Examples of chemotherapeutic agents include erlotinib (such as TARCEVA®, Genentech / OSI Pharm.), bortezomib (such as VELCADE®, Millenium Pharm.), fulvestrant (such as FASLODEX®, AstraZeneca), sutent (such as SU11248, Pfizer), letrozole (such as FEMARA®, Novartis), imatinib mesylate (such as GLEEVEC®, Novartis), PTK787 / ZK222584 (Novartis), oxaliplatin (such as ELOXATIN®, Sanofi), 5-fluorouracil (5-FU), leucovorin, rapamycin (also known as sirolimus) (such as RAPAMUNE®, Wyeth), lapatinib (such as TYKERB®, GSK572016, GlaxoSmithKline), lonafamib (such as SCH 66336), sorafenib (such as BAY43-9006, Bayer Labs.), capecitabine (such as XELODA®, Roche), docetaxel (such as TAXOTERE®), and gefitinib (such as IRESSA®, Astrazeneca), AG1478, AG1571 (such as SU 5271; Sugen Inc.), alkylating agents such as thiotepa and cyclosphosphamide (such as CYTOXAN®); alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylomelamine; acetogenins (especially bullatacin and bullatacinone); a camptothecin (including the synthetic analogue topotecan); bryostatin; callystatin; CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues); cryptophycins (particularlyPatent Application cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogues, KW-2189 and CB1-TM1); eleutherobin; pancrati statin; a sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlomaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechl or ethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; antibiotics such as the enediyne antibiotics (e.g., calicheamicin, particularly calicheamicin γi1and calicheamicin θi1); dynemicin, including dynemicin A; bisphosphonates, such as clodronate; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores), aclacinomysins, actinomycin, anthramycin, azaserine, bleomycins, cactinomycin, carabicin, carminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (such as ADRIAMYCIN®, including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5 -fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfomithine; elliptinium acetate; an epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; Trametes Versicolor polysaccharide-K (Krestin, PSK) (JHS Natural Products, Eugene, OR); razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2', 2''-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; cytarabinePatent Application (cytosine arabinoside, “Ara-C”); cyclophosphamide; thiotepa; taxoids, e.g., paclitaxel (such as TAXOL®, Bristol-Myers Squibb Oncology, Princeton, N. J.), ABRAXANE™ (a Cremophor-free, albumin-engineered nanoparticle formulation of paclitaxel (American Pharmaceutical Partners, Schaumberg, IL)), and doxetaxel (such as TAXOTERE®, Rhone-Poulenc Rorer, Antony, France); chloranbucil; gemcitabine (such as GEMZAR®); 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP- 16); ifosfamide; mitoxantrone; vincristine; vinorelbine (such as NAVELBINE®); novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda; ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylomithine (DMFO); retinoids such as retinoic acid; capecitabine; and pharmaceutically acceptable salts, acids or derivatives of any of the above.

[0365] Chemotherapeutic agents, as used herein, also refers to (i) anti-hormonal agents that act to regulate or inhibit hormone action on tumors such as anti-estrogens and selective estrogen receptor modulators (SERMs), including, for example, tamoxifen (including NOLVADEX® tamoxifen), raloxifene, droloxifene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY 117018, onapristone, and toremifene (such as FARESTON®); (ii) aromatase inhibitors that inhibit the enzyme aromatase, which regulates estrogen production in the adrenal glands, such as, for example, 4(5)-imidazoles, aminoglutethimide, megestrol acetate (such as MEGASE®), exemestane (such as AROMASIN®), formestanie, fadrozole, vorozole (such as RIVISOR®), letrozole (such as FEMARA®), and anastrozole (such as ARIMIDEX®); (iii) anti-androgens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; as well as troxacitabine (a 1,3-dioxolane nucleoside cytosine analog); (iv) aromatase inhibitors; (v) protein kinase inhibitors; (vi) lipid kinase inhibitors; (vii) antisense oligonucleotides, particularly those which inhibit expression of genes in signaling pathways implicated in aberrant cell proliferation, such as, for example, PKC-alpha, Ralf and H-Ras; (viii) VEGF receptor and angiogenesis inhibitors (including ribozymes such as ANGIOZYME®) and a HER2 expression inhibitor; (ix) vaccines such as gene therapy vaccines, for example, ALLOVECTIN-7® vaccine (plasmid / lipid complex containing the DNA sequences encoding HLA-B7 and ß2 microglobulin), LEUVECTIN® vaccine (plasmid DNA expression vector encoding interleukin-2 (IL-2) complexed with a lipid delivery vehicle (DMRIE / DOPE)), and VAXID® vaccine (patient-specific naked DNA vaccine); IL-2 or aldesleukin (such as PROLEUKIN®); topoisomerase 1 inhibitors (such as TOPOTECAN®); gonadotropin-releasing hormone antagonists (such as ABARELIX®); (x) anti-angiogenic agentsPatent Application such as bevacizumab (such as AVASTIN®, Genentech); and (xi) pharmaceutically acceptable salts, acids or derivatives of any of the above.

[0366] In some instances, the treatment methods provided herein may further comprise administering an immunotherapy agent such as an immune checkpoint inhibitor as part of the method. These treatments work by “taking the brakes off’ the immune system (are immune-activating), allowing it to mount a stronger and more effective attack against cancer and other disorders. Several different types of checkpoint inhibitors, targeting different checkpoints or “brakes” on immune cells, are currently in use. Exemplary immunotherapy agents are PD-1 inhibitors (such as nivolumab and pembrolizumab), PD-L1 inhibitors (such as atezolizumab, durvalumab, and avelumab), and CTLA-4 inhibitors (such as ipilimumab). In one example, the second form of therapy comprises a PD-L1 inhibitor, a PD-1 inhibitor, or a CTLA-4 inhibitor. In some instances, combinations of such inhibitors can be administered. In some instances, the PD-L1 inhibitor, the PD-1 inhibitor, and / or the CTLA-4 inhibitor may be an inhibitory antibody that binds specifically to PD-L1, PD-1, or CTLA-4, respectively.

[0367] In some instances, the treatment methods provided herein may further comprise administering radiation therapy to the subject. Radiation therapy uses high-energy radiation to shrink tumors and kill cancer cells. X-rays, gamma rays, and charged particles are types of radiation used for cancer treatment. The radiation may be delivered by a machine outside the body (external -beam radiation therapy), or it may come from radioactive material placed in the body near cancer cells (internal radiation therapy, also called brachytherapy). Systemic radiation therapy uses radioactive substances, such as radioactive iodine, that travel in the blood to kill cancer cells.EXAMPLES EXAMPLE 1

[0368] A library was used to discover sequences with specific and potent binding to the TERT540 peptide in the context of HLA-A*02:01, also known as the TERT540 pMHC or TERT540 pHLA. The library was screened by multiple rounds of panning with positive antigen to enrich for specific binders and with irrelevant pHLAs to eliminate non-specific TERT540 pHLA binders as well as binders towards the HLA itself.

[0369] Following initial biophysical and functional characterization, binders were chosen for further optimization.Patent Application

[0370] Promising, optimized binders with improved potency were identified by biophysical and functional characterization, and tested for specificity. Chosen binders were more potent than the parent binders and maintained no binding to irrelevant pHLAs. These optimized binders were reformatted into mammalian vectors for expression and further characterization.

[0371] The anti-HLA-A2 / TERTs4o binder sequences were produced and incorporated into TCR mimetic (TCRm) T-cell Engager antibody constructs. The TCRms included one or more binder specific for the TERT540 pHLA complex and a binder specific for CD3 expressed by an immune cell.FIGS. 1-4 provide schematics of TCRms of the disclosure. FIGS. 1-2 provide examples of TERT TCRms of the disclosure, which are immune cell engagers that target the TERT540-PHLA complex on a target cell while engaging an immune cell via the immune cell receptor CD3.

[0372] Certain exemplary and / or candidate TCRms of the disclosure, including some of those described in the examples and for which amino sequences are provided herein, are named in a manner that incorporates their respective TCRm TCE formats. For example, the TCRm referred to herein as TERT(1+1) refers to a specific anti-TERT TCRm, described in Tables la, lb, Id, and Table 2, which is in the 1+1 TCRm TCE format. Similarly, the TCRm referred to herein as TERT(2+1) is a specific anti-TERT TCRm TCE, described herein, which is in the 2+1 TCRm TCE format.

[0373] The schematics provided in FIGS. 1-2 are examples of TCRms, in the 1+1 TCRm TCE format (such as in the TCRm TERT(1+1) (FIG. 1)) and in the 2+1 TCRm TCE format (TERT(2+1) (FIG. 2)).

[0374] FIG. 3 diagrams exemplary TCRms of the disclosure in the TERT 1+1 TCRm TCE format. The TCRm TERT(1+1), described herein, has a 1+1 TCRm TCE format, exemplified as embodiment 601. This exemplary 1+1 TCRm TCE includes a first antigen binding domain 603 (e.g., for a TERT540-PHLA complex) on a first peptide and a second binding domain 605 (e.g., for an immune cell receptor such as CD3). The antigen binding domains are bound to an Fc region 609 via a hinge. In certain preferred aspects, the second antigen binding domain 605 comprises a Fab. The Fc region may include modifications (shown as “X”) as described herein, including a label, linker, or other point of attachment. Preferably, the modifications and / or substitutions provide Fc silencing and heterodimerization (knob-into-hole Fc). In certain aspects, the TCRm includes a third polypeptide, which is a light chain.Patent Application

[0375] Modifications of the TERT 1+1 TCRm TCE format are contemplated and embodied by the present disclosure. As shown in FIG. 3, the TERT TCRm TERT(1+1)’, as provided herein, exemplified as embodiment 621, is in the TERT 1+1 TCRm TCE format like TERT(1+1). However, as shown in FIG. 3, TERT(1 +1)’ incorporates a C-terminal lysine. Further variations are contemplated by the present disclosure. For example, the first antigen binding domain (603) may be a binder other than the exemplified scFv, such as a Fab or Fab fragment. Likewise, the second antigen binding domain may be a binder other than the exemplified Fab, e.g., an scFv.

[0376] FIGS. 4A-4B diagram exemplary anti-TERT TCRm TCEs of the disclosure in the TERT 2+1 TCRm TCE format. A schematic for the format of the anti-TERT TCRm TCE referred to as TERT(2+1) herein, is provided in FIG. 4A. This TCRm (611) exemplifies the general TERT 2+1 TCRm TCE antibody format. This exemplary TCRm includes a first antigen binding domain 603 (e.g., for a TERT54o-pHLA complex) on a first peptide and a second binding domain 607 (e g., for an immune cell receptor such as CD3) and third binding domain 603b on a second peptide, which may be the same binder and / or bind to the same target as the first antigen binding domain 603. In certain aspects, the second binding domain comprises an scFv. The antigen binding domains are bound to an Fc region 609 via a hinge. The Fc region may include modifications as described herein, including a label, linker, or other point of attachment. Preferably, the modifications and / or substitutions provide Fc silencing and heterodimerization (knob-into-hole Fc).

[0377] In certain aspects, in the 2+1 format, exemplified as embodiment 631, the first antigen binding domain 603 and the third antigen binding domain 603b each target an antigen (e.g., on a target cell / an HLA-peptide complex) on a target cell, while the remaining, second antigen binding domain binds 607 to an immune cell (e.g., as an immune cell engager or activator). The first and third antigen binding domains 603 / 603b may bind to the same antigen (e.g., an HLA-peptide complex). The first and third antigen binding domains 603 / 603b may bind to different targets. For example, the first and third antigen binding domains 603 / 603b may bind to different antigen epitopes, different TERT peptides presented by HLA, different HLA-TERT peptide complexes (e.g., presented by different HLA alleles), and / or bound or unbound TERT peptides, and the like, while the third antigen binding domain binds to CD3 on a T cell.

[0378] TCRm TCEs comprising variations relative the TERT 2+1 TCRm TCE format exemplified by TERT(2+1) are contemplated by the present disclosure. For example, a schematic for the TERT TCRm TCE, referred to herein as TERT(2+1)’, is provided in FIG. 4 A as 631. As shown, thePatent Application format of this TCRm differs from that of TERT(2+1) by the addition of a C-terminal lysine that was added to each of the heavy chains of TERT(2+1)’, relative to TERT(2+1), which improves protein expression during manufacturing. C-terminal lysines in the 2+1 format exemplified by TERT(2+1)’ are cleaved off during manufacturing by carboxypeptidase, and this cleavage should not alter structural stability, biological properties, or pharmacokinetics of the molecule. The amino acid sequence of the TCRm TERT(2+1)’ as used in the Examples is provided in Tables Id and 2.

[0379] FIG. 4B provides schematics of additional, exemplary TERT 2+1 TCRm TCEs, with variations of the 2+1 format relative to TERT(2+1). As shown, a schematic for the TERT TCRm TERT(2+1)” is provided as 613 and includes a first antigen binding domain 603 (e.g., for a TERT54o-pHLA complex) on a first peptide, a second binding domain 607 (e.g., for an immune cell receptor such as CD3) on a second peptide, and third binding domain 603b on the first peptide, which may be the same binder and / or bind to the same target as the first antigen binding domain 603. In certain aspects, the second binding domain 607 comprises an scFv. The antigen binding domains are bound to an Fc region 609 via a hinge. The Fc region may include modifications as described herein, including a label, linker, or other point of attachment. Preferably, the modifications and / or substitutions provide Fc silencing and heterodimerization (knob-into-hole Fc). As exemplified, in the 2+1 format of TERT(2+1), all antigen binding domains are in an scFv format. However, different types of antigen binding domains, e.g., a Fab or Fab fragment, are contemplated by the disclosure.

[0380] As also shown in FIG. 4B, the TCRm referred to herein as TERT(2+1)*, exemplified as embodiment 615, is in a 2+1 format that includes a first binding domain (e.g., for a TERT54o-pHLA complex) on a first peptide 603, and a second binding domain 605 (e.g., for an immune cell receptor such as CD3) and third binding domain 603b on a second peptide, which may be the same binder and / or bind to the same target as the first antigen binding domain 603. The 2+1 format used by the TCRm TERT(2+1)* exemplifies an immune cell binder (605) as a Fab or Fab fragment as exemplified in the TERT(1+1) format provided in FIG. 3.

[0381] In certain aspects, in a TERT 2+1 TCRm TCE format, the first antigen binding domain 603 and the third antigen binding domain 603b bind to a target cell (each target an antigen, e.g., on a target cell / an HLA-peptide complex), while the remaining, second antigen binding domain binds to an immune cell (e.g., as an immune cell engager or activator). The first and third antigen binding domains 603 / 603b may bind to the same antigen (e.g., an HLA-peptide complex). The first andPatent Application third antigen binding domains 603 / 603b may bind to different targets. For example, the first and third antigen binding domains 603 / 603b may bind to different antigen epitopes, different TERT peptides presented by HLA, different HLA-TERT peptide complexes (e.g., presented by different HLA alleles), and / or bound or unbound TERT peptides, and the like, while the second antigen binding domain binds to CD3 on a T cell.

[0382] The exemplary TCRm in the TERT 1+1 TCRm TCE format as used throughout this Example, incorporates an anti-TERT54o / HLA-A*O2 binder into a TCRm comprising three peptide chains, the sequences of which are provided in Table Id and Table 2. The exemplary TCRm used throughout this Example, in the TERT 2+1 TCRm TCE format incorporates an anti-TERT54O / HLA-A*O2 binder into a TCRm comprising two peptide chains, the sequences of which are provided in Table Id and Table 2. The exemplary TCRm used throughout this Example, in the TERT 2+1 TCRm TCE format incorporates an anti-TERTs4o / HLA-A*O2 binder into a TCRm comprising three peptide chains, the sequences of which are provided in Table Id and Table 2. The amino acid sequences of TERT(1+1), TERT(1+1)’, TERT(2+1), TERT(2+1)’, TERT(2+1)” and TERT(2+1)* are used in the present Examples are provided in Table Id and Table 2.

[0383] TERT (1+1)’ and TERT(2+1)’ differs from TERT(1+1) and TERT(2+1) by the addition of a C-terminal lysine that was added to each of the heavy chains of TERT(1+1)’ and TERT(2+1)’, relative to TERT(1+1) and TERT(2+1), to improve protein expression during manufacturing. C-terminal lysines in TERT(1+1)’ and TERT(2+1)’ are cleaved off during manufacturing by carboxypeptidase, and this cleavage should not alter structural stability, biological properties, or pharmacokinetics of the molecule. In certain aspects, the peptide chains lack C-terminal lysines, as provided in Table Id (SEQ ID NOS: 54-57) and Table 2. In certain aspects, the peptide chains comprise one or more linkers, which are exemplified as glycine-serine based linkers. These linkers are non-limiting, and different glycine-serine linkers are contemplated by the present disclosure as are other linkers known in the art.

[0384] To assess the potency of TERT(1+1) and TERT(2+1), their binding affinity and avidity for the TERT54o-pHLA complex was evaluated. FIGS. 5A-5D provide affinity and avidity data for TERT(1+1) and TERT(2+1), obtained via Surface Plasmon Resonance (SPR) using a Carterra instrument. Desirably, the data shows that TERT TCRms have sub-nM affinity for the TERT540-pHLA complex. As such, when TERT TCRms bind to an immune cell, their higher affinity for thePatent Application TERTs4o-pH A peptide complex will help assure it engages an immune cell along with a target cell expressing the TERT540-pHLA complex.

[0385] FIGS. 6A-6C show in vitro cytotoxicity of TERT TCRms against target-positive cancer cells with presumed varying levels of target expression, with target-negative cancer cells being used as negative controls. The following cells were incubated with TERT TCRms at increasing concentrations: U937-A2 (AML derived cells that are U937 cells, which are HLA-A*03:01-positive, transduced with HLA-A*02:01); ML2 (AML derived cells); THP-1 (AML derived cells); AML14 (AML derived cells); EM2 (CML in blast crisis-derived cells); NCI-H1755 (NSCLC derived cells); COV413 (ovarian cancer derived cells); T98G (glioblastoma derived cells with TERT promoter mutation C250T); U937 WT (AML derived cells, which are negative for HLA-A*02:01); and U2OS (osteosarcoma derived cells, which are negative for TERT expression). As shown in FIGS. 6A-6B, despite some of the targets in this assay being presumed to have low target copy numbers per cell, cytotoxicity against target-positive cells was readily detectable with both TERT(1+1) and TERT(2+1). For target-negative cells, U937 WT cells (which are negative for HLA-A*02:01) and U2OS cells (which are negative for TERT expression) were used, as shown in FIG. 6C. As expected, TERT TCRms did not show appreciable cytotoxicity against TERT540-pHLA negative cells. Overall, the potent (sub-nM EC50) and specific activity exhibited by TERT TCRms against myeloid and solid tumor cell lines in vitro highlights their therapeutic potential in oncology.

[0386] As shown in FIGS. 7A-7B, both TERT(1+1) and TERT(2+1) induce potent (sub-nM) TERT54o-pHLA-specific T-cell activation against target-positive tumor cells (THP-1). There is no T cell activation in HLA-A*02:01 negative PBMCs only without target cells. These results are consistent with the predicted, on-target activity of TERT TCRms.

[0387] As shown in FIGS. 8A-8B in a spheroid tumor model assay, TERT(1+1) and TERT(2+1) selectively exhibited cytotoxicity against target-positive COR-L23-A2 cells (NSCLC derived cells, transduced with HLA-A*02:01). In this model, TERT TCRms did not show appreciable cytotoxicity against COR-L23 WT cells (which are negative for HLA-A*02:01), consistent with the on-target activity profile of TERT TCRms. The spheroid tumor model provides a three-dimensional model for solid tumors such as NSCLC. These results indicate the ability of TERT TCRms to not only selectively kill target-positive cells, but to do so in the context of a three-Patent Application dimensional tumor model that is expected to better recapitulate tumor microanatomy than two-dimensional models, which further supports exploration of their potential as oncologic therapies.

[0388] The pharmacokinetics (PK) of TERT TCRms were evaluated in humanized FcRn mice, which can be used to predict how monoclonal antibodies (mAbs) and other therapeutics will behave in humans. As shown in FIGS. 9A-9B, in human FcRn mice, both TERT(1+1) and TERT(2+1) exhibited a PK profile comparable to a conventional IgG-like antibody, with a halflife of 10 days or greater across all dose levels characterized. For both TERT TCRms, multiple dose levels resulted in serum concentrations higher than the EC90 values obtained from in vitro cytotoxicity experiments with AML cell lines and solid tumor cell lines exposed to TERT TCRms. This suggests that administration of TERT TCRms at these dose levels is likely to result in antitumor activity in in vivo experiments with mouse tumor models.

[0389] FIGS. 10A- 1 OB show that TERT TCRms demonstrate in vivo efficacy in a pilot experiment with a mouse tumor model of disseminated AML (U937-A2 injected i.v. in PBMC-reconstituted NSG mice). As shown in FIG. 10A, the decrease in tumor bioluminescence seen on day 30 of the study in mice treated with either TERT(1+1) or TERT(2+1) compared to mice given vehicle control indicates activity of TERT TCRms against tumor cells in the context of a living organism. TERT TCRms were administered i.v. at a dose of 0.1 mg / kg, once weekly, beginning at day 2 of the study. Human PBMCs were injected on day 1 of the study, and tumor cells were injected on day 14 of the study. Mice with fewer than 0.75% circulating human T-cells (as evaluated by flow cytometry) were excluded from the efficacy readout. As shown in FIG. 10B, repeated weekly administration of TERT TCRms was well tolerated in this study, with no effects on body weight observed in TERT TCRm-treated mice compared to vehicle control.

[0390] FIGS. 11 A-l 1C shows data from multi-positional X-scan analysis, in which the specificity for TERT TCRms were assessed relative to the amino acid sequence of the TERT540 peptide as presented in a pHLA complex. As shown in FIGS. 11A-11C, both TERT(1+1) and TERT(2+1) are highly specific for the TERT540 peptide. Surprisingly, in the TERT 1+1 TCRm TCE format, residues at positions 6 and 8 of TERT540 are important for TCRm binding, whereas they are not in the TERT 2+1 TCRm TCE format. However, both TERT(1+1) and TERT(2+1) TCRms rely on 3 or more positions of TERT540 for binding.

[0391] To further assess the safety profile of TERT TCRms and their PHLA-TERT540 binders, the cytotoxic potential of TERT TCRms towards cells pulsed with potentially cross-reactive peptidesPatent Application predicted by in silico algorithms was evaluated. The 20 peptides computationally determined to pose the highest cross-reactive risk were tested ...

Claims

Patent Application CLAIMS WHAT IS CLAIMED IS:

1. An antibody or antigen binding fragment thereof, wherein said antibody or antigen binding fragment thereof binds to an HLA-TERT540peptide complex on a target cell, wherein said antibody or antigen binding fragment thereof comprisesa heavy chain variable region that comprises an HC CDR1 sequence comprising an amino acid sequence as set forth in any one of SEQ ID NOS: 3 and 58-72, an HC CDR2 sequence comprising an amino acid sequence as set forth in any one of SEQ ID NOS: 4 and 73-90, and an HC CDR3 sequence comprising an amino acid sequence as set forth in any one of SEQ ID NOS: 5 and 91-118; anda light chain variable region that comprises an LC CDR1 sequence comprising an amino acid sequence as set forth in any one of SEQ ID NOS: 6 and 119-143, an LC CDR2 sequence comprising an amino acid sequence as set forth in any one of SEQ ID NOS: 7 and 144-171, and an LC CDR3 sequence comprising an amino acid sequence as set forth in any one of SEQ ID NOS: 8 and 172-199.

2. The antibody or antigen binding fragment of claim 1, wherein the heavy chain variable region comprises an amino acid sequence that is at least 90% identical to any one of SEQ ID NOS: 1 and 200-227 and the light chain variable region comprises an amino acid sequence that is at least 90% identical to any one of SEQ ID NOS: 2 and 228-255.

3. The antibody or antigen binding fragment thereof of claim 1 or claim 2, wherein the HC variable region sequence comprises an amino acid sequence having at least 95% similarity to any one of SEQ ID NOS: 1 and 200-227.

4. The antibody or antigen binding fragment thereof of claim 1 or claim 2, wherein the HC variable region sequence comprises an amino acid sequence having at least 98% similarity to any one of SEQ ID NOS: 1 and 200-227.

5. The antibody or antigen binding fragment thereof of claim 1 or claim 2, wherein the HC variable region sequence comprises an amino acid sequence as set forth in any one of SEQ ID NOS: 1 and 200-227.Patent Application 6. The antibody or antigen binding fragment thereof of any one of claims 1-5, wherein the LC variable region sequence comprises an amino acid sequence having at least 95% similarity to any one of SEQ ID NOS: 2 and 228-255.

7. The antibody or antigen binding fragment thereof of any one of claims 1-5, wherein the LC variable region sequence comprises an amino acid sequence having at least 98% similarity to any one of SEQ ID NOS: 2 and 228-255.

8. The antibody or antigen binding fragment thereof of any one of claims 1-5, wherein the LC variable region sequence comprises an amino acid sequence as set forth in any one of SEQ ID NOS: 2 and 228-255.

9. The antibody or antigen binding fragment thereof, wherein the antibody or fragment is a T-cell receptor (TCR) mimetic (TCRm) antibody or fragment thereof, wherein said TCR mimetic (TCRm) antibody or fragment thereof comprises: a first binding domain that binds to the HLA-TERT540peptide complex on a target cell and a second binding domain that binds to an immune cell receptor.

10. The antibody or antigen binding fragment thereof of claim 9, wherein the TCRm comprises a first polypeptide chain, a second polypeptide chain, and a third polypeptide chain, wherein the first polypeptide chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 12, the second polypeptide chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 13, and the third polypeptide chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 14.

11. The antibody or antigen binding fragment thereof of claim 10 wherein the first polypeptide chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 12, the second polypeptide chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 13, and the third polypeptide chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 14.

12. The antibody or antigen binding fragment thereof of claim 11, wherein the first polypeptide chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 12, the second polypeptide chain comprises an amino acid sequence that is at least 98% identical to SEQPatent Application ID NO: 13, and the third polypeptide chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 14.

13. The antibody or antigen binding fragment thereof of claim 12, wherein the first polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 12, the second polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 13, and the third polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 14.

14. The antibody or antigen binding fragment thereof of claim 9, wherein the TCRm comprises a first polypeptide chain, a second polypeptide chain, and a third polypeptide chain, wherein the first polypeptide chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 56, the second polypeptide chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 57, and the third polypeptide chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 14.

15. The antibody or antigen binding fragment thereof of claim 14, wherein the first polypeptide chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 56, the second polypeptide chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 57, and the third polypeptide chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 14.

16. The antibody or antigen binding fragment thereof of claim 15, wherein the first polypeptide chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 56, the second polypeptide chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 57, and the third polypeptide chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 14.

17. The antibody or antigen binding fragment thereof of claim 16, wherein the first polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 56, the second polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 57, and the third polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 14.Patent Application 18. The antibody or antigen binding fragment thereof of claim 9, wherein the TCRm comprises a third binding domain, wherein the third binding domain binds to an HLA-TERT peptide complex.

19. The antibody or antigen binding fragment thereof of claim 18, wherein the third binding domain binds to an HLA-TERT540peptide complex.

20. The antibody or antigen binding fragment thereof of claim 18 or 19, wherein the TCRm comprises a first polypeptide and a second polypeptide, wherein the first polypeptide chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 10 and the second polypeptide chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 11.

21. The antibody or antigen binding fragment thereof of claim 20, wherein the first polypeptide chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 10 and the second polypeptide chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 11.

22. The antibody or antigen binding fragment thereof of claim 21, wherein the first polypeptide chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 10 and the second polypeptide chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 11.

23. The antibody or antigen binding fragment thereof of claim 22, wherein the first polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 10 and the second polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 11.

24. The antibody or antigen binding fragment thereof of claim 18 or 19, wherein the TCRm comprises a first polypeptide and a second polypeptide, wherein the first polypeptide chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 54 and the second polypeptide chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 55.Patent Application 25. The antibody or antigen binding fragment thereof of claim 24, wherein the first polypeptide chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 54 and the second polypeptide chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 55.

26. The antibody or antigen binding fragment thereof of claim 25, wherein the first polypeptide chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 54 and the second polypeptide chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 55.

27. The antibody or antigen binding fragment thereof of claim 26, wherein the first polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 54 and the second polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 55.

28. The antibody or antigen binding fragment thereof of claim 9, wherein the TCRm comprises a polypeptide having an amino acid sequence that is at least 90% identical to SEQ ID NO: 9.

29. The antibody or antigen binding fragment thereof of claim 28, wherein the TCRm comprises a polypeptide having an amino acid sequence that is at least 95% identical to SEQ ID NO: 9.

30. The antibody or antigen binding fragment thereof of claim 29, wherein the TCRm comprises a polypeptide having an amino acid sequence that is at least 98% identical to SEQ ID NO: 9.

31. The antibody or antigen binding fragment thereof of claim 30, wherein the TCRm comprises a polypeptide having an amino acid sequence as set forth in SEQ ID NO: 9.

32. The antibody or antigen binding fragment thereof of claim 18 or claim 19, wherein the TCRm comprises a first polypeptide chain, a second polypeptide chain, and a third polypeptide chain, wherein the first polypeptide chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 10, the second polypeptide chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 258 and the third polypeptide chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 14.Patent Application 33. The antibody or antigen binding fragment thereof of claim 32, wherein the first polypeptide chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 10, the second polypeptide chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 258, and the third polypeptide chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 14.

34. The antibody or antigen binding fragment thereof of claim 33, wherein the first polypeptide chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 10, the second polypeptide chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 258, and the third polypeptide chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 14.

35. The antibody or antigen binding fragment thereof of claim 34, wherein the first polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 10, the second polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 258, and the third polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 14.

36. The antibody or antigen binding fragment thereof of claim 18 or 19, wherein the TCRm comprises a first polypeptide and a second polypeptide, wherein the first polypeptide chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 256 and the second polypeptide chain comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 257.

37. The antibody or antigen binding fragment thereof of claim 36, wherein the first polypeptide chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 256 and the second polypeptide chain comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 257.

38. The antibody or antigen binding fragment thereof of claim 37, wherein the first polypeptide chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 256 and the second polypeptide chain comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 257.Patent Application 39. The antibody or antigen binding fragment thereof of claim 38, wherein the first polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 256 and the second polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 257.

40. The antibody or antigen binding fragment thereof of any one of claims 9-39, wherein the immune cell receptor is CD3 and the second binding domain that binds to the immune cell receptor comprises an amino acid sequence having 90%, 95%, 98%, 99% similarity to or comprises an amino acid sequence as set forth in SEQ ID NO: 259.

41. The antibody or antigen binding fragment thereof of any one of claims 1-39, wherein the antibody or antigen binding fragment thereof does not have a substantial binding affinity to the HLA class I molecule in the absence of TERT540peptide or does not have substantial binding affinity to TERT540peptide in the absence of the HLA class I molecule.

42. The antibody or antigen binding fragment thereof of any one of claims 1-41, wherein the TERT540peptide comprises ILAKFLHWL (SEQ ID NO.: 15).

43. A pharmaceutical preparation comprising:(a) a pharmaceutically acceptable carrier; and(b) antibody or antigen binding fragment thereof of any one of claims 1-42.

44. A method for treating a subject having a cancer, comprising administering to a subject in need thereof a therapeutically effective amount of the pharmaceutical preparation of claim 4345. The method of claim 44, wherein the cancer is selected from pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, lung cancer, colon cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease, nonHodgkin's lymphoma, cancer of the esophagus, cancer of the gastroesophageal junction, cancer of the small intestine, ampullary cancer, hepatocellular carcinoma, cholangiocarcinoma, intrahepatic cholangiocarcinoma, gallbladder cancer, biliary tract cancer, mesothelioma, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, chronic or acutePatent Application leukemias including acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, solid tumors of childhood, lymphocytic lymphoma, myelodysplastic syndromes, myeloproliferative neoplasms, multiple myeloma, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, neoplasm of the central nervous system (CNS), glioma, high-grade glioma, glioblastoma, astrocytoma, oligodendroglioma, ependymoma, medulloblastoma, neuroblastoma, primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma, environmentally induced cancers including those induced by asbestos, and combinations of said cancers.

46. A method for treating an autoimmune disease, allergy, and / or disorder associated with chronic inflammation in a subject, wherein the method comprises providing to a subject in need a therapeutically effective amount of the pharmaceutical preparation of claim 43.

47. The method of claim 46, wherein the autoimmune disease, allergy, or disorder associated with chronic inflammation is selected from chronic obstructive pulmonary disease, diabetes, systemic atherosclerosis, allergies, rheumatoid arthritis, inflammatory bowel disease (IBD), alcoholic liver disease, chronic renal disease, and Systemic Lupus Erythematosus (SLE).

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