Multivalent single-partite antibodies for dual-antigen restricted immunotherapy

Abstract

The present invention relates to antibody constructs and such constructs for use as a medicament and in immunotherapy to treat cancer, or mixed tumours, benign tumours, or allergic diseases, or infection diseases, or autoimmune diseases. Moreover, the present invention relates to nucleic acid molecule(s) encoding said antibody constructs. The present invention also relates to vectors comprising the nucleic acid molecule(s). Furthermore, the present invention relates to pharmaceutical compositions comprising said antibody constructs. Moreover, the present invention relates to diagnostic compositions in vivo comprising said antibody constructs. Furthermore, the present invention relates to a diagnostic kit in vitro. Moreover, the present invention relates to a kit comprising said antibody constructs.

Description

[0001] Multivalent single-partite antibodies for dual-antigen restricted immunotherapy

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to antibody constructs and such constructs for use as a medicament and in immunotherapy to treat cancer. Moreover, the present invention relates to nucleic acid molecule(s) encoding said antibody constructs. The present invention also relates to vectors comprising the nucleic acid molecule(s). Furthermore, the present invention relates to pharmaceutical compositions comprising said antibody constructs. Moreover, the present invention relates to a kit comprising said antibody constructs.

[0004] BACKGROUND

[0005] Competitive dual and / or multi antigen restricted Immunotherapies

[0006] There is a large diversity of different dual- or multi-antigen targeting antibodies, antibody fragments, polypeptides, and genetically engineered immune cells (1 , 2, 6, 7, 8, 67, 68). The large majority of them are not restricted to a combination of two or more target antigens. Therefore, to gain on specificity, different dual- or multi-antigen restricted T cell-engaging formats are under development.

[0007] (i) For the CAR T cell format, a dual-promotor technique is claiming to be dual antigen specific (9). This format is using two different plasmid reporters to activate the CAR T cells, putting the promotor for the CAR receptor under the control of a synthetic Notch receptor. Once this dual targeting CAR T cell is activated, in a sequel of binding to two different antigens and promotor activation, it can bind and kill also cells expressing only one of the target antigens, not really discriminating between dual and double antigen positive cells.

[0008] (ii) Another multi targeting technology is combining a modular protein system with different antigen specific binders, using artificial latching orthogonal cage-key proteins (LOCKR), in combination with CAR T cells or antibody formats, for restricted multi target binding (10).

[0009] (iii) Using an antibody format, the hemibody technology is the most advanced dual antigen restricted immunotherapy (11 , 12, 13). Two hemibody constructs bind and reconstitute a tri-specific antibody complex on the surface of a dual antigen positive target cell. Each hemibody is composed of an antigen binding single-chain variable fragment (scFvl or scFv2) fused to either the variable heavy (VH) or the variable light chain (VL) domain of a T cell-activating anti-CD3 antibody. When a complementary pair of hemibodies binds the respective antigens on the surface of a target cell, the VH and VL domains become aligned, re-associate and reconstitute the original CD3- binding site. This way, CD3-positive T lymphocytes become activated and retargeted for target cell destruction. In the patent application WO 94 / 09131 are different antibody formats reported, targeting two different antigens, but also using two separate antibody fragments similar to the hemibodies.

[0010] (iv) An alternative antibody format is the Prodrug-Activating Chain Exchange (PACE) (54). This technique is using the antibody-domain exchange reaction, to redirect two inactive antibodies against two different target antigens. When the two antibodies are close together, the non-functional anti-CD3 Fvs fused to lgG-CH3 domains drive CH3 chain-exchange reaction between them, forming now an active anti-CD3 binding site to recruit T cells for target cell destruction. The drawback of this PACE format is, after the lgG-CH3 chain-exchange, the active antibody dimercomplex is in the tissue and bloodstream, with the ability to bind to single antigen positive cells causing significant toxicity.

[0011] (v) The antibody format of the Demibodies is using at least two molecules, each specific for a different antigen on a target cell. Each Demibody has a dimerization domain like a leucine zipper, receptor-ligand (like cytokine and cytokine receptor), actin and an actin-binding protein or DNA aptamer. The binding to the target antigens at the cell surface and the dimerization is required in order to form an active complex (73). The drawbacks are, at high enough concentration the dimerization can happen without binding to target antigens, and after the active Demibody dimercomplex is formed, it can bind to single antigen positive cells causing serious side effects.

[0012] (vi) The Precision Guided Antibody Tumor Engager (PrecisionGATE™) platform (Revitope Oncology, Inc.) is similar to the hemibody technology, using two antibodies directed against two different antigens with a split anti-CD3 effector domain (55). In the Revitope format, each construct with a split anti-CD3 VH or VL domain is connected via a linker, containing a specific protease cutting site, to a blocking peptide and therefore cannot activate T cells by themselves. Once the blocking peptide is removed by proteases, the two PrecisionGATE constructs function in the same way as the hemibodies do, recruiting effector T cells against dual antigen positive target cells (56).

[0013] (vii) There are several more antibody formats using conditional T cell activation by proteolytic cleavage to extend the therapeutic window for targeting antigens also expressed on healthy vital tissues (57, 58, 59). These formats are using single constructs, or two and more separate constructs directed against single, double or multi target antigens. All constructs are able to bind to target antigens, but the T cell recruiting domain is inactive. In the tumor micro-environment, with high protease activity, the inhibitory or blocking anti-CD3 domains are removed, allowing now the constructs to bind and activate T cells.

[0014] Drawbacks of all current dual- or multi antigen restricted immunotherapies

[0015] For all current dual and / or multi-antigen restricted immunotherapies there are several major drawbacks.

[0016] All formats, which are using an irreversible activation step, like antibody-domain exchange reactions, protease cleaving of a blocking peptide, or dimerization domains can bind after activation to single antigen positive cells, causing "on-target off tumor” site effects.

[0017] For formats using two or more GMP ("Good Manufacturing Practice") certified constructs, this will increase production cost, environmental resources and carbon food print (14). For clinical testing, each of the constructs must be tested and validating separate and in combination. This will increase the cost for the clinical trials, the time and regulatory requirements (15, 16). If there are negative side effects in the patient under therapy, it will be difficult to clearly distinguish what construct caused the negative effect, compromising the combinatorial nature of the formats.

[0018] The open and exposed hydrophobic amino acid residues within the hemibody split effector variable heavy (VH) and the variable light chain (VL) domains are likely in contact with the surrounding aqueous solvent and are thus potentially contributing to protein aggregation. This in turn is recognized by the production host and leads to intracellular degradation of aggregates thereby lowering recombinant GMP production yields. Also, due to unfavorable charge patches in the hemibodies and LOCKR proteins, the binding and transport properties of these constructs in vivo is negatively affected and that can cause a short plasma half-life (17). Also, the non-natural surface of the hemibody split and exposed VH I VL interface, or the LOCKR protein helices, might be recognized by the patient immune system as foreign or dangerous entities. There is a possible presence of T cell epitopes in the hemibody and LOCKR protein format. This could cause a drug immune response, via neutralizing antibodies, jeopardizing the therapeutic effect (18).

[0019] All current antigen restricted formats, the hemibodies and the LOCKR CAR T cells, are only targeting a dual-antigen signature.

[0020] SUMMARY OF THE INVENTION

[0021] It is one object of the present invention to provide improved means to specifically identify and / or eliminate specific types of cells. Moreover, it is an object of the present invention to provide improved means to specifically identify and / or eliminate cells that have a specific combination of two and / or three and / or four specific antigens on their cell surface. It is also another object of the present invention to provide improved means to target a heterogeneous cell population and / or different sub clones with loss of single or multi target antigens.

[0022] Furthermore, it is an object of the present invention to provide improved means to specifically identify and / or eliminate cancerous cells. Furthermore, it is an object of the present invention to provide improved means to specifically identify and / or eliminate cells that belong to a specific cell lineage.

[0023] It is another object of the present invention to provide antibody constructs which can be produced in an improved way. Furthermore, it is an object of the present invention to provide means for improved (e.g.: more efficacious and / or safer) T cell therapy.

[0024] The present invention achieves the above objects by providing the antibody constructs of the invention.

[0025] The present invention also relates to antibody constructs and such constructs for use as a medicament and in immunotherapy to treat cancer, or mixed tumours, benign tumours, or allergic diseases, or infection diseases, or autoimmune diseases. Moreover, the present invention relates to nucleic acid molecule(s) encoding said antibody constructs. The present invention also relates to vectors comprising the nucleic acid molecule(s). Furthermore, the present invention relates to pharmaceutical compositions comprising said antibody constructs. Moreover, the present invention relates to diagnostic compositions in vivo comprising said antibody constructs. Furthermore, the present invention relates to a diagnostic kit in vitro. Moreover, the present invention relates to a kit comprising said antibody constructs. To improve T cell mediated immunotherapy, the inventors designed the antibody constructs of the invention (which are also referred to herein as "KeyLock antibodies”), where different antigen targeting domains are integrated into a single construct. In a KeyLock antibody, the variable heavy (VH) and light chain (VL) of an anti-CD3 effector antibody are separated / split from each other by a protein structure comprising at least one target antigen single chain variable fragments (scFv) (see, for instance, Figure 1A). In the absence of target antigen, the adjacent VH and VL domains interact with each other, termed swift-swapping of antibody variable domains, blocking the anti- CD3 VH and VL domains from correctly pairing with each other and therefore preventing the binding and activation of effector T cells (Figure 1 B). Only when all target antigen scFvs are bound to their respective antigens, the blocking of the separated / split anti-CD3 VH and VL domains is abrogated, allowing them to form the original anti CD3-bindi ng site. This way, CD3-positive T lymphocytes become activated and retargeted for target cell lysis in a highly specific manner (Figure 1C). It is expected that this specificity will result in improved safety of the antibody constructs of the invention.

[0026] In addition, a KeyLock antibody can be directed against three different antigens (see, for instance, Figure 2A) or four different antigens (see, for instance, Figure 2B). In the absence of the third or fourth target antigen, the adjacent VH and VL domains swift-swapp with each other, blocking the anti-CD3 VH and VL domains from correctly pairing and therefore preventing the binding and activation of effector T cells. Only when all three target antigens (Figure 20), or four target antigens (Figure 2D), are expressed on the same cell and all target antigen directed scFvs are bound to their respective antigens, the blocking of the separated / split anti-CD3 VH and VL domains is abrogated, allowing them to bind and activate the CD3 receptor complex on T cells for target cell killing.

[0027] An advantage of the invention is, the different target antigen binding domains and the and the split effector binding fragment VH and VL domains, can be arranged in the KeyLock format in different order and orientations from the peptide amino-terminus (N- or NF -terminus) to the carboxyl-terminus (C- or COOH-terminus) (see, for instance, Figure 3). This allows an optimization of KeyLock antibodies depending on target antigen density, target antigen extra cellular size and structure, antibody affinity to the respective antigen and the interaction to the connected other target antigen VH or VL domains.

[0028] Also an advantage of the invention is, a large serum protein can be integrated in the format (see, for instance, Figure 4). This will increase the serum half live in vivo, or the tissue penetration, or the protease stability. Thus, in a preferred embodiment of the invention, the antibody constructs of the invention further comprise a domain containing a serum protein.

[0029] In addition, different KeyLock single chain variable fragments (scFv) can be attached to an antibody constant domain (see, for instance, Figure 5). The Ig constant domains will also increase the serum half live in vivo. This also allows to combine two or up to six different KeyLock chains in a single antibody construct, (see, for instance, Figure 6). If six different KeyLock chains, against 4 different target antigens, are fused to pairing immunoglobulin constant chains, a single KeyLock antibody with six different dual-antigen specificities is created (see, for instance, Figure 6D). It is expected that such antibody constructs will allow efficacious targeting of a heterogeneous malignant cell population. The target antigen loss under immunotherapy, like the loss of BCMA on the malignant plasma cells following anti-BCMA CAR T cell therapy, is a major drawback of all current single antigen restricted therapies (77). A KeyLock antibody with six different KeyLock chains, against 4 different target antigens, is still functional against 10 different single or double antigen-loss scenarios.

[0030] Advantageously, the antibody constructs of the invention can be produced at high yields. Furthermore, they are stable against proteases and exhibit a favourable serum stability and thermostability profile (see, for instance, Figures 30 and 32). Furthermore, they have no or low unspecific T cell activation capability in the absence of their respected target antigens, reducing the risk for cytokine storm in tissue with no target cells (see, for instance, Figure 33). It is expected that this specificity will result in improved safety of the antibody constructs of the invention.

[0031] The present invention comprises the following preferred embodiments:

[0032] 1 . An antibody construct comprising

[0033] (i) at least one variable heavy chain (VH) of an anti-CD3 antibody,

[0034] (ii) at least one variable light chain (VL) of the anti-CD3 antibody, and

[0035] (iii) at least two single chain variable fragments (scFvs), wherein the antibody construct comprises the same number of (i) variable heavy chain(s) (VH) of the anti- CD3 antibody and (ii) variable light chain(s) (VL) of the anti-CD3 antibody, wherein each of the variable heavy chain(s) (VH) and the variable light chain(s) (VL) of the anti-CD 3 antibody are connected to each other by a protein structure comprising at least one or more of the single chain variable fragments (scFvs), and wherein in the antibody construct, at least two of the single chain variable fragments (scFvs) are against different target epitopes of the same target antigen or against different target antigens.

[0036] 2. The antibody construct according to item 1 , wherein at least one VH and VL of the anti-C D3 antibody form an anti-CD3 binding domain in the presence of said different target epitopes or target antigens on a cell surface.

[0037] 3. The antibody construct according to item 1 or 2, wherein the antibody construct is a single covalently linked construct.

[0038] 4. The antibody construct according to any one of items 1 to 3, wherein the antibody construct or the protein structure further comprises an immunoglobulin Fc region and / or an immunoglobulin constant region.

[0039] 5. The antibody construct according to item 4, wherein said immunoglobulin Fc region and / or immunoglobulin constant region are an immunoglobulin, preferably an IgG. The antibody construct according to any one of items 1 to 5, wherein the antibody construct or the protein structure further comprises albumin, single albumin-domains, albumin-binding domains, and / or other plasma / serum proteins such as part of membrane proteins cleaved and detected in the plasma / serum. The antibody construct according to any one of items 1 to 6, wherein said different target epitopes or target antigens are tumor and / or cancer epitopes or antigens. The antibody construct according to any one of items 1 to 7, wherein said different target epitopes or target antigens are prostate tissue epitopes or antigens. The antibody construct according to any one of items 1 to 8, wherein said different target epitopes or target antigens are cell type and / or cell lineage specific epitopes or antigens. The antibody construct according to any one of items 1 to 9, wherein said different target epitopes or target antigens are allergens and or auto-epitopes or auto-antigens. The antibody construct according to item any one of items 1 to 10 wherein at least one, two, or all target epitopes or target antigens are selected from the group consisting of CD38, SLAMF7 / CS1 / CD319, ERBB2 / Her2 / NEU / CD340, FGFR1 / CD331 , EGFR, EpCAM / CD326, ERBB3, ERBB4, MET, PDGFRA, PDGFRB, FGFR2, FGFR3, FGFR4, KIT, FLT3, IGF1 R, RET, ROS1 , ALK, TGFBR1 , TGFBR2, ACVR2A, ACVR1 B, LRP5, LRP6, E-Cadherin I CDH1 , N-Cadherin I CDH2, AQP3, AQP1 , AQP4, AQP5, AQP9, CD45 1 PTPRC, CD34, CD1381 SDC1 , CD901 THY1 , CD1231 IL3RA, IL1 RAP, IL2RB I CD122, CD117 / Kit, CD244, CD49F I ITGA6, ABCG2, ESAM, CD10 I MME, CD81 , GHR, PRLR, PSMA, PSCA, CD133, CD166, CD44v6 I CD44, CD29 I ITGB1 , CD24, Lgr5, CEA / CEACAM5, VEGFR2 I KDR, CXCR4 / CD33, CD56 / NCAM1 , CD19, CD20, B7H3 / CD276, L1CAM, CD37, LFA-1 / ITGB2 / CD18, ROR1 , ROR2, CD70, CD123 / IL3RA, IGF1R / CD221, CCR7 I CD197, CCR4 I CD194, TNFRSF9 I 4-1 BB, CTLA4 I CD152, GPC3 I Glypican-3, CADM1 , CD4, B7-h4 I VTCN1 , TYRP1 , PDL1 / CD274, PDCD1 / PD-1, PDCD1 LG2 / PD-L2, BTLA, DLL3, CEACAM1 / CD66a, LFA- 1 I ITGB2, CD22, NRP1 1 CD304, CHRNA1 , GD2 I Disialoganglioside, CTAG1A / NY-ESO-1 , CD70, CD30 I TNFRSF8, CD98hc I SLC3A2, CD52, CD64 I FCGR1A, CD46 I MCP, CD6, CD10 I MME, CD25 I IL2RA, CD40, CD47, CD49a I ITGA1 , CD49c I ITGA3, CD49e I ITGA5, CD51 1 ITGAV, CD55, CD571 B3GAT1 , CD59, CD61 I ITGB3, CDw75 I ST6GAL1 , CD79a, CD82, CD95 I FAS, CD99, CD105 I ENG, CD107a I LAMP1 , CD107b I LAMP2, CD131 I CSF2RB, CD147 I BSG, CD151 , AGTR1 , AGTR2, KRAS, NRAS, HRAS, and BRAF or combinations thereof. The antibody construct according to any one of items 1 to 11, wherein the anti-CD3 antibody is a T cellactivating antibody. The antibody construct according to any one of items 1 to 12, wherein (I) the variable heavy chain (VH) of the anti-CD3 antibody comprises the complementarity determining regions (CDRs) 1 , 2 and 3 comprising the amino acid sequences of SEQ ID NOs: 274, 275, and 276, respectively, and wherein (ii) the variable light chain (VL) of the anti-CD3 antibody comprises the complementarity determining regions (CDRs) 1 , 2 and 3 comprising the amino acid sequences of SEQ ID NOs: 277, 278, and 279, respectively, or wherein (I) the variable heavy chain (VH) of the anti-CD3 antibody comprises the complementarity determining regions (CDRs) 1 , 2 and 3 comprising the amino acid sequences of SEQ ID NOs: 280, 281 , and 282, respectively, and wherein (ii) the variable light chain (VL) of the anti-CD3 antibody comprises the complementarity determining regions (CDRs) 1 , 2 and 3 comprising the amino acid sequences of SEQ ID NOs: 283, 284, and 285, respectively. The antibody construct according to any one of items 1 to 13, wherein (I) the variable heavy chain (VH) of the anti-CD3 antibody comprises the amino acid sequence of SEQ ID NO: 270 or an amino acid sequence at least 95%, at least 98% or at least 99% identical thereto, and wherein (ii) the variable light chain (VL) of the anti- CD3 antibody comprises the amino acid sequence of SEQ ID NO: 271 or an amino acid sequence at least 95%, at least 98% or at least 99% identical thereto, or wherein (I) the variable heavy chain (VH) of the anti- CD3 antibody comprises the amino acid sequence of SEQ ID NO: 272 or an amino acid sequence at least 95%, at least 98% or at least 99% identical thereto, and wherein (ii) the variable light chain (VL) of the anti- CD3 antibody comprises the amino acid sequence of SEQ ID NO: 273 or an amino acid sequence at least 95%, at least 98% or at least 99% identical thereto. The antibody construct according to any one of items 1 to 14, wherein the anti-CD3 binding domain is capable of binding to CD3 on CD3 positive T cells and activate said cells. The antibody construct according to any one of items 1 to 15, wherein the antibody construct is a construct which forms, at concentration range 0.0001 nM to 0.001 nM, range 0.001 nM to 0.01 nM, range 0.01 nM to 0.1 nM, range 0.1 nM to 1 nM, range 1 nM to 10 nM, range 10 nM to 100 nM, range 100 nM to 1000 nM a functional anti-CD3 binding domain in the absence of said different target antigens, 10 fold (1 -log), 100 fold (2-log) or 1000 fold (3-log) lower as in the presence of said different target antigens. The antibody construct according to any one of items 1 to 16, wherein the antibody construct comprises not more than one VH and not more than one VL of the anti-CD3 antibody and not more than two single chain variable fragments (scFvs) against different target antigens. The antibody construct according to any one of items 1 to 16, wherein the antibody construct comprises two VH and two VL, three VH and three VL, or four VH and four VL of the anti-CD3 antibody. The antibody construct according to any one of items 1 to 16, wherein the antibody construct comprises two, three or four single chain variable fragments (scFvs) against different target epitopes or target antigens. The antibody construct according to any one of items 1 to 16, wherein the antibody construct comprises two, three or four single chain variable fragments (scFvs) against different target antigens and comprises two, three or four single chain variable fragments (scFvs) as pairs or triplicates, direct connected to each other or separated at least by one single chain variable fragments (scFvs) against a different target. The antibody construct according to any one of items 1 to 20, wherein the target antigens are ERBB2 / Her2 / NEU / CD340 and EpCAM / CD326 or CD38 and SLAMF7 / CS1 / CD319. The antibody construct according to any one of items 1 to 21, wherein all of the at least one VH and at least one VL of the anti-CD3 antibody are linked to one of said single chain variable fragments (scFvs) comprised by the protein structure by a peptide linker. The antibody construct according to any one of items 1 to 22, wherein the peptide linker allows formation of an anti-CD3 binding domain from at least one VH and VL of the anti-CD3 antibody in the presence of said different target epitopes or target antigens. The antibody construct according to any one of items 1 to 23, wherein the antibody construct is selected from the following (a) to (q):

[0040] (a) an antibody construct comprising the amino acid sequence of SEQ ID NO: 293 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 293,

[0041] (b) an antibody construct comprising the amino acid sequence of SEQ ID NO: 294 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 294,

[0042] (c) an antibody construct comprising the amino acid sequence of SEQ ID NO: 295 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 295,

[0043] (d) an antibody construct comprising the amino acid sequence of SEQ ID NO: 296 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 296,

[0044] (e) an antibody construct comprising the amino acid sequence of SEQ ID NO: 297 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 297,

[0045] (f) an antibody construct comprising the amino acid sequence of SEQ ID NO: 298 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 298,

[0046] (g) an antibody construct comprising the amino acid sequence of SEQ ID NO: 299 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 299,

[0047] (h) an antibody construct comprising the amino acid sequence of SEQ ID NO: 300 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 300,

[0048] (I) an antibody construct comprising the amino acid sequence of SEQ ID NO: 301 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 301 ,

[0049] (j) an antibody construct comprising the amino acid sequence of SEQ ID NO: 302 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 302,

[0050] (k) an antibody construct comprising the amino acid sequence of SEQ ID NO: 303 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 303,

[0051] (l) an antibody construct comprising rises the amino acid sequence of SEQ ID NO: 304 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 304,

[0052] (m) an antibody construct comprising the amino acid sequence of SEQ ID NO: 305 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 305,

[0053] (n) an antibody construct comprising the amino acid sequence of SEQ ID NO: 306 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 306,

[0054] (o) an antibody construct comprising the amino acid sequence of SEQ ID NO: 307 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 307, and

[0055] (p) an antibody construct comprising the amino acid sequence of SEQ ID NO: 308 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 308. The antibody construct according to any one of items 1 to 24, further comprising an additional protein-binding group selected from a single-domain antibody (also known as a Nanobody or VHH), a monobody (derived from fibronectin type III), an anticalin (derived from lipocalins), an affibody (derived from immunoglobulin- binding protein A) and a DARPin (Designed Ankyrin Repeat Protein). A nucleic acid molecule, or a set of nucleic acid molecules, encoding the antibody construct according to any one of items 1 to 25. A recombinant vector, or a set of recombinant vectors, comprising the nucleic acid molecule or set of nucleic acid molecules according to item 26. A recombinant host cell comprising the recombinant vector or set of recombinant vectors according to item 27 and optionally expressing the antibody construct according to any one of items 1 to 25. A pharmaceutical composition comprising one or more antibody constructs according to any one of items 1 to 25. A kit comprising one or more antibody constructs according to any one of items 1 to 25. An antibody construct according to any one of items 1 to 25 for use as a medicament. An antibody construct according to any one of items 1 to 25 for use in immunotherapy to treat cancer. The antibody construct for use according to item 31 or 32, wherein the antibody construct is to be administrated into the patient intra venous, sub cutaneous, intra thecal or intra peritoneal, via injections, short infusion (1 minute to 8 hours) or via a continuous infusion (> 8 hours). The antibody construct for use according to any one of items 31 to 33, wherein the antibody construct is to be administrated locally into a benign tumor or a cancer via injections, to increase local concentrations of the antibody construct and reduce systemic serum levels of the antibody construct. 35. The antibody construct for use according to any one of items 31 to 34, wherein the antibody construct is to be administrated locally into a benign tumor or a cancer via injections, with the antibody construct embedded into a semi solid matrix to delay the antibody construct release over time and to reduce a wash out from the local injection site.

[0056] BRIEF DESCRIPTION OF THE DRAWINGS

[0057] Figure 1 shows the principle of the invention. (A) Design of a KeyLock single chain polypeptide, targeting two different antigens (x and y), with two antibody variable domains anti x (T 1 black spotted) and anti y (T2 black waves) in VL-VH orientation (from the NH2-terminus to COOH terminus), covalently connected with a linker (gray solid line). The anti-CD3 receptor complex directed effector binding domain (F), from an anti-CD3 antibody, is split / separated into the VH fragment (F1 light solid gray) at the NH2-terminus and into the VL fragment (F2 dark solid gray) at the COOH terminus with a short 6 amino acid His-tag (H). The linker, the position and the orientation of the different variable domains allows swift-swapping (SWSW) between these domains, causing the blocking of the F1 and F2 fragments, to pair and build a functional F domain. (B) If a cell expresses one target antigen, only one target antigen specific domain is bound (left T 1 and right T2), and only one oCD3 domain (left F1 and right F2) is not blocked any more. The single F1 and F2 domains are not able to restore the biological effector function. (C) If a cell expresses both antigens (X and Y) at its cell surface, simultaneous binding of both T1 and T2 domains to the surface of the cell is abrogating the blocking of both F1 and F2 fragments, bringing them in close proximity, causes association of fragments F1 and F2 and restoration of the biological function of the F domain by complementation.

[0058] Figure 2 shows the principle of the invention against a triple- or quadruple-antigen signature. (A) Tri-specific KeyLock antibody. For a tri-specific KeyLock antibody, in a single chain polypeptide format, three different target antigen binding domains (T1 black spots, T2 black waves and T3 black mesh) are connected with the split / separated anti-CD3 receptor complex variable F1 (light solid gray) and F2 (dark solid gray) fragments. The F1 fragment is at the NH2-terminus and the F2 fragment is between the T2 and T3 domains. (B) Quadruple-specific KeyLock antibody. For a quadruple-specific KeyLock antibody, in a single chain polypeptide format, four different target antigen binding domains (T1 black spots, T2 black waves, T3 black mesh and T4 whites spotted) are connected with the split / separated anti-CD3 receptor complex variable F1 (light solid gray) and F2 (dark solid gray) fragments. The F1 fragment is between the T4 and T1 domains and the F2 fragment is between the T2 and T3 domains. If a cell expresses all three (C) or four antigens (D) at its cell surface, simultaneous binding of T 1 +T2+T3 (C) or T1 +T2+T3+T4 (D) domains to the surface of this cell is abrogating the blocking of the F1 and F2 fragments, bringing them in close proximity, causes association of fragments F1 and F2 and restoration of the biological function of the F domain by complementation.

[0059] Figure 3 shows alternative combinations of the antigen binding domains in the antibody construct of the invention. At position C1 and C3 are the split / separated effector domain binding variable F1 (light solid gray) and F2 (dark solid gray) fragments. To the F1 and F2 fragments, different target antigen binding domains (T) can be connected. No T connected, one T connected, two same T connected, two different T connected. At position C2, between the F1 and F2 fragments, different connected target antigen binding domains (T) are possible. Two different T connected, three or more T connected with the same or different target antigen specificity. For a KeyLock antibody, different arranged C1 , C2 and C3 variants are possible. Each C1 peptide can directly connect to each C3 peptide (C1-C3). Each C1 peptide can directly connect to each C2 peptide (C1-C2). Each C3 peptide can directly connect to each C2 peptide (C2-C3). Each C1 peptide can directly connect to each C2 and C3 peptide (C1-C2-C3). Each C1 , C2 and C3 peptide can be connected alone, and / or with a C2 and / or with a C3 peptide, to a serum protein. The serum proteins can be albumin, immunoglobulins (IgG, IgA, IgD, IgM), lipoproteins, receptors, cytokines, hormones.

[0060] Figure 4 shows alternative formats of the invention in connection with serum proteins. (A) At position P1 , P2, P3, P4 and / or P5 can a peptide, full length serum protein, part of a serum protein or serum protein binding domain be connected. (B) The protein is between target antigen binding domains. (C) The protein is at the NH2-terminus (N) connected. (D) The protein is at the COOH terminus (C) connected. The serum proteins can be albumin, immunoglobulins (IgG, IgA, IgD, IgM), lipoproteins, receptors, cytokines, hormones.

[0061] Figure 5 shows examples for alternative immunoglobulin formats of the invention. Top left, sketch of an IgG immunoglobulin with the constant domains, constant light (CL) and constant heavy (CH). Disulfide bridges (black solid lines). Different peptides from C1 , C2 and / or C3 (Figure 3), can be connected to the CH or CL domains. (A) The split / separated anti-CD3 receptor complex variable F1 (light solid gray) and F2 (dark solid gray) fragments are on the same peptide chain, connected to CL, CH1 and / or CH2 domains. (B) The F1 and F2 fragments are on separate / different peptide chains, connected to a CH1 and / or CL domain, of the same antibody. The formation of a correct heterodimer, with pairing F1 and F2 fragments, is building the final KeyLock antibody. (C) The F1 and F2 fragments are on separate / different peptide chains, connected to CH2 domains of the same antibody. The formation of a correct hetero-dimer, with pairing F1 and F2 fragments, is building the final KeyLock antibody.

[0062] Figure 6 shows examples for escaping target antigen-loss by the invention. (A) Against four different target antigens (triangle, pentagon, circle and square) are four specific target antigen binding domains directed (antitriangle = black waves, anti-pentagon = black spots, anti-circle = white spots and anti-square = black mesh). If a target cell is losing one or two of the target antigens on its cell surface, up to ten different antigen loss scenarios are possible. These single or double antigen negative cells can still be targeted with a KeyLock antibody, if different KeyLock chains are connected to an antibody construct. (B) Two different KeyLock binders connected to the CH2 domain. (C) Four different KeyLock binders connected to the CH1 and CL domains. (D) Six different KeyLock binders connected to the CH1, CL and CH3 domains. Specific mutants (black circle) in the CH and CL domains allow for correct pairing and formation of the respected heterodimers. A Key Lock-anti body with six different binders, is able to target all 10 different antigen-loss scenarios, significantly reducing the risk of tumor escape under therapy.

[0063] Figure 7 shows alignment of different antibody VH (A) and VL (B) sequences. Two anti-CD3 receptor complex binders (diL2K and UCHT1), anti-Erbb2 (HER2 / neu), anti-EpCAM, anti-CD38 and anti-SLAMF7 binders. With a grey box the constant frame work (FR) 1 to 4, and with a black box the three CDR regions, of the two anti-CD3 binders are marked. The programm Jalview (© EMBL 2022 | EMBL-EBI) was used. * (asterisk) indicates positions which have a single, fully conserved residue, (colon) indicates conservation between groups of strongly similar properties, roughly equivalent to scoring > 0.5 in the Gonnet PAM 250 matrix, (period) indicates conservation between groups of weakly similar properties, roughly equivalent to scoring =< 0.5 and > 0 in the Gonnet PAM 250 matrix

[0064] Figure 8 shows different orientation of within connected antibody variable domains, VHA / L or VLA / H (polypeptide orientation from the NH2-terminus to the COOH terminus), has an impact on the antibody dissociation constant KD against the respected target antigen. (A) ScFv fragments from the anti CD3 clone UCHT 1 , VH / VL (top, black circles) and VL / VH (bottom, black squares), against the human CD3 epsilon-gamma dimer (EG). For the ELISA assay, a recombinant protein IgG fused with the human CD3 epsilon and CD3 gamma chain, was labeled to a 96-well cell culture plate. After incubating the plates with different concentrations of the anti-CD3-VHA / L and anti-CD3- VL / VH ScFv constructs, washing with PBS, the binding of the ScFv was detected using an HRP-labeled secondary antibody against the 6x His-tag of the ScFv constructs. (B) Quantification of the results shown in (A).

[0065] Figure 9 shows KeyLock antibodies targeting HER2 / neu and EpCAM. Different antigen targeting varaible domains (VH / L). OCD3 VH (light solid gray) and VL domain (dark solid gray), anti-HER2 / neu domain (black spots), anti- EpCAM domain (black waves), flexible glycine-serine linkers (gray solid line), GS-6x histidine tag (H). Constructs T05.2, T06.2, T07.2 and T08.2 with oCD3 sequence from the UCHT 1 clone.

[0066] Figure 10 shows KeyLock antibodies targeting CD38 and SLAMF7. Different antigen targeting varaible domains (VH / VL). OCD3 VH (light solid gray) and VL (dark solid gray) domain, anti-CD38 domain (black spots), anti-SLAMF7 domain (black waves), flexible glycine-serine linkers (gray solid line), GS-6x histidine tag (H). Constructs T01 , T02, T03 and T04 with oCD3 sequence from the diL2K clone.

[0067] Figure 11 shows the recombinant protein production of the KeyLock antibody T05.2 targeting HER2 / neu and EpCAM using eukaryote cells. Expi-HEK293 cells are transfected with pCEP4 Mammalian Expression Vector (Invitrogen ™) carrying the respective KeyLock antibody sequence. Five days after transfection, the cell culture supernatant is harvested, dialyzed against 1 x PBS (pH 7.4 at 4 °C overnight) and the KeyLock antibody, carrying a His-tag at the C terminus, are extracted and purified with CO-IMAC beads (at 4 °C for 2 h in rotation). The fraction not binding to the CO-IMAC beads is the flow through (FT). The protein-bead complex is washed with CO-IMAC Loading buffer with 20 column volumes. Samples from the consecutive washing steps are the wash 1-2 (W1 and W2). For eluting the KeyLock antibodies, CO-IMAC Elution buffer with two column volumes is used for 3 consecutive times (E1, E2 and E3). The different wash and elution fractions are loaded onto a 10% SDS-PAGE Gel and Coomassie Brilliant Blue (CBB) stained. Abbreviations: FT = Flow through, W1-2 = Wash 1-2, E1-3 = Elution 1-3, kDa = Kilodalton; Figure 12 shows the recombinant protein production of the KeyLock antibody T06.2 targeting HER2 / neu and EpCAM using eukaryote Expi-HEK293 cells. Abbreviations: FT = Flow through, W1-2 = Wash 1-2, E1-3 = Elution 1-3, kDa = Kilodalton;

[0068] Figure 13 shows the recombinant protein production of the KeyLock antibody T07.2 targeting HER2 / neu and EpCAM using eukaryote Expi-HEK293 cells. Abbreviations: FT = Flow through, W1-2 = Wash 1-2, E1-3 = Elution 1-3, kDa = Kilodalton;

[0069] Figure 14 shows the recombinant protein production of the KeyLock antibody T08.2 targeting HER2 / neu and EpCAM using eukaryote Expi-HEK293 cells.

[0070] Figure 15 shows the recombinant protein production of the KeyLock antibodies T01 and T02 targeting CD38 and SLAMF7 using eukaryote Expi-HEK293 cells. Abbreviations: FT = Flow through, W1-2 = Wash 1-2, E1-3 = Elution 1-3

[0071] Figure 16 shows the recombinant protein production of the KeyLock antibodies T03 and T04 targeting CD38 and SLAMF7 using eukaryote Expi-HEK293 cells. Abbreviations: FT = Flow through, W1-2 = Wash 1-2, E1-3 = Elution 1-3

[0072] Figure 17 shows the final purified KeyLock antibodies, targeting HER2 / neu and EpCAM, used for in vitro experiments. After CO-IMAC extraction and elution of the KeyLock proteins, the elution fractions are desalted and the protein was concentrated and buffer exchanged into storage buffer. The final KeyLock protein purity and concentration was analysed, loading them onto a 10% SDS-PAGE Gel, with a bovine serum albumin protein standard (BSA) and CBB stained.

[0073] Figure 18 shows the final purified KeyLock antibodies, targeting CD38 and SLAMF7, used for in vitro experiments.

[0074] Figure 19 pCold_T02 bacterial clone screening. Fig 19 shows recombinant production of KeyLock antibody T02, targeting CD38 and SLAMF7, in prokaryote E. coll cells. SHuffle® T7 Express Competent E. coll cells are transfected with pCOLD™ IV bacterial Expression Vector (TaKaRa Bio Inc) carrying the respective KeyLock antibody sequence. (A) For each bacteria clone, the cell pellet (P) and cell supernatant after cell disruption (S), a fraction was loaded onto a 10% SDS-PAGE Gel (reducing) and total protein load was analysed with a CBB stain (top row). (B) To analyze what protein bands are from the KeyLock antibodies, a Western blot was performed with unstained 10% SDS-PAGE Gels and protein lysate from the different clones (bottom row). The Western blot membranes are stained with an anti-His-tag antibody (1 :3000 anti-6xHis) to detect the KeyLock antibodies. The colorimetric detection system used was alkaline phosphatase.

[0075] Figure 20 A to C shows extracted KeyLock antibodies T02, T03 and T04, targeting CD38 and SLAMF7, produced in E. coll cells. For each KeyLock antibody, a bacteria clone is growing in 2x YT medium supplemented with kananmycin and carbenicilin and 0.2% glucose. KeyLock protein production was induced adding 0.1 mM IPTG and incubated at 16°C for 18 - 20 hours. Afterward from the E. coli cell pellet the antibodies are extracted using cobalt beads. After incubation with the cobal beads at 4° C over night, the beads are loaded onto a purification column and washed with CO-IMAC Loading, washing and elution Buffer. All fractions were loaded onto a SDS PAGE gel to check the KeyLock protein production. (A) KeyLock antibody T02 production, (B) KeyLock antibody T03 production and (C) KeyLock antibody T04 production.

[0076] Figure 21 shows the final purified KeyLock antibodies T02, T03 and T04, targeting CD38 and SLAMF7, produced in E. coli cells, used for in vitro experiments. From the column elution fractions, the fractions with the highest and most clean KeyLock protein concentrations are pooled and loaded onto a 10% SDS PAGE gel with a BSA standard to determine total protein concentrations. The production yield per 1 liter (L) bacterial culture, with an optical density (OD) between 1.0 and 2.0, and the yield per E. coli cells, was calculated.

[0077] Figure 22 shows KeyLock antibodies targeting HER2 / neu and EpCAM, linked to human albumin. (A) Sketch of single chain antibody format with the full-length human albumin (P02768, amino acid 19 - 601) covalent attached at the NH2-terminus, via a long (58 amino acids) flexible glycine-serine linker (gray solid line). (B) Recombinant production yields of Albumin-KeyLock fusion antibodies in eukaryote cells. Final purified protein was loaded onto a 10% SDS-PAGE Gel and CBB stained.

[0078] Figure 23 shows for different KeyLock antibodies (i.e. T05.2, T06.2 and T07.2 according to the invention), no engagement of T cells against single antigen positive cells. Briefly, three different KeyLock antibodies, produced in eukaryote cells, with antibody variable domains in different orientations, were titrated to co-cultures of HER2 / neu (A) or EpCAM (B) single positive and luciferase positive CHO-cells and T cells as described. After two days coculture, the viability of the CHO cells was measured, using a luciferase substrate. As positive and negative controls were used, two bi-specific antibodies (BITE_Her2 = aCD3-aHER2 / neu and BITE_EpCAM = aCD3-aEpCAM), a final 5% Dimethylsulfoxid (DMSO) solution, T cells and target cells combination and target cells only. The data represent the means of three independent experiments. These results show that, using antigen specific VL and VH domains and to connect them with split / separated anti-CD3 VL and VH domains in a single antibody format, is blocking FvCD3 formation, if not both target antigens are expressed on the target cells. The BITE antibodies used in this Figure were as follows: Figure 23A: BITE_Her2 = SEQ ID NO: 325 + SEQ ID NO: 329; BITE_EpCAM = SEQ ID NO: 325 + SEQ ID NO: 328; Figure 23B: BITE_Her2 = SEQ ID NO: 325 + SEQ ID NO: 329; BITE_EpCAM = SEQ ID NO: 325 + SEQ ID NO: 328.

[0079] Figure 24 shows specific killing of dual antigen positive target cells with different KeyLock antibodies in a dose dependent manner (i.e. T05.2, T06.2 and T07.2 according to the invention). Briefly, three different KeyLock antibodies, produced in eukaryote cells, two different bi-specific antibodies (BITE_Her2 = aCD3-aHER2 / neu and BITE_EpCAM = aCD3-aEpCAM) and a final 5% Dimethylsulfoxid (DMSO) solution, were titrated to co-cultures of HER2 / neu and EpCAM double positive and luciferase positive CHO-cells and T cells as described. After two days co-culture, the viability of the CHO cells was measured, using a luciferase substrate. The KeyLock-antibodies T05.2 (aCD3VH-aHER2 / neuVL / VH-aEpCAMVL / VH-aCD3VL), T06.2 (aCD3VH-aEpCAMVL / VH-aHER2 / neuVL / VH-aCD3VL) and T07.2 (aCD3VL-aHER2 / neuVH / L-aEpCAMVH / L-aCD3VH) are dual-antigen-restricted. These results from Figures 23 and 24 show that, using antigen specific VL and VH domains and to connect them with split / separated anti-CD3 VL and VH domains in a single antibody format, is blocking FvCD3 formation. Only in the presence of both respective target antigens, the FvCD3 are able to cooperate in order to reconstitute or complement T cell engaging function. Importantly, the scFvHER2 / neu and scFvEpCAM targeting moieties of the KeyLock antibodies could be switched from (VL-VH) to the (VH-VL) orientation, and the anti-CD3 VL and VH domains could be switched from the NH2-terminus to the COOH terminus, clearly indicating the modular character of the invention. The BITE antibodies used in this Figure were as follows: BITE_Her2 = SEQ ID NO: 325 + SEQ ID NO: 329; BiTE_EpCAM = SEQ ID NO: 325 + SEQ ID NO: 328.

[0080] Figure 25 shows for the KeyLock antibodies (i.e. T01 , T02, T03 and T04 according to the invention) no engagement of T cells against single antigen positive cells. Briefly, four different KeyLock antibodies, produced in eukaryote cells, with different variable domain orientation / arrangement, were titrated to co-cultures of CD38 (A) or SLAMF7 (B) single positive and luciferase positive CHO-cells and T cells as described. After two days co-culture, the viability of the OHO cells was measured, using a luciferase substrate. As positive and negative controls were used, a final 5% D imethylsulfoxid (DMSO) solution, T cells and target cells combination and target cells only. The data represent the means of three independent experiments.

[0081] Figure 26 shows specific killing of dual antigen positive target cells for different KeyLock antibody in a dose dependent manner (i.e. T01 , T02, T03 and T04 according to the invention). Briefly, four different KeyLock antibodies, produced in eukaryote cells and a final 5% Dimethylsulfoxid (DMSO) solution, were titrated to cocultures of CD38 and SLAMF7 double positive and luciferase positive CHO-cells and T cells as described. After two days co-culture, the viability of the OHO cells was measured, using a luciferase substrate. The KeyLock- antibodies T02 (aCD3VH-aSLAMF7VL / VH-aCD38VL / VH-aCD3VL), T03 (aCD3VL-aCD38VH / VL-aSLAMF7VH / VL- OCD3VH) and T04 (OCD3VL- OSLAMF7VH / L-CICD38VH / L-CICD3VH) are dual-antigen-restricted. These results from Figures 25 and 26 show that, using antigen specific VL and VH domains and to connect them with split / separated anti-CD3 VL and VH domains in a single antibody format, is blocking FvCD3 formation. Only in the presence of both respective target antigens, the FvCD3 are able to cooperate in order to reconstitute or complement T cell engaging function. Importantly, the scFvCD38 and scFvSLAMF7 targeting moieties could be switched from (VL-VH) to the (VH-VL) orientation / configuration, and the anti-CD3 VL and VH domains could be switched from the NH2-terminus to the COOH terminus, clearly indicating that the modular character of the constructs allows replacement of a targeting moiety by another targeting moiety with desired specificity.

[0082] Figure 27 shows specific killing of dual antigen positive target cells with different KeyLock antibodies in a dose dependent manner (i.e. T02, T03 and T04 according to the invention) recombinant produced in prokaryotic cells. Briefly, three different KeyLock antibodies, produced in E. coll cells, a bi-specific antibody produced in eukaryotic HEK293 cells (KiH-aCD3-aCD38), and a final 5% Dimethylsulfoxid (DMSO) solution, were titrated to co-cultures of CD38 and SLAMF7 single (A and B) and double (C) positive and luciferase positive CHO-cells and T cells as described. After two days co-culture, the viability of the CHO cells was measured, using a luciferase substrate. At low nM concentration, the KeyLock-antibodies T03 (OCD3VL-CICD38VH / L-CISLAMF7VH / L-CICD3VH) and T04 (OCD3VL- OSLAMF7VH / VL-CICD38VH / VL-CICD3VH) are dual-antigen-restricted. The BiTE antibodies used in this Figure were as follows: Figure 27A: KiH_aCD38 = SEQ ID NO: 325 + SEQ ID NO: 327; Figure 27B: KiH_aCD38 = SEQ ID NO: 325 + SEQ ID NO: 327; Figure 270: KiH_aCD38 = SEQ ID NO: 325 + SEQ ID NO: 327.

[0083] Figure 28 shows side by side comparison of specific killing of dual antigen positive target cells for eukaryotic and prokaryotic produced KeyLock antibodies in a dose dependent manner (i.e. T03 and T04 according to the invention). Briefly, two different KeyLock antibodies, produced in prokaryotic E. coli cells (pColdJ and eukaryotic HEK293 cells (pCEP4_), a bi-specific antibody produced in eukaryotic HEK293 cells (KiH-aCD3-aCD38), and a final 5% Dimethylsulfoxid (DMSO) solution, were titrated to co-cultures of CD38 and SLAMF7 single (A, B) and double (C) positive and luciferase positive CHO-cells and T cells as described. After two days co-culture, the viability of the CHO cells was measured, using a luciferase substrate. At low nM concentration the KeyLock- antibodies T03 (OCD3VL-CICD38VH / VL-CISLAMF7VH / L-CICD3VH) and T04 (oCD3VL- OSLAMF7VH / L-CICD38VH / L- OCD3VH) are dual-antigen-restricted. These results show that, independent of the cellular origin of the KeyLock antibody production, the KeyLock antibodies are dual-antigen-restricted. This clearly indicates the overall stability of the constructs and no eukaryotic post-translational modification is necessary for the KeyLock antibody function. The BiTE antibodies used in this Figure were as follows: Figure 28A: KiH_aCD38 = SEQ ID NO: 325 + SEQ ID NO: 327; Figure 28B: KiH_aCD38 = SEQ ID NO: 325 + SEQ ID NO: 327; Figure 28C: KiH_aCD38 = SEQ ID NO: 325 + SEQ ID NO: 327.

[0084] Figure 29 shows side by side comparison of specific killing of dual antigen positive target cells for different KeyLock antibody formats, with or without human albumin attached, in a dose dependent manner (i.e. T02 and T04 according to the invention). Briefly, two KeyLock antibodies in a single chain scFv format (T02 (OCD3VH-CISLAMF7VL / H- OCD38VL / VH-CICD3VL) and T04 (OCD3VL- OSLAMF7VH / L-OCD38VH / L-CICD3VH), and two KeyLock antibodies in a single chain scFv format with the human albumin attached at the NH2-terminus (A1 +T02 (Albumin-oCD3VH- OSLAMF7VL / H-CICD38VL / H-CICD3VL) and A1 +T04 (Albumin-oCD3VL- OSLAMF7VH / L-OCD38VH / L-CICD3VH), two bi-specific antibodies as controls (BiTE_SLAMF7 = aCD3-aSLAMF7, (BiTE_CD38 = aCD3-aCD38), and a final 5% Dimethylsulfoxid (DMSO) solution, were titrated to co-cultures of CD38 and SLAMF7 single (A and B) and double (C) positive and luciferase positive CHO-cells and T cells as described. After two days co-culture, the viability of the CHO cells was measured, using a luciferase substrate. At low nM concentration KeyLock-antibodies with and without the human albumin attached are dual-antigen restricted. These results show that, integrating large protein structures not directly involved in target antigen binding, has no negative impact on the KeyLock antibody dual-antigen-specificity. This clearly indicates the overall robustness and versatility of the KeyLock antibody format. The BiTE antibodies used in this Figure were as follows: Figure 29A: BiTE_CD38 = SEQ ID NO: 325 + SEQ ID NO: 327; BiTE_SLAMF7 = SEQ ID NO: 325 + SEQ ID NO: 326; Figure 29B: BiTE_CD38 = SEQ ID NO: 325 + SEQ ID NO: 327; BiTE_SLAMF7 = SEQ ID NO: 325 + SEQ ID NO: 326; Figure 290: BITE_CD38 = SEQ ID NO: 325 + SEQ ID NO: 327; BITE_SLAMF7 = SEQ ID NO: 325 + SEQ ID NO: 326.

[0085] Figure 30 shows serum stability of different KeyLock antibodies (i.e. T02, T03 and T04 according to the invention). Briefly, three different KeyLock antibodies, produced in eukaryotic cells, were incubated in fresh human serum at 36°C for 48 hours. Afterward these constructs (-Serum), together with constructs not incubated with human serum, were titrated to co-cultures of CD38 and SLAMF7 single (A, B) and double (C) positive and luciferase positive CHO- cells and T cells as described. As control a bi-specific antibody (KiH-aCD3-aCD38), final 5% Dimethylsulfoxid (DMSO) solution and target cells only or target cells with T cells as described. After two days co-culture, the viability of the CHO cells was measured, using a luciferase substrate. At low nM concentration the incubation with fresh human serum had no negative effect on the dual-antigen-restricted function of the KeyLock-antibodies T02 (OCD3VH-CISLAMF7VL / H-CICD38VL / H-CICD3VL), T03 (aCD3VL-aCD38VH / L-aSLAMF7VH / VL-aCD3VH) and T04 (OCD3VL- OSLAMF7VH / VL-CICD38VH / VL-CICD3VH). These results show that, fresh human serum with all its varius proteases has no negative effect on the KeyLock format with its long flexible protein linkers. This clearly indicates the overall stability of the KeyLock antibodies. The BiTE antibodies used in this Figure were as follows: Figure 30A: KiH_aCD38 = SEQ ID NO: 325 + SEQ ID NO: 327; Figure 30B: KiH_aCD38 = SEQ ID NO: 325 + SEQ ID NO: 327; Figure 30C: KiH_aCD38 = SEQ ID NO: 325 + SEQ ID NO: 327.

[0086] Figure 31 shows size exclusion chromatography (SEC) profiles of four different KeyLock antibodies targeting HER2 / neu and EpCAM, produced in eukaryotic cells. (A) to (D): For each of the constructs next to the SEC profile the protein SDS-gel with CBB staining is shown (small insert pictures). T05.2 (aCD3VH-aHER2 / neuVL / n- a EpCAM VLA / H-aCD3VL), T06.2 (aCD3VH-aEpCAMVL / VH-aHER2 / neuVL / H-aCD3VL), T07.2 (aCD3VL- aHER2 / neuVHA / L-aEpCAMVH / VL-aCD3VH) and T08.2 (aCD3VL-aEpCAMVH / VL-aHER2 / neuVH / VL -aCD3VH) show a mostly monomer fraction in the elution peak. These results shows for different KeyLock antibodies in physiological solution, with different target antigen scFv (VL-VH) and (VH-VL) orientations and split anti-CD3 VL and VH domains at the NH2-terminus or the COOH terminus, the majority is in the monomere fraction. This clearly indicates the overall low protein-aggregation rate of the KeyLock antibody format.

[0087] Figure 32 shows the thermostability of different KeyLock antibodies (i.e. T02, T03 and T04 according to the invention). Three different constructs, produced in prokaryotic cells (pColdJ and eukaryotic cells (pCEP4_) were analyzed. As control the anti-CD38 IgG antibody Daratumumab (Darzalex®) was used. The thermostability of the KeyLock antibodies is high, 54.5°C, independent if they are produced in prokaryotic or eukaryotic cells. All three constructs had similar thermostability (A) T02 (OCD3VH-OSLAMF7VL / H-CICD38VL / H-CICD3VL), (B) T03 (OCD3VL- OCD38VH / L-CISLAMF7VH / L-CICD3VH) and (C) T04 (oCD3VL- OSLAMF7VH / L-CICD38VH / L-CICD3VH). These results shows the thermostability for different KeyLock antibodies in physiological solution, with different target antigen scFv (VL-VH) and (VH-VL) orientations and split anti-CD3 VL and VH domains at the NH2-terminus or the COOH terminus. This clearly indicates the overall stability of the KeyLock antibody format. Figure 33 shows no unspecific T cell activation of different KeyLock antibodies (i.e. T01 , T02, T03, T04, T05.2, T06.2, T07.2 and T08.2 according to the invention). (A) Eight different KeyLock antibodies were tested for their ability to activate cytotoxic T cells in the absence of their respective target antigens at a final concentration of 10 nM. As positive control for T cell activation, a bi-specific antibody aCD3-aCD28 was used. None of the KeyLock constructs were able to activate T cells, in contrast to the positive control. (B) Quantification of the results shown in (A). The data represent the mean of two independent experiments. These are important data, showing that different KeyLock antibodies, against different antigens, with different target antigen scFv (VL-VH) and (VH-VL) orientations and split anti-CD3 VL and VH domains at the NH2-terminus or the COOH terminus, have no unspecific T cell activation in the absence of target cells. This clearly indicates the overall reduced risk of the KeyLock antibody format for unspecific T cell activation and unwanted cytokine release syndrome.

[0088] Figure 34 shows that KeyLock antibodies (i.e. T02 and T05.2 according to the invention) have to bind all their respective target antigens on the surface of a cell to release the blocked antiCD3 domain and complement T cell engaging functions. It is not possible for a KeyLock antibody to be activated via binding with only one arm to a target cell and with the second arm to an antigen in solution. Briefly, KeyLock antibodies at different concentrations, together with titrated soluble human target antigens were co-cultured with single antigen positive and Luziferase positive CHO cells and T cells as described and CHO cell viability was determined through the Luziferase signal after 24 hours cell culture. For soluble target antigens, only the extracellular domain of the respective human antigen was fused the human Ig constant Fc domain, expressed and purified in HEK293 cells. As positive and negative controls were used a final 5% Dimethylsulfoxid (DMSO) solution, T cells and target cells combination and target cells only. A bi-specific antibody against CD38 (aCD3-aCD38) and EpCAM (aCD3-aEpCAM) was used as control for T cell activation and target cell killing. The KeyLock antibody T02 against CD38 and SLAMF7 (OCD3VH- OSLAMF7VL / H-OCD38VL / H-CICD3VL) was not able to redirect T cells against single antigen positive cells in the presence of both (CD38 and SLAMF7) soluble target antigens (A, target cells CD38+) and (B, target cells SLAMF7+), or in the presence of only one target antigen in culture (C, target cells CD38+ and soluble antigen SLAMF7) and (D, target cells SLAMF7+ and soluble antigen CD38). The same imperative, dual antigen expression on target cells for KeyLock antibody effector function, is the case for a different KeyLock antibody T05.2 against Her2 and EpCAM (aCD3VH-aHER2 / neuVL / H-aEpCAMVL / H-aCD3VL). The T05.2 KeyLock antibody is not able to redirect T cells against single antigen positive cells in the presence of both (Her2 / neu and EpCAM) soluble target antigens (E, target cells Her2 / neu+) and (F, target cells EpCAM+), or in the presence of only one target antigen in culture (G, target cells Her2 / neu+ and soluble antigen EpCAM) and (H, target cells EpCAM+ and soluble antigen Her2 / neu). These results clearly indicate that a KeyLock antibody has to bind first with all its target antigen binding domains onto the surface of a cell in order to release the blocked antiCD3 effector domains to restore T cell engaging function. The BITE antibodies used in this Figure were as follows: Figure 34A: BITE_anti-CD38 = SEQ ID NO: 325 + SEQ ID NO: 327; Figure 34B: BITE_anti-CD38 = SEQ ID NO: 325 + SEQ ID NO: 327; Figure 34C: BITE_anti-CD38 = SEQ ID NO: 325 + SEQ ID NO: 327; Figure 34D: BITE_aSLAMF7 = SEQ ID NO: 325 + SEQ ID NO: 326; Figure 34G: BiTE_aHer2 = SEQ ID NO: 325 + SEQ ID NO: 329; Figure 34H: BITE_aEpCAM = SEQ ID NO: 325 + SEQ ID NO: 328.

[0089] Figure 35 shows competitive blocking experiments for the invention. For this KeyLock antibodies (i.e. T02 and T05.2 according to the invention) at different concentrations, together with titrated soluble human target antigens were co-cultured with dual antigen positive and Luziferase positive OHO cells and T cells and OHO viability was determined through the Luziferase signal after 24 hours cell culture. As positive and negative controls were used, a final 5% D imethy Isulfoxid (DMSO) solution, T cells and target cells combination and target cells only, a bi-specific antibody against CD38 (aCD3-aCD38) and a bi-specific antibody against EpCAM (aCD3-aEpCAM). The T cell mediated target cell killing was blocked, in a concentration dependent manner, by soluble target antigen (A, target cells CD38+ and SLAMF+, the soluble antigen CD38) and (B, target cells Her2 / neu+ and EpCAM+, the soluble antigen EpCAM). These results clearly indicate again, that KeyLock antibodies have to bind with all their target antigens binding domains onto the surface of a cell in order to release the blocked antiCD3 effector domains to restore T cell engaging function and target cell killing. Importantly, these results again show that the targeting module can be easily replaced by another module with appropriate specificity. The BITE antibodies used in this Figure were as follows: Figure 35A: BITE_aCD38 = SEQ ID NO: 325 + SEQ ID NO: 327; Figure 35B: BITE_aEpCAM = SEQ ID NO: 325 + SEQ ID NO: 328.

[0090] DETAILED DESCRIPTION OF THE INVENTION

[0091] Definitions and General Techniques

[0092] Unless otherwise defined below, the terms used in the present invention shall be understood in accordance with their common meaning known to the person skilled in the art.

[0093] Literature references referred to herein may be cited by using the full reference, or by using an abbreviation such as a number, for instance, "(76)”, and by specifying the corresponding full reference in the "References” section. All literature reference referred to herein are incorporated by reference for all purposes in their entirety.

[0094] The term "antibody” as used herein refers to any functional antibody that is capable of specific binding to the antigen of interest, as generally outlined in chapter 7 of Paul, W.E. (Ed.).: Fundamental Immunology 2nd Ed. Raven Press, Ltd., New York 1989, which is incorporated herein by reference. Without particular limitation, the term "antibody” encompasses antibodies from any appropriate source species, including chicken and mammalian such as mouse, goat, non-human primate and human. Preferably, the antibody is a humanized antibody. The antibody is preferably a monoclonal antibody which can be prepared by methods well-known in the art. The term "antibody” encompasses an lgG-1 , -2, -3, or -4, IgE, IgA, IgM, or IgD isotype antibody. The term "antibody” encompasses monomeric antibodies (such as IgD, IgE, IgG) or oligomeric antibodies (such as IgA or IgM). The term "antibody” also encompasses - without particular limitations - isolated antibodies and modified antibodies such as genetically engineered antibodies, e.g. chimeric antibodies. Single-chain variable fragments (scFvs) are known in the art. A single-chain variable fragment (scFv) is a fusion protein comprising the variable regions of the heavy (VH) and light chains (VL) of immunoglobulins, connected with a linker peptide of preferably 10 to 25 amino acids. It is understood that in accordance with the invention, the protein N-to-C-terminal order of the VH and VL chains of the scFvs is not particularly limited, i.e., scFvs may comprise the variable regions of the heavy (VH) and light chains (VL) of immunoglobulins in a VH to VL order or in a VL to VH order.

[0095] The nomenclature of the domains of antibodies follows the terms as known in the art. Each monomer of an antibody comprises two heavy chains and two light chains, as generally known in the art. Of these, each heavy and light chain comprises a variable domain (termed V|_] for the heavy chain and V|_ for the light chain) which is important for antigen binding. These heavy and light chain variable domains comprise (in an N-terminal to C-terminal order) the regions FR1 , CDR1 , FR2, CDR2, FR3, CDR3, and FR4 (FR, framework region; CDR, complementarity determining region which is also known as hypervariable region). The identification and assignment of the above- mentioned antibody regions within the antibody sequence is generally in accordance with Kabat et al. (Sequences of proteins of immunological interest, U.S. Dept, of Health and Human Services, Public Health Service, National Institutes of Health, Bethesda, Md. 1983), or Chothia et al. (Conformations of immunoglobulin hypervariable regions. Nature. 1989 Dec 21 -28;342(6252):877-83.), or may be performed by using the IMGT / V-QUEST software described in Giudicelli et al. (IMGT / V-QUEST, an integrated software program for immunoglobulin and T cell receptor V-J and V-D-J rearrangement analysis. Nucleic Acids Res. 2004 Jul 1 ;32(Web Server issue): W435-40.), which is incorporated herein by reference. Preferably, the antibody regions indicated above are identified and assigned by using the IMGTA / -QUEST software.

[0096] As used herein, the terms "binding” or "bind” refer to specific binding to the antigen of interest. Preferably, the Kd value is less than 100 nM, or less than 50 nM, or less than 10 nM, or less than 5 nM, or less than 1 nM, or less than 0.1 nM, or less than 0.01 nM, or less than 0.001 nM or less than 0.0001 nM.

[0097] In the context of the present invention, binding or competitive binding of antibodies or antigen-binding portions or scFvs to the antigen of interest can be measured by using surface plasmon resonance measurements as a reference standard assay, as described below, by using Enzyme-linked Immunosorbent Assay (ELISA) or by other measurements such as cell-based assays.

[0098] The terms "KD” or "KD value” relate to the equilibrium dissociation constant as known in the art. In the context of the present invention, these terms relate to the equilibrium dissociation constant of an antibody or scFv with respect to a particular antigen of interest. The equilibrium dissociation constant is a measure of the propensity of a complex (e.g. an antigen-antibody or antigen-scFv complex) to reversibly dissociate into its components (e.g. the antigen and the antibody or scFv). For the antibody constructs according to the invention and the antibodies and scFvs contained therein, KD are preferably determined by using surface plasmon resonance measurements. The terms "cancer” and "cancer cell” is used herein in accordance with their common meaning in the art (see for instance Weinberg R. et al.: The Biology of Cancer. Garland Science: New York 2006. 850p.).

[0099] The cancers to the treated according to the present invention are not particularly limited. Such cancers are preferably selected from myeloid and or lymphoid origin (e.g., acute myeloid leukemia (AML), AML with defining genetic abnormalities, AML defined by differentiation, myeloid sarcoma, myelodysplastic neoplasms (MDS), myeloproliferative neoplasms (MPN), mastocytosis, chronic myelomonocytic leukemia (CMML), acute leukemia of ambiguous lineage (ALAL), acute leukemia of mixed phenotype (MPAL), histiocyte / dendritic cell neoplasms, precursor B cell neoplasms, b cell lymphoblastic leukaemias / lymphomas, mature B cell neoplasms, neoplastic small lymphocytic proliferation, splenic B cell lymphomas and leukemias, lymphoplastic lymphoma, marginal zone lymphoma, follicular lymphoma, cutaneous follicle centre lymphoma, mantel cell lymphoma, transformed indolent B cell lymphoma, large B cell lymphomas, Burkitt lymphoma, KSHV / HHV8 associated B cell lymphoid prolifarations and lymphoma, lymphoid proliferations and lymphomas associated with immune deficiency and dysregulation, Hodgkin lymphoma, T cell plasma cell dyscrasias (e.g., Multiple Myeloma, plasmacytoma, plasma cell leukemia and amyloidosis), precursor T cell neoplasms, T lymphoblastic leukaemia / lymphoma, mature T cell and NK cell leukaemias, primary cutaneous T cell lymphomas, intestinal T cell and NK cell lymphomas, hepatosplenic T cell lymphoma, anaplastic large cell lymphoma, nodal T follicular helper cell lymphoma, peripheral T cell lymphoma, EBV positive NK / T cell lymphomas, mesenchymal dendritic cell neoplasms, myofibroblastic tumour, spleen vascular stromal tumours) or solid tissue origin (e.g. melanoma, basal cell carcinoma, Merkel cell carcinoma, squamous cell carcinoma of the skin / lung / mouth / nasal cavity / nasopharynx / throat / esophagus or vagina, gastric cancer, colon cancer, pancreatic cancer, liver cancer, rectal cancer, prostate cancer, urothelial cancer, testis cancer, ovarian cancer, endometrial cancer, cervical cancer, brain cancer, breast cancer, renal cell carcinoma, Ewing's sarcoma, non-small cell lung cancer and small cell lung cancer, neuroendocrine carcinoma, soft tissue sarcomas, osteosarcomas, gallbladder carcinomas, papillary adenocarcinoma, Ewing sarcoma).

[0100] The cancers to be treated according to the present invention can be selected, for instance, from solid cancers and hematopoietic malignancies. A "solid cancer” is a cancer which forms one or more solid tumors. Such solid cancers forming solid tumors are generally known in the art. The term "solid cancer” encompasses both a primary tumor formed by the cancer and possible secondary tumors, which are also known as metastases.

[0101] Preferred solid cancers to be treated according to the invention are selected from the group consisting of melanoma, colorectal cancer, prostate cancer, head and neck cancer, urothelial cancer, stomach cancer, pancreatic cancer, liver cancer, testis cancer, ovarian cancer, endometrial cancer, cervical cancer, brain cancer, breast cancer, gastric cancer, renal cell carcinoma, Ewing's sarcoma, non-small cell lung cancer and small cell lung cancer.

[0102] Preferred hematopoietic malignancies cancers to be treated according to the invention are selected from the group consisting of leukemias (e.g., acute myeloid leukemias (AML), myelodysplastic neoplasms (MDS), T cell acute lymphoblastic leukemia (T-ALL), large granular lymphocyte leukemia, and B-cell chronic lymphocytic leukemia (B- CLL)), lymphomas (e.g., Natural killer / T cell lymphoma), and plasma cell dyscrasias (e.g., Multiple Myeloma and amyloidosis).

[0103] The term "cancer immunotherapy” is known in the art and generally relates to a treatment of cancer in which the immune system of the patient is used to treat the cancer. Cancer cells harbor genomic mutations which give rise to cancer cell antigens that are specific to the cancer cells and different from the antigens of non-cancerous cells. Thus, in a preferred aspect of cancer immunotherapy in accordance with the present invention, a cancer immunotherapy is a cancer immunotherapy wherein such cancer cell antigens are recognized by the immune system (e.g., by T cells or NK-cells), and wherein cancer cells expressing these antigens are killed by the immune system. Preferred methods to assess cancer immunotherapy are the methods according to Gouttefangeas C et al.: "Flow Cytometry in Cancer Immunotherapy: Applications, Quality Assurance and Future.” (2015) In: Cancer Immunology: Translational Medicine from Bench to Bedside (N. Rezaei editor). Springer. Chapter 25: pages 471 - 486; and the methods according to Van der Burg SH, et al.: "Immunoguiding, the final frontier in the immunotherapy of cancer.” (2014) In Cancer Immunotherapy meets oncology (CM Britten, S Kreiter, M. Diken & HG Rammensee eds). Springer International Publishing Switzerland p37-51 ISBN: 978-3-319-05103-1.

[0104] As used herein, a "cancer immunotherapy” optionally encompasses a treatment where in addition to the immune system which is used to treat the cancer, additional mechanisms of cancer treatment are used. According to the present invention, a cancer immunotherapy in human patients can also encompass additional treatment effects which are independent from the immune system. Another example of a cancer immunotherapy where additional mechanisms of cancer treatment can be used is a combination therapy with known chemotherapeutic agent(s) and / or small molecule inhibitors (inhibiting enzyme function via reducing / blocking their catalytic activity and / or stabilizing an enzyme inactive state and / or increasing enzyme degradation by binding to them). Such combination therapy with known chemotherapeutic agent(s) or small molecule inhibitors may, for instance, not only include the treatment of cancer in which the immune system is used to treat the cancer but also include a treatment of cancer in which the cancer cells are killed by said chemotherapeutic agent(s) or small molecule inhibitors directly. Another combination therapy could be immune check point inhibitors, blocking inhibitory signals on cancer cells and / or immune cells, to activate T cells and / or to abrogate immune cells inhibitory signals for cancer immunotherapy. Another combination therapy could be small molecules (like all trans retinoic acid) to upregulate antigen expression on cancers cells for immunotherapy. Another combination therapy could be chimeric antigen receptor (CAR) T cells. Another combination therapy could be different types of radiation therapy (e.g. 3D conformal radiation therapy, intensity-modulated radiation therapy (IMRT), volumetric modulated radiation therapy (VMAT), image-guided radiation therapy (IGRT), stereotactic radiosurgery (SRS), brachytherapy, superficial x-ray radiation therapy (SXRT) or intraoperative radiation therapy (IORT)).

[0105] Terms such as "wherein at least one VH and VL of the anti-CD3 antibody form an anti-CD3 binding domain in the presence of said different target epitopes or target antigens on a cell surface” in connection with the invention will be understood and mean that in the presence of said different target epitopes or target antigens on a cell surface, the antibody construct of the invention will bind to said different target epitopes or target antigens on the cell surface, such that least one VH and VL of the anti-CD3 antibody form an anti-CD3 binding domain.

[0106] The variable heavy chain(s) (VH) of the anti-CD3 antibody and variable light chain(s) (VL) of the anti-CD3 antibody are functional variable heavy chain(s) (VH) and functional variable light chain(s) (VL) of the anti-CD3 antibody. Functional VH and VL of the anti-CD3 antibody means that the VH and VL of the anti-CD3 antibody are able to form a functional anti-CD3 binding domain in the presence of the respective target antigens. Said functional anti- CD3 binding domain can bind and activate the CD3 receptor, thereby activating T cells.

[0107] In accordance with the present invention, each occurrence of the term "comprising” may optionally be substituted with the term "consisting of”.

[0108] KeyLock antibody technology

[0109] To improve T cell mediated immunotherapy, the inventors designed a novel antibody based format, termed KeyLock antibodies, where different antigen targeting domains are integrated into a single construct. In a single chain KeyLock antibody the variable heavy (VH) and light chain (VL) of an anti-CD3 effector antibody are separated / split from each other by at least two target antigen single chain variable fragments (scFv) (Figure 1). In an IgG based hetero-dimer KeyLock antibody the variable heavy (VH) and light chain (VL) of an anti-CD3 effector antibody are separated / split from each other by at least one target antigen single chain variable fragments (scFv) (Figure 5B and 5C).

[0110] In the absence of target antigen, the adjacent VH and VL domains interact with each other, termed swift-swapping of antibody variable domains, blocking the anti-CD3 VH and VL domains from correctly pairing with each other and therefore preventing the binding and activation of effector T cells. Only when all target antigen scFvs are bound to their respective antigens, the blocking of the separated / split anti-CD3 VH and VL domains is abrogated, allowing them to align, re-associate, and reconstitute the original anti CD3-binding site. This way, CD3-positive T lymphocytes become activated and retargeted for target cell lysis. Using individual target antigen scFv domains against different antigens, KeyLock antibodies against a dual-, triple- or quadruple-antigen signature can be designed (Figure 1 and 2).

[0111] In the KeyLock antibody format, the individual VH and VL domains are joined by peptide linkers via genetic engineering. Different scFv moieties can be employed as antigen specific modules in different order, relative to the separated / split anti-CD3 VH and VL domains (Figure 3 and Figure 5). Stability of the KeyLock antibodies can be optimized by adding constant immunoglobulin domains, albumin, single albumin-domains, albumin binding domains or plasma proteins in the antibody format in order to mimic naturally occurring antibodies or serum proteins and to simultaneously enhance pharmacokinetic properties like prolonged half-life in serum and protection from proteolytic digestion by proteases (Figure 4). Moreover, stability of the KeyLock format can be enhanced by optimizing the recombinant production. Production cell lines have been established that first produce two or three polypeptides that can be easily reassembled in order to generate a functional drug. Such techniques utilize directed disulphide- bridges, crosslinking reagents to covalently join two different polypeptides, hetero- or homo-dimerization domains like leucine-zipper domains, Fc-domains, coiled-coil domains and others like IgG knob into hole technologies (19, 20, 21 , 22, 23, 24) (Figure 5 and Figure 6). A single KeyLock chain can target at least two different antigens. Two different KeyLock chains, against four different targets, can be fused to a constant antibody Ig chain, creating a single antibody with dual-antigen specificity against two different antigen combinations / antigen signatures (Figure 6B). If six different KeyLock chains, against 4 different target antigens, are fused to pairing immunoglobulin constant chains, a single KeyLock antibody with six different dual-antigen specificities is created (Figure 6D). Such an antibody would be of great importance targeting a heterogeneous malignant cell population. The target antigen loss under immunotherapy, like the loss of BCMA on the malignant plasma cells following anti-BCMA CAR T cell therapy, is a major drawback of all current single antigen restricted therapies (77). A KeyLock antibody with six different KeyLock chains, against 4 different target antigens, is still functional against 10 different single or double antigen-loss scenarios.

[0112] The KeyLock technology is using circular permutation for changing the order of the effector anti CD3 VH and VL domains, but with preserved ability for the separated / split anti CD3 VH and VL domains to interact correctly and to bind and activate T cells. Circular permutation is known to function in a variety of proteins when the original protein N and C termini are fairly close in space, as it is true for the KeyLock format using scFvs and specific linkers, connecting the different antibody VH and VL domains with each other. Circular permutation describes a type of relationship between proteins, whereby the proteins have a changed order of amino acids in their protein sequence (25). Circular permutation can be done by artificially engineering individual antibody binders into a single chain format. The result is a single structure with different connectivity, but overall similar function. The high homology between antibodies VH and VL sequences, caused by the similar amino acid sequences in the framework constant regions (Figure 7), allows the VH and VL domains of antibodies with different antigen specificity to interact with each other, as we term it swift-swapping (SWSW).

[0113] Depending on the location in the polypeptide chain, and the connecting linkers, different SWSW of the individual VH and VL domains is possible. This allows different VH and VL pairs to form (26, 27, 28). Pairing is possible between:

[0114] (I) The variable domains from the same antibody-clone can interact and form heterodimeric VH / VL or VL / VH pairs. These pairs will sustain their original ability to bind their respective target antigen, but with possible changes in their affinity (Figure 8).

[0115] (ii) The variable domains from two different antibody-clones can SWSW and form heterodimeric VH / VL or VL / VH pairs, as well as homodimeric VL / VL pairs or even VHA / H pairs. These pairs will lost their ability to bind their respective target antigen (28, 29). Development of the multivalent KeyLock format

[0116] It is reported for a long time, it is possible to covalently connect split / separated variable domains from different antibody clones with each other, targeting different antigens, to form multivalent sFvs, Diabodies, Demibodies or hemibodies (11 , 29, 30, 50, 73, 74). The split / separated VH and VL domains sustain their ability to pair with their corresponding split / separated VH and / or VL domain, even if they are not on the same construct, and to bind their respective antigens. The original split / separated VH and VL format of the sFvs, bispecific binders and Diabodies was designed to improve recombinant production and not for therapeutic use in vivo against target cells with a dualantigen signature (30, 50, 51 , 74).

[0117] The multivalent Fvs and the Diabodies both are taking advantage of the intermolecular pairing of VH and VL domains. Interestingly only Diabodies with a short (5 amino acid) or no linker connecting the VH and VL domains on the same chain, form functional and bivalent dimers. If a long linker (15 amino acids) is used, the Diabodies form mostly monomers, due to intermolecular pairing of the two VH and VL domains connected to each other on the same chain. These monomers are not able to bind their respective antigens. For the multivalent Fvs it is shown, there is a rapid equilibrium between the interconversion of nonbinding monomers into functional multivalent dimers (29). In the presence of antigen there is a shift towards more functional dimer formation. This demonstrates (i) The interconversion of non-functional into functional VH and VL pairs is transient and reversible, (ii) The presence of an antigen is causing the non-functional VH and VL monomer to disassociate, following the rearrangement of the respective VH and VL domains from two antibody fragments and the formation of a functional dimer. The antigen has generally contact with both VH and VL domains, increasing for the antigen bound VH / VL pair the activation energy barrier to disassociate, (iii) The linker between the VH and VL domains is important for correct pairing.

[0118] Therefore, for novel multivalent dual-antigen restricted binders for immunotherapy, the inventors combined different ideas in a single-partite antibody format: (i) Separating the VH from the VL domain from an antiCD3 effector antibody, (ii) using the connected VH and VL domains from target antigen directed scFvs as blocking domains of the antiCD3 Fv domain, in the absence of their respective target antigens, (iii) abandon proteolytic cleavage sites, allowing flexible linkers to reverse the blocking and release of the antiCD3 Fv effector domain.

[0119] The different ideas behind this new design are: (i) The effector cell directed VH and VL domains are not directly connected to each other (separated / split), but they are still able to successfully pair and reconstitute the original antibody binding site, (ii) The effector cell directed VH and VL domains are covalently connected (in a single chain format, Figure 1 , 2, 3 and 4), or in close proximity (in a IgG format, Figure 5 and 6), to a target antigen directed antibody VH and / or VL domain, (iii) Antibody variable domains against different target antigens can be expressed in the VH-VL and / or VL-VH orientation, (iv) Specific linkers between the different VH and VL domains allow for an optimized SWSW. This improves blocking the effector cell specific VH and VL domains if no target antigen is there for binding, (v) Binding to the target antigens is stabilizing the antigen specific VH and VL pairs, reducing the SWSW with the connected and / or in close proximity effector cell directed VH and VL domains, (vi) Only when all target antigen specific VH and VL domains are bound to a cell expressing all the respective antigens, the effector cell binding domains are free for correct pairing, to recruit effector cells for target cell killing.

[0120] All antibody constructs in accordance with the invention can be obtained by methods known in the art. Such methods include methods for the production of recombinant polypeptides. The antibody constructs of the invention can be expressed in recombinant host cells according to the invention. Recombinant host cells of the invention are preferably mammalian cells such as CHO, Sp2 / 0, NSO, PER. C6 and HEK cells. Recombinant host cells can also be prokaryote cells as Escherichia coll or Corynebacterium.

[0121] For recombinant production it will be understood that the antibody constructs of the invention are meant to optionally include a secretion signal peptide sequence, which is optimized for each producer cell line and antibody construct which is cleaved from the final antibody in the cellular secretion process, into the cell culture medium or into the bacteria periplasm. Similarly, the antibody constructs of the invention are meant to also optionally include affinity tags, e.g. in order to facilitate purification and / or detection, and optional protease cleavage sites between the tag and the polypeptide, e.g. in order to facilitate removal of the tags by protease cleavage in the production process.

[0122] It is also understood that any reference to amino acid sequences referred to herein is meant to encompass not only the unmodified amino acid sequence but also typical posttranslational modifications of these amino acid sequences (e.g., glycosylation, phosphorylation, S-nitrosylation, methylation, deamidation or the clipping of particular amino acids) occurring in cellular expression systems known in the art, including mammalian cells such as CHO and HEK cells.

[0123] Target antigens and scFvs capable of binding to target antigens

[0124] The antibody construct of the present invitations comprises at least two of the single chain variable fragments, wherein at least two of the single chain variable fragments (scFvs) are against different target epitopes of the same target antigen or against different target antigens, i.e., are capable of binding to said different target epitopes of the same antigen or to binding to said different target antigens.

[0125] In one embodiment, said different target epitopes or target antigens may be cancer antigens.

[0126] The target antigens and / or cancer antigens may be selected from CD38 (e.g., UniProtKB - P28907, Gene ID: 952), SLAMF7 / CS1 / CD319 (e.g., UniProtKB - Q9NQ25, Gene ID: 57823), ERBB2 / Her2 / NEU / CD340 (e.g., UniProtKB - P04626, Gene ID: 2064), FGFR1 / CD331 (e.g., UniProtKB - P11362, Gene ID: 2260), EGFR (e.g., UniProtKB - P00533, Gene ID: 1956), EpCAM / CD326 (e.g., UniProtKB - P16422, Gene ID: 4072), ERBB3 (e.g., UniProtKB - P21860, Gene ID: 2065), ERBB4 (e.g., UniProtKB - Q15303, Gene ID: 2066), MET (e.g., UniProtKB - P08581, Gene ID: 4233), PDGFRA (e.g., UniProtKB - P16234, Gene ID: 5156), PDGFRB (e.g., UniProtKB - P09619, Gene ID: 5159), FGFR2 (e.g., UniProtKB - P21802, Gene ID: 2263), FGFR3 (e.g., UniProtKB - P22607, Gene ID: 2261), FGFR4 (e.g., UniProtKB - P22455, Gene ID: 2264), KIT (e.g., UniProtKB - P10721 , Gene ID:3815), FLT3 (e.g., UniProtKB - P36888, Gene ID: 2322), IGF1 R (e.g., UniProtKB - P08069, Gene ID: 3480), RET (e.g., UniProtKB - P07949, Gene ID: 5979), ROS1 (e.g., UniProtKB - P08922, Gene ID: 6098), ALK (e.g., UniProtKB - Q9UM73, Gene ID: 238), TGFBR1 (e.g, UniProtKB - P36897, Gene ID: 7046), TGFBR2 (e.g, UniProtKB - P37173, Gene ID: 7048), ACVR2A (e.g., UniProtKB - P27037, Gene ID: 92), ACVR1 B (e.g., UniProtKB - P36896, Gene ID: 91), LRP5 (e.g., UniProtKB - 075197, Gene ID: 4041), LRP6 (e.g., UniProtKB - 075581, Gene ID: 4040), E-Cadherin I CDH1 (e.g., UniProtKB - P12830, Gene ID: 999), N-Cadherin I CDH2 = (e.g., UniProtKB - P19022, Gene ID: 1000), AQP3 (e.g., UniProtKB - Q92482, Gene ID: 360), AQP1 (e.g., UniProtKB - P29972, Gene ID: 358), AQP4 (e.g., UniProtKB - P55087, Gene ID: 361), AQP5 (e.g., UniProtKB - P55064, Gene ID: 362), AQP9 (e.g., UniProtKB - 043315, Gene ID: 366), CD45 I PTPRC (e.g., UniProtKB - P08575, Gene ID: 5788), CD34 (e.g., UniProtKB - P28906, Gene ID: 947), CD138 1 SDC1 (e.g., UniProtKB - P18827, Gene ID: 6382), CD90 I THY1 (e.g., UniProtKB - P04216, Gene ID: 7070), CD1231 IL3RA (e.g., UniProtKB - P26951, Gene ID: 3563), IL1RAP (e.g., UniProtKB - Q9NPH3, Gene ID: 3556), IL2RB / CD122 (e.g., UniProtKB - P14784, Gene ID: 3560), CD117 I Kit (e.g., UniProtKB - P10721,Gene ID: 3815), CD244 (e.g., UniProtKB - Q9BZW8, Gene ID: 51744), CD49F I ITGA6 (e.g., UniProtKB - P23229, Gene ID: 3655), ABCG2 (e.g., UniProtKB - Q9UNQ0, Gene ID: 9429), ESAM (e.g., UniProtKB - Q96AP7, Gene ID: 90952), CD10 I MME (e.g., UniProtKB - P08473, Gene ID: 4311), CD81 (e.g., UniProtKB - P60033, Gene ID: 975), GHR (e.g., UniProtKB - P10912, Gene ID: 2690), PRLR (e.g., UniProtKB - P16471, Gene ID: 5618), PSMA (e.g., UniProtKB - Q04609, Gene ID: 2346), PSCA (e.g., UniProtKB - 043653, Gene ID: 8000), CD133 (e.g., UniProtKB - 043490, Gene ID: 8842), CD166 (e.g., UniProtKB - Q13740, Gene ID: 214), CD44v6 I CD44 (e.g., UniProtKB - P16070, Gene ID: 960), CD29 I ITGB1 (e.g., UniProtKB - P05556, Gene ID: 3688), CD24 (e.g., UniProtKB - P25063, Gene ID: 100133941), Lgr5 (e.g., UniProtKB - 075473, Gene ID: 8549), CEA / CEACAM5 (e.g., UniProtKB - P06731, Gene ID: 1048), VEGFR2 I KDR (e.g., UniProtKB - P35968, Gene ID: 3791), CXCR4 (e.g., UniProtKB - P61073, Gene ID: 7852), CD33 (e.g., UniProtKB - P20138, Gene ID: 945), CD56 I NCAM1 (e.g., UniProtKB - P13591, Gene ID: 4684), CD19 (e.g., UniProtKB - P15391, Gene ID: 930), CD20 (e.g., UniProtKB - P11836, Gene ID: 931), B7H3 / CD276 (e.g., UniProtKB - Q5ZPR3, Gene ID: 80381), L1CAM (e.g., UniProtKB - P32004, Gene ID: 3897), CD37 (e.g., UniProtKB - P11049, Gene ID: 951), LFA-1 1 ITGB2 1 CD18 (e.g., UniProtKB - P05107, Gene ID: 3689), R0R1 (e.g., UniProtKB - Q01973, Gene ID: 4919), R0R2 (e.g., UniProtKB - Q01974, Gene ID: 4920), CD70(e.g., UniProtKB - P32970, Gene ID: 970), CD123 I IL3RA (e.g., UniProtKB - P26951, Gene ID: 3563), IGF1R / CD221 (e.g., UniProtKB - P08069, Gene ID: 3480), CCR71 CD197 (e.g., UniProtKB - P32248, Gene ID: 1236), CCR41 CD194 (e.g., UniProtKB - P51679, Gene ID: 1233), TNFRSF914-1 BB (e.g., UniProtKB - Q07011, Gene ID: 3604), CTLA41 CD152 (e.g., UniProtKB - P16410, Gene ID: 1493), GPC3 I Glypican-3 (e.g., UniProtKB - P51654, Gene ID: 2719), CADM1 (e.g., UniProtKB - Q9BY67, Gene ID: 23705), CD4 (e.g., UniProtKB - P01730, Gene ID: 920), B7-h4 I VTCN1 (e.g., UniProtKB - Q7Z7D3, Gene ID: 79679), TYRP1 (e.g., UniProtKB - P17643, Gene ID: 7360), CTLA4 I (e.g., UniProtKB - P16410, Gene ID: 1493), PDL1 I CD274 (e.g., UniProtKB - Q9NZQ7, Gene ID: 29126), PDCD1 I PD-1 (e.g., UniProtKB - Q15116, Gene ID: 5133), PDCD1LG2 / PD-L2 (e.g., UniProtKB - Q9BQ51, Gene ID: 80380), BTLA (e.g., UniProtKB - Q7Z6A9, Gene ID: 151888), DLL3 (e.g., UniProtKB - Q9NYJ7, Gene ID: 10683), CEACAM1 1 CD66a (e.g., UniProtKB - P13688, Gene ID: 634), LFA-1 I ITGB2 (e.g., UniProtKB - P05107, Gene ID: 3689), CD22 (e.g., UniProtKB - P20273, Gene ID: 933), NRP1 I CD304 (e.g., UniProtKB - 014786, Gene ID: 8829), CHRNA1 (e.g., UniProtKB - P02708, Gene ID: 1134), GD2 1 Disialoganglioside (CAS Registry Number® 65988- 71-8), CTAG1A / NY-ESO-1 (e.g., UniProtKB - P78358, Gene ID: 246100), CD70 (e.g., UniProtKB - P32970, Gene ID: 970), CD30 I TNFRSF8 (e.g., UniProtKB - P28908, Gene ID: 943), CD98hc I SLC3A2 (e.g., UniProtKB - P08195, Gene ID: 6520), CD52 (e.g., UniProtKB - P31358, Gene ID: 1043), CD641 FCGR1 A (e.g., UniProtKB - P12314, Gene ID: 2209), and CD461 MCP (e.g., UniProtKB - P15529, Gene ID: 4179).

[0127] The target antigens and / or cancer antigens may be intracellular proteins, aberrantly expressed on extra cellular part of the cell plasma membrane domain. These target antigens and / or cancer antigens may be selected from KRAS (e.g., UniProtKB - P01116, Gene ID: 3845), NRAS (e.g., UniProtKB - P01111, Gene ID: 4893), HRAS (e.g., UniProtKB - P01112, Gene ID: 3265), KRAs isoform KRAS4A (NM_033360.4), KRAS isoform KRAS4B (NM_004985.5), HSPD1 (e.g., UniProtKB - P10809, Gene ID: 3329), HSPA5 (e.g., UniProtKB - P11021, Gene ID: 3309), ATP5A1 (e.g., UniProtKB - P25705, Gene ID: 11946), CALR (e.g., UniProtKB - P27797, Gene ID: 811), HSP90B1 (e.g., UniProtKB -P14625, Gene ID: 7184), GANAB (e.g., UniProtKB - Q14697, Gene ID: 23193), EEF1A1P5 (e.g., UniProtKB - Q5VTE0, Gene ID: 158078), TXNDC5 (e.g., UniProtKB - Q8NBS9, Gene ID: 81567), RPS27A (e.g., UniProtKB - P62979, Gene ID: 6233), RAF1 (e.g., UniProtKB - P04049, Gene ID: 5894).

[0128] Preferably, the target antigens and / or cancer antigens may be selected from CD38 (e.g., UniProtKB - P28907, Gene ID: 952), SLAMF7 / CS1 / CD319 (e.g., UniProtKB - Q9NQ25, Gene ID: 57823), ERBB2 / Her2 / NEU / CD340 (e.g., UniProtKB - P04626, Gene ID: 2064) and EpCAM / CD326 (e.g., UniProtKB - P16422, Gene ID: 4072). These target antigens and / or cancer antigens may comprise the following amino acid sequences, respectively:

[0129] SEQ ID NO: 1 - CD38 antigen = P28907

[0130] MANCEFSPVSGDKPCCRLSRRAQLCLGVSILVLILVWLAVWPRWRQQWSGPGTTKRFPETVLARCVKYTEIHP EMRHVDCQSVWDAFKGAFISKHPCNITEEDYQPLMKLGTQTVPCNKILLWSRIKDLAHQFTQVQRDMFTLEDTLL GYLADDLTWCGEFNTSKINYQSCPDWRKDCSNNPVSVFWKTVSRRFAEAACDWHVMLNGSRSKIFDKNSTFG SVEVHNLQPEKVQTLEAWVIHGGREDSRDLCQDPTIKELESIISKRNIQFSCKNIYRPDKFLQCVKNPEDSSCTSEI

[0131] SEQ ID NO: 2 - SLAMF7 antigen = Q9NQ25

[0132] MAGSPTCLTLIYILWQLTGSAASGPVKELVGSVGGAVTFPLKSKVKQVDSIVWTFNTTPLVTIQPEGGTIIVTQNRN RERVDFPDGGYSLKLSKLKKNDSGIYYVGIYSSSLQQPSTQEYVLHVYEHLSKPKVTMGLQSNKNGTCVTNLTCC MEHGEEDVIYTWKALGQAANESHNGSILPISWRWGESDMTFICVARNPVSRNFSSPILARKLCEGAADDPDSSM VLLCLLLVPLLLSLFVLGLFLWFLKRERQEEYIEEKKRVDICRETPNICPHSGENTEYDTIPHTNRTILKEDPANTVYS TVEIPKKMENPHSLLTMPDTPRLFAYENVI

[0133] SEQ ID NO: 3 - ERBB2 antigen = HER2 / neu = P04626 MELAALCRWGLLLALLPPGAASTQVCTGTDMKLRLPASPETHLDMLRHLYQGCQVVQGNLELTYLPTNASLSFL QDIQEVQGYVLIAHNQVRQVPLQRLRIVRGTQLFEDNYALAVLDNGDPLNNTTPVTGASPGGLRELQLRSLTEILK GGVLIQRNPQLCYQDTILWKDIFHKNNQLALTLIDTNRSRACHPCSPMCKGSRCWGESSEDCQSLTRTVCAGGC ARCKGPLPTDCCHEQCAAGCTGPKHSDCLACLHFNHSGICELHCPALVTYNTDTFESMPNPEGRYTFGASCVTA CPYNYLSTDVGSCTLVCPLHNQEVTAEDGTQRCEKCSKPCARVCYGLGMEHLREVRAVTSANIQEFAGCKKIFG SLAFLPESFDGDPASNTAPLQPEQLQVFETLEEITGYLYISAWPDSLPDLSVFQNLQVIRGRILHNGAYSLTLQGLG ISWLGLRSLRELGSGLALIHHNTHLCFVHTVPWDQLFRNPHQALLHTANRPEDECVGEGLACHQLCARGHCWGP GPTQCVNCSQFLRGQECVEECRVLQGLPREYVNARHCLPCHPECQPQNGSVTCFGPEADQCVACAHYKDPPF CVARCPSGVKPDLSYMPIWKFPDEEGACQPCPINCTHSCVDLDDKGCPAEQRASPLTSIISAWGILLVWLGWF GILIKRRQQKIRKYTMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKI PVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVRENRGRLGSQDLLN WCMQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFT HQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSE FSRMARDPQRFWIQNEDLGPASPLDSTFYRSLLEDDDMGDLVDAEEYLVPQQGFFCPDPAPGAGGMVHHRHR SSSTRSGGGDLTLGLEPSEEEAPRSPLAPSEGAGSDVFDGDLGMGAAKGLQSLPTHDPSPLQRYSEDPTVPLPS

[0134] ETDGYVAPLTCSPQPEYVNQPDVRPQPPSPREGPLPAARPAGATLERPKTLSPGKNGWKDVFAFGGAVENPE YLTPQGGAAPQPHPPPAFSPAFDNLYYWDQDPPERGAPPSTFKGTPTAENPEYLGLDVPV

[0135] SEQ ID NO: 4 - EpCAM antigen = P16422

[0136] MAPPQVLAFGLLLAAATATFAAAQEECVCENYKLAVNCFVNNNRQCQCTSVGAQNTVICSKLAAKCLVMKAEMN GSKLGRRAKPEGALQNNDGLYDPDCDESGLFKAKQCNGTSMCWCVNTAGVRRTDKDTEITCSERVRTYWIIIEL KHKAREKPYDSKSLRTALQKEITTRYQLDPKFITSILYENNVITIDLVQNSSQKTQNDVDIADVAYYFEKDVKGESLF HSKKMDLTVNGEQLDLDPGQTLIYYVDEKAPEFSMQGLKAGVIAVIWWIAWAGIWLVISRKKRMAKYEKAEIK EMGEMHRELNA

[0137] In another embodiment, the target antigens may be antigens for targeting prostate exocrine and fibromuscular tissue, including PSMA (e.g., UniProtKB — Q04609, Gene ID: 2346), PSCA(e.g., UniProtKB - 043653, Gene ID: 8000), FGFR1 / CD331 (e.g., UniProtKB - P11362, Gene ID: 2260), FGFR2 (e.g., UniProtKB - P21802, Gene ID: 2263), FGFR3 (e.g., UniProtKB - P22607, Gene ID: 2261), FGFR4 (e.g., UniProtKB - P22455, Gene ID: 2264), CD6 (e.g., UniProtKB - P30203, Gene ID: 923), CD10 / MME (e.g., UniProtKB - P08473, Gene ID: 4311), CD24 (e.g., UniProtKB - P25063, Gene ID: 100133941), CD25 / IL2RA (e.g., UniProtKB - P01589, Gene ID: 3559), CD29 / ITGB1 (e.g., UniProtKB - P05556, Gene ID: 3688), CD38 (e.g., UniProtKB - P28907, Gene ID: 952), CD40 (e.g., UniProtKB - P25942, Gene ID: 958), CD44 (e.g., UniProtKB - P16070, Gene ID: 960), CD46 (e.g., UniProtKB - P15529, Gene ID: 4179), CD47 (e.g., UniProtKB - Q08722, Gene ID: 961), CD49a / ITGA1 (e.g., UniProtKB - P56199, Gene ID: 3672), CD49c / ITGA3 (e.g., UniProtKB - P26006, Gene ID: 3675), CD49e / ITGA5 (e.g., UniProtKB - P08648, Gene ID: 3678), CD51 / ITGAV (e.g., UniProtKB - P06756, Gene ID: 3685), CD55 (e.g., UniProtKB - P08174, Gene ID: 1604), CD56 / NCAM1 (e.g., UniProtKB - P13591 , Gene ID: 4684), CD57 / B3GAT1 (e.g., UniProtKB - Q9P2W7, Gene ID: 27087), CD59 (e.g., UniProtKB - P13987, Gene ID: 966), CD61 / ITGB3 (e.g., UniProtKB - P05106, Gene ID: 3690), CDw75 / ST6GAL1 (e.g., UniProtKB - P15907, Gene ID: 6480), CD79a (e.g., UniProtKB - P11912, Gene ID: 973), CD81 (e.g., UniProtKB - P60033, Gene ID: 975), CD82 (e.g., UniProtKB -P27701, Gene ID: 3732), CD90 / THY1 (e.g., UniProtKB

[0138] - P04216, Gene ID: 7070), CD95 / FAS (e.g., UniProtKB - P25445, Gene ID: 355), CD99 (e.g., UniProtKB

[0139] - P14209, Gene ID: 4267), CD105 / ENG (e.g., UniProtKB - P17813, Gene ID: 2022), CD107a / LAMP1 (e.g., UniProtKB - P11279, Gene ID: 3916), CD107b / LAMP2 (e.g., UniProtKB - P13473, Gene ID: 3920), CD117 / KIT (e.g., UniProtKB - P10721, Gene ID:3815), CD123 / IL3RA (e.g., UniProtKB - P26951, Gene ID: 3563), CD131 / CSF2RB (e.g., UniProtKB - P32927, Gene ID: 1439), CD147 / BSG (e.g., UniProtKB - P35613, Gene ID: 682), CD151 (e.g., UniProtKB - P48509, Gene ID: 977), AGTR1 (e.g., UniProtKB - P30556, Gene ID: 185), AGTR2 (e.g., UniProtKB - P50052, Gene ID: 186), TGFBR2 (e.g., UniProtKB - P37173, Gene ID: 7048) and EpCAM / CD326 (e.g., UniProtKB - P16422, Gene ID: 4072).

[0140] The antibody construct of the present invention comprises single chain variable fragments (scFvs) capable of binding to the above target antigens / cancer antigens. Preferably, the antibody construct comprises scFvs comprising the VH and VL sequences of the following antibodies:

[0141] CD38 (e.g. , UniProtKB - P28907, Gene ID: 952)

[0142] Anti_CD38 antibody - Daratumumab

[0143] VH - SEQ ID NO: 5

[0144] EVQLLESGGGLVQPGGSLRLSCAVSGFTFNSFAMSWVRQAPGKGLEWVSAISGSGGGTYYADSVKGRFTISRD NSKNTLYLQMNSLRAEDTAVYFCAKDKILWFGEPVFDYWGQGTLVTVSS

[0145] VL - SEQ ID NO: 6

[0146] EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPA RFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPTFGQGTKVEIK

[0147] Anti_CD38 antibody - Morphosys Ag 2004

[0148] VH - SEQ ID NO: 7

[0149] QVQLVESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVSNIYSDGSNTFYADSVKGRFTISRD NSKNTLYLQMNSLRAEDTAVYYCARNMYRWPFHYFFDYWGQGTLVTVSS VL - SEQ ID NO: 8

[0150] DIELTQPPSVSVAPGQTARISCSGDNIGNKYVSWYQQKPGQAPVWIYGDNNRPSGIPERFSGSNSGNTATLTIS

[0151] GTQAEDEADYYCSSYDSSYFVFGGGTKLTVL

[0152] Anti_CD38 antibody - Isatuximab

[0153] VH - SEQ ID NO: 9

[0154] QVQLVQSGAEVAKPGTSVKLSCKASGYTFTDYWMQWVKQRPGQGLEWIGTIYPGDGDTGYAQKFQGKATLTA

[0155] DKSSKTVYMHLSSLASEDSAVYYCARGDYYGSNSLDYWGQGTSVTVSS

[0156] VL -SEQ ID NO: 10

[0157] DIVMTQSHLSMSTSLGDPVSITCKASQDVSTWAWYQQKPGQSPRRLIYSASYRYIGVPDRFTGSGAGTDFTFTIS

[0158] SVQAEDLAVYYCQQHYSPPYTFGGGTKLEIK

[0159] SLAMF7 / CS1 / CD319 (e.g., UniProtKB - Q9NQ25, Gene ID: 57823)

[0160] Anti_SLAMF7 antibody - Elotuzumab

[0161] VH -SEQ ID NO: 11

[0162] EVQLVESGGGLVQPGGSLRLSCAASGFDFSRYWMSWVRQAPGKGLEWIGEINPDSSTINYAPSLKDKFIISRDNA

[0163] KNSLYLQMNSLRAEDTAVYYCARPDGNYWYFDVWGQGTLVTVSS

[0164] VL -SEQ ID NO: 12

[0165] DIQMTQSPSSLSASVGDRVTITCKASQDVGIAVAWYQQKPGKVPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTI

[0166] SSLQPEDVATYYCQQYSSYPYTFGQGTKVEIK

[0167] ERBB2 / Her2 / NEU / CD340 (e.g., UniProtKB - P04626, Gene ID: 2064)

[0168] Anti_Her2 antibody - Trastuzumab

[0169] VH -SEQ ID NO: 13

[0170] EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTS

[0171] KNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS

[0172] VL -SEQ ID NO: 14

[0173] DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTIS

[0174] SLQPEDFATYYCQQHYTTPPTFGQGTKVEIKR Anti_Her2 antibody - Pertuzumab

[0175] VH -SEQ ID NO: 15

[0176] EVQLVESGGGLVQPGGSLRLSCAASGFTFTDYTMDWVRQAPGKGLEWVADVNPNSGGSIYNQRFKGRFTLSVD

[0177] RSKNTLYLQMNSLRAEDTAVYYCARNLGPSFYFDYWGQGTLVTVSS

[0178] VL -SEQ ID NO: 16

[0179] DIQMTQSPSSLSASVGDRVTITCKASQDVSIGVAWYQQKPGKAPKLLIYSASYRYTGVPSRFSGSGSGTDFTLTIS

[0180] SLQPEDFATYYCQQYYIYPYTFGQGTKVEIKR

[0181] EGFR (e.q„ UniProtKB - P00533, Gene ID: 1956)

[0182] Anti_EGFR antibody - Cetuximab

[0183] VH -SEQ ID NO: 17

[0184] QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNS

[0185] KSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSA

[0186] VL -SEQ ID NO: 18

[0187] DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSV

[0188] ESEDIADYYCQQNNNWPTTFGAGTKLELKR

[0189] Anti_EGFR antibody - Matuzumab

[0190] VH -SEQ ID NO: 19

[0191] QVQLVQSGAEVKKPGASVKVSCKASGYTFTSHWMHWVRQAPGQGLEWIGEFNPSNGRTNYNEKFKSKATMTV

[0192] DTSTNTAYMELSSLRSEDTAVYYCASRDYDYDGRYFDYWGQGTLVTVSS

[0193] VL -SEQ ID NO: 20

[0194] DIQMTQSPSSLSASVGDRVTITCSASSSVTYMYWYQQKPGKAPKLLIYDTSNLASGVPSRFSGSGSGTDYTFTIS

[0195] SLQPEDIATYYCQQWSSHIFTFGQGTKVEIK

[0196] Anti_EGFR antibody - Necitumumab

[0197] VH -SEQ ID NO: 21

[0198] QVQLQESGPGLVKPSQTLSLTCTVSGGSISSGDYYWSWIRQPPGKGLEWIGYIYYSGSTDYNPSLKSRVTMSVD

[0199] TSKNQFSLKVNSVTAADTAVYYCARVSIFGVGTFDYWGQGTLVTVSS

[0200] VL -SEQ ID NO: 22 EIVMTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTIS

[0201] SLEPEDFAVYYCHQYGSTPLTFGGGTKAEIKR

[0202] Anti_EGFR antibody - Nimotuzumab

[0203] VH -SEQ ID NO: 23

[0204] QVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADE

[0205] SSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSS

[0206] VL -SEQ ID NO: 24

[0207] DIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTD

[0208] FTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLEIK

[0209] Anti_EGFR antibody - Panitumumab

[0210] VH -SEQ ID NO: 25

[0211] QVQLQESGPGLVKPSETLSLTCTVSGGSVSSGDYYWTWIRQSPGKGLEWIGHIYYSGNTNYNPSLKSRLTISIDT

[0212] SKTQFSLKLSSVTAADTAIYYCVRDRVTGAFDIWGQGTMVTVSS

[0213] VL -SEQ ID NO: 26

[0214] DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYDASNLETGVPSRFSGSGSGTDFTFTIS

[0215] SLQPEDIATYFCQHFDHLPLAFGGGTKVEIKR

[0216] Anti_EGFRvlll antibody

[0217] VH -SEQ ID NO: 27

[0218] LQQSGGGLVKPGASLKLSCVTSGFTFRKFGMSWVRQTSDKRLEWVASISTGGYNTYYSDNVKGRFTISRENAKN

[0219] TLYLQMSSLKSEDTALYYCTRGYSSTSYAMDYWGQGTTVTVSS

[0220] VL - SEQ ID NO: 28

[0221] DIELTQSPASLSVATGEKVTIRCMTSTDIDDDMNWYQQKPGEPPKFLISEGNTLRPGVPSRFSSSGTGTDFVFTIE

[0222] NTLSEDVGDYYCLQSFNVPLTFGDGTKLEIK

[0223] EpCAM / CD326 (e.q„ UniProtKB - P16422, Gene ID: 4072) anti_EpCAM_1 antibody

[0224] VH - SEQ ID NO: 29 EVQLLEQSGAELVRPGTSVKISCKASGYAFTNYWLGWVKQRPGHGLEWIGDIFPGSGNIHYNEKFKGKATLTADK

[0225] SSSTAYMQLSSLTFEDSAVYFCARLRNWDEPMDYWGQGTTVTVSS

[0226] VL - SEQ ID NO: 30

[0227] ELVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGS

[0228] GTDFTLTISSVQAEDLAVYYCQNDYSYPLTFGAGTKLEIK anti_EpCAM_2 antibody

[0229] VH - SEQ ID NO: 31

[0230] DIVMTQSPLSLPVTPGEPASISCRSTKSLLHSDGITYLYWYLQKPGQSPQLLIYQLSNLASGVPDRFSSSGSGTDF

[0231] TLKISRVEAEDEGVYYCAQNLEIPRTFGCGTKLEIKRT

[0232] VL - SEQ ID NO: 32

[0233] QVQLVQSGAEVKKPGESVKISCKASGYTFTNYGMNWVRQQPGQCLKWMGWINTYTGESTYADDFKGRFAFSL

[0234] DTSASTAYLQLSSLRSEDTAVYFCARFAIKGDYWGQGTLVTVSS anti_EpCAM_3 antibody

[0235] VH - SEQ ID NO: 33

[0236] MQVKLQQSGAELVRPGASVKLSCKASGYTFTNYWINWVKQRPGQGLEWIGNIYPSYIYTNYNQEFKDKVTLTVD

[0237] ESSSTAYMQLSSPTSEDSAVYYCTRSPYGYDEYGLDYWGQGTTVTVSS

[0238] VL - SEQ ID NO: 34

[0239] DIELTQSPSSLTVTAGEKVTMNCKSSQSLLNSRNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSG

[0240] TDFTLTISSVQAEDLAVYYCQNDYVYPLTFGAGTKLEIKR

[0241] KIT (e.g., UniProtKB - P10721 , Gene ID:3815)

[0242] Anti_KIT antibody

[0243] VH - SEQ ID NO: 35

[0244] QVQLVQSGAAVKKPGESLKISCKGSGYRFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISAGK

[0245] SISTAYLQWSSLKASDTAMYYCARHGRGYNGYEGAFDIWGQGTMVTVSS

[0246] VL - SEQ ID NO: 36

[0247] AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISS

[0248] LQPEDFATYYCQQFNSYPLTFGGGTKVEIK FLT3 (e.q., UniProtKB - P36888, Gene ID: 2322)

[0249] Anti_FLT3 = Klon-Seq-19+22

[0250] VH - SEQ ID NO: 37

[0251] EVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTR

[0252] DTSTSTVYMELSSLRSEDTAVYYCARGVGAHDAFDIWGQGTTVTVSS

[0253] VL - SEQ ID NO: 38

[0254] DWMTQSPLSLPVTPGEPASISCRSSQSLLHSNGNNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSDT

[0255] DFTLQISRVEAEDVGVYYCMQGTHPAISFGQGTRLEIK

[0256] Anti_FLT3 = Klon-Seq-21+24

[0257] VH - SEQ ID NO: 39

[0258] EVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWARQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTR

[0259] DTSTSTVYMELSSLRSEDTAVYYCARWAAAVADYWGQGTLVTVSS

[0260] VL - SEQ ID NO: 40

[0261] DWMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGT

[0262] DFTLKISRVEAEDVGVYYCMQSLQTPFTFGPGTKVDIK

[0263] IGF1R (e.q., UniProtKB - P08069, Gene ID: 3480)

[0264] Anti_IGF1R

[0265] VH - SEQ ID NO: 41

[0266] EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKS

[0267] TSTAYMELSSLRSEDTAVYYCARAPLRFLEWSTQDHYYYYYMDVWGKGTTVTVS

[0268] VL - SEQ ID NO: 42

[0269] ELTQDPAVSVALGQTVRITCQGDSLRSYYATWYQQKPGQAPILVIYGENKRPSGIPDRFSGSSSGNTASLTITGAQ

[0270] AEDEADYYCK SRDGSGQHLV FGGGTKLTVL

[0271] N-Cadherin I CDH2 = (e.q., UniProtKB - P19022, Gene ID: 1000)

[0272] Anti_N-Cadherin

[0273] VH - SEQ ID NO: 43 MAQVQLVQSGGGLAQPGGSLRLSCAASGFTFSRHAMIWVRQAPGKGLEWVSSISGSSDSTSYADSVKGRFTIS

[0274] RDNSKNTLYLQMNTLRAEDTAVYYCAKATGYSYYYGMD VWGPGTTVTVSSVL

[0275] VL - SEQ ID NO: 44

[0276] DIQMTQSPSSLSASLGDRVTITCRASQGISNYLAWYQQKPGKAPKLLIYGATTLQHGVPSRFSGSGSGTDFSLTIS

[0277] SLQPEDF AIYFC QQAHSFPPTFGGGTKLEK

[0278] CD451 PTPRC (e.g., UniProtKB - P08575, Gene ID: 5788)

[0279] Anti_CD45 = Fred Hutchinson Cancer Center

[0280] VH - SEQ ID NO: 45

[0281] QVQLVESGGGLVQPGGSLKLSCAASGFDFSRYWMSWVRQAPGKGLEWIGEINPTSSTINFTPSLKDKVFISRDN

[0282] AKNTLYLQMSKVRSEDTALYYCARGNYYRYGDAMDYWGQGTSVTVS

[0283] VL - SEQ ID NO: 46

[0284] DIVLTQSPASLAVSLGQRATISCRASKSVSTSGYSYLHWYQQKPGQPPKLLIYLASNLESGVPARFSGSGSGTDF

[0285] TLNIHPVEEEDAATYYCQHSRELPFTFGSGTKLEIK

[0286] Anti_CD45 - RORB_31_7

[0287] VH - SEQ ID NO: 47

[0288] EVQLVESGAEVKKPGASVKVSCKASGYTFTNYIIHWVKQEPGQGLEWIGYFNPYNHGTKYNEKFKGRATLTADK

[0289] SISTAYMELSSLRSEDTAVYYCARSGPYAWFDTWGQGTTVTVSS

[0290] VL - SEQ ID NO: 48

[0291] DILLTQSPATLSLSPGERATFSCRASQNIGTSIQWYQQKTNGAPRLLIRSSSESISGIPSRFSGSGSGTDFTLTISSL

[0292] EPEDFAVYYCQQSNTWPFTFGQGTKLEIK

[0293] Anti_CD45 - RORB_31_8

[0294] VH - SEQ ID NO: 49

[0295] EVQLVESGAEVKKPGASVKVSCKASGYTFTNYIIHWVKQEPGQGLEWIGYFNPYNHGTKYNEKFKGRATLTADK

[0296] SISTAYMELSSLRSEDTAVYYCARSGPYAWFDTWGQGTTVTVSS

[0297] VL - SEQ ID NO: 50

[0298] DILLTQSPATLSLSPGERATLSCRASQNIGTSIQWYQQKPGQAPRLLIRSSSESISGIPSRFSGSGSGTDFTLTISSL

[0299] EPEDFAVYYCQQSNTWPFTFGQGTKLEIK Anti_CD45 - RORB_32_7

[0300] VH - SEQ ID NO: 51

[0301] QVQLVESGAEVKKPGASVKVSCKASGYTFTNYIIHWVKQEPGQGLEWIGYFNPYNHGTKYNEKFKGRATLTADK

[0302] SISTAYMELSSLRSEDTAVYYCARSGPYAWFDTWGQGTTVTVSS

[0303] VL - SEQ ID NO: 52

[0304] DILLTQSPATLSLSPGERATFSCRASQNIGTSIQWYQQKTNGAPRLLIRSSSESISGIPSRFSGSGSGTDFTLTISSL

[0305] EPEDFAVYYCQQSNTWPFTFGQGTKLEIK

[0306] Anti_CD45 - RORB_32_8

[0307] VH - SEQ ID NO: 53

[0308] QVQLVESGAEVKKPGASVKVSCKASGYTFTNYIIHWVKQEPGQGLEWIGYFNPYNHGTKYNEKFKGRATLTADK

[0309] SISTAYMELSSLRSEDTAVYYCARSGPYAWFDTWGQGTTVTVSS

[0310] VL - SEQ ID NO: 54

[0311] DILLTQSPATLSLSPGERATLSCRASQNIGTSIQWYQQKPGQAPRLLIRSSSESISGIPSRFSGSGSGTDFTLTISSL

[0312] EPEDFAVYYCQQSNTWPFTFGQGTKLEIK

[0313] Anti_CD45 - RA_China

[0314] VH - SEQ ID NO: 55

[0315] QVQLKQSGAELAKPGASVKMSCKASGSTFTTYfflHWVKQRPGQGLEffIGYINPNTGYTEYNQKFKAKATLTADKS

[0316] SSTAYMQLSSLTSEDSAVYYCVRFITWGGffGQGTTLTVSS

[0317] VL - SEQ ID NO: 56

[0318] QIVLTQSPAIMSAYLGERVTMTCTASSSVSSSHLHWYQQKPGSSPKLfflYSTSNLASGVPARFSGSGSGTSYSLTI

[0319] SSMETEDAATYYCHQYHRSPLTFGAGTKLELKR

[0320] Anti_CD45 - RO_ Milteniy

[0321] VH - SEQ ID NO: 57

[0322] QITLKESGPTLVKPTQTLTLTCTFSGFSLTTYGIGVGWIRQPPGKALEWLTHIWWNDNKYYSPSLKSRLTITKDSSK

[0323] NQWLTMTNMDPVDTATYYCLYGYTYWGQGTLVTVSA

[0324] VL - SEQ ID NO: 58

[0325] DWMTQTPLSLPVTLGQPASISCRSSQSLLYSNGNTYLHWYQQRPGQSPRLLIYKLSNRFSGVPDRFSGSGSGT

[0326] DFTLKISRVEAEDVGVYYCSQSTHVPWTFGGGTKLEIK CD34 (e.q„ UniProtKB - P28906, Gene ID: 947)

[0327] Anti_CD34 - China

[0328] VH - SEQ ID NO: 59

[0329] QVQLVQSGSELKKPGASVKVSCKASGYTFTNYGMNWVRQAPGQGLEWMGWINTNTGEPKYAEEFKGRFVFSL

[0330] DTSVSTAYLQISSLKAEDTAVYYCARGYGNYARGAWLAYWGQGTLVTVSS

[0331] VL - SEQ ID NO: 60

[0332] DWMTQSPLSLPVTLGQPASISCRSSQTIVHSNGNTYLEWFQQRPGQSPRRLIYQVSNRFSGVPDRFSGSGSGT

[0333] DFTLKISRVEAEDVGVYYCFQGSHVPRTFGGGTKVEIKR

[0334] CD1381 SDC1 (e.g., UniProtKB - P18827, Gene ID: 6382) anti_CD138 antibody - Indatuximab

[0335] VH - SEQ ID NO: 61

[0336] QVQLQQSGSELMMPGASVKISCKATGYTFSNYWIEWVKQRPGHGLEWIGEILPGTGRTIYNEKFKGKATFTADIS

[0337] SNTVQMQLSSLTSEDSAVYYCARRDYYGNFYYAMDYWGQGTSVTVSS

[0338] VL - SEQ ID NO: 62

[0339] DIQMTQSTSSLSASLGDRVTISCSASQGINNYLNWYQQKPDGTVELLIYYTSTLQSGVPSRFSGSGSGTDYSLTIS

[0340] NLEPEDIGTYYCQQYSKLPRTFGGGTKLEIK

[0341] Anti_CD138 - China_Klone58

[0342] VH - SEQ ID NO: 63

[0343] QIQLVQSGPELKKPGETVKISCKASGYTFTDYGMNWVKQAPGKGLKWMVWINTYTGAPTFADDFKGRFALSLET

[0344] SASTAYLQINNLKNEDTATYFCAKSYGWYFDVWGAGTTVTVSS

[0345] VL - SEQ ID NO: 64

[0346] DVLMTQTPLSLPVSLQASISCRSSQSILHSNGNTYLEWYLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFTL

[0347] KISRVEAEDLGIYYCFQGSHVPWTFGGGTKLEIK

[0348] Anti_CD138 - China_Klone59166

[0349] VH - SEQ ID NO: 65 QIQLVQSGPELKKPGETVKISCKASGYTFTDYGMNWVKQAPGKGLKWMVWINTYTGAPTFADDFKGRFALSLET

[0350] SASTAYLQINNLKNEDTATYFCAKSYGWYFDVWGAGTTVTVSS

[0351] VL - SEQ ID NO: 66

[0352] DIQMTQTTSSLSASLGDRVTISCRASQDISNYLNWYQQKPDGTVKLLIYYTSRLHSGVPSRFSGSGSGTDYSLTIS

[0353] NLEPEDIATYYCQQYSKRPffTFGGGTKLEIK

[0354] Anti_CD138 - Klon-58208

[0355] VH - SEQ ID NO: 67

[0356] QIQLVQSGPELKKPGETVKISCKASGYTFTDYGMNWVKQAPGKGLKWMVWINTYTGAPTFADDFKGRFALSLET

[0357] SASTAYLQINNLKNEDTATYFCAKSYGWYFDVWGAGTTVTVSS

[0358] VL - SEQ ID NO: 68

[0359] DVLMTQTPLSLPVSLQASISCRSSQSILHSNGNTYLEWYLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGTDFTL

[0360] KISRVEAEDLGIYYCFQGSHVPWTFGGGTKLEIK

[0361] Anti_CD138 antibody = Ravtansine

[0362] VH - SEQ ID NO: 69

[0363] QVQLQQSGSELMMPGASVKISCKATGYTFSNYWIEWVKQRPGHGLEWIGEILPGTGRTIYNEKFKGKATFTADIS

[0364] SNTVQMQLSSLTSEDSAVYYCARRDYYGNFYYAMDYWGQGTSVTVSS

[0365] VL - SEQ ID NO: 70

[0366] DIQMTQSTSSLSASLGDRVTISCSASQGINNYLNWYQQKPDGTVELLIYYTSTLQSGVPSRFSGSGSGTDYSLTIS

[0367] NLEPEDIGTYYCQQYSKLPRTFGGGTKLEIK

[0368] CD123 / IL3RA (e.g„ UniProtKB - P26951, Gene ID: 3563)

[0369] Anti_CD123 antibody

[0370] VH - SEQ ID NO: 71

[0371] QIQLVQSGPELKKPGETVKISCKASGYIFTNYGMNWVKQAPGKSFKWMGWINTYTGESTYSADFKGRFAFSLET

[0372] SASTAYLHINDLKNEDTATYFCARSGGYDPMDYWGQGTSVTVSS

[0373] VL - SEQ ID NO: 72

[0374] DIVLTQSPASLAVSLGQRATISCRASESVDNYGNTFMHWYQQKPGQPPKLLIYRASNLESGIPARFSGSGSRTDF

[0375] TLTINPVEADDVATYYCQQSNEDPPTFGAGTKLELK CD117 / Kit (e.q„ UniProtKB - P10721 ,Gene ID: 3815)

[0376] Anti_CD117 antibody

[0377] VH - SEQ ID NO: 73

[0378] QVQLVQSGAAVKKPGESLKISCKGSGYRFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISAGK

[0379] SISTAYLQWSSLKASDTAMYYCARHGRGYNGYEGAFDIWGQGTMVTVSS

[0380] VL - SEQ ID NO: 74

[0381] AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISS

[0382] LQPEDFATYYCQQFNSYPLTFGGGTKVEIK

[0383] PSMA (e.g., UniProtKB - Q04609, Gene ID: 2346)

[0384] Anti_PSMA antibody - J591_HC_Seq 1 +10

[0385] VH - SEQ ID NO: 75

[0386] EVQLVQSGAEVKKPGASVKISCKTSGYTFTEYTIHWVKQASGKGLEWIGNINPNNGGTTYNQKFEDRATLTVDKS

[0387] TSTAYMELSSLRSEDTAVYYCAAGWNFDYWGQGTTVTVSS

[0388] VL - SEQ ID NO: 76

[0389] DIVMTQSPSSLSASVGDRVTITCKASQDVGTAVDWYQQKPGKAPKLLIYWASTRHTGVPDRFTGSGSGTDFTLTI

[0390] SSLQPEDFADYFCQQYNSYPLTFGGGTKLEIK

[0391] Anti_PSMA antibody - J591 _2P_Seq2+11

[0392] VH - SEQ ID NO: 77

[0393] EVQLVQSGPEVKKPGATVKISCKTSGYTFTEYTIHWVKQAPGKGLEWIGNINPNNGGTTYNQKFEDKATLTVDKS

[0394] TDTAYMELSSLRSEDTAVYYCAAGWNFDYWGQGTLLTVSS

[0395] VL - SEQ ID NO: 78

[0396] DIQMTQSPSSLSTSVGDRVTLTCKASQDVGTAVDWYQQKPGQSPKLLIYWASTRHTGIPSRFSGSGSGTDFTLTI

[0397] SSLQPEDFADYYCQQYNSYPLTFGAGTKVD I K

[0398] Anti_PSMA antibody - J591_CysDB_Seq9+15

[0399] VH - SEQ ID NO: 79 EVQLVQSGAEVKKPGASVKISCKTSGYTFTEYTIHWVKQASGKGLEWIGNINPNNGGTTYNQKFEDRATLTVDKS

[0400] TSTAYMELSSLRSEDTAVYYCAAGWNFDYWGQGTTVTVSS

[0401] VL - SEQ ID NO: 80

[0402] DIVMTQSPSSLSASVGDRVTITCKASQDVGTAVDWYQQKPGKAPKLLIYWASTRHTGVPDRFTGSGSGTDFTLTI

[0403] SSLQPEDFADYFCQQYNSYPLTFGGGTKLEIK

[0404] PSCA (e.g., UniProtKB - 043653, Gene ID: 8000)

[0405] Anti_PSCA - 2B3parental

[0406] VH - SEQ ID NO: 81

[0407] EVQLVESGGGLVQPGGSLRLSCAASGFNIKDYYIHWVRQAPGKGLEWVAWIDPENGDTEFVPKFQGRATISADT

[0408] SKNTAYLQMNSLRAEDTAVYYCKTGGFWGQGTLVTVSS

[0409] VL - SEQ ID NO: 82

[0410] DIQLTQSPSSLSASVGDRVTITCSASSSVRFIHWYQQKPGKAPKRLIYDTSKLASGVPSRFSGSGSGTDFTLTISSL

[0411] QPEDFATYYCQQWSSSPFTFGQGTKVEIK

[0412] AntLPSCA - 2B3_A2

[0413] VH - SEQ ID NO: 83

[0414] EVQLVESGGGLVQPGGSLRLSCAASGFNIKDYYIHWVRQAPGKGLEWVAWIDPEYGDSEFVPKFQGRATMSAD

[0415] TSKNTAYLQMNSLRAEDTAVYYCKTGGFWGRGTLVTVSS

[0416] VL - SEQ ID NO: 84

[0417] DIQLTQSPSSLSASVGDRVTITCSASSSVRFIHWYQQKPGKAPKRLIYDTSKLASGVPSRFSGSGSGTDFTLTISSL

[0418] QPEDFATYYCQQWGSSPFTFGQGTKVEIK

[0419] AntLPSCA - 2B3_A11

[0420] VH - SEQ ID NO: 85

[0421] EVQLVEYGGGLVQPGGSLRLSCAASGFNIKDYYIHWVRQAPGKGLEWVAWIDPENGDTEFVPKFQGRATMSAD

[0422] TSKNTAYLQMNSLRAEDTAVYYCKTGGFWGQGTLVTVSS

[0423] VL - SEQ ID NO: 86

[0424] DIQLTQSPSTLSASMGDRVTITCSASSSVRFIHWYQQKPGKAPKRLIYDTSKLASGVPSRFSGSGSGTDFTLTISSL

[0425] QPEDFATYYCQQWGSSPFTFGQGTKVEIK Anti_PSCA - 2B3_C5

[0426] VH - SEQ ID NO: 87

[0427] EVQLVESGGGLVQPGGSLRLSCAASGFNIKDYYIHWVRQAPGKGLEWVAWIDPENGDTEFVPKFQGRATISADT

[0428] SKNTVYLQMNSLRAKDTAVYYCKTGGFWGQGTLVTVSS

[0429] VL - SEQ ID NO: 88

[0430] DIQLIQSPSSLSASVGDRVTITCSASSSVRFIHWYQQKPGKAPKRLIYDTSKLASGVPSRFSGSGSGTDFTLTISSL

[0431] QPEDFATYYCQQWSSSPFTFGQGTKVEIK

[0432] CD133 (e.q„ UniProtKB - 043490, Gene ID: 8842)

[0433] Anti_CD133 - AC133

[0434] VH - SEQ ID NO: 89

[0435] QVQLQQSGAELVRPGASVKLSCKASGYTFSDFEMHWVKQTPVHGLEWIGDIDPGTGDTAYNLKFKGKATLTTDK

[0436] SSSTAYMELRSLTSEDSAVYYCTLGAFVYWGQGTLVTVS

[0437] VL - SEQ ID NO: 90

[0438] DVWTQTPLSLPVSFGDQVSISCRSSQSLANSYGNTYLSWYLHKPGQSPQLLIYGISNRFSGVPDRFSGSGSGTD

[0439] FTLKISTIKPEDLGMYYCLQGTHQPYTFGGGTKLEIKRADAAA

[0440] Anti_CD133 - clone /

[0441] VH - SEQ ID NO: 91

[0442] LEVKLVESGPELKKPGETVKISCKASGYTFTDYSMHWVNQAPGKGLKWMGWINTETGEPSYADDFKGRFAFSLE

[0443] TSASTAYLQINNLKNEDTATYFCATDYGDYFDYWGQGTTLTVSSAKTTPPSVTSGQAGQ

[0444] VL - SEQ ID NO: 92

[0445] AQAAELDIVLSQSPAIMSASPGEKVTISCSASSSVSYMYWYQQKPGSSPKPWIYRTSNLASGVPARFSGSGSGTS

[0446] YSLTISSMEAEDAATYYCQQYHSYPPTFGAGTKLELK

[0447] CD166 (e.q„ UniProtKB - Q13740, Gene ID: 214)

[0448] Anti_CD166 antibody

[0449] VH - SEQ ID NO: 93 QITLKESGPTLVKPTQTLTLTCTFSGFSLSTYGMGVGWIRQPPGKALEWLANIWWSEDKHYSPSLKSRLTITKDTS

[0450] KNQWLTMTNMDPVDTATYYCVQIDYGNDYAFTYWGQGTLVTVSS

[0451] VL - SEQ ID NO: 94

[0452] DIVMTQSPLSLPVTPGEPASISCRSSKSLLHSNGITYLYWYLQKPGQSPQLLIYQMSNLASGVPDRFSGSGSGTDF

[0453] TLKISRVEAEDVGVYYCAQNLELPYTFGQGTKLEIK

[0454] CD44v6 1 CD44 (e.g., UniProtKB - P16070, Gene ID: 960)

[0455] Anti_CD44v6 - BIWA 4 / 8aa+ VL BIWA 4

[0456] VH - SEQ ID NO: 95

[0457] EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVSTISSGGSYTYYLDSIKGRFTISRDN

[0458] AKNSLYLQMNSLRAEDTAVYYCARQGLDYWGRGTLVTVSS

[0459] VL - SEQ ID NO: 96

[0460] EIVLTQSPATLSLSPGERATLSCSASSSINYIYWYQQKPGQAPRLLIYLTSNLASGVPARFSGSGSGTDFTLTISSLE

[0461] PEDFAVYYCLQWSSNPLTFGGGTKVEIK

[0462] Anti_CD44v6 - BIWA 4 / 8aa+VL BIWA 8 aa

[0463] VH - SEQ ID NO: 97

[0464] EVQLVESGGGLVKPGGSLRLSCAASGFTFSSYDMSWVRQAPGKGLEWVSTISSGGSYTYYLDSIKGRFTISRDN

[0465] AKNSLYLQMNSLRAEDTAVYYCARQGLDYWGRGTLVTVSS

[0466] VH - SEQ ID NO: 98

[0467] EIVLTQSPATLSLSPGERATLSCSASSSINYIYWLQQKPGQAPRILIYLTSNLASGVPARFSGSGSGTDFTLTISSLE

[0468] PEDFAVYYCLQWSSNPLTFGGGTKVEIK

[0469] CD29 1 ITGB1 (e.g., UniProtKB - P05556, Gene ID: 3688)

[0470] Anti_CD29 antibody

[0471] VH - SEQ ID NO: 99

[0472] EVQLQQSGPEVGRPGSSVKISCKASGYTFTGYILSWVKQSPGQGLEWIGWVDPEYGSTDSAEKFKKRATLTADI

[0473] SSNTAYIQLSSLTSEDTATYFCTRYYDGYYRRWFAYWGQGTLVTVSS

[0474] VL - SEQ ID NO: 100 DIQMTQSPASLSASLGDIVSIECLASEGISNNLAWHQQKPGKSPQLLIYGAHSLHDGVPSRFSGSGSGTQYSLKIS

[0475] GMQPEDEGVYYCQQGYKYPITFGGGTKLELK

[0476] CD24 (e.q„ UniProtKB - P25063, Gene ID: 100133941)

[0477] Anti_CD24 antibody

[0478] VH-SEQIDNO: 101

[0479] MADVHLQESGPDLVKPSQSLSLTCTVTGYSITSGYTWHWIRQFPGNTVEWMGYIQYTGSTRYNPALRGRLSISR

[0480] DTSKNQFFLQLISVTTADTGTYFCARGTTASFDYWGQGTTLTVASAGSA

[0481] VL-SEQIDNO: 102

[0482] DIVMSQSPSSLNVSVGEKVTMRCRSSQSLLYSSDQKNYLTWYQQKPGQSPKLLISWASTRASGVPDRFTGSGS

[0483] GTDFTLTISSVKAEDLGVYYCQQYFIYPLTFGVG

[0484] Lgr5 (e.g., UniProtKB - 075473, Gene ID: 8549)

[0485] Anti_Lgr5 antibody

[0486] VH-SEQIDNO: 103

[0487] MEWSWVFLFFLSVTTGVHSEVQLVQSGAEVKKPGESLRISCKGSGYSFTAYWIEWVRQAPGKGLEWIGEILPGS

[0488] DSTNYNEKFKGHVTISADKSISTAYLQWSSLKASDTAVYYCARSGYYGSSQYWGQGTLVTVSS

[0489] VL-SEQIDNO: 104

[0490] MSVPTQVLGLLLLWLTDARCDIVLTQSPASLAVSPGQRATITCRASESVDSYGNSFMHWYQQKPGQPPKLLIYLT

[0491] SNLESGVPDRFSGSGSGTDFTLTINPVEANDAATYYCQQNAEDPRTFGGGTKLEIK

[0492] CEA / CEACAM5 (e.g„ UniProtKB -P06731, Gene ID: 1048)

[0493] Anti_CEA_antibody

[0494] VH-SEQIDNO: 105

[0495] EVQLVESGGGWQPGRSLRLSCSSSGFDFTTYWMSWVRQAPGKGLEWVAEIHPDSSTINYAPSLKDRFTISRDN

[0496] SKNTLFLQMDSLRPEDTGVYFCASLYFGFPWFAYWGQGTPVTVSS

[0497] VL-SEQIDNO: 106 DIQLTQSPSSLSASVGDRVTITCKASQDVGTSVAWYQQKPGKAPKLLIYWTSTRHTGVPSRFSGSGSGTDFTFTI

[0498] SSLQPEDIATYYCQQYSLYRSFGGGTKVGIK

[0499] VEGFR21 KDR (e.q„ UniProtKB - P35968, Gene ID: 3791)

[0500] Anti_VEGFR2 antibody

[0501] VH - SEQ ID NO: 107

[0502] QEQLMESGGGLVTPGGTLTLTCTASGFTISSYYMSWVRQAPGKGLEYIGLIKTDGTTYYANWAKGRVTISRTSTT

[0503] VDLKMTSLTTEDTATYFCVRDAAYSVGRDDFDPWGPGTLVTISS

[0504] VH - SEQ ID NO: 108

[0505] ELVMTQTPASVSEPVGGTVTIKCQASQNIYSGLAWYQQKPGQPPKLLIYQSSTLASGVPSRFKGSGSGTQFTLTI

[0506] SGVQREDAATYYCQSGANSNDITFGGGTELEIL

[0507] Anti_VEGFR2 antibody - Ramucirumab

[0508] VH - SEQ ID NO: 109

[0509] EVQLVQSGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSSISSSSSYIYYADSVKGRFTISRDN

[0510] AKNSLYLQMNSLRAEDTAVYYCARVTDAFDIWGQGTMVTVSS

[0511] VL - SEQ ID NO: 110

[0512] DIQMTQSPSSVSASIGDRVTITCRASQGIDNWLGWYQQKPGKAPKLLIYDASNLDTGVPSRFSGSGSGTYFTLTIS

[0513] SLQAEDFAVYFCQQ AKAFPPTFGGGTKVDIK

[0514] CXCR4 (e.g., UniProtKB - P61073, Gene ID: 7852)

[0515] Anti_CXCR4 - BMS

[0516] VH - SEQ ID NO: 111

[0517] QVQLVQSGGGLVQPGGSLRLSCAAAGFTFSSYSMNWVRQAPGKGLEWVSYISSRSRTIYYADSVKGRFTISRDN

[0518] AKNSLYLQMNSLRDEDTAVYYCARDYGGQPPYYYYYGMDVWGQGTTVTVSS

[0519] VH - SEQ ID NO: 112

[0520] AIRMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSLIYAASSLQSGVPSRFSGSGSGTDFTLTIS

[0521] SLQPEDFVTYYCQQYNSYPRTFGQGTKVEIK Anti_CXCR4 antibody - Pfizer

[0522] VH-SEQIDNO: 113

[0523] EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYYMSWVRQAPGKGLEWVGFIRHKANFETTEYSTSVKGRFTISR

[0524] DDSKNSLYLQMNSLKTEDTAVYYCARDLPGFAYWGQGTLVTVSS

[0525] VL-SEQIDNO: 114

[0526] DIVMTQSPDSLAVSLGERATINCKSSQSLFNSHTRKNYLAWYQQKPGQPPKLLIYWASARGSGVPDRFSGSGSG

[0527] TDFTLTISSLQAEDVAVYYCKQSFNLRTFGGGTKVEIK

[0528] Anti_CXCR4 antibody - Medarex

[0529] VH-SEQIDNO: 115

[0530] QVQLVQSGGGLVQPGGSLRLSCAAAGFTFSSYSMNWVRQAPGKGLEWVSYISSRSRSIYYADSVKGRFTISRDN

[0531] AKNSLYLQMNSLRDEDTAVYYCARDYGGQPPYYYYYGMDVWGQGTTVTVSS

[0532] VL-SEQIDNO: 116

[0533] EIVLTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSLIYAASSLQSGVPPRFSGSGSGTDFTLTIS

[0534] SLQPEDFATYYCQQYNSYPRTFGQGTKVEIK

[0535] PDGFRB (e.q„ UniProtKB - P09619, Gene ID: 5159)

[0536] Anti_PDGFRb antibody - XB1511 + VL_D8

[0537] VH-SEQIDNO: 117

[0538] QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADE

[0539] STSTAYMELSSLRSEDTAVYYCAIHGGDRSYWGQGTLVTVSS

[0540] VL-SEQIDNO: 118

[0541] EIVMTQSPGTLTLSPGEGATLSCRASQSVTSNYLAWYQQRPGQAPRLLIYDASNRATGIPDRFSGSGFGTDFTLTI

[0542] SRLEPEDFAVYYCQQYVNSRTFGQGTKVEIK

[0543] Anti_PDGFRb antibody - XB2202+VL_A4

[0544] VH-SEQIDNO: 119

[0545] QVQLVQSGAEVKKPGSSVRVSCKASGGTFSRHAISWVRQAPGQGLEWIGGILPILKTPNYAQRFQGRVTINADES

[0546] TSTVYMEMSSLRSEDTAVYYCATHGGDRSYWGQGTLVTVSS

[0547] VL-SEQIDNO: 120 DWMTQSPSSLSASVGDRVTITCQASQDISNWLNWYQQKPGKAPKLLIYEASNLETGVPSRFSGSGSGTDFTFTI

[0548] SSLQPEDIATYYCQQYNNVLRTFGQGTKVEIK

[0549] CD33 (e.q„ UniProtKB - P20138, Gene ID: 945)

[0550] Anti_CD33 antibody

[0551] VH-SEQIDNO: 121

[0552] QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYDINWVRQAPGQGLEWMGWMNPNSGNTGFAQKFQGRVTMT

[0553] RDTSTSTVYMELSSLRSEDTAVYYCARDRANTDYSLGMDVWGQGTLVTVSS

[0554] VL-SEQIDNO: 122

[0555] QSVLTQPPSASGTPGQRVTISCSGSRSNIGSNTVNWYQQLPGTAPKLLIYGNNQRPSGVPDRFSGSKSGTSASL

[0556] AISGLQSEDEADYYC ATWDDSLIGWVFGGGTKLTVL

[0557] CD56 / NCAM1 (e.g., UniProtKB - P13591, Gene ID: 4684)

[0558] Anti_CD56 antibody

[0559] VH-SEQIDNO: 123

[0560] QVQLVESGGGWQPGRSLRLSCAASGFTFSWVRQAPGKGLEWVAYADSVKGRFTISRDNSKNTLYLQMNSLRA

[0561] EDTAVYYCARWGQGTLVTVSS

[0562] VL-SEQIDNO: 124

[0563] DWMTQSPLSLPVTLGQPASISCWFQQRPGQSPRRLIYGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFGQG

[0564] TKVEIK

[0565] CD19 (e.q„ UniProtKB - P15391, Gene ID: 930)

[0566] Anti_CD19 antibody - cA19

[0567] VH-SEQIDNO: 125

[0568] QVQLQESGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYNGKFKGKATLTA

[0569] DESSSTAYMQLSSLRSEDSAVYSCARRETTTVGRYYYAMDYWGQGTTVTVSS

[0570] VL-SEQIDNO: 126 DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGTDFT

[0571] LNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLQIKR

[0572] Anti_CD 19 antibody - hA19

[0573] VH - SEQ ID NO: 127

[0574] QVQLQQSGAEVKKPGSSVKVSCKASGYAFSSYWMNWVRQRPGQGLEWIGQIWPGDGDTNYNGKFKGRATITA

[0575] DESTNTAYMELSSLRSEDTAFYSCARRETTTVGRYYYAMDYWGQGTTVTVSS

[0576] VL - SEQ ID NO:128

[0577] DIQLTQSPSSLSASVGDRVTITCKASQSVDYDGDSYLNWYQQIPGKAPKLLIYDASNLVSGVPSRFSGSGSGTDY

[0578] TFTISSLQPEDIATYHCQQSTEDPWTFGGGTKLQIKR

[0579] Anti_CD19 antibody - Blincyto

[0580] VH - SEQ ID NO: 129

[0581] QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYNGKFKGKATLTA

[0582] DESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAMDYWGQGTTVTVSS

[0583] VL - SEQ ID NO: 130

[0584] DIQLTQSPASLAVSLGQRATISCKASQSVDYDDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGTDF

[0585] TLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIK

[0586] CD20 (e.q„ UniProtKB - P11836, Gene ID: 931)

[0587] Anti_CD20 antibody - Genmab

[0588] VH - SEQ ID NO: 131

[0589] AVQLVESGGGLVQPGRSLRLSCAASGFTFGDYTMHWVRQAPGKGLEWVSGISWNSGSIGYADSVKGRFTISRD

[0590] NAKNSLYLQMNSLRAEDTALYYCTKDNQYGSGSTYGLGVWGQGTLVTVSS

[0591] VL - SEQ ID NO: 132

[0592] EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTIS

[0593] SLEPEDFAVYYCQQRSNWPLTFGGGTKVEIK

[0594] L1CAM (e.q„ UniProtKB - P32004, Gene ID: 3897)

[0595] Anti_L1CAM antibody

[0596] VH - SEQ ID NO: 133 DVHSQVQLQQPGAELVKSGASVNLSCRASGYTFTRYWMLWVRQRPGHGLEWVGEINPRNDRTNYNEKFKTKA

[0597] TLTVDRSSSTAYMQLTSLTSEDSAVYFCALGGGYAMDYWGQGTSVTVSS

[0598] VL-SEQIDNO: 134

[0599] DIQMTQTTSSLSAFLGDRVTISCRASQDISNYLNWYQQKPDGTVKLLIYYTSRLHSGVPSRFSGSGSGTDYSLTIS

[0600] NLEQEDFATYFCQQGNTLPWTFGGGTKLEIK

[0601] LFA-11 ITGB21 CD18 (e.g., UniProtKB - P05107, Gene ID: 3689)

[0602] Anti_LFA1 antibody

[0603] VH-SEQIDNO: 135

[0604] EVQLVESGGGLVQPGGSLRLSCATSGYTFTEYTMHWMRQAPGKGLEWVAGINPKNGGTSHNQRFMDRFTISVD

[0605] KSTSTAYMQMNSLRAEDTAVYYCARWRGLNYGFDVRYFDVWGQGTLVTVSS

[0606] VL-SEQIDNO: 136

[0607] DIQMTQSPSSLSASVGDRVTITCRASQDINNYLNWYQQKPGKAPKLLIYYTSTLHSGVPSRFSGSGSGTDYTLTIS

[0608] SLQPEDFATYYCQQGNTLPPTFGQGTKVEIKR

[0609] R0R1 (e.g., UniProtKB -Q01973, Gene ID: 4919)

[0610] Anti_ROR1 antibody

[0611] VH-SEQIDNO: 137

[0612] QVQLQQSGAELVRPGASVTLSCKASGYTFSDYEMHWVIQTPVHGLEWIGAIDPETGGTAYNQKFKGKAILTADKS

[0613] SSTAYMELRSLTSEDSAVYYCTGYYDYDSFTYWGQGTLVTVSA

[0614] VL-SEQIDNO: 138

[0615] DIVMTQSQKIMSTTVGDRVSITCKASQNVDAAVAWYQQKPGQSPKLLIYSASNRYTGVPDRFTGSGSGTDFTLTI

[0616] SNMQSEDLADYFCQQYDIYPYTFGGGTKLEIK

[0617] CD123 / IL3RA (e.q., UniProtKB - P26951, Gene ID: 3563)

[0618] Anti_CD123 antibody

[0619] VH-SEQIDNO: 139 QIQLVQSGPELKKPGETVKISCKASGYIFTNYGMNWVKQAPGKSFKWMGWINTYTGESTYSADFKGRFAFSLET

[0620] SASTAYLHINDLKNEDTATYFCARSGGYDPMDYWGQGTSVTVSS

[0621] VL-SEQIDNO: 140

[0622] DIVLTQSPASLAVSLGQRATISCRASESVDNYGNTFMHWYQQKPGQPPKLLIYRASNLESGIPARFSGSGSRTDF

[0623] TLTINPVEADDVATYYCQQSNEDPPTFGAGTKLELK

[0624] IGF1R / CD221 (e.g., UniProtKB - P08069, Gene ID: 3480)

[0625] Anti_IGF1R antibody - 2C6

[0626] VH-SEQIDNO: 141

[0627] EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSGISWNSGSKGYVDSVKGRFTISRD

[0628] NAKNSLYLQMNSLRAEDTALYYCAKDIRIGVAASYYFGEDVWGHGTTVTVSS

[0629] VL-SEQIDNO: 142

[0630] AIQLTQSPSSLSASVGDRVTITCRASQGISSVLAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISS

[0631] LQPEDFATYYCQQFNSYPYTFGQGTKLEIK

[0632] Anti_IGF1R antibody - 9H2

[0633] VH-SEQIDNO: 143

[0634] QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYVMHWVRQAPGQRLEWMGWINAGNGNTRYSQKFQGRVTITR

[0635] DTSASTVYMELSSLRSEDTAVYYCARGGMPVAGPGYFYYYGMDVWGQGTTVTVSS

[0636] VL-SEQIDNO: 144

[0637] EIVLTQSPGTLSLSPGERATLSCRASQSVSRSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTI

[0638] SRLEPEDFAVYCCQQYGSSPWTFGQGTKVEIKRT

[0639] CCR71 CD197 (e.g., UniProtKB - P32248, Gene ID: 1236)

[0640] Anti_CCR7 antibody - R735

[0641] VH-SEQIDNO: 145

[0642] EVQLLESGGGLVQPGGPLRLSCAASGFTFSNYNMHWVRQAPGKGLEWVSGIGPRRGRTYYADSVKGRFTISRD

[0643] NSKNTLYLQMNSLRAEDTAVYYCARSYAYQYRGLDYWGQGTLVTVSS

[0644] VL-SEQIDNO: 146 EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTI

[0645] SRLEPEDFAVYYCQQGSPVTFGQGTKVEIK

[0646] Anti_CCR7 antibody - R707BI

[0647] VH - SEQ ID NO: 147

[0648] EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYAIHWVRQAPGKGLEWVSAITPRGGYTYYADSVKGRFTISRDN

[0649] SKNTLYLQMNSLRAEDTAVYYCARGLTISYTPGFDYWGQGTLVTVSS

[0650] VL - SEQ ID NO:148

[0651] EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTI

[0652] SRLEPEDFAVYYCQQSYSSPITFGQGTKVEIK

[0653] CCR41 CD194 (e.q„ UniProtKB - P51679, Gene ID: 1233)

[0654] Anti_CCR4 antibody

[0655] VH - SEQ ID NO: 149

[0656] EVQLVESGGDLVQPGRSLRLSCAASGFIFSNYGMSWVRQAPGKGLEWVATISSASTYSYYPDSVKGRFTISRDN

[0657] AKNSLYLQMNSLRVEDTALYYCGRHSDGNFAFGYWGQGTLVTVSS

[0658] VL - SEQ ID NO:150

[0659] DVLMTQSPLSLPVTPGEPASISCRSSRNIVHINGDTYLEWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTD

[0660] FTLKISRVEAEDVGVYYCFQGSLLPWTFGQGTKVEIK

[0661] TNFRSF914-1 BB (e.g., UniProtKB - Q07011 , Gene ID: 3604)

[0662] Anti_4-1 BB antibody

[0663] VH - SEQ ID NO: 151

[0664] EVQLVQSGAEVKKPGESLRISCKGSGYSFSTYWISWVRQMPGKGLEWMGKIYPGDSYTNYSPSFQGQVTISADK

[0665] SISTAYLQWSSLKASDTAMYYCARGYGIFDYWGQGTLVTVSS

[0666] VH - SEQ ID NO: 152

[0667] SYELTQPPSVSVSPGQTASITCSGDNIGDQYAHWYQQKPGQSPVLVIYQDKNRPSGIPERFSGSNSGNTATLTIS

[0668] GTQAMDEADYYCATYTGFGSLAVFGGGTKLTVL CTLA41 CD152 (e.q„ UniProtKB - P16410, Gene ID: 1493)

[0669] Anti_CTLA4 antibody - 8H5

[0670] VH-SEQIDNO: 153

[0671] QVQLVQSGAELKKPGASVKVSCKASGYTFTSYWTNWTRQAPGQGLEWIGRIAPGSGTTYYNEVFKGRVTITVDK

[0672] STSTAYMELSSLRSEDTAVYFCARGDYGSYWGQGTLVTVSS

[0673] VL-SEQIDNO: 154

[0674] EIVLTQSPATLSLSPGERATLSCSASSSISYMHWFQQRPGQSPRRWIYDTSKLASGVPARFSGSGSGTDYTLTIS

[0675] SLEPEDFATYYCHQRTSYPLTFGQGTKLEIK

[0676] Anti_CTLA4 anitbody - 3B10

[0677] VH-SEQIDNO: 155

[0678] QVQLVQSGAELKKPGASVKVSCKASGYTFTDYNMDWVRAQPGQGLEWIGNINPNSESTSYNQKFKGRVTITVDK

[0679] STSTAYMELRSLRSDDTAVYYCTRDGNRYDAWFAYWGQGTLVTVSS

[0680] VL-SEQIDNO: 156

[0681] EIVLTQSPATLSLSPGERATLSCSASSSVTYMHWFQQKPGKAPKLWIYSTSILASGVPARFSGSGSGTDYTLTISR

[0682] LEPEDFATYYCQQRTSYPLTFGQGTKLEIK

[0683] Anti_CTLA4 antibody - INN 8568

[0684] VH-SEQIDNO: 157

[0685] QVQLVESGGGWQPGRSLRLSCAASGFTFSSYTMHWVRQAPGKGLEWVTFISYDGNNKYYADSVKGRFTISRD

[0686] NSKNTLYLQMNSLRAEDTAIYYCARTGWLGPFDYWGQGTLVTVSS

[0687] VL-SEQIDNO: 158

[0688] EIVLTQSPGTLSLSPGERATLSCRASQSVGSSYLAWYQQKPGQAPRLLIYGAFSRATGIPDRFSGSGSGTDFTLTI

[0689] SRLEPEDFAVYYCQ QYGSSPWTFGQGTKVEIK

[0690] GPC31 Glypican-3 (e.q„ UniProtKB - P51654, Gene ID: 2719)

[0691] Anti_Glypican-3 antibody

[0692] VH-SEQIDNO: 159

[0693] EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSTIQKQGLPTQYADSVKGRFTISRD

[0694] NSKNTLYLQMNSLRAEDTAVYYCAKNRAKFDYWGQG TLVTVSS VL-SEQIDNO: 160

[0695] DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYNASMLQSGVPSRFSGSGSGTDFTLTIS

[0696] SLQPEDFATYYCQQNRGFPLTFGQGTKVEIK

[0697] CADM1 (e.g., UniProtKB- Q9BY67, Gene ID: 23705)

[0698] Anti_CADM1 antibody

[0699] VH-SEQIDNO: 161

[0700] QITLKESGPTLVKPTQTLTLTCTFSGFSLSTSGVGVGWIRQPPGKALEWLALIYWDDDKRYSPSLKSRLTITKDTSK

[0701] NQWLTMTNMDPVDTAIYYCAHRRVEWFALAGNWFDPWGQGSLVTVSS

[0702] VL-SEQIDNO: 162

[0703] DIQMTQSPSSLSASVGDRVTITCRAWLAWYQQKPEKAPKSLIYAASSLQSSGSGSGTDFTLTISSLQPEDFATYYY

[0704] PLTFGGGTKVEIK

[0705] CD4 (e.g., UniProtKB - P01730, Gene ID: 920)

[0706] Anti_CD4 antibody

[0707] VH-SEQIDNO: 163

[0708] QVQLQQSGPEWKPGASVKMSCKASGYTFTSYVIHWVRQKPGQGLDWIGYINPYNDGTDYDEKFKGKATLTSDT

[0709] STSTAYMELSSLRSEDTAVYYCAREKDNYATGAWFAYWGQGTLVTVSS

[0710] VL-SEQIDNO: 164

[0711] DIVMTQSPDSLAVSLGERVTMNCKSSQSLLYSTNQKNYLAWYQQKPGQSPKLLIYWASTRESGVPDRFSGSGS

[0712] GTDFTLTI SSVQAEDVAVYYCQQYYSYRTFGGGTKLEI K

[0713] B7-h4 / VTCN1 (e.g., UniProtKB - Q7Z7D3, Gene ID: 79679)

[0714] Anti_B7-h4 antibody - c56

[0715] VH-SEQIDNO: 165

[0716] QVQLQQWGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPPGKGLEWIGSIYYSGSTYYNPSLKSRVTISVDT

[0717] SKNQFSLKLSSVTAADTAVYYCARLSKGYSSSWAYSYYGPDAWGQGTMVTVSS

[0718] VL-SEQIDNO: 166 QPVLTQSHSVSESPGKTVTITCTGSGGNIATQYVQWYQQRPGSGPTTVIYDDDQRPSGVPDRFTGSIDSSSNSA

[0719] SLTISGLKTEDEADYYCQSYDSTNHGVFGGGAKVTVL

[0720] Anti_B7-h4 antibody - c26

[0721] VH - SEQ ID NO: 167

[0722] QVTLKESGAEVKNPGSSVKVSCKASGGTFSSYAISWLRQAPGQGLEWMGWINPNSGGSDYAQRFQGRVTMTR

[0723] DTSINTVYMELSRLRSDDTAVYYCARVEKRGYYYGMDVWGQGTMVTVSS

[0724] VL - SEQ ID NO: 168

[0725] DIVMTQTPATLSVSPGKRATLSCRASQSVSSNYLAWYQQKPGQAPRLLIYGASRRATGIPDRFSGSGSGTDFTLTI

[0726] SSLQAEDVAVYYCQQYGSSPRTFGQGTKLEIK

[0727] Anti_B7-h4 antibody - c3#68

[0728] VH - SEQ ID NO: 169

[0729] QVTLKESGAEVKNPGSSVKVSCKASGGTFSSYAISWLRQAPGQGLEWMGWINPNSGGSDYAQRFQGRVTMTR

[0730] DTSINTVYMELSRLRSDDTAVYYCARVEKRGYYYGMDVWGQGTMVTVSS

[0731] VL - SEQ ID NO:170

[0732] QPGLTQPPSASGTPGQRVTISCSGSSSNIGSNYVYWYQQFPGTAPKLLIYRNNQRPSGVPDRFSGSKSGTSASL

[0733] AISGLRSEDEADYYCAAWDDSLSVWVFGGGTKVTVL

[0734] TYRP1 (e.q„ UniProtKB - P17643, Gene ID: 7360)

[0735] Anti_TYRP1 antibody

[0736] VH - SEQ ID NO: 171

[0737] QVQLVQSGSELKKPGASVKISCKASGYTFTSYAMNWVRQAPGQGLESMGWINTNTGNPTYAQGFTGRFVFSMD

[0738] TSVSTAYLQISSLKAEDTAIYYCAPRYSSSWYLDY WGQGTLVTVSS

[0739] VL - SEQ ID NO: 172

[0740] EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTIS

[0741] SLEPEDFAVYYCQQRSNWLMYTFGQGTKLEIK

[0742] PDL1 1 CD274 (e.g., UniProtKB - Q9NZQ7, Gene ID: 29126)

[0743] Anti_PD-L1 antibody - Atezolizumab VH-SEQIDNO: 173

[0744] EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISAD

[0745] TSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSS

[0746] VL-SEQIDNO: 174

[0747] DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTIS

[0748] SLQPEDFATYYCQQYLYHPATFGQGTKVEIK

[0749] Anti_PD-L1 antibody - Durvalumab

[0750] VH-SEQIDNO: 175

[0751] EVQLVESGGGLVQPGGSLRLSCAASGFTFSRYWMSWVRQAPGKGLEWVANIKQDGSEKYYVDSVKGRFTISRD

[0752] NAKNSLYLQMNSLRAEDTAVYYCAREGGWFGELAFDYWGQGTLVTVSS

[0753] VL-SEQIDNO: 176

[0754] EIVLTQSPGTLSLSPGERATLSCRASQRVSSSYLAWYQQKPGQAPRLLIYDASSRATGIPDRFSGSGSGTDFTLTI

[0755] SRLEPEDFAVYYCQ QYGSLPWTFGQGTKVEIKR

[0756] Anti_PD-L1 antibody - Nivolumab

[0757] VH-SEQIDNO: 177

[0758] QVQLVESGGGWQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISRD

[0759] NSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVTVSS

[0760] VL-SEQIDNO: 178

[0761] EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTIS

[0762] SLEPEDFAVYYCQQ SSNWPRTFGQGTKVEIK

[0763] PDCD11 PD-1 (e.g„ UniProtKB - Q15116, Gene ID: 5133)

[0764] Anti_PD-1 antibody - Nivolumab

[0765] VH-SEQIDNO: 179

[0766] QVQLVESGGGWQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISRD

[0767] NSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVTVSS

[0768] VL-SEQIDNO: 180 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTIS

[0769] SLEPEDFAVYYCQQ SSNWPRTFGQGTKVEIK

[0770] Anti_PD-1 antibody - Pembrolizumab

[0771] VH - SEQ ID NO: 181

[0772] QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGINPSNGGTNFNEKFKNRVTLTT

[0773] DSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMGFDYWGQGTTVTVSS

[0774] VL - SEQ ID NO: 182

[0775] EIVLTQSPATLSLSPGERATLSCRASKGVSTSGYSYLHWYQQKPGQAPRLLIYLASYLESGVPARFSGSGSGTDF

[0776] TLTISSLEPEDFAVYYCQHSRDLPLTFGGGTKVEIKR

[0777] PDCD1LG2 1 PD-L2 (e.q„ UniProtKB - Q9BQ51 , Gene ID: 80380)

[0778] Anti_PD-L2 antibody

[0779] VH - SEQ ID NO: 183

[0780] QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYTMHWVRQAPGQGLEWIGYINPRSGYTEYNQKFKDRTTLTAD

[0781] KSTSTAYMELSSLRSEDTAVYYCARPWFAYWGQGTLVTVSS

[0782] VL - SEQ ID NO: 184

[0783] DIVMTQSPAFLSVTPGEKVTITCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGT

[0784] DFTLTISSLQAEDVAVYYCQNDYSYPLTFGQGTKLEIK

[0785] BTLA (e.g., UniProtKB - Q7Z6A9, Gene ID: 151888)

[0786] Anti_BTLA antibody

[0787] VH - SEQ ID NO: 185

[0788] QVQLQESGPGLVKPSETLSLTCTVHGGSINHYYWSWIRQPPGKGLEWIGYIYYSGSTKYNPSLKSRVSISVDTSK

[0789] NQFSLKLTSVTAADTAVYYCAREWPYYYYEMDVWGQGTTVTVSS

[0790] VL - SEQ ID NO: 186

[0791] EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTI

[0792] SRLEPEDFAVYYCQQYGSSFRTFGQGTKVEIK DLL3 (e.q„ UniProtKB - Q9NYJ7, Gene ID: 10683)

[0793] Anti_DLL3 antibody - Rovalpituzumab

[0794] VH-SEQIDNO: 187

[0795] QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQAPGQGLEWMGWINTYTGEPTYADDFKGRVTMTT

[0796] DTSTSTAYMELRSLRSDDTAVYYCARIGDSSPSDYWGQGTLVTVSS

[0797] VL-SEQIDNO: 188

[0798] EIVMTQSPATLSVSPGERATLSCKASQSVSNDWWYQQKPGQAPRLLIYYASNRYTGIPARFSGSGSGTEFTLTIS

[0799] SLQSEDFAVYYCQQDYTSPWTFGQGTKLEIK

[0800] CEACAM11 CD66a (e.q„ UniProtKB - P13688, Gene ID: 634)

[0801] Anti_CEACAM1 antibody - clone 6G5J

[0802] VH-SEQIDNO: 189

[0803] KPGETVKISCKTSGYIFRNYGMKWVKQAPGKGLKWMGWINTYTGEPTYADDFKGRFAFSLETSASTAYLQINNLK

[0804] NEDMATYFCARRGMITTSNYALDNWGQGTSVTVSS

[0805] VL-SEQIDNO: 190

[0806] SSLSASLGDRVTISCRASQDISNYLNWYQQKPDGTIKLLIYYTSKLHSGVPSRFSGSGSGTDYSLTISNLDQEDIAT

[0807] YFCQQGNTLPWTFGGGTKLEIK

[0808] Anti_CEACAM1 antibody - clone 18-20

[0809] VH-SEQIDNO: 191

[0810] KPGETVKISCKASGYTFTVYGMNWVKQAPGKDLKWMGWINTYTGEPTYADDFKGRFAFSLETSASTAYLQINNL

[0811] KNEDMATYFCARKAFYRYDGGMDYWGQGTSVTVSS

[0812] VL-SEQIDNO: 192

[0813] SSLSASLGGKVTITCKASQDINKFLAWYQHKPGKGPRLLIHYTSTLQPGIPSRFSGSGSGRDYSFSISNLEPEDIAT

[0814] YYCLQYDNLYTFGGGTKLEIK

[0815] LFA-11 ITGB2 (e.q„ UniProtKB - P05107, Gene ID: 3689)

[0816] Anti_LFA-1 antibody - Efalizumab

[0817] VH-SEQIDNO: 193 EVQLVESGGGLVQPGGSLRLSCAASGYSFTGHWMNWVRQAPGKGLEWVGMIHPSDSETRYNQKFKDRFTISV

[0818] DKSKNTLYLQMNSLRAEDTAVYYCARGIYFYGTTYFDYWGQGTLVTVSS

[0819] VL-SEQIDNO: 194

[0820] DIQMTQSPSSLSASVGDRVTITCRASKTISKYLAWYQQKPGKAPKLLIYSGSTLQSGVPSRFSGSGSGTDFTLTIS

[0821] SLQPEDFATYYCQQHNEYPLTFGQGTKVEIK

[0822] CD22 (e.g., UniProtKB - P20273, Gene ID: 933)

[0823] Anti_CD22 antibody - Pinatuzumab

[0824] VH-SEQIDNO: 195

[0825] EVQLVESGGGLVQPGGSLRLSCAASGYEFSRSWMNWVRQAPGKGLEWVGRIYPGDGDTNYSGKFKGRFTISA

[0826] DTSKNTAYLQMNSLRAED TAVYYCARDGSSWDWYFDVWGQGTLVTVSS

[0827] VH-SEQIDNO: 196

[0828] DIQMTQSPSSLSASVGDRVTITCRSSQSIVHSVGNTFLEWYQQKPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTD

[0829] FTLTISSLQPEDFATYYCFQGSQFPYTFGQGTKVEIK

[0830] NRP11 CD304 (e.g., UniProtKB - 014786, Gene ID: 8829)

[0831] Anti_NRP1 antibody

[0832] VH-SEQIDNO: 197

[0833] EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSQISPAGGYTNYADSVKGRFTISAD

[0834] TSKNTAYLQMNSLRAEDTAVYYCARGELPYYRMSKVMDVWGQGTLVTVSS

[0835] VL-SEQIDNO: 198

[0836] DIQMTQSPSSLSASVGDRVTITCRASQYFSSYLAWYQQKPGKAPKLLIYGASSRASGVPSRFSGSGSGTDFTLTI

[0837] SSLQPEDFATYYCQQYLGSPPTFGQGTKVEIKR

[0838] CHRNA1 (e.g., UniProtKB - P02708, Gene ID: 1134)

[0839] Anti_AChR_alpha antibody

[0840] VH-SEQIDNO: 199 ESGGDLVQPGGSLRVSCVASGFTFRTYVMNWVRQAPGKGLEWVAHISPEGTEEYYADPVKGRFTISRDNAKNS

[0841] VFLQMNSLRGEDTAVYYCARVRRYGPSTLSPFTWKDNHYAMDVWGQGTTVTVSP

[0842] VL - SEQ ID NO: 200

[0843] ELVMTQSPSSLSASVGDRVTITCRASQTISNYLNWYQQKPGKAPKLLIYGASSLQSGVPSRFSGSGSGTDFTLTIS

[0844] SLQPEDFATYYCQQSYSTPPTYTFGQGTKLEIK

[0845] GD21 Disialoganglioside (CAS Registry Number® 65988-71 -8)

[0846] Anti_GD2 antibody

[0847] VH - SEQ ID NO: 201

[0848] EVQLVQSGAEVEKPGASVKISCKASGSSFTGYNMNWVRQNIGKSLEWIGAIDPYYGGTSYNQKFKGRATLTVDK

[0849] STSTAYMHLKSLRSEDTAVYYCVSGMEYWGQGT

[0850] VL - SEQ ID NO: 202

[0851] DWMTQTPLSLPVTPGEPASISCRSSQSLVHRNGNTYLHWYLQKPGQSPKLLIHKVSNRFSGVPDRFSGSGSGT

[0852] DFTLKISRVEAEDLGVYFCSQSTHVPPLTFGAGTKLELKS

[0853] CTAG1A / NY-ESO-1 (e.g„ UniProtKB - P78358, Gene ID: 246100)

[0854] Anti_CTAGE1 antibody

[0855] VH - SEQ ID NO: 203

[0856] QVQLVQSGGGWRPGGSLRLSCAASGFSFIDYGMSWVRQVPGKGLEWVAGMNWSGDKKGHAESVKGRFIISR

[0857] DNAKNTLYLEMSSLRVEDTALYFCARGEYSNRFDPRGRGTLVTVSS

[0858] VL - SEQ ID NO: 204

[0859] DIVMTQTPLSLPVTLGQPASLSCRSSQSLVFTDGNTYLNWFQQRPGQSPRRLIYKVSSRDPGVPDRFSGTGSGT

[0860] DFTLEISRVEAEDIGVYYCMQGTHWPPI FGQGTKVEIK

[0861] CD30 1 TNFRSF8 (e.q. , UniProtKB - P28908, Gene ID: 943)

[0862] Anti_CD30 antibody - 2H9

[0863] VH - SEQ ID NO: 205 QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTKYTPSLKSRVTISVDTS

[0864] KHQFSLKLSSVTAADTAVYYCARETVYYFDLWGRGTLVTVSS

[0865] VL - SEQ ID NO: 206

[0866] EIVLTQSPATLSLSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYDASNRATGIPARLSGSGSGTDFTLTIS

[0867] SLEPEDFAVYYCQQRSNWPWTFGQGTKVEIK

[0868] CD98hc I SLC3A2 (e.g., UniProtKB - P08195, Gene ID: 6520)

[0869] Anti_CD98hc antibody - EJ212 / 076-CI 10

[0870] VH - SEQ ID NO: 207

[0871] QVQLVESGGGLVKPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSSISSSSSYIYYADSVKGRFTISRDN

[0872] AKNSLYLQMNSLRAEDTAVYYCARSSGWYDGEFDPWGQGTLVTVSS

[0873] VL - SEQ ID NO: 208

[0874] SYVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQLPGTAPKLLIYSNDQRPSGVPDRFSGSKSGTSASLA

[0875] ISGLQSEDEADYYCAAWDDSLNGWFGGGTKLTVL

[0876] CD52 (e.g., UniProtKB - P31358, Gene ID: 1043)

[0877] Anti_CD52 antibody - 2E8 Humanised

[0878] VH - SEQ ID NO: 209

[0879] EVHLVESGGGLVQPGGSLRLSCAASGFTFSRYGMSWVRQAPGKGLELVAMMKTKGGRTYYPDSVKGRFTISRD

[0880] NAKNSLYLQMNSLRAEDTAIYYCASDGYYWGQGTTVTVSS

[0881] VL - SEQ ID NO: 210

[0882] DWMTQTPLSLSVTLGQPASISCKSSQSLLHSDGKTYLNWLQQRPGQSPRRLIYLVSKLDSGVPDRFSGSGSGTD

[0883] FTLKIS RVEAEDVGIYYCWQGTH LWTFGGGTKVEIK

[0884] CD641 FCGR1A (e.g., UniProtKB - P12314, Gene ID: 2209)

[0885] Anti_CD64 antibody

[0886] VH - SEQ ID NO: 211 QVQLVESGGGWQPGRSLRLSCAASGFIFSGYGMHWVRQAPGKGLEWVTVIWYDGSNKYYADSVKGRFTISRD

[0887] NSKNTLYLQMNSLRAEDTAVYYCARDTGDRFFDYWGQGTLVTVSS

[0888] VL - SEQ ID NO: 212

[0889] EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASSRATGIPARFGGSGSGTDFTLTIS

[0890] SLEPEDFAVYYCQLRSNWPPYTFGQGTKLEIK

[0891] CD46 1 MCP (e.g., UniProtKB - P15529, Gene ID: 4179)

[0892] Anti_CD46 antibody

[0893] VH - SEQ ID NO: 213

[0894] QVQLVQSGAEVKRPGASMKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGWINPNSGATNYAQKFQGRVTMTR

[0895] DTSTSTVYMELSSLRSEDTAVYYCAKSWVSWFASWGQGTLVTVSS

[0896] VL - SEQ ID NO: 214

[0897] DIVMTQSPDSLAVSLGERATINCKSSQSVLSSSNNKNYLNWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSG

[0898] TDFTLTISSLQAEDVAVYYCQQHFNTPFAFGQGTKLEIK

[0899] More preferably, the antibody construct comprises scFvs comprising the VH and VL sequences of the following antibodies: antiCD38 - Morphosys Ag 2004

[0900] SEQ ID NO: 215 - antiCD38 VH = Morphosys Ag 2004

[0901] QVQLVESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVSNIYSDGSNTFYADSVKGR

[0902] FTISRDNSKNTLYLQMNSLRAEDTAVYYCARNMYRWPFHYFFDYWGQGTLVTVSS

[0903] SEQ ID NO: 216 - antiCD38 VL = Morphosys Ag 2004

[0904] DIELTQPPSVSVAPGQTARISCSGDNIGNKYVSWYQQKPGQAPVWIYGDNNRPSGIPERFSGSNSGNT

[0905] ATLTISGTQAEDEADYYCSSYDSSYFVFGGGTKLTVL antiSLAMF7 = Elotuzumab

[0906] SEQ ID NO: 217 - antiSLAMF7 VH = Elotuzumab

[0907] EVQLVESGGGLVQPGGSLRLSCAASGFDFSRYWMSWVRQAPGKGLEWIGEINPDSSTINYAPSLKDKF

[0908] IISRDNAKNSLYLQMNSLRAEDTAVYYCARPDGNYWYFDVWGQGTLVTVSS SEQ ID NO: 218 - antiSLAMF7 VL = Elotuzumab

[0909] DIQMTQSPSSLSASVGDRVTITCKASQDVGIAVAWYQQKPGKVPKLLIYWASTRHTGVPDRFSGSGSGT DFTLTI SSLQPEDVATYYCQQYSSYPYTFGQGTKVEI K

[0910] Anti-Erbb2 - Trastuzumab

[0911] SEQ ID NO: 219 - Anti-Erbb2-VH (Trastuzumab )

[0912] EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRF TISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS

[0913] SEQ ID NO: 220 - Anti-Erbb2-VL (Trastuzumab )

[0914] DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGT DFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIK

[0915] Anti-EpCAM antibody

[0916] SEQ ID NO: 221 - Anti-EpCAM VH

[0917] EVQLLEQSGAELVRPGTSVKISCKASGYAFTNYWLGWVKQRPGHGLEWIGDIFPGSGNIHYNEKFKGK ATLTADKSSSTAYMQLSSLTFEDSAVYFCARLRNWDEPMDYWGQGTTVTVSS

[0918] SEQ ID NO: 222 - Anti-EpCAM VL

[0919] ELVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRF TGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYPLTFGAGTKLEIK

[0920] Linkers used in the present invention

[0921] Linkers which can be used to connect VH and VL domains, and / or VH and VH domains, and / or VL and VL domains, and / or VH with human serum protein, and / or VL with human serum protein, and / or VH with Ig constant CH 1 domain, and / or VL with Ig constant CH1 domain, and / or VH with Ig constant CH2 domain, and / or VL with Ig constant CH2 domain, and / or VH with Ig constant CH3 domain, and / or VL with Ig constant CH3 domain, and / or VH with Ig constant CL domain, and / or VL with Ig constant CL domain with each other in the antibody construct of the present invention or which can be used to connect any human protein to a KeyLock antibody may include the following linkers:

[0922] SEQ ID NO: 223 A preferred linkers connecting the antibody VH - VL domains, VL - VH domains, VH - VH domains and VL - VL domains in the antibody constructs of the invention is as follows:

[0923] GGGGSGGGGSGGGGSGGGGS

[0924] SEQ ID NO: 224

[0925] Alternative linker connecting antibody variable domains

[0926] GGGGS

[0927] SEQ ID NO: 225

[0928] Alternative linker connecting antibody variable domains

[0929] GGGGSGGGGS

[0930] SEQ ID NO: 226

[0931] Alternative linker connecting antibody variable domains

[0932] GGGGSGGGGSGGGG

[0933] SEQ ID NO: 227

[0934] Alternative linker connecting antibody variable domains

[0935] GGGGSGGGGSGGGGGGGGSGGGGS

[0936] SEQ ID NO: 228

[0937] Alternative linker connecting antibody variable domains

[0938] GGGGSGGGGSGGGGGGGGSGGGGSGGGGS

[0939] SEQ ID NO: 229

[0940] Linker connecting human albumin to a KeyLock antibody SGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGG

[0941] SEQ ID NO: 230

[0942] Alternative linker connecting antibody variable domains

[0943] GSGGGGSGGGGSGGGGS

[0944] SEQ ID NO: 231

[0945] Alternative linker connecting antibody variable domains

[0946] GSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGS

[0947] SEQ ID NO: 232

[0948] Alternative linker connecting antibody variable domains

[0949] GSGGSGNSSGGGS

[0950] SEQ ID NO: 233

[0951] Alternative linker connecting antibody variable domains

[0952] GSGGSGNSSGGGSGGGGS

[0953] SEQ ID NO: 234

[0954] Alternative linker connecting antibody variable domains

[0955] GGGGSGSGGSGNSSGGGSGGGGS

[0956] SEQ ID NO: 235

[0957] Alternative linker connecting antibody variable domains

[0958] GSGGGGNSSGGGGNSSGGGS SEQ ID NO: 236

[0959] Alternative linker connecting antibody variable domains

[0960] GGGGSGSGGGGNSSGGGGNSSGGGSGGGGS

[0961] SEQ ID NO: 237

[0962] Alternative linker connecting antibody variable domains

[0963] AEAWYNLGNAYYKQGDYQKAIEYYQKALELDPN

[0964] SEQ ID NO: 238

[0965] Alternative linker connecting antibody variable domains

[0966] PVPSTPPTPSPSTPPTPSPS

[0967] SEQ ID NO: 239

[0968] Alternative linker connecting antibody variable domains

[0969] EPKSADKTHTAPPAPAPELLGGP

[0970] SEQ ID NO: 240

[0971] Alternative linker connecting antibody variable domains

[0972] EAAAKEAAAKEAAAK

[0973] SEQ ID NO: 241

[0974] Alternative linker connecting antibody variable domains

[0975] APAPAPAPAPAPAP

[0976] SEQ ID NO: 242

[0977] Alternative linker connecting antibody variable domains GKSSGSGSESKS

[0978] SEQ ID NO: 243

[0979] Alternative linker connecting antibody variable domains

[0980] TSGSGKSSEGKG

[0981] SEQ ID NO: 244

[0982] Alternative linker connecting antibody variable domains

[0983] GSTSGSGKSSEGSGSTKG

[0984] SEQ ID NO: 245

[0985] Alternative linker connecting antibody variable domains

[0986] GSTSGKPSEGKG

[0987] SEQ ID NO: 246

[0988] Alternative linker connecting antibody variable domains

[0989] KESGSVSSEQLAQFRSLD

[0990] SEQ ID NO: 247

[0991] Alternative linker connecting antibody variable domains

[0992] DHVPLT

[0993] SEQ ID NO: 248

[0994] Alternative linker connecting antibody variable domains

[0995] DEYDPTIEDSY SEQ ID NO: 249

[0996] Alternative linker connecting antibody variable domains

[0997] TAGQEEYSAMRDQYM

[0998] SEQ ID NO: 250

[0999] Alternative linker connecting antibody variable domains

[1000] PKRNKPTVYGVSPNYDKWEMERTD

[1001] SEQ ID NO: 251

[1002] Alternative linker connecting antibody variable domains

[1003] RNKPTVYGVSPNYDKWEMERT

[1004] SEQ ID NO: 252

[1005] Alternative linker connecting antibody variable domains

[1006] LITTLHYPAPKRNKPTVYGVSPNYDKWEMERTD

[1007] SEQ ID NO: 253

[1008] Alternative linker connecting antibody variable domains

[1009] APKRNKPTVYGVSPNYDK

[1010] Alternative second linkers for the present invention

[1011] A second flexible linker can be used to covalently connect the linkers from Sequence 223 to 253 to the antibody variable domains, serum proteins and / or Ig constant domains, using standard techniques, in a single polypeptide chain, at the N-terminal and / or C-terminal end. Only one set or multiple set of linkers can be used in various combinations in one Keylock construct. KeyLock constructs with same or different linkers can be connected to any serum protein or constant Ig domains. The flexible linkers may include the following linkers:

[1012] SEQ ID NO: 254 Alternative linker connecting linkers with antibody variable domains, serum proteins and / or Ig constant domains GS

[1013] SEQ ID NO: 255

[1014] Alternative linker connecting linkers with antibody variable domains, serum proteins and / or Ig constant domains SG

[1015] SEQ ID NO: 256

[1016] Alternative linker connecting linkers with antibody variable domains, serum proteins and / or Ig constant domains AS

[1017] SEQ ID NO: 257

[1018] Alternative linker connecting linkers with antibody variable domains, serum proteins and / or Ig constant domains SA

[1019] SEQ ID NO: 258

[1020] Alternative linker connecting linkers with antibody variable domains, serum proteins and / or Ig constant domains GGG

[1021] SEQ ID NO: 259

[1022] Alternative linker connecting linkers with antibody variable domains, serum proteins and / or Ig constant domains

[1023] GGS

[1024] SEQ ID NO: 260

[1025] Alternative linker connecting linkers with antibody variable domains, serum proteins and / or Ig constant domains

[1026] AAGAATAA SEQ ID NO: 261

[1027] Alternative linker connecting linkers with antibody variable domains, serum proteins and / or Ig constant domains

[1028] SGGS

[1029] SEQ ID NO: 262

[1030] Alternative linker connecting linkers with antibody variable domains, serum proteins and / or Ig constant domains

[1031] SGGGS

[1032] SEQ ID NO: 263

[1033] Alternative linker connecting linkers with antibody variable domains, serum proteins and / or Ig constant domains

[1034] GSGSGS

[1035] SEQ ID NO: 264

[1036] Alternative linker connecting linkers with antibody variable domains, serum proteins and / or Ig constant domains

[1037] GGSGG

[1038] SEQ ID NO: 265

[1039] Alternative linker connecting linkers with antibody variable domains, serum proteins and / or Ig constant domains GGSSG

[1040] SEQ ID NO: 266

[1041] Alternative linker connecting linkers with antibody variable domains, serum proteins and / or Ig constant domains

[1042] GT

[1043] SEQ ID NO: 267 Alternative linker connecting linkers with antibody variable domains, serum proteins and / or Ig constant domains

[1044] GSGGGTGGGSG

[1045] SEQ ID NO: 268

[1046] Alternative linker connecting linkers with antibody variable domains, serum proteins and / or Ig constant domains

[1047] GGSGGSGGSGG

[1048] SEQ ID NO: 269

[1049] Alternative linker connecting linkers with antibody variable domains, serum proteins and / or Ig constant domains

[1050] GGGGG

[1051] Anti-CD3 antibody

[1052] In a preferred embodiment, the antibody construct of the present invention may comprise the following VH and VL sequences of the following anti-CD3 antibodies: antiCD3 dil_2K antibody

[1053] VH - SEQ ID NO: 270

[1054] DVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTMHWVRQAPGQGLEWIGYINPSRGYTNYADSVKGRFTITTDK

[1055] STSTAYMELSSLRSEDTATYYCARYYDDHYCLDYWGQGTTVTVSS

[1056] VL - SEQ ID NO: 271

[1057] DIVLTQSPATLSLSPGERATLSCRASQSVSYMNWYQQKPGKAPKRWIYDTSKVASGVPARFSGSGSGTDYSLTIN

[1058] SLEAEDAATYYCQQWSSNPLTFGGGTKVEIK

[1059] OR antiCD3 humanized UCHT1 antibody

[1060] VH - SEQ ID NO: 272 EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYTMNWVRQAPGKGLEWVALINPYKGVSTYNQKFKDRFTISVD

[1061] KSKNTAYLQMNSLRAEDTAVYYCARSGYYGDSDWYFDVWGQGTLVTVSS

[1062] VL- SEQ ID NO: 273

[1063] DIQMTQSPSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPKLLIYYTSRLESGVPSRFSGSGSGTDYTLTIS

[1064] SLQPEDFATYYCQQGNTLPWTFGQGTKVEIK

[1065] In a preferred embodiment, the VH and VL of the anti-CD3 antibody in the antibody construct comprise the following CDRs (A) or (B):

[1066] (A)

[1067] SEQ ID NO: 274 - UCHT1 VH CDR1-IMGT (27 - 38)

[1068] GYSFTGYT

[1069] SEQ ID NO: 275 - UCHT1 VH CDR2-IMGT (56 - 65)

[1070] INPYKGVS

[1071] SEQ ID NO: 276 - UCHT1 VH CDR3-IMGT (105 - 117)

[1072] ARSGYYGDSDWYFDV

[1073] SEQ ID NO: 277 - UCHT1 VL CDR1-IMGT (27 - 38)

[1074] QDIRNY

[1075] SEQ ID NO: 278 - UCHT1 VL CDR2-IMGT (56 - 65)

[1076] YTS

[1077] SEQ ID NO: 279 - UCHT1 VL CDR3-IMGT (105 - 117)

[1078] QQGNTLPWT

[1079] OR

[1080] (B)

[1081] SEQ ID NO: 280 - diL2K VH CDR1-IMGT (27 - 38)

[1082] GYTFTRYT SEQ ID NO: 281 - dil_2K VH CDR2-IMGT (56 - 65)

[1083] INPSRGYT

[1084] SEQ ID NO: 282 - diL2K VH CDR3-IMGT (105 - 117)

[1085] ARYYDDHYCLDY

[1086] SEQ ID NO: 283 - diL2K VL CDR1-IMGT (27 - 38)

[1087] QSVSY

[1088] SEQ ID NO: 284 - diL2K VL CDR2-IMGT (56 - 65)

[1089] DTS

[1090] SEQ ID NO: 285 - diL2K VL CDR3-IMGT (105 - 117)

[1091] QQWSSNPLT

[1092] Albumin

[1093] The antibody construct of the present invention or the protein structure of the antibody construct of the present invention may further comprise albumin, single albumin-domains, albumin-binding domains and / or other plasma / serum proteins such as part of membrane proteins cleaved and detected in the plasma / serum.

[1094] In a specific embodiment, the following albumin sequence may be used in accordance with the invention:

[1095] SEQ ID NO: 286 - human albumin (excluding the signal peptide and propeptide sequence)

[1096] DAHKSEVAHRFKDLGEENFKALVLIAFAQYLQQCPFEDHVKLVNEVTEFAKTCVADESAENCDKSLHTLFGDKLC TVATLRETYGEMADCCAKQEPERNECFLQHKDDNPNLPRLVRPEVDVMCTAFHDNEETFLKKYLYEIARRHPYF YAPELLFFAKRYKAAFTECCQAADKAACLLPKLDELRDEGKASSAKQRLKCASLQKFGERAFKAWAVARLSQRFP

[1097] KAEFAEVSKLVTDLTKVHTECCHGDLLECADDRADLAKYICENQDSISSKLKECCEKPLLEKSHCIAEVENDEMPA DLPSLAADFVESKDVCKNYAEAKDVFLGMFLYEYARRHPDYSWLLLRLAKTYETTLEKCCAAADPHECYAKVFD EFKPLVEEPQNLIKQNCELFEQLGEYKFQNALLVRYTKKVPQVSTPTLVEVSRNLGKVGSKCCKHPEAKRMPCAE

[1098] DYLSWLNQLCVLHEKTPVSDRVTKCCTESLVNRRPCFSALEVDETYVPKEFNAETFTFHADICTLSEKERQIKKQ TALVELVKHKPKATKEQLKAVMDDFAAFVEKCCKADDKETCFAEEGKKLVAASQAALGL

[1099] Additional sequences which may be used in accordance with the invention include: SEQ ID NO: 287 - Secretion leader signal peptide for human albumin for KeyLock-albumin fusion constructs

[1100] MAKWVTFISLLFLFSSAYS

[1101] SEQ ID NO: 288 - Propeptide at N terminal end of human albumin. Propeptide, which is a part of a protein that is cleaved during maturation or activation. Once cleaved, a propeptide generally has no independent biological function.

[1102] RGVFRR

[1103] Immunoglobulin domains

[1104] The antibody construct of the present invention may further comprise an immunoglobulin Fc region and / or an immunoglobulin constant region.

[1105] In a specific embodiment, an immunoglobulin Fc region and / or an immunoglobulin constant region may comprise any one of the following sequences:

[1106] SEQ ID NO: 289 - FC_Knop_WDN

[1107] GGGGGSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNA KTKPREEQYASTYRWSVLTVLHQDWLNGKEYKCKVSNKALAAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQ VSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK

[1108] SEQ ID NO: 290 - FC_hole_WDN

[1109] METDTLLVFVLLVWVPAGNGDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKF NWYVDGVEVHNAKTKPREEQYASTYRWSVLTVLHQDWLNGKEYKCKVSNKALAAPIEKTISKAKGQPREPQVC TLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGK

[1110] SEQ ID NO: 291 - Human IGHG1 = CH1-CH2-CH3 (AS is for the Nhel restriction site)

[1111] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSS LGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRWSVLTVLHQDWLNGKEYKCKVSNKALAAPIEKTISKAKG QPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSP SEQ ID NO: 292 - Human kappa_light_chain = CL (AS is for the Nhel restriction site)

[1112] ASAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK

[1113] ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[1114] Examples of the antibody construct of the present invention

[1115] In a specific embodiment, the antibody construct may comprise any one of the following sequences:

[1116] SEQ ID NO: 293 - T01

[1117] DVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTMHWVRQAPGQGLEWIGYINPSRGYTNYADSVKGRFTITTDK

[1118] STSTAYMELSSLRSEDTATYYCARYYDDHYCLDYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSDIELTQP

[1119] PSVSVAPGQTARISCSGDNIGNKYVSWYQQKPGQAPVWIYGDNNRPSGIPERFSGSNSGNTATLTISGTQAEDE

[1120] ADYYCSSYDSSYFVFGGGTKLTVLGGGGSGGGGSGGGGSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGF

[1121] TFSSYGMHWVRQAPGKGLEWVSNIYSDGSNTFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARNM

[1122] YRWPFHYFFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCKASQDV

[1123] GIAVAWYQQKPGKVPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATYYCQQYSSYPYTFGQGTK

[1124] VEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFDFSRYWMSWVRQAPGKGLE

[1125] WIGEINPDSSTINYAPSLKDKFIISRDNAKNSLYLQMNSLRAEDTAVYYCARPDGNYWYFDVWGQGTLVTVSSGG

[1126] GGSGGGGSGGGGSGGGGSDIVLTQSPATLSLSPGERATLSCRASQSVSYMNWYQQKPGKAPKRWIYDTSKVA

[1127] SGVPARFSGSGSGTDYSLTINSLEAEDAATYYCQQWSSNPLTFGGGTKVEIKGSHHHHHH

[1128] SEQ ID NO: 294 - T02

[1129] DVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTMHWVRQAPGQGLEWIGYINPSRGYTNYADSVKGRFTITTDK

[1130] STSTAYMELSSLRSEDTATYYCARYYDDHYCLDYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQ

[1131] SPSSLSASVGDRVTITCKASQDVGIAVAWYQQKPGKVPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISSLQPE

[1132] DVATYYCQQYSSYPYTFGQGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAAS

[1133] GFDFSRYWMSWVRQAPGKGLEWIGEINPDSSTINYAPSLKDKFIISRDNAKNSLYLQMNSLRAEDTAVYYCARPD

[1134] GNYWYFDVWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIELTQPPSVSVAPGQTARISCSGDNIGNKYVS

[1135] WYQQKPGQAPVWIYGDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCSSYDSSYFVFGGGTKLTVLG

[1136] GGGSGGGGSGGGGSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVSNI

[1137] YSDGSNTFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARNMYRWPFHYFFDYWGQGTLVTVSSGG

[1138] GGSGGGGSGGGGSGGGGSDIVLTQSPATLSLSPGERATLSCRASQSVSYMNWYQQKPGKAPKRWIYDTSKVA

[1139] SGVPARFSGSGSGTDYSLTINSLEAEDAATYYCQQWSSNPLTFGGGTKVEIKGSHHHHHH SEQ ID NO: 295 - T03

[1140] DIVLTQSPATLSLSPGERATLSCRASQSVSYMNWYQQKPGKAPKRWIYDTSKVASGVPARFSGSGSGTDYSLTIN

[1141] SLEAEDAATYYCQQWSSNPLTFGGGTKVEIKGGGGSGGGGSGGGGSGGGGSQVQLVESGGGLVQPGGSLRL

[1142] SCAASGFTFSSYGMHWVRQAPGKGLEWVSNIYSDGSNTFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY

[1143] YCARNMYRWPFHYFFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIELTQPPSVSVAPGQTARISCS

[1144] GDNIGNKYVSWYQQKPGQAPVWIYGDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCSSYDSSYFVF

[1145] GGGTKLTVLGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFDFSRYWMSWVRQAP

[1146] GKGLEWIGEINPDSSTINYAPSLKDKFIISRDNAKNSLYLQMNSLRAEDTAVYYCARPDGNYWYFDVWGQGTLVT

[1147] VSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCKASQDVGIAVAWYQQKPGKVPKLLIY

[1148] WASTRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATYYCQQYSSYPYTFGQGTKVEIKGGGGSGGGGSGGGGS

[1149] GGGGSDVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTMHWVRQAPGQGLEWIGYINPSRGYTNYADSVKGR

[1150] FTITTDKSTSTAYMELSSLRSEDTATYYCARYYDDHYCLDYWGQGTTVTVSSGSHHHHHH

[1151] SEQ ID NO: 296 - T04

[1152] DIVLTQSPATLSLSPGERATLSCRASQSVSYMNWYQQKPGKAPKRWIYDTSKVASGVPARFSGSGSGTDYSLTIN

[1153] SLEAEDAATYYCQQWSSNPLTFGGGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRL

[1154] SCAASGFDFSRYWMSWVRQAPGKGLEWIGEINPDSSTINYAPSLKDKFIISRDNAKNSLYLQMNSLRAEDTAVYY

[1155] CARPDGNYWYFDVWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCKASQ

[1156] DVGIAVAWYQQKPGKVPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATYYCQQYSSYPYTFGQG

[1157] TKVEIKGGGGSGGGGSGGGGSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGL

[1158] EWVSNIYSDGSNTFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARNMYRWPFHYFFDYWGQGTLV

[1159] TVSSGGGGSGGGGSGGGGSGGGGSDIELTQPPSVSVAPGQTARISCSGDNIGNKYVSWYQQKPGQAPVWIY

[1160] GDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCSSYDSSYFVFGGGTKLTVLGGGGSGGGGSGGGGS

[1161] GGGGSDVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTMHWVRQAPGQGLEWIGYINPSRGYTNYADSVKGR

[1162] FTITTDKSTSTAYMELSSLRSEDTATYYCARYYDDHYCLDYWGQGTTVTVSSGSHHHHHH

[1163] SEQ ID NO: 297 - T05

[1164] DVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTMHWVRQAPGQGLEWIGYINPSRGYTNYADSVKGRFTITTDK

[1165] STSTAYMELSSLRSEDTATYYCARYYDDHYCLDYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQ

[1166] SPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPED

[1167] FATYYCQQHYTTPPTFGQGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGF

[1168] NIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGG

[1169] DGFYAMDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSELVMTQSPSSLTVTAGEKVTMSCKSSQSLLNS

[1170] GNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYPLTFGA

[1171] GTKLEIKGGGGSGGGGSGGGGSGGGGSEVQLLEQSGAELVRPGTSVKISCKASGYAFTNYWLGWVKQRPGHG LEWIGDIFPGSGNIHYNEKFKGKATLTADKSSSTAYMQLSSLTFEDSAVYFCARLRNWDEPMDYWGQGTTVTVS

[1172] SGGGGSGGGGSGGGGSGGGGSDIVLTQSPATLSLSPGERATLSCRASQSVSYMNWYQQKPGKAPKRWIYDTS

[1173] KVASGVPARFSGSGSGTDYSLTINSLEAEDAATYYCQQWSSNPLTFGGGTKVEIKGSHHHHHH

[1174] SEQ ID NO: 298 - T06

[1175] DVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTMHWVRQAPGQGLEWIGYINPSRGYTNYADSVKGRFTITTDK

[1176] STSTAYMELSSLRSEDTATYYCARYYDDHYCLDYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSELVMTQ

[1177] SPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLT

[1178] ISSVQAEDLAVYYCQNDYSYPLTFGAGTKLEIKGGGGSGGGGSGGGGSGGGGSEVQLLEQSGAELVRPGTSVKI

[1179] SCKASGYAFTNYWLGWVKQRPGHGLEWIGDIFPGSGNIHYNEKFKGKATLTADKSSSTAYMQLSSLTFEDSAVY

[1180] FCARLRNWDEPMDYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRAS

[1181] QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQG

[1182] TKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLE

[1183] WVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVS

[1184] SGGGGSGGGGSGGGGSGGGGSDIVLTQSPATLSLSPGERATLSCRASQSVSYMNWYQQKPGKAPKRWIYDTS

[1185] KVASGVPARFSGSGSGTDYSLTINSLEAEDAATYYCQQWSSNPLTFGGGTKVEIKGSHHHHHH

[1186] SEQ ID NO: 299 - T07

[1187] DIVLTQSPATLSLSPGERATLSCRASQSVSYMNWYQQKPGKAPKRWIYDTSKVASGVPARFSGSGSGTDYSLTIN

[1188] SLEAEDAATYYCQQWSSNPLTFGGGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRL

[1189] SCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYY

[1190] CSRWGGDGFYAMDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRAS

[1191] QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQG

[1192] TKVEIKGGGGSGGGGSGGGGSGGGGSEVQLLEQSGAELVRPGTSVKISCKASGYAFTNYWLGWVKQRPGHGL

[1193] EWIGDIFPGSGNIHYNEKFKGKATLTADKSSSTAYMQLSSLTFEDSAVYFCARLRNWDEPMDYWGQGTTVTVSS

[1194] GGGGSGGGGSGGGGSGGGGSELVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPK

[1195] LLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYPLTFGAGTKLEIKGGGGSGGGGSGG

[1196] GGSGGGGSDVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTMHWVRQAPGQGLEWIGYINPSRGYTNYADSV

[1197] KGRFTITTDKSTSTAYMELSSLRSEDTATYYCARYYDDHYCLDYWGQGTTVTVSSGSHHHHHH

[1198] SEQ ID NO: 300 - T08

[1199] DIVLTQSPATLSLSPGERATLSCRASQSVSYMNWYQQKPGKAPKRWIYDTSKVASGVPARFSGSGSGTDYSLTIN

[1200] SLEAEDAATYYCQQWSSNPLTFGGGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLLEQSGAELVRPGTSVKI

[1201] SCKASGYAFTNYWLGWVKQRPGHGLEWIGDIFPGSGNIHYNEKFKGKATLTADKSSSTAYMQLSSLTFEDSAVY FCARLRNWDEPMDYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSELVMTQSPSSLTVTAGEKVTMSCKS

[1202] SQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYS

[1203] YPLTFGAGTKLEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQ

[1204] APGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQ

[1205] GTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAP

[1206] KLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKGGGGSGGGGSGG

[1207] GGSGGGGSDVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTMHWVRQAPGQGLEWIGYINPSRGYTNYADSV

[1208] KGRFTITTDKSTSTAYMELSSLRSEDTATYYCARYYDDHYCLDYWGQGTTVTVSSGSHHHHHH

[1209] SEQ ID NO: 301 - T05.2

[1210] EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYTMNWVRQAPGKGLEWVALINPYKGVSTYNQKFKDRFTISVD

[1211] KSKNTAYLQMNSLRAEDTAVYYCARSGYYGDSDWYFDVWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDI

[1212] QMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISS

[1213] LQPEDFATYYCQQHYTTPPTFGQGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSC

[1214] AASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCS

[1215] RWGGDGFYAMDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSELVMTQSPSSLTVTAGEKVTMSCKSSQ

[1216] SLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYP

[1217] LTFGAGTKLEIKGGGGSGGGGSGGGGSGGGGSEVQLLEQSGAELVRPGTSVKISCKASGYAFTNYWLGWVKQ

[1218] RPGHGLEWIGDIFPGSGNIHYNEKFKGKATLTADKSSSTAYMQLSSLTFEDSAVYFCARLRNWDEPMDYWGQGT

[1219] TVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPKL

[1220] LIYYTSRLESGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPWTFGQGTKVEIKGSHHHHHH

[1221] SEQ ID NO: 302 - T06.2

[1222] EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYTMNWVRQAPGKGLEWVALINPYKGVSTYNQKFKDRFTISVD

[1223] KSKNTAYLQMNSLRAEDTAVYYCARSGYYGDSDWYFDVWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSE

[1224] LVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSG

[1225] TDFTLTISSVQAEDLAVYYCQNDYSYPLTFGAGTKLEIKGGGGSGGGGSGGGGSGGGGSEVQLLEQSGAELVR

[1226] PGTSVKISCKASGYAFTNYWLGWVKQRPGHGLEWIGDIFPGSGNIHYNEKFKGKATLTADKSSSTAYMQLSSLTF

[1227] EDSAVYFCARLRNWDEPMDYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRV

[1228] TITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTP

[1229] PTFGQGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQA

[1230] PGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQG

[1231] TLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPK

[1232] LLIYYTSRLESGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQGNTLPWTFGQGTKVEIKGSHHHHHH SEQ ID NO: 303 - T07.2

[1233] DIQMTQSPSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPKLLIYYTSRLESGVPSRFSGSGSGTDYTLTIS

[1234] SLQPEDFATYYCQQGNTLPWTFGQGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRL

[1235] SCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYY

[1236] CSRWGGDGFYAMDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRAS

[1237] QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQG

[1238] TKVEIKGGGGSGGGGSGGGGSGGGGSEVQLLEQSGAELVRPGTSVKISCKASGYAFTNYWLGWVKQRPGHGL

[1239] EWIGDIFPGSGNIHYNEKFKGKATLTADKSSSTAYMQLSSLTFEDSAVYFCARLRNWDEPMDYWGQGTTVTVSS

[1240] GGGGSGGGGSGGGGSGGGGSELVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPK

[1241] LLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYPLTFGAGTKLEIKGGGGSGGGGSGG

[1242] GGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGYSFTGYTMNWVRQAPGKGLEWVALINPYKGVSTYNQKF

[1243] KDRFTISVDKSKNTAYLQMNSLRAEDTAVYYCARSGYYGDSDWYFDVWGQGTLVTVSSGSHHHHHH

[1244] SEQ ID NO: 304 - T08.2

[1245] DIQMTQSPSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPKLLIYYTSRLESGVPSRFSGSGSGTDYTLTIS

[1246] SLQPEDFATYYCQQGNTLPWTFGQGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLLEQSGAELVRPGTSVKI

[1247] SCKASGYAFTNYWLGWVKQRPGHGLEWIGDIFPGSGNIHYNEKFKGKATLTADKSSSTAYMQLSSLTFEDSAVY

[1248] FCARLRNWDEPMDYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSELVMTQSPSSLTVTAGEKVTMSCKS

[1249] SQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYS

[1250] YPLTFGAGTKLEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQ

[1251] APGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQ

[1252] GTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAP

[1253] KLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKGGGGSGGGGSGG

[1254] GGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGYSFTGYTMNWVRQAPGKGLEWVALINPYKGVSTYNQKF

[1255] KDRFTISVDKSKNTAYLQMNSLRAEDTAVYYCARSGYYGDSDWYFDVWGQGTLVTVSSGSHHHHHH

[1256] In another embodiment, the antibody construct of the present invention or the protein structure may comprise albumin, single albumin-domains, albumin-binding domains or plasma / serum proteins.

[1257] Such an antibody construct may have the following sequence:

[1258] SEQ ID NO: 305 - A1 + T01 MAKWVTFISLLFLFSSAYSRGVFRRDAHKSEVAHRFKDLGEENFKALVLIAFAQYLQQCPFEDHVKLVNEVTEFAK

[1259] TCVADESAENCDKSLHTLFGDKLCTVATLRETYGEMADCCAKQEPERNECFLQHKDDNPNLPRLVRPEVDVMCT

[1260] AFHDNEETFLKKYLYEIARRHPYFYAPELLFFAKRYKAAFTECCQAADKAACLLPKLDELRDEGKASSAKQRLKCA

[1261] SLQKFGERAFKAWAVARLSQRFPKAEFAEVSKLVTDLTKVHTECCHGDLLECADDRADLAKYICENQDSISSKLK

[1262] ECCEKPLLEKSHCIAEVENDEMPADLPSLAADFVESKDVCKNYAEAKDVFLGMFLYEYARRHPDYSWLLLRLAK

[1263] TYETTLEKCCAAADPHECYAKVFDEFKPLVEEPQNLIKQNCELFEQLGEYKFQNALLVRYTKKVPQVSTPTLVEVS

[1264] RNLGKVGSKCCKHPEAKRMPCAEDYLSWLNQLCVLHEKTPVSDRVTKCCTESLVNRRPCFSALEVDETYVPKE

[1265] FNAETFTFHADICTLSEKERQIKKQTALVELVKHKPKATKEQLKAVMDDFAAFVEKCCKADDKETCFAEEGKKLVA

[1266] ASQAALGLSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGG

[1267] GSGGGADVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTMHWVRQAPGQGLEWIGYINPSRGYTNYADSVKG

[1268] RFTITTDKSTSTAYMELSSLRSEDTATYYCARYYDDHYCLDYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGG

[1269] SDIELTQPPSVSVAPGQTARISCSGDNIGNKYVSWYQQKPGQAPVWIYGDNNRPSGIPERFSGSNSGNTATLTIS

[1270] GTQAEDEADYYCSSYDSSYFVFGGGTKLTVLGGGGSGGGGSGGGGSGGGGSQVQLVESGGGLVQPGGSLRL

[1271] SCAASGFTFSSYGMHWVRQAPGKGLEWVSNIYSDGSNTFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY

[1272] YCARNMYRWPFHYFFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITC

[1273] KASQDVGIAVAWYQQKPGKVPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATYYCQQYSSYPYTF

[1274] GQGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFDFSRYWMSWVRQAP

[1275] GKGLEWIGEINPDSSTINYAPSLKDKFIISRDNAKNSLYLQMNSLRAEDTAVYYCARPDGNYWYFDVWGQGTLVT

[1276] VSSGGGGSGGGGSGGGGSGGGGSDIVLTQSPATLSLSPGERATLSCRASQSVSYMNWYQQKPGKAPKRWIYD

[1277] TSKVASGVPARFSGSGSGTDYSLTINSLEAEDAATYYCQQWSSNPLTFGGGTKVEIKGSHHHHHH

[1278] SEQ ID NO: 306 - A1 + T02

[1279] MAKWVTFISLLFLFSSAYSRGVFRRDAHKSEVAHRFKDLGEENFKALVLIAFAQYLQQCPFEDHVKLVNEVTEFAK

[1280] TCVADESAENCDKSLHTLFGDKLCTVATLRETYGEMADCCAKQEPERNECFLQHKDDNPNLPRLVRPEVDVMCT AFHDNEETFLKKYLYEIARRHPYFYAPELLFFAKRYKAAFTECCQAADKAACLLP

[1281] KLDELRDEGKASSAKQRLKCASLQKFGERAFKAWAVARLSQRFPKAEFAEVSKLVTDLTKVHTECCHGDLLECA

[1282] DDRADLAKYICENQDSISSKLKECCEKPLLEKSHCIAEVENDEMPADLPSLAADFVESKDVCKNYAEAKDVFLGMF

[1283] LYEYARRHPDYSWLLLRLAKTYETTLEKCCAAADPHECYAKVFDEFKPLVEEPQNLIKQNCELFEQLGEYKFQNA

[1284] LLVRYTKKVPQVSTPTLVEVSRNLGKVGSKCCKHPEAKRMPCAEDYLSWLNQLCVLHEKTPVSDRVTKCCTESL

[1285] VNRRPCFSALEVDETYVPKEFNAETFTFHADICTLSEKERQIKKQTALVELVKHKPKATKEQLKAVMDDFAAFVEK

[1286] CCKADDKETCFAEEGKKLVAASQAALGLSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGG

[1287] GGSGGGGSGGGGSGGGGSGGGADVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTMHWVRQAPGQGLE

[1288] WIGYINPSRGYTNYADSVKGRFTITTDKSTSTAYMELSSLRSEDTATYYCARYYDDHYCLDYWGQGTTVTVSSGG

[1289] GGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCKASQDVGIAVAWYQQKPGKVPKLLIYWASTRH TGVPDRFSGSGSGTDFTLTISSLQPEDVATYYCQQYSSYPYTFGQGTKVEIKGGGGSGGGGSGGGGSGGGGS

[1290] EVQLVESGGGLVQPGGSLRLSCAASGFDFSRYWMSWVRQAPGKGLEWIGEINPDSSTINYAPSLKDKFIISRDNA

[1291] KNSLYLQMNSLRAEDTAVYYCARPDGNYWYFDVWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIELTQP

[1292] PSVSVAPGQTARISCSGDNIGNKYVSWYQQKPGQAPVWIYGDNNRPSGIPERFSGSNSGNTATLTISGTQAEDE

[1293] ADYYCSSYDSSYFVFGGGTKLTVLGGGGSGGGGSGGGGSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGF

[1294] TFSSYGMHWVRQAPGKGLEWVSNIYSDGSNTFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARNM

[1295] YRWPFHYFFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIVLTQSPATLSLSPGERATLSCRASQSVS

[1296] YMNWYQQKPGKAPKRWIYDTSKVASGVPARFSGSGSGTDYSLTINSLEAEDAATYYCQQWSSNPLTFGGGTKV EIKGSHHHHHH

[1297] SEQ ID NO: 307 - A1 + T03

[1298] MAKWVTFISLLFLFSSAYSRGVFRRDAHKSEVAHRFKDLGEENFKALVLIAFAQYLQQCPFEDHVKLVNEVTEFAK

[1299] TCVADESAENCDKSLHTLFGDKLCTVATLRETYGEMADCCAKQEPERNECFLQHKDDNPNLPRLVRPEVDVMCT

[1300] AFHDNEETFLKKYLYEIARRHPYFYAPELLFFAKRYKAAFTECCQAADKAACLLPKLDELRDEGKASSAKQRLKCA

[1301] SLQKFGERAFKAWAVARLSQRFPKAEFAEVSKLVTDLTKVHTECCHGDLLECADDRADLAKYICENQDSISSKLK

[1302] ECCEKPLLEKSHCIAEVENDEMPADLPSLAADFVESKDVCKNYAEAKDVFLGMFLYEYARRHPDYSWLLLRLAK

[1303] TYETTLEKCCAAADPHECYAKVFDEFKPLVEEPQNLIKQNCELFEQLGEYKFQNALLVRYTKKVPQVSTPTLVEVS

[1304] RNLGKVGSKCCKHPEAKRMPCAEDYLSWLNQLCVLHEKTPVSDRVTKCCTESLVNRRPCFSALEVDETYVPKE

[1305] FNAETFTFHADICTLSEKERQIKKQTALVELVKHKPKATKEQLKAVMDDFAAFVEKCCKADDKETCFAEEGKKLVA

[1306] ASQAALGLSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGG

[1307] GSGGGADIVLTQSPATLSLSPGERATLSCRASQSVSYMNWYQQKPGKAPKRWIYDTSKVASGVPARFSGSGS

[1308] GTDYSLTINSLEAEDAATYYCQQWSSNPLTFGGGTKVEIKGGGGSGGGGSGGGGSGGGGSQVQLVESGGGLV

[1309] QPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVSNIYSDGSNTFYADSVKGRFTISRDNSKNTLYLQMNS

[1310] LRAEDTAVYYCARNMYRWPFHYFFDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIELTQPPSVSVAP

[1311] GQTARISCSGDNIGNKYVSWYQQKPGQAPVWIYGDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCSS

[1312] YDSSYFVFGGGTKLTVLGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFDFSRYWM

[1313] SWVRQAPGKGLEWIGEINPDSSTINYAPSLKDKFIISRDNAKNSLYLQMNSLRAEDTAVYYCARPDGNYWYFDVW

[1314] GQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCKASQDVGIAVAWYQQKPGK

[1315] VPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATYYCQQYSSYPYTFGQGTKVEIKGGGGSGGGG

[1316] SGGGGSGGGGSDVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTMHWVRQAPGQGLEWIGYINPSRGYTNYA

[1317] DSVKGRFTITTDKSTSTAYMELSSLRSEDTATYYCARYYDDHYCLDYWGQGTTVTVSSGSHHHHHH

[1318] SEQ ID NO: 308 - A1 + T04 MAKWVTFISLLFLFSSAYSRGVFRRDAHKSEVAHRFKDLGEENFKALVLIAFAQYLQQCPFEDHVKLVNEVTEFAK

[1319] TCVADESAENCDKSLHTLFGDKLCTVATLRETYGEMADCCAKQEPERNECFLQHKDDNPNLPRLVRPEVDVMCT

[1320] AFHDNEETFLKKYLYEIARRHPYFYAPELLFFAKRYKAAFTECCQAADKAACLLPKLDELRDEGKASSAKQRLKCA

[1321] SLQKFGERAFKAWAVARLSQRFPKAEFAEVSKLVTDLTKVHTECCHGDLLECADDRADLAKYICENQDSISSKLK

[1322] ECCEKPLLEKSHCIAEVENDEMPADLPSLAADFVESKDVCKNYAEAKDVFLGMFLYEYARRHPDYSWLLLRLAK

[1323] TYETTLEKCCAAADPHECYAKVFDEFKPLVEEPQNLIKQNCELFEQLGEYKFQNALLVRYTKKVPQVSTPTLVEVS

[1324] RNLGKVGSKCCKHPEAKRMPCAEDYLSWLNQLCVLHEKTPVSDRVTKCCTESLVNRRPCFSALEVDETYVPKE

[1325] FNAETFTFHADICTLSEKERQIKKQTALVELVKHKPKATKEQLKAVMDDFAAFVEKCCKADDKETCFAEEGKKLVA

[1326] ASQAALGLSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGG

[1327] GSGGGADIVLTQSPATLSLSPGERATLSCRASQSVSYMNWYQQKPGKAPKRWIYDTSKVASGVPARFSGSGS

[1328] GTDYSLTINSLEAEDAATYYCQQWSSNPLTFGGGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLV

[1329] QPGGSLRLSCAASGFDFSRYWMSWVRQAPGKGLEWIGEINPDSSTINYAPSLKDKFIISRDNAKNSLYLQMNSLR

[1330] AEDTAVYYCARPDGNYWYFDVWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRV

[1331] TITCKASQDVGIAVAWYQQKPGKVPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATYYCQQYSSY

[1332] PYTFGQGTKVEIKGGGGSGGGGSGGGGSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVR

[1333] QAPGKGLEWVSNIYSDGSNTFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARNMYRWPFHYFFDY

[1334] WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIELTQPPSVSVAPGQTARISCSGDNIGNKYVSWYQQKPG

[1335] QAPVWIYGDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCSSYDSSYFVFGGGTKLTVLGGGGSGGG

[1336] GSGGGGSGGGGSDVQLVQSGAEVKKPGASVKVSCKASGYTFTRYTMHWVRQAPGQGLEWIGYINPSRGYTNY

[1337] ADSVKGRFTITTDKSTSTAYMELSSLRSEDTATYYCARYYDDHYCLDYWGQGTTVTVSSGSHHHHHH

[1338] Additional Sequences

[1339] When producing the antibody construct of the present invention, the following sequences may be used:

[1340] SEQ ID NO: 309 - Eukaryote secretion leader

[1341] METDTLLVFVLLVWVPAGNG

[1342] SEQ ID NO: 310 - Fusion prokaryote production leader protein Thioredoxin-1 :E. coli, P0AA25

[1343] MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDEIADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPT

[1344] LLLFKNGEVAATKVGALSKGQLKEFLDANLAGGSGGGGGSLEVLFQGPM

[1345] SEQ ID NO: 311 - Linker for connecting prokaryote Thioredoxin-1 :E. coli, P0AA25, to 3C protease domain GGSGGGGGS

[1346] SEQ ID NO: 312 - 3C protease domain for cutting off the prokaryote Thioredoxin-1 from the KeyLock antibody

[1347] LEVLFQGPM

[1348] SEQ ID NO: 313 - Amino acid linker for Kasl restriction site, connecting secretion leader and / or human albumin with KeyLock antibodies

[1349] GA

[1350] Histidine tags which may be used in accordance with the present invention include:

[1351] SEQ ID NO: 314 - Preferred Histidin tag = 6x His tag

[1352] GSHHHHHH

[1353] SEQ ID NO: 315 - Alternative Histidin tag = 8x His tag

[1354] GSHHHHHHHH

[1355] SEQ ID NO: 316 - Alternative Histidin tag = 6x His tag

[1356] HHHHHH

[1357] SEQ ID NO: 317 - Alternative Histidin tag = 4x His tag

[1358] HHHH

[1359] SEQ ID NO: 318 - Alternative Histidin tag = 4x His tag

[1360] GSHHHH SEQ ID NO: 319 - Alternative Histidin tag = 10x His tag

[1361] GSHHHHHHHHHH

[1362] BITE antibodies which were used as controls for the target cell killing experiments include the following antibodies. The BiTE antibodies are IgG antibodies in the knob-hole format. The “Fc_Knob_antiCD3_UCHT1” (SEQ ID NO: 325) was paired with the "Fc_hole” constructs. The "Fc_hole” constructs are against SLAMF7, CD38, EpCAM and Her2, respectively:

[1363] SEQ ID NO: 325 Fc_Knob_antiCD3_UCHT1

[1364] GP

[1365] EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYTMNWVRQAPGKGLEWVALINPYKGVSTYNQKFKDRFTISVD KSKNTAYLQMNSLRAEDTAVYYCARSGYYGDSDWYFDVWGQGTLVTVSSGGGGSGGGGSGGGGSDIQMTQS PSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPKLLIYYTSRLESGVPSRFSGSGSGTDYTLTISSLQPEDF ATYYCQQGNTLPWTFGQGTKVEIK GS

[1366] GGGGGSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNA KTKPREEQYASTYRWSVLTVLHQDWLNGKEYKCKVSNKALAAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQ VSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK

[1367] SEQ ID NO: 326 Fc_hole_anti_SLAMF7

[1368] DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYASTYRWSVLTVLHQDWLNGKEYKCKVSNKALAAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSL SPGK GS GGS

[1369] EVQLVESGGGLVQPGGSLRLSCAASGFDFSRYWMSWVRQAPGKGLEWIGEINPDSSTINYAPSLKDKFIISRDNA KNSLYLQMNSLRAEDTAVYYCARPDGNYWYFDVWGQGTLVTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLS ASVGDRVTITCKASQDVGIAVAWYQQKPGKVPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATYY CQQYSSYPYTFGQGTKVEIKGSHHHHHH

[1370] SEQ ID NO: 327 Fc_hole_anti_CD38

[1371] DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYASTYRWSVLTVLHQDWLNGKEYKCKVSNKALAAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSL SPGK GS GGS

[1372] QVQLVESGGGLVQPGGSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVSNIYSDGSNTFYADSVKGRFTISRD

[1373] NSKNTLYLQMNSLRAEDTAVYYCARNMYRWPFHYFFDYWGQGTLVTVSSGGGGSGGGGSGGGGSDIELTQPP

[1374] SVSVAPGQTARISCSGDNIGNKYVSWYQQKPGQAPVWIYGDNNRPSGIPERFSGSNSGNTATLTISGTQAEDEA

[1375] DYYCSSYDSSYFVFGGGTKLTVLGQGSHHHHHH

[1376] SEQ ID NO: 328 Fc_hole_anti_EpCAM

[1377] DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREE

[1378] QYASTYRWSVLTVLHQDWLNGKEYKCKVSNKALAAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAV

[1379] KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSL

[1380] SPGK GS GGS

[1381] EVQLLEQSGAELVRPGTSVKISCKASGYAFTNYWLGWVKQRPGHGLEWIGDIFPGSGNIHYNEKFKGKATLTADK

[1382] SSSTAYMQLSSLTFEDSAVYFCARLRNWDEPMDYWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSELVMTQ

[1383] SPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLT

[1384] ISSVQAEDLAVYYCQNDYSYPLTFGAGTKLEIKGSHHHHHH

[1385] SEQ ID NO: 329 Fc_hole_anti_Her2

[1386] DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREE

[1387] QYASTYRWSVLTVLHQDWLNGKEYKCKVSNKALAAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAV

[1388] KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSL

[1389] SPGK GS GGS

[1390] EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTS

[1391] KNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMT

[1392] QSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPE

[1393] DFATYYCQQHYTTPPTFGQGTKVEIKGSHHHHHH

[1394] Preparation of Compositions of the Invention

[1395] Compositions in accordance with the present invention are prepared in accordance with known standards for the preparation of pharmaceutical compositions.

[1396] For instance, the compositions are prepared in a way that they can be stored and administered appropriately, e.g. by using pharmaceutically acceptable components such as carriers, excipients or stabilizers. Such pharmaceutically acceptable components are not toxic in the amounts used when administering the pharmaceutical composition to a patient. The pharmaceutical acceptable components added to the pharmaceutical compositions may depend, for instance, on the particular intended use of the pharmaceutical compositions and the route of administration.

[1397] In general, the pharmaceutically acceptable components used in connection with the present invention are used in accordance with knowledge available in the art, e.g. from Remington's Pharmaceutical Sciences, Ed. AR Gennaro, 20th edition, 2000, Williams & Wilkins, PA, USA.

[1398] EXAMPLES

[1399] Example 1 - Selecting antigen targets on solid tumours

[1400] The first KeyLock antibodies were designed against well-known solid tumour antigens, used in clinic for immunotherapy before. Breast cancer is one of the most common malignant tumours in women, with great medical need to optimize treatment of late stage or metastasised tumours (33). Gastric cancer is the fifth most common cancer and the third most common cause of cancer death globally (34). The protein HER2 / neu (ERBB2) is overexpressed on malignant cells of 20-30% of breast cancer patients, 4.4 - 53.4% with a mean range of 17.9% of gastric- and gastroesophageal cancer patients and on a variety of different solid tumours (salivary-, vaginal-, bladder-, endometrial-, cervical-, hepatobiliary-, ampullary-, ovarian-, colorectal, non-small cell lung- and pancreatic cancer) (35, 36). Anti-HER2 targeting therapies in clinic or clinical development are using most often antibodies in the IgG format, like trastuzumab (Herceptin®), or different antibody-drug conjugates (37). Due to the co-expression of HER2 / neu on healthy vital tissues, using high potent T cell redirecting immunotherapies like CAR T cells against single antigen HER2 / neu positive cells, can cause "on-target / off-tumor” recognition with severe and life threatening site effects (38, 39). The cell adhesion antigen EpCAM is found on normal and malignant epithelia cells and is frequently overexpressed in different solid tumours. It plays an important role in EpCAM positive malignant cell proliferation and survival (40, 41). Due to the co-expression of EpCAM on healthy epithelia, using a T cell redirecting therapy like the bi-specific EpCAM-CD3 antibody solitomab, can cause severe side effects (42).

[1401] Therefore, the spatiotemporal patterning of HER2 / neu and EpCAM in healthy tissues, and the high co-expression on malignant cells, makes a dual antigen restricted HER2 / neu and EpCAM targeting strategy highly promising for treating different solid tumours (41 , 43, 52, 53).

[1402] Example 2 - Selecting antigen targets on hematopoietic malignancies

[1403] The second group of KeyLock antibodies were designed against the antigens CD38 and SLAMF7 (CD319), expressed on different hematopoietic malignancies. These are: (i) T cell acute lymphoblastic leukemia (T-ALL) (60, 61 , 62). (ii) Natural killer / T cell lymphoma and large granular lymphocyte leukemia (63). (iii) B-cell chronic lymphocytic leukemia (B-CLL) (64). (iv) Multiple Myeloma and other plasma cell dyscrasias like amyloidosis (65, 69). The combination of CD38 and SLAMF7 is also found on tumor-associated macrophages (TAMs). TAMs are important for the tumour microenvironment. They play a role in the conversion of infiltrating T cells to an exhaustion phenotype, allowing the tumours to evade the host immune response and continue the unchecked tumour growth (66). The unique subset of SLAMF7h'9hCD38h'9hTAMs show a strong correlation with exhausted T cells and are independent prognostic factor for poor survival in different solid tumours. Therefore, targeting TAMs is a promising way to optimize immunotherapy of solid tumours.

[1404] Example 3 - Generation of single and dual antigen positive firefly luciferase-positive CHO cells

[1405] The firefly luciferase (ffluc) and the genes coding for full-length CD38 (Uniprot P28907, Gene ID 952), and / or SLAMF7 (Uniprot Q9NQ25, Gene ID 57823), and / or Her2 / neu (Uniprot P04626, Gene ID 2064), and / or EpCAM (Uniprot P16422, Gene ID 4072) were cloned in PiggyBac (PB) cDNA Cloning and expression Vector, PB-CMV- MCS-EF1a-RedPuro cDNA Cloning and Expression Vector (PB514B-2, SystemBiosciences, LLC, Palo Alto, CA, USA). The Chinese hamster ovary (CHO, DSMZ, Braunschweig, Germany, DSMZ no.: ACC 110) cells were transfected with one or two PB vectors, for the expression of the respective single or dual antigens. Briefly, a total of 0.3 x 106adherent CHO cells in 2 ml F-12K media were transfected by 250 pl DNA-lipid complex consisting of 12.5 pl LipofectamineTM 2000 Transfection Reagent (Thermo Fisher Scientific, MA, USA), 1 pg PB transposase, and 1.5 pg of PB vectors in Opti-MEM medium at 37 °C in a humidified incubator supplemented with 5% CO2 for 48 h. Single or dual antigen-positive CHO cells were screened and selected by the expressed antigen levels using the fluorescence-activated cell sorting (FACS). The sequences used for CD38, SLAMF7, Her2 and EpCAM were obtained from the National Center for Biotechnology Information website (www.ncbi.nlm.nih). The cells were cultivated in F12-K medium supplemented with 200 pM L-glutamine, 10% FBS, penicillin (200 U / ml), and streptomycin (200 pg / ml) (Thermo Fisher Scientific, MA, USA). All cell lines were tested negative for mycoplasma. Authentication of the cells was performed by the provider.

[1406] Example 4 - Isolation of human CD8+ T cells from blood healthy donors

[1407] CD8+ T cells, from healthy human donors, were used as effector cells to test the in vitro activity of the KeyLock antibodies. First, the mononuclear cells were separated from the peripheral blood through Ficoll density gradient centrifugation (Ficoll-Paque™ Plus, Cytivia) at 3000 rpm with acceleration at 5 and deceleration at 1 for 18 minutes. Afterward the separated peripheral blood mononuclear cells (PBMCs) were collected and washed with 40ml PBS. The PBMCs were then subjected to red blood cell lysis with 10 ml of Gibco's ACK lysing buffer for 15 minutes at room temperature. Then, the PBMCs were washed one time with 40 ml PBS and proceed for magnetic selection according to the user manual of the magnetic beads provided by the company (CD8+ T Cell Isolation Kit, human, Miltenyi). A negative selection was used to isolate the CD8+ T cell population from the CD8 negative PBMC cells. Briefly, the number of PBMNCs was first determined and 1x 108PBMNCs were resuspended in 400 pil of MACS buffer (PBS at pH 7.2 supplemented with 0.5% BSA and 2 mM EDTA). Then, 200 pil of MicroBeads (to deplete CD8 negative cells) was added to the PBMNCs suspension, mixed well and incubated for 15 minutes and then washed with 50 ml MACS buffer and resuspended in 500 pl of MACS buffer. The labelled PBMNC were then loaded onto their respective columns according to the cell number. All magnetic labelled CD8-negative cells are captured on to the magnetic column while the CD8-positive cells were eluted by washing the column with MACS buffer. The CD8-positive cells were then collected and cultivated in RPMI 1640 supplemented with 10% (v / v) FBS, 1 % (v / v) GlutaMAX™, 1 % (v / v) penicillin-streptomycin-neomycin antibiotic mixture and 50 lU / ml human IL-2. In order to expand the cells, Gibco™ Dynabeads™ Human T-Activator CD3 / CD28 for T Cell Expansion and Activation was added to the CD8+ T cells with one bead to five cells ratio and the expansion was carried out for five days. After the 5 days, the CD3 / CD28 Dynabeads™ were captured and removed from the proliferating CD8 positive T cells, using a magnet. The CD8+ T cells were used directly for in vitro killing assays or cryopreserved in 50% (v / v) complete RPMI 1640 supplemented with 45% (v / v) FBS and 10% (v / v) DMSO for future use.

[1408] Example 5 - Recombinant production of KeyLock antibodies targeting HER2 / neu - EpCAM and CD38 - SLAMF7 in eukaryote and prokaryote cells

[1409] To generate different KeyLock antibody DNA sequences, custom gene synthesis services including chemical synthesis was used (GeneArt®, Thermo Fisher Scientific Inc.). Each DNA plasmid had their specific restriction enzyme cloning sides at the end for subcloning into the expression plasmids. The final sequences were cloned into an expression plasmid (pCEP4 Mammalian Expression Vector, Invitrogen™) and / or (pCOLD™ IV bacterial Expression Vector (TaKaRa Bio Inc). Standard methods of molecular biology were used (see, e.g. Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, New York (2001)).

[1410] The expression plasmids carrying the respective individual antibody sequence were transformed, separate for each sequence, into E. coli Maehl cells (Invitrogen, C862003). After short thawing E. coli cells on ice, 10 piL of antibody plasmid (10-100 ng DNA) was mixed with 90piL of cells by gentle pipetting and incubated for 20 minutes on ice. Afterwards, a heat-shock was performed at 42°C for 90 seconds followed by incubation on ice for 5 minutes. The cells were then transferred into 600 piL of 2xYT medium (Carl Roth, X966.3) and incubated at 37°C and 230 rpm for one hour. After a subsequent centrifugation at 13000 rpm for two minutes, 500 piL of the supernatant was discarded. The cells were resuspended in the remaining 200 piL of medium and plated out on LB (Carl Roth, 6673.1) agar plates containing a plasmid specific selection antibiotic (100 pig / mL Carbenicillin). The plates were then incubated upside down at 37°C overnight. On the next day, a single cell colony was picked and inoculated in 5 mL of 2xYT medium (100 pig / mL Carbenicillin) and incubated at 37°C overnight. An aliquot of each over-night culture was stored at 4°C for short-term preservation. The over-night cultures were centrifuged at room temperature and 4500 g for 10 minutes after which the antibody plasmids were isolated according to the QIAprep Spin Miniprep Kit (QIAGEN, 27115). For the elution step, 20 pil pre-warmed UltraPure Water (Invitrogen, 11538646) was added to the middle of the column membrane and incubated at room temperature for two minutes before centrifugation at 13000 rpm for three minutes. The elution step was repeated one more time to maximize the final plasmid DNA yield. To determine the concentration and purity of the plasmids, the products were analyzed photometrically at 260nm and 280 nm using the NanoDrop® 2000 (Thermo Scientific, ND-2000). The plasmids were stored at -20°C until further use.

[1411] To ensure the correct pCEP4 antibody plasmids were picked and produced, and no cross contamination happened in the production process, 20 l (300 - 800 ng DNA) of each plasmid was send for DNA sequencing by LGC Genomics (Berlin, Germany), using the sequencing primers pCEP4_Fwd (5- GCGTGTACGGTGGGAGGTCTAT ATA-3' - SEQ I D NO 320), SV40_Rev (5'-CACTGCATTCTAGTTGTGGTT-3' -

[1412] - SEQ I D NO 321) and pGEX3_Rev (5'-CCGGGAGCTGCATGTGTCAGAGG-3' - SEQ I D NO 322).

[1413] To ensure the correct pCOLD IV antibody plasmids were picked and produced, and no cross contamination happened in the production process, 20 l (300 - 800 ng DNA) of each plasmid was send for DNA sequencing by LGC Genomics (Berlin, Germany), using the sequencing primers pCold Fwd (5'-ACGCCATATCGCCGAAAGG-3'

[1414] - SEQ I D NO 323)and pCold Rev (5'-GGCAGGGATCTTAGATTCTG-3' - SEQ ID NO 324).

[1415] For large-scale plasmid amplifications, short-term preserved E. coll Maehl cultures, positive for the respectife antibody plasmid, were inoculated in 500 mL of 2xYT medium (100 pig / mL Carbenicillin) and incubate at 37°C and 230 rpm overnight. An aliquot (1 ml) of each overnight culture was mixed with the same volume of LB-medium (+ 30% glycerol) and stored at -80°C for long-term preservation. The overnight cultures (499 ml) were centrifuged at 5000 g and 4°C for 30 minutes after which the plasmids were isolated according to the NucleoBond Xtra Maxi Kit (Maschery Nagel, 740414.50) while considering low copy numbers. The final purified pCEP4 antibody plasmids were reconstituted in UltraPure Water (Invitrogen, 11538646). To determine the concentration and purity of the plasmids, the products were analyzed photometrically at 260nm and 280 nm using the NanoDrop® 2000 (Thermo Scientific, ND-2000). The plasmids were stored at -20°C until further use.

[1416] Recombinant KeyLock production in eukaryote cells:

[1417] The DNA sequences of different anti CD38 - SLAMF7 and anti HER2 / neu - EpCAM KeyLock antibodies were cloned into the pCEP4 Mammalian Expression Vector (Invitrogen™). The pCEP4 vector is for transient high protein expression from a CMV promotor, also including the EBNA-1 gen for episomal expression in eukaryote cells. As expressing cells the inventors used the Expi-HEK293 cell system (Expi293™ Expression System Kit, Gibco™).

[1418] The cells were seeded on the day of transfection in fresh Freestyle™ 293 Expression Medium (Gibco, 12338018) at a concentration of 2*106 cells / mL in an orbital shaker flask. The transfection composite was prepared by mixing one part of plasmid with three parts of PEI in fresh Freestyle™ 293 Expression Medium according to the following scheme: Transfection Composite for Transient Transfection: Volume of Culture 100 mL, Freestyle Medium 10 mL, Amount of DNA 200 g of plasmid, Volume of 1 mg / mL PEI 600 piL, Volume for 0,5% TN1 2,5 mL, Volume of flask 500 mL.

[1419] The transfection composite was incubated at room temperature for 15 minutes to allow complex formation and was then added to the cells. The cell cultures were incubated at 37°C, 5% CO2 and constant shaking at 110 rpm for five days or until cell viability dropped below 60%. After the first day of transfection, Tryptone N1 (PanReac AppliChem, A1553.1000) was added to a final concentration of 0,5% (v / v) to each cell culture. Five days after KeyLock-plasmid transfection, we harvested the cell culture supernatant for protein purification. The supernatant was first dialyzed against 1 x PBS, pH 7.4 at 4 °C, overnight. The next day the supernatant was mixed with equilibrated CO-IMAC beads and incubate at 4 °C for 2 h in rotation. Afterward the protein-bead complex were washed with CO-IMAC Loading buffer 20CV, followed from eluting the proteins with CO-IMAC Elution buffer 2CV for 4 times. The different wash and elution fractions were loaded onto a 10% SDS-PAGE Gel and Coomassie Brilliant Blue (CBB) stained (Figures 11-16). After elution the protein fractions were desalted, using HiPrep™ 26 / 10 desalting columns (Sigma-Aldrich®). Afterward the protein was concentrated and buffer exchanged into the final storage buffer using Amicron® Ultra-4 30K tubes (Millipore®). The final KeyLock protein purity and concentration was analysed, loading them onto a 10% SDS-PAGE Gel, with a bovine serum albumin protein standard (BSA) and CBB stained (Figures 17, 18).

[1420] Recombinant KeyLock production in prokaryote cells:

[1421] The DNA sequences of the three different anti CD38 - SLAMF7 KeyLock antibodies (T02, T03 and T04) were cloned into the pCOLD™ IV bacterial Expression Vector (TaKaRa Bio Inc). The pCOLD system is using the coldshock Protein A (cspA) promoter for expression of high-purity, high-yield recombinant protein in E. coll. The vector selectively induces target protein synthesis at a low temperature (16°C), a condition which suppresses the expression of host proteins and decreases protease activity. This results in high yields of target proteins.

[1422] A second plasmid was used to co-express the viral 3C protease in the bacteria, to cut off the N-terminal Thioredoxin- 1 leader protein from the final KeyLock antibodies.

[1423] For each KeyLock antibody, four different transfected bacteria clones were picked, and analysed for KeyLock antibody expression, using an anti-His-tag antibody (SHuffle® T7 Competent E. coll cells, New England Biolabs Inc.) (Figure 19). The highest expressing clones were used for following bacterial fermentation and production in a larger volume, using 2x YT medium supplemented with kananmycin and carbenicilin and 0.2% glucose. KeyLock protein production was induced adding 0.1 mM IPTG and incubated at 16°C for 18 - 20 hours. Afterward from the E. coll cell pellet the antibodies were extracted using cobalt beads. After incubation with the cobal beads at 4° C overnight, the beads were loaded onto a purification column and washed with CO-IMAC Loading, washing and elution Buffer. All fractions were loaded onto a SDS PAGE gel to check the KeyLock protein production (Figure 20). The cleanest elution fractions with the highest KeyLock protein concentration were pooled for each construct and loaded onto a SDS PAGE gel with a BSA standard to determine total protein concentrations (Figure 21).

[1424] Recombinant production of KeyLock-Albumin fusion constructs:

[1425] The DNA sequences of three different anti-HER2 / neu - EpCAM - KeyLock antibodies (T06.2, T07.2 and T08.2) was fused to the human Albumin sequence (A1-T06.2, A1-T07.2 and A1-T08.2), and cloned into the pCEP4 Mammalian Expression Vector (Invitrogen ™). The expression and purification of the constructs was the same as for the different KeyLock constructs without Albumin fusion. The final purified KeyLock-Albumin proteins were loaded onto 10% SDS-PAGE Gel and CBB stained (Figures 22).

[1426] The expression data (Figure 11 to Figure 22) show, different KeyLock antibodies, using different antiCD3 effector binding domains (UCHT 1 and diL2K), different target antigen binding domains (anti-HER2 / neu, -EpCAM, -SLAMF7 and -CD38), different orientations of the variable domains (VH-VL and VL-VH), different attachment of the split / separated antiCD3 VH and VL domains (at the N-terminal and C-terminal ends), with or without connected human albumin in the KeyLock antibody format, all can be expressed, purified, and concentrated for a final protein in solution. This is possible in prokaryote and eukaryote cells, using different expression plasmids (pCEP4 and pCOLD IV) and different expressing cells (Expi-HEK293 and SHuffle® T7 cells).

[1427] Example 6 - Testing the in vitro activity of KeyLock antibodies against single and double antigen positive target cells

[1428] The single- and double-antigen positive CHO cells were positive for the luziferase fire fly protein, expressed by viable cells only. The CHO cells were seeded into 96-well cell culture plates (7500 cells in 50 piL per well for 150 cells per piL) and incubated at 37°C and 5% CO2 for 24 hours. The next day, fresh human CD3+ / CD8+ T-cells were added to each well, for a final CHO cell to T cells ratio of 1 to 3. A dilution series of the KeyLock antibodies was prepared and 20 piL was added to each respective well. As positive control for target cell killing with a T cell recruiting antibody, a bi-specific antibody against Her2 (SEQ ID No xx), EpCAM (SEQ ID No xx), CD38 (SEQ ID No xx) and SLAMF7 (SEQ ID No xx) was used at a respective final concentration (Figures 23, 24, 27, 28, 29, 30, 34 and 35).

[1429] As positive control for target cell killing independent of the T cell function, a DMSO solution with a final concentration of 5% in the cell culture medium was used (Figures 23 to 30, and 34 to 35).

[1430] As reference control, to control for unspecific donor T cell activation against the target cells, target cells only and target cells mixed with T cells was used (Figures 23 to 30, and 34 to 35).

[1431] The plates were incubated at 37°C and 5% CO2 for 48 hours. Then 20 piL of 3.5 mM luciferin solution was added to each well (final concentration 0.5 mM) and the plates were incubated at 37°C for 30 minutes. Afterwards, the bioluminescence of the firefly-luciferase was measured, using a Tecan Spark plate reader (with an absorbance scan from 350 nm to 700 nm with an integration time interval of 1000 ms). Example 7 - Testing the protease stability of KeyLock antibodies in human serum

[1432] The possible degradation of the KeyLock antibodies by protease in the serum is a risk for their therapeutic efficacy. Therefore, we tested the serum stability of the KeyLock antibodies, produced in eukaryote CHO cells (pCEP4) (Figure 30). The KeyLock antibodies were incubated for 24 hours at 36°C in fresh human serum. For human serum, the peripheral blood from a healthy human donor was collected in a S-Monovette® (Sarstedt, Germany). The sample was centrifuged at 3.000 rpm for 5 minutes and the serum supernatend was pipetted into a fresh tube. The serum was not heat inactivated and used directly for the KeyLock incubation. The next day the serum treated antibodies were used for target cell killing experiments, side by side with KeyLock antibodies not incubated in serum.

[1433] Example 8 - Testing the size-exclusion chromatography profile of the KeyLock antibodies

[1434] For analyzing if KeyLock antibodies form monomers, dimers or multimers in solution, KeyLock antibodies were purified by immobilized metal affinity chromatography (IMAC), desalted and resuspended in physiologic phosphate buffer solution (PBS). Then, the KeyLock antibodies were analyzed by size-exclusion chromatography (SEC) using fast protein liguid chromatography (FPLC). In physiologic PBS solution, in the absence of any stabilizers, some KeyLock antibodies are essentially monomeric (Figure 31). This finding is important in light of the split / separated hydrophobic interfaces of the V domains which in some cases are responsible for poor solubility and the propensity to aggregate (72).

[1435] Example 9 - Testing the thermostability profile of the KeyLock antibodies

[1436] The thermal stability of an antibody is important for its solubility, aggregation and production (75). Therefore, we tested the thermal stability of different KeyLock antibodies, produced in eukaryote and prokaryote cells (Figure 32). We used the IgG antibody Daratumumab as control for antibody thermostability. The heat stability of the KeyLock antibodies, in a single chain polypeptide, is 54.5°C, similar to the hemibodies (13).

[1437] Example 10 - Testing the unspecific T cell activation by the KeyLock antibodies in vitro

[1438] With the increased use of different T cell engaging therapies in clinic comes the increasing awareness of their inherent and potentially fatal adverse effects, most notably the cytokine release syndrome (CRS) (76). For antibodies with a functional anti-CD3 domain, it is possible to bind and activate cytotoxic T cells in the absence of target cells, causing a CRS. Therefore, we tested if KeyLock antibodies are able to activate cytotoxic T cells if no target antigen is there to bind (Figure 33). We co-incubated high dense fresh isolated human T cells in vitro, with high concentration of different anti-CD38-SLAMF7 and anti-HER2 / neu-EpCAM KeyLock antibodies (10 nM). For T cell activation, the production of Interferon-gamma was measured in the culture supernatant using standard IFN- gamma ELISA assay. In contrast to the bi-specific CD3-CD28 construct, used as positive control, none of the KeyLock antibodies were able to significantly active T cells by themselves.

[1439] Example 11 - Testing to block the KeyLock antibody binding on single and double antigen positive target cells When co-incubated with single or double antigen positive Chinese Hamster Ovary (CHO) cells, different KeyLock antibodies (T02 and T05.2) were not able to activate cytotoxic T cells to lyse single or double antigen positive target cells, if the cells are not expressing both antigens on their cell surface (Figure 34) and if the binding of one KeyLock antibody targeting scFv domain was blocked from binding to its respective antigen epitope (Figure 35). We coincubated human CD8 positive T cells in vitro, with different concentrations of anti-CD38-SLAMF7 and anti- HER2 / neu-EpCAM KeyLock antibodies (T02 5nM, 10nM 20nM and 30nM, and for T05.2 0.00195nM, 0.0039nM, 0.0078nM and 0.01563nM). For target cell killing the CHO cell viability was determined through the Luziferase signal after 24 hours cell culture. For soluble target antigens, only the extracellular domain of the respective human antigen (CD38 (Uniprot P28907, Gene ID 952), SLAMF7 (Uniprot Q9NQ25, Gene ID 57823), Her2 / neu (Uniprot P04626, Gene ID 2064), EpCAM (Uniprot P16422, Gene ID 4072) was fused the human Ig constant Fc domain, expressed and purified in HEK293 cells.

[1440] Example 12 - Measuring KeyLock antibody concentration in solution

[1441] To determine the final KeyLock antibody concentration, the Pierce™ BCA Protein Assay Kit (Thermo Scientific™, #23225) was used after the manufactures protocol. In short, protein-standard and antibody samples were pipette into separate test tubes. Then working reagent was added to each tube, mixed and incubate at room temperature for 30-60 minutes in the dark. With a spectrophotometer set to 562 nm the absorbance of all samples was measured within 10 minutes. The average 562 nm absorbance measurement of the Blank standard was subtracted from the 562nm absorbance measurement of all other individual standards and antibody samples, to generate a standard curve to determine the protein concentration.

[1442] INDUSTRIAL APPLICABILITY

[1443] The constructs of the invention can be manufactured industrially and sold, e.g., in the form of pharmaceutical compositions or medicaments for the treatment of diseases such as, for instance, cancer. Thus, the invention is industrially applicable.

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Claims

CLAIMS1 . An antibody construct comprising(i) at least one variable heavy chain (VH) of an anti-CD3 antibody,(ii) at least one variable light chain (VL) of the anti-CD3 antibody, and(ill) at least two single chain variable fragments (scFvs), wherein the antibody construct comprises the same number of (I) variable heavy chain(s) (VH) of the anti- CD3 antibody and (ii) variable light chain(s) (VL) of the anti-CD3 antibody, wherein each of the variable heavy chain(s) (VH) and the variable light chain(s) (VL) of the anti-CD 3 antibody are connected to each other by a protein structure comprising at least one or more of the single chain variable fragments (scFvs), and wherein in the antibody construct, at least two of the single chain variable fragments (scFvs) are against different target epitopes of the same target antigen or against different target antigens.

2. The antibody construct according to claim 1 , wherein at least one VH and VL of the anti-CD3 antibody form an anti-CD3 binding domain in the presence of said different target epitopes or target antigens on a cell surface.

3. The antibody construct according to claim 1 or 2, wherein the antibody construct is a single covalently linked construct.

4. The antibody construct according to any one of claims 1 to 3, wherein the antibody construct or the protein structure further comprises an immunoglobulin Fc region and / or an immunoglobulin constant region.

5. The antibody construct according to claim 4, wherein said immunoglobulin Fc region and / or immunoglobulin constant region are an immunoglobulin, preferably an IgG.

6. The antibody construct according to any one of claims 1 to 5, wherein the antibody construct or the protein structure further comprises albumin, single albumin-domains, albumin-binding domains, and / or other plasma / serum proteins such as part of membrane proteins cleaved and detected in the plasma / serum.

7. The antibody construct according to any one of claims 1 to 6, wherein said different target epitopes or target antigens are tumor and / or cancer epitopes or antigens.

8. The antibody construct according to any one of claims 1 to 7, wherein said different target epitopes or target antigens are prostate tissue epitopes or antigens.

9. The antibody construct according to any one of claims 1 to 8, wherein said different target epitopes or target antigens are cell type and / or cell lineage specific epitopes or antigens.

10. The antibody construct according to any one of claims 1 to 9, wherein said different target epitopes or target antigens are allergens and or auto-epitopes or auto-antigens.

11. The antibody construct according to claim any one of claims 1 to 10 wherein at least one, two, or all target epitopes or target antigens are selected from the group consisting of CD38, SLAMF7 / CS1 / CD319, ERBB2 / Her2 / NEU / CD340, FGFR1 / CD331 , EGFR, EpCAM / CD326, ERBB3, ERBB4, MET, PDGFRA, PDGFRB, FGFR2, FGFR3, FGFR4, KIT, FLT3, IGF1 R, RET, R0S1 , ALK, TGFBR1 , TGFBR2, ACVR2A, ACVR1 B, LRP5, LRP6, E-Cadherin I CDH1 , N-Cadherin I CDH2, AQP3, AQP1 , AQP4, AQP5, AQP9, CD45 1 PTPRC, CD34, CD1381 SDC1 , CD901 THY1 , CD1231 IL3RA, IL1 RAP, IL2RB I CD122, CD117 / Kit, CD244, CD49F I ITGA6, ABCG2, ESAM, CD10 I MME, CD81 , GHR, PRLR, PSMA, PSCA, CD133, CD166, CD44v6 I CD44, CD29 I ITGB1 , CD24, Lgr5, CEA / CEACAM5, VEGFR2 I KDR, CXCR4 / CD33, CD56 / NCAM1 , CD19, CD20, B7H3 / CD276, L1CAM, CD37, LFA-1 / ITGB2 / CD18, R0R1 , R0R2, CD70, CD123 / IL3RA, IGF1R / CD221, CCR7 I CD197, CCR4 I CD194, TNFRSF9 I 4-1 BB, CTLA4 I CD152, GPC3 I Glypican-3, CADM1 , CD4, B7-h4 I VTCN1 , TYRP1 , PDL1 / CD274, PDCD1 / PD-1, PDCD1 LG2 / PD-L2, BTLA, DLL3, CEACAM1 / CD66a, LFA- 1 I ITGB2, CD22, NRP1 1 CD304, CHRNA1 , GD2 I Disialoganglioside, CTAG1A / NY-ESO-1 , CD70, CD30 I TNFRSF8, CD98hc I SLC3A2, CD52, CD64 I FCGR1A, CD46 I MCP, CD6, CD10 I MME, CD25 I IL2RA, CD40, CD47, CD49a I ITGA1 , CD49c I ITGA3, CD49e I ITGA5, CD51 1 ITGAV, CD55, CD571 B3GAT1 , CD59, CD61 I ITGB3, CDw75 I ST6GAL1 , CD79a, CD82, CD95 I FAS, CD99, CD105 I ENG, CD107a I LAMP1 , CD107b I LAMP2, CD131 I CSF2RB, CD147 I BSG, CD151 , AGTR1 , AGTR2, KRAS, NRAS, HRAS, and BRAF or combinations thereof.

12. The antibody construct according to any one of claims 1 to 11 , wherein the anti-CD3 antibody is a T cellactivating antibody.

13. The antibody construct according to any one of claims 1 to 12, wherein (i) the variable heavy chain (VH) of the anti-CD3 antibody comprises the complementarity determining regions (CDRs) 1 , 2 and 3 comprising the amino acid sequences of SEQ ID NOs: 274, 275, and 276, respectively, and wherein (ii) the variable light chain (VL) of the anti-CD3 antibody comprises the complementarity determining regions (CDRs) 1 , 2 and 3 comprising the amino acid sequences of SEQ ID NOs: 277, 278, and 279, respectively, or wherein (i) the variable heavy chain (VH) of the anti-CD3 antibody comprises the complementarity determining regions (CDRs) 1 , 2 and 3 comprising the amino acid sequences of SEQ ID NOs: 280, 281 , and 282, respectively, and wherein (ii) the variable light chain (VL) of the anti-CD3 antibody comprises the complementarity determining regions (CDRs) 1 , 2 and 3 comprising the amino acid sequences of SEQ ID NOs: 283, 284, and 285, respectively.

14. The antibody construct according to any one of claims 1 to 13, wherein (i) the variable heavy chain (VH) of the anti-CD3 antibody comprises the amino acid sequence of SEQ ID NO: 270 or an amino acid sequence at least 95%, at least 98% or at least 99% identical thereto, and wherein (ii) the variable light chain (VL) of the anti- CD3 antibody comprises the amino acid sequence of SEQ ID NO: 271 or an amino acid sequence at least 95%, at least 98% or at least 99% identical thereto, or wherein (i) the variable heavy chain (VH) of the anti- CD3 antibody comprises the amino acid sequence of SEQ ID NO: 272 or an amino acid sequence at least 95%, at least 98% or at least 99% identical thereto, and wherein (ii) the variable light chain (VL) of the anti- CD3 antibody comprises the amino acid sequence of SEQ ID NO: 273 or an amino acid sequence at least 95%, at least 98% or at least 99% identical thereto.

15. The antibody construct according to any one of claims 1 to 14, wherein the anti-CD3 binding domain is capable of binding to CD3 on CD3 positive T cells and activate said cells.

16. The antibody construct according to any one of claims 1 to 15, wherein the antibody construct is a construct which forms, at concentration range 0.0001 nM to 0.001 nM, range 0.001 nM to 0.01 nM, range 0.01 nM to 0.1 nM, range 0.1 nM to 1 nM, range 1 nM to 10 nM, range 10 nM to 100 nM, range 100 nM to 1000 nM a functional anti-CD3 binding domain in the absence of said different target antigens, 10 fold (1 -log), 100 fold (2-log) or 1000 fold (3-log) lower as in the presence of said different target antigens.

17. The antibody construct according to any one of claims 1 to 16, wherein the antibody construct comprises not more than one VH and not more than one VL of the anti-CD3 antibody and not more than two single chain variable fragments (scFvs) against different target antigens.

18. The antibody construct according to any one of claims 1 to 16, wherein the antibody construct comprises two VH and two VL, three VH and three VL, or four VH and four VL of the anti-CD3 antibody.

19. The antibody construct according to any one of claims 1 to 16, wherein the antibody construct comprises two, three or four single chain variable fragments (scFvs) against different target epitopes or target antigens.

20. The antibody construct according to any one of claims 1 to 16, wherein the antibody construct comprises two, three or four single chain variable fragments (scFvs) against different target antigens and comprises two, three or four single chain variable fragments (scFvs) as pairs or triplicates, direct connected to each other or separated at least by one single chain variable fragments (scFvs) against a different target.21 . The antibody construct according to any one of claims 1 to 20, wherein the target antigens are ERBB2 / Her2 / NEU / CD340 and EpCAM / CD326 or CD38 and SLAMF7 / CS1 / CD319.

22. The antibody construct according to any one of claims 1 to 21, wherein all of the at least one VH and at least one VL of the anti-CD3 antibody are linked to one of said single chain variable fragments (scFvs) comprised by the protein structure by a peptide linker.

23. The antibody construct according to any one of claims 1 to 22, wherein the peptide linker allows formation of an anti-CD3 binding domain from at least one VH and VL of the anti-CD3 antibody in the presence of said different target epitopes or target antigens.

24. The antibody construct according to any one of claims 1 to 23, wherein the antibody construct is selected from the following (a) to (q):(a) an antibody construct comprising the amino acid sequence of SEQ ID NO: 293 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 293,(b) an antibody construct comprising the amino acid sequence of SEQ ID NO: 294 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 294,(c) an antibody construct comprising the amino acid sequence of SEQ ID NO: 295 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 295,(d) an antibody construct comprising the amino acid sequence of SEQ ID NO: 296 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 296,(e) an antibody construct comprising the amino acid sequence of SEQ ID NO: 297 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 297,(f) an antibody construct comprising the amino acid sequence of SEQ ID NO: 298 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 298,(g) an antibody construct comprising the amino acid sequence of SEQ ID NO: 299 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 299,(h) an antibody construct comprising the amino acid sequence of SEQ ID NO: 300 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 300,(I) an antibody construct comprising the amino acid sequence of SEQ ID NO: 301 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 301 ,(j) an antibody construct comprising the amino acid sequence of SEQ ID NO: 302 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 302,(k) an antibody construct comprising the amino acid sequence of SEQ ID NO: 303 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 303,(l) an antibody construct comprising rises the amino acid sequence of SEQ ID NO: 304 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 304,(m) an antibody construct comprising the amino acid sequence of SEQ ID NO: 305 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 305,(n) an antibody construct comprising the amino acid sequence of SEQ ID NO: 306 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 306,(o) an antibody construct comprising the amino acid sequence of SEQ ID NO: 307 or an amino acid sequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 307, and(p) an antibody construct comprising the amino acid sequence of SEQ ID NO: 308 or an amino acidsequence at least 90%, at least 95%, at least 98% or at least 99% identical thereto, wherein the antibody construct preferably has the CDRs of the amino acid sequence of SEQ ID NO: 308.

25. The antibody construct according to any one of claims 1 to 24, further comprising an additional proteinbinding group selected from a single-domain antibody (also known as a Nanobody or VHH), a monobody (derived from fibronectin type III), an anticalin (derived from lipocalins), an affibody (derived from immunoglobulin-binding protein A) and a DARPin (Designed Ankyrin Repeat Protein).

26. A nucleic acid molecule, or a set of nucleic acid molecules, encoding the antibody construct according to any one of claims 1 to 25.

27. A recombinant vector, or a set of recombinant vectors, comprising the nucleic acid molecule or set of nucleic acid molecules according to claim 26.

28. A recombinant host cell comprising the recombinant vector or set of recombinant vectors according to claim 27 and optionally expressing the antibody construct according to any one of claims 1 to 25.

29. A pharmaceutical composition comprising one or more antibody constructs according to any one of claims 1 to 25.

30. A kit comprising one or more antibody constructs according to any one of claims 1 to 25.31 . An antibody construct according to any one of claims 1 to 25 for use as a medicament.

32. An antibody construct according to any one of claims 1 to 25 for use in immunotherapy to treat cancer.

33. The antibody construct for use according to claim 31 or 32, wherein the antibody construct is to be administrated into the patient intra venous, sub cutaneous, intra thecal or intra peritoneal, via injections, short infusion (1 minute to 8 hours) or via a continuous infusion (> 8 hours).

34. The antibody construct for use according to any one of claims 31 to 33, wherein the antibody construct is to be administrated locally into a benign tumor or a cancer via injections, to increase local concentrations of the antibody construct and reduce systemic serum levels of the antibody construct.

35. The antibody construct for use according to any one of claims 31 to 34, wherein the antibody construct is to be administrated locally into a benign tumor or a cancer via injections, with the antibody construct embedded into a semi solid matrix to delay the antibody construct release over time and to reduce a wash out from the local injection site.

Images