Modified α1-α2 domains of non-natural NKG2D ligands that bind to non-natural NKG2D receptors

Abstract

The present disclosure generally relates to the preparation of polypeptides consisting of modified α1-α2 domains of NKG2D ligands that specifically bind to non-native extracellular domains of non-native NKG2D receptors, and wherein heterologous molecules are attached to the modified 1-2 domains of NKG2D ligands. The present disclosure also relates to modified α1-α2 domains of NKG2D ligands attached to heterologous molecules including polypeptides, and in some embodiments antibodies or antibody fragments. The present disclosure also relates to modified forms of NKG2D receptors that are engineered to provide a combination of enhanced and reduced binding to non-native and native forms of NKG2D ligands, respectively.

Description

Technical Field

[0001] This application generally relates to the preparation of polypeptides consisting of α1-α2 domains of NKG2D ligands that specifically bind to the non-natural extracellular domain of a non-natural NKG2D receptor, and wherein a heterologous molecule is linked to the α1-α2 domains of the NKG2D ligands. Background Technology

[0002] NKG2D is an activating receptor expressed as a type II homodimer integrase on the surface of natural killer (NK) cells and certain T cells. When NKG2D binds to one of eight native ligands primarily expressed on the surface of damaged cells, NKG2D-activated NK cells kill stress cells, or when NKG2D binds to T cells, the ligand-occupied NKG2D co-stimulates T cells to perform their effector functions. The three-dimensional structures of the extracellular domain of human native NKG2D, several of its soluble native ligands, and in some cases, the binding complexes of the soluble ligand and receptor extracellular domains have been resolved. The monomeric α1-α2 domain of the NKG2D ligand specifically binds to both extracellular domains of the native NKG2D homodimer. Summary of the Invention

[0003] This disclosure relates to modified α1-α2 domains of NKG2D ligands linked to heterologous molecules, including peptides, antibodies, or antibody fragments in some embodiments. The invention also relates to modified forms of the NKG2D receptor engineered to provide combinations of enhanced and weakened binding to non-natural and natural forms of NKG2D ligands, respectively. Attached Figure Description

[0004] Figure 1A : Comparison of the natural NKG2D.wt extracellular domain (SEQ ID NO:17) with the non-natural variants NKG2D.YA (SEQ ID NO:18) and NKG2D.AF (SEQ ID NO:25). The positions of Y152 and Y199 are shown, and the mutated residues present in the non-natural variants are highlighted in gray.

[0005] Figure 1B: Comparison of the α1-α2 domain of natural / wild-type ULBP2 (SEQ ID NO:4) and non-natural variants of ULBP2 (including ULBP2.R80W) (SEQ ID NO:108). Key residues for binding of non-natural ULBP2 variants to non-natural NKG2D.YA or NKG2D.AF receptors are highlighted in gray. The locations of residue R80 and the M154-F159 region explored for the literal variants binding to NKG2D.YA (ULBP2.S3, SEQ ID NO:127) or NKG2D.AF (ULBP2.C, SEQ ID NO:111; ULBP2.R, SEQ ID NO:113; ULBP2.AA, SEQ ID NO:115; and ULBP2.AB, SEQ ID NO:117) are shown.

[0006] Figure 2 Summary of mutational and protein aggregation characteristics of candidate non-natural Fc-eNKG2D variants determined by size exclusion chromatography (SEC), such as Figure 3 and Figure 4 shown in.

[0007] Figure 3 Size exclusion chromatography comparison of non-natural Fc-NKG2D fusion proteins analyzed on an Akta HiLoad 16 / 600 Superdex 200 column. Migration of correctly assembled material was exemplified by discrete, symmetrical peaks eluted at higher volumes, while aggregated material eluted more rapidly at lower volumes. The sites and properties of these modifications were indicated by amino acid numbers Y152, Y199, or both (SEQ ID NOs: 48, 43, 42, 58, 41, and 40, starting from the top of the figure).

[0008] Figure 4 Size exclusion chromatograms of non-natural Fc-eNKG2D variants with one or two amino acid variations were analyzed on an Akta Superdex 200 Increase 10 / 300 GL column. Migration of correctly assembled material was exemplified by discrete, symmetrical peaks eluted at higher volumes, while aggregated material eluted at lower volumes was characterized by broad, low-amplitude peaks or peak series. The letters in parentheses represent amino acids at positions 152 and 199, respectively (SEQ ID NO: 57, 56, 55, 54, 53, 52, 51, 50, 49, 48, 47, 46, 45, 44, and 40, from top to bottom).

[0009] Figure 5The percentage saturation (Rmax) of the eNKG2D variant normalized to wild-type NKG2D was determined by binding to MICwed-MicAbody or binding to MIC25-MicAbody. Wild-type Fc-NKG2D and each Fc-eNKG2D receptor were captured on an AHC biosensor and then exposed to 20 nM of trastuzumab-specific MicAbody. Dissociation kinetics were monitored and the Rmax values ​​of the Fc-eNKG2D fusions were ranked. Untested samples (nt) were due to severe aggregation after SEC grading or insufficient amounts of expressed or recovered material.

[0010] Figure 6 ELISA binding of ULBP2 wild-type, MICwed-, and MIC25-rituximab-MicAbody with Fc-eNKG2D candidates. Legend is shown at the top of the figure, but individual curves are also labeled in each figure due to the overlap of many curves.

[0011] Figure 7 : Figure 4 The EC50 values ​​(nM) for the Fc-eNKG2D ELISA are shown. nt = not tested; nb = no binding or very low binding even at 300 nM, therefore the EC50 value is not calculated.

[0012] Figure 8 Binding of eNKG2D variants to wild-type ligands. Wild-type ligands (all in Fc-fusion form) were captured onto the Octet AHC biosensor, and each natural NKG2D, NKG2D.Y152A, or eNKG2D5 (Y152A / Y199F) as an Fc-fusion was titrated from 300 nM to 0.41 nM. The maximum binding response was quantified by Octet. (Note that the ordinate of each graph is different).

[0013] Figure 9 Validated by dotted ELISA, a subset of combined mutants within ULBP2, relative to the native NKG2D.wt, resulted in selective binding of phage clones to NKG2D.AF. The mutants were graded based on their frequency of occurrence in the selected phages.

[0014] Figure 10 Titration ELISA of each phage variant was used to confirm selective binding to Fc-NKG2D.AF and to reduce or eliminate binding to Fc-NKG2D.wt. Mutations are detailed in [link to relevant documentation]. Figure 11 .

[0015] Figure 11The specificity of the selected ULBP2 variants of NKG2D.AF was preserved by quantitative ELISA in the form of rituximab-MicAbody, maintaining their binding to NKG2D.AF. Specific amino acid modifications of each ULBP2 variant were shown as the ratio of its binding to the Fc-NKG2D.wt fusion vessel to that to the Fc-NKG2D.AF fusion vessel. The amino acid residue positions of ULBP2 are... Figure 1B As shown.

[0016] Figure 12 Selected mutations at the indicated amino acid positions in ULBP2.R80W ( Figure 1B Y152A-specific phage clones were generated (SEQ ID NO:108).

[0017] Figure 13 ELISA data of four non-natural α1-α2ULBP2 variants of MicAbody binding to NKG2D.wt, NKG2D.YA, and NKG2D.AF. The Fc-NKG2D variant was used as the capture agent. MicAbody was titrated and detected with an HRP-conjugated anti-human κ antibody.

[0018] Figure 14 Using the NetMHC 4.0 server, queries were performed against HLA supertype representatives to detect changes in the peptide-MHCI immunogenicity of ULBP2.C (SEQ ID NO:111), ULBP2.R (SEQ ID NO:113), ULBP2.AA (SEQ ID NO:115), and ULBP2.AB (SEQ ID NO:117) relative to wild-type ULBP2 (SEQ ID NO:4). For the input sequence, the variable region of each variant (according to...) was entered. Figure 1B The comparison involved residues 154-159 in the 9-mer peptide window, along with nine upstream and nine downstream residues (a total of 24 residues), and the predicted immunogenicity of the 9-mer peptide window was detected. Dark gray boxes correspond to peptides strongly predicted to bind to the MHCI pocket (defined as % grade < 0.5), thus having a high chance of presentation. Light gray boxes correspond to predicted weak binding (% grade < 2). See Example 5 for details.

[0019] Figure 15Using the NetMHCII 2.3 server, queries were performed against HLA-DR, HLA-DQ, and HLA-DP to detect changes in the peptide-MHC class II immunogenicity of ULBP2.C (SEQ ID NO:111), ULBP2.R (SEQ ID NO:113), ULBP2.AA (SEQ ID NO:115), and ULBP2.AB (SEQ ID NO:117) relative to wild-type ULBP2 (SEQ ID NO:4). For the input sequence, the variable region of each variant (according to...) was entered. Figure 1B The comparison involved residues 154-159 in the 15-mer peptide window, as well as 15 residues upstream and downstream (a total of 36 residues), and the predicted immunogenicity of the 15-mer peptide window was detected. Dark gray boxes correspond to peptides that are strongly predicted to bind to the MHCII pocket and are therefore likely to be presented and immunogenic. Light gray boxes correspond to predicted weak binding.

[0020] Figure 16A and Figure 16B Rituximab-MicAbody, composed of ULBP2.wt (wild-type), ULBP2.R80W (which has enhanced affinity for wild-type NKG2D), ULPB2.S3 (NKG2D.YA-selected orthogonal variant), or ULBP2.R (NKG2D.AF-selected orthogonal variant), as measured by ELISA, binds to natural NKG2D-wt, NKG2D-YA, and NKG2D-AF. Figure 16A ELISA curve. Due to precipitation of the reagents developed for the TMB-Ultra ELISA, the reduced absorption at 458 nm in some assays at higher concentrations is an artifact often observed in high-affinity binders at higher concentrations. Figure 16B :according to Figure 16A The curve in the figure represents the EC50 value (in nM) determined in GraphPad Prism. nd = is uncertain because there is no relationship between increased concentration and binding.

[0021] Figure 17A , Figure 17B and Figure 17C In an in vitro cell lysis assay using CD8 effector cells (CD8 effector cells consist of a CD8a hinge / transmembrane domain, intracellular 4-1BB, and CD3ζ signaling domain) transduced with untransduced or NKG2D.wt, NKG2D.YA, or NKG2D.AF CAR constructs, target cells were preloaded with calcein and exposed to effector cells at an increased effector-to-target (E:T) ratio. Calcein release was quantified after 5 hours. Figure 17A Cell lysis of HeLa cells. Figure 17BLysis of HeLa cells transfected to overexpress ULBP1 on the surface. Figure 17C Cell lysis of HeLa cells expressing non-native ULBP2.R. Error bars correspond to the standard deviation of the technical replicas in the experiment.

[0022] Figure 18A and Figure 18B MicAbody induces cell lysis of tumor cell lines via NKG2D-CAR CD8 T cell targeting. Figure 18A With increasing concentrations of ULBP2.S2 or ULBP2.R rituximab-Micabody, Ramos cells expressing the rituximab target, CD20, were preloaded with calcein and exposed to NKG2D.AF- or NKG2D.YA-CAR cells at an E:T ratio of 20:1. Cell lysis levels were quantified after 2 hours of incubation. Figure 18B The mouse tumor line CT26, transfected to express human Her2, was used as a target for cell lysis parallel to Ramos cells. NKG2D.AF-CAR CD8 T cells were pre-armed with saturated concentrations (5 nM) of rituximab-ULBP2.R, trastuzumab-ULBP2.R, or an equimolar mixture of two MicAbody. Unbound MicAbody was removed by washing, and CD8 cells were added to the target cells at two different E:T ratios. Cell lysis was measured 2 hours later. Detailed Implementation

[0023] Natural killer (NK) cells and certain (CD8+αβ and γδ) T cells of the immune system play an important role as first-line natural defenses against tumor cells and infected cells in humans and other mammals (Cerwenka, A., and L.L. Lanier. 2001. NK cells, viruses and cancer. Nat. Rev. Immunol. 1:41-49). NK cells and some T cells display NKG2D on their surface, a prominent homodimer surface immune receptor responsible for recognizing target cells and activating the natural defense against pathological cells (Lanier, LL, 1998. NK cell receptors. Ann. Rev. Immunol. 16:359-393; Houchins JP et al. 1991. DNA sequence analysis of NKG2, a family of related cDNA clones encoding type II integral membrane proteins on human NK cells. J. Exp. Med. 173:1017-1020; Bauer, S et al., 1999. Activation of NK cells and T cells by NKG2D, a receptor for stress-inducible MICA. Science 285:727-730).The human NKG2D molecule possesses a C-type lectin-like (extracellular) domain that binds to eight different homologous ligands. The most studied ligands are the monomers MICA and MICB, which share 84% sequence identity or homology. These are polymorphic analogs of major histocompatibility complex (MHC) class I chain-associated glycoproteins (MICs) (Weis et al. 1998. The C-type lectin superfamily of the immune system. Immunol. Rev. 163:19-34; Bahram et al. 1994. A second lineage of mammalian MHC class I genes. PNAS 91:6259-6263; Bahram et al. 1996a. Nucleotide sequence of the human MHC class I MICA gene. Immunogenetics 44:80-81; Bahram and Spies TA. 1996. Nucleotide sequence of human MHC class I MICB). cDNA. Immunogenetics 43:230-233). Non-pathological expression of MICA and MICB is limited to intestinal epithelium, keratinocytes, endothelial cells, and monocytes. However, aberrant surface expression of these MIC proteins is a response to various cellular stresses, including proliferation, oxidation, and heat shock, and marks pathological expression in cells (Groh et al. 1996. Cell stress-regulated human MHC class I gene expressed in GI epithelium. PNAS 93:12445-12450; Groh et al. 1998. Recognition of stress-induced MHC molecules by intestinal γδT cells. Science 279:1737-1740; Zwirner et al. 1999. Differential expression of MICA by endothelial cells, fibroblasts, keratinocytes, and monocytes. Human Immunol. 60:323-330).Pathological expression of MIC proteins also appears to be associated with certain autoimmune diseases (Ravetch, JV and Lanier LL. 2000. Immune Inhibitory Receptors. Science 290:84-89; Burgess, SJ. 2008. Immunol. Res. 40:18-34). Different regulation of NKG2D ligands, such as polymorphic MICA and MICB, is important for providing the immune system with a means to recognize and respond to a wide range of emergency signals while still protecting healthy cells from unwanted attacks (Stephens HA, (2001) MICA and MICB genes: can the enigma of their polymorphism be resolved? Trends Immunol. 22:378-85; Spies, T. 2008. Regulation of NKG2D ligands: a purposeful but delicate affair. Nature Immunol. 9:1013-1015).

[0024] Viral infection is a common trigger for MIC protein expression, and it has been determined that virus-infected cells are attacked by NK or T cells (Groh et al. 1998; Groh et al. 2001. Co-stimulation of CD8+αβT cells by NKG2D via engagement by MIC induced on virus-infected cells. Nat. Immunol. 2:255-260; Cerwenka, A., and LLLanier. 2001). In fact, to avoid such attacks on their host cells, cytomegaloviruses and other viruses have evolved mechanisms to prevent the expression of MIC proteins on the surface of the cells they infect, thus evading the targeting of the innate immune system (Lodoen, M., K. Ogasawara, J.A. Hamerman, H. Arase, J.P. Choukins, E.S. Mocarski, and L.L. Anier. 2003. NKG2D-mediated NK cell protection against cytomegalovirus is impaired by gp40 modulation of RAE-1 molecules. J. Exp. Med. 197: 1245-1253; Stern-Ginossar et al., (2007) Host immune system gene targeting by viral miRNA. Science 317: 376-381; Stern-Ginossar et al., (2008) Human microRNAs regulate stress-induced immune responses mediated by the receptor NKG2D. Nature Immunology 9: 1065-73; Slavuljica, IA Busche, MBabic, M Mitrovic, I Cekinovic,E Markova Car,EP Pugel,A Cikovic,VJLisnic,WJ Britt,U Koszinowski,M Messerle,A Krmpotic and SJonjic.2010.Recombinant mouse cytomegalovirus expressing a ligand for theNKG2D receptor is attenuated and has improved vaccineproperties.J.Clin.Invest.120:4532-4545)。

[0025] The expression of MIC proteins is also induced in many tumor cells, and the presence of MIC proteins can make them sensitive to NK cell targeting and lysis. Unsurprisingly, the lack of expression of MIC proteins is a way for many malignant cells, such as lung cancer and glioblastoma brain cancer, to evade the innate immune system (Busche, A et al. 2006, NK cell-mediated rejection of experimental human lung cancer by genetic overexpression of MHC class I chain-related gene A. Human Gene Therapy 17:135-146; Doubrovina, ES, MM Doubrovin, E Vider, RB Sisson, RJ O'Reilly, B Dupont, and YMVyas, 2003. Evasion from NK Cell Immunity by MHC Class I Chain-Related Molecules Expressing Colon Adenocarcinoma (2003) J. Immunology 6891-99; Friese, M. et al. 2003. MICA / NKG2D-mediated immunogene therapy of experimental gliomas. Cancer Research 63:8996-9006; Fuertes, MB, MV Girart, LL). Molinero, CIDomaica, LE Rossi, MM Barrio, J Mordoh, GA Rabinovich and NW Zwirner. (2008) Intracellular Retention of the NKG2D Ligand MHC Class I Chain-Related Gene Ain Human Melanomas Confers Immune Privilege and Prevents NK Cell-Mediated Cytotoxicity. J. Immunology, 180:4606-4614).

[0026] The high-resolution structure of human MICA binding to NKG2D has been resolved, demonstrating that the α3 domain of MICA does not directly interact with NKG2D (Li et al. 2001. Complex structure of the activating immunoreceptor NKG2D and its MHC class I-like ligand MICA. Nature Immunol. 2:443-451; Protein Data Bank accession code 1HYR). Similar to MICB, the α3 domain of MICA connects to the α1-α2 platform domain via a short, flexible linker and naturally positions itself as a "spacer" between the platform and the surface of MIC-expressing cells. The three-dimensional structures of the α3 domains of human MICA and MICB are almost identical (root mean square distance over 94 C-αα′ < Furthermore, they are functionally interchangeable (Holmes et al. 2001. Structural Studies of Allelic Diversity of the MHC Class I Homolog MICB, a Stress-Inducible Ligand for the Activating Immunoreceptor NKG2D. J Immunol. 169:1395-1400).

[0027] It has been described that the α1-α2 domains of certain modified non-natural NKG2D ligands have a higher affinity for binding to the natural human NKG2D receptor than the natural α1-α2 domain (Candice S. E. Engyel, Lindsey J. Willis, Patrick Mann, David Baker, Tanja Kortemme, Roland K. Strong and Benjamin J. McFarland. Mutations Designed to Destabilize the Receptor-Bound Conformation Increase MICA-NKG2D Association Rate and Affinity. Journal of Biological Chemistry Vol. 282, no. 42, pp. 30658–30666, 2007; Samuel H. Henager, Melissa A. Hale, Nicholas J. Maurice, Erin C. Dunnington, Carter J. Swanson, Megan J. Peterson, Joseph J. Ban, David J. Culpepper, Luke). D.Davies,Lisa K.Sanders,and BenjaminJ.McFarland.Combining different design strategies for rational affinitymaturation of the MICA-NKG2D interface.Protein Science 2012VOL21:1396-1402).Here we describe the non-natural α1-α2 domains of NKG2D ligands that have been modified to bind to non-natural NKG2D receptors, which themselves are mutated at two specific sites, each of which individually results in impaired or lost binding to the natural α1-α2 domains of all currently known human NKG2D ligands (David J. Culpepper, Michael K. Maddox, Andrew B. Caldwell, and Benjamin J. McFarland. Systematic mutation and thermodynamic analysis of central tyrosine pairs in polyspecific NKG2D receptor interactions. (Mol Immunol. 2011 January; 48(4):516–523).

[0028] The bispecific molecule created in this invention consists of (a) a specifically modified non-natural α1-α2 domain and (b) a specifically targeted heterologous molecule, including but not limited to heterologous peptides or polypeptides. The bispecific molecule can bind to a chimeric antigen receptor (CAR), wherein the CAR receptor is composed of a non-natural NKG2D receptor extracellular domain that binds to its homologously modified α1-α2 domain with a higher affinity than its natural α1-α2 domain. The second specificity of the bispecific molecule consists of the heterologous component that specifically binds to its corresponding target molecule. Then, genetically engineered immune system cells composed of such CARs and homologous bispecific molecules can overcome many drawbacks, including known severe systemic toxicity, antigen escape, and the limited and uncontrolled persistence of current CAR-T and CAR-NK cell therapies (Kalos M, Levine, BL, Porter, DL, Katz, S, Grupp, SA, Bagg, A and June, C.. T cells with chimeric antigen receptors have potent antitumor effects and can establish memory in patients with advanced leukemia. Sci Transl Med 2011; 3:95ra73; Morgan RA, Yang JC, Kitano M, Dudley ME, Laurencot CM, Rosenberg SA. Case report of a serious adverse event following the administration of T cells transduced with a chimeric antigen receptor recognizing ERBB2. Mol Ther 2010, 18:843–851).

[0029] Gene transfer technology can be used to modify T cells and NK cells to directly and stably express transmembrane signal transduction receptors on their surface. These transmembrane signal transduction receptors confer new antigen specificity to T cells through antibody-derived binding domains (Saar Gill & Carl H. June. Going viral:chimeric antigen receptor T cell therapy for hematological malignancies. Immunological Reviews 2015. Vol. 263:68–89; Wolfgang Glienke, Ruth Esser, Christoph Priesner, Julia D. Suerth, Axel Schambach, Winfried S. Wels, Manuel Grez, Stephan Kloess, Lubomir Arseniev and Ulrike Koehl. 2015. Advantages and applications of CAR-expressing natural killer cells. Front. Pharmacol. doi:10.3389 / fphar.2015.00021). This type of T cell expressing a chimeric antigen receptor (CAR-T cell) is an application of a method that combines the antigen recognition domain of a specific antibody with the intracellular domain of the CD3-ζ chain to form a single chimeric protein and a cytoplasmic polypeptide sequence from co-stimulatory molecules such as CD27, CD28, ICOS, 4-1BB, or OX40. The intracellular domain of the CD3-ζ chain is a primary neurotransmitter for signaling from the endogenous T cell antigen receptor (TCR). The CAR constructed in this way can trigger T cell activation when it binds to the target antigen in a manner similar to the endogenous T cell receptor but independent of the major histocompatibility complex (MHC).

[0030] As used in this article, "soluble MIC protein," "soluble MICA," and "soluble MICB" refer to MIC proteins that contain the α1, α2, and α3 domains of MIC proteins but lack transmembrane or intracellular domains. NKG2D ligands and ULBP1-6 naturally do not possess an α3 domain (Cerwenka A, Lanier LL. 2004. NKG2D ligands: unconventional MHC class I-like molecules exploited by viruses and cancer. Tissue Antigens 61(5):335–43. doi:10.1034 / j.1399-0039.2003.00070.x.PMID 12753652). The "α1-α2 domain" of NKG2D ligands refers to the protein domain of the ligand that binds to the NKG2D receptor.

[0031] In some embodiments, the α1-α2 domain of the non-natural NKG2D ligand protein of the present invention has at least 80% identity or homology with the native or natural α1-α2 domain of the NKG2D ligand (SEQ ID NO: 1-9 are MICA, MICB, ULBP1, ULBP2, ULBP3, ULBP4, ULBP5, ULBP6, and OMCP, respectively). In other embodiments, the modified α1-α2 domain has at least 85% identity with the native or natural α1-α2 domain of the NKG2D ligand. In other embodiments, the modified α1-α2 domain has at least 90% identity with the native or natural α1-α2 domain of the natural NKG2D ligand protein and binds to non-natural NKG2D.

[0032] Preferably, the modified or non-natural α1-α2 domain of the non-natural MIC protein of the present invention has at least 80% identity or homology with the native or natural α1-α2 domain of OMPC (SEQ ID NO: 9) or one of the eight known human NKG2D ligand proteins (SEQ ID NO: 1-8), and binds the non-natural NKG2D extracellular domain. In some embodiments, the non-natural α1-α2 domain has at least 85% identity with the native or natural α1-α2 domain of the NKG2D ligand protein and binds the non-natural NKG2D. In other embodiments, the non-natural α1-α2 platform domain has at least 90%, 95%, 96%, 97%, 98%, or 99% identity with the native or natural α1-α2 platform of the human natural α1-α2 domain protein and binds the non-natural NKG2D.

[0033] In some embodiments, a heterologous molecular tag may be fused to the N-terminus or C-terminus of a non-natural α1-α2 domain of a soluble MIC protein to aid in the purification of the soluble MIC protein. Tag sequences include peptides such as polyhistidine, myc-peptides, FLAG tags, streptavidin-like tags, or small molecules such as biotin. This tag can be removed after the MIC molecule has been isolated using methods known to those skilled in the art.

[0034] In other embodiments of the invention, the α1-α2 domain of the NKG2D ligand can be specifically mutated to produce a non-natural α1-α2 domain that binds to a non-natural NKG2D receptor, which is itself engineered to have a reduced affinity for the natural NKG2D ligand. This can be accomplished, for example, through genetic engineering. Such modified non-natural NKG2D receptors can be used to generate NKG2D-based chimeric antigen receptors (CARs) on the surface of NK cells, T cells, or other cells of the immune system, which preferentially bind to and are activated by molecules composed of the non-natural α1-α2 domain of the invention. As described below, these non-natural NKG2D receptor pairs and their invented homologous non-natural NKG2D ligands will offer significant safety, efficacy, and manufacturing advantages for the treatment of cancer and viral infections compared to current CAR-T cells and CAR-NK cells.

[0035] CRA-engineered T cells have emerged as a promising approach for adoptive T-cell therapy in cancer, and CRAs targeting many different molecules have been tested in CAR-T cells as therapeutic agents for malignancies (Porter DL, Levine BL, Kalos M, Bagg A, June CH., 2011. Chimeric antigen receptor-modified T cells in chronic lymphoid leukemia. N Engl J Med. 365:725–733.). While significant clinical efficacy has been observed in hundreds of patients receiving adoptive transfer of T cells expressing CD19-specific chimeric antigen receptors, the conventional process of engineering CARs—targeting specific antigens, isolating autologous T cells from patients, genetically engineering autologous T cells to express personalized CARs, expanding modified cells in vitro, and controlling the quality of their production—is cumbersome and expensive.

[0036] Once autologous CAR-T cells are reinfused into a donor patient, their proliferation in the body becomes uncontrollable, resulting in a treatment known as “live therapy” that does not show a dose-response relationship with efficacy (Gill and June, 2015). Furthermore, tumor escape from CAR-T cells may occur through the loss of antigens on tumor cells (Stephan A. Grupp, MD, Ph.D., Michael Kalos, Ph.D., David Barrett, MD, Ph.D., Richard Aplenc, MD, Ph.D., David L. Porter, MD, Susan R. Rheingold, MD, David T. Teachey, MD, Anne Chew, Ph.D., Bernd Hauck, Ph.D., J. Fraser Wright, Ph.D., Michael C. Milone, MD, Ph.D., Bruce L. Levine, Ph.D., and Carl H. June, MD: Chimeric Antigen Receptor–Modified T Cells for Acute Lymphoid Leukemia. N. Engl. J. Med. 2013; 368:1509–1518). While such escape pathways can be addressed through sequential therapy with CAR-T cells targeting different targets or through an initial infusion of a T-cell product containing two or more specific CARs, such steps represent a significant additional expansion in terms of manufacturing processes, quality control, and cost.

[0037] In addition to CAR-T cells that target tumors with single-chain antibody-binding domains (scFv), CAR-T cells utilizing the ligand-binding domain of the NKG2D receptor have been studied in animals and recently in humans (Sentman CL, Meehan KR. NKG2D CARs as cell therapy for cancer. Cancer J. 2014 Mar-Apr; 20(2):156-9. doi:10.1097 / PPO.0000000000000029; Manfred Lehner, Gabriel Julia Proff, Niels Schaft, Jan floral 2012|PLOS ONE 10.1371 / journal.pone.0031210; www.clinicaltrials.gov NCT02203825). Since the expression of NKG2D ligands is increased on the surface of stressed cells (such as tumor cells), this natural NKG2D ligand family is significant as a target for cancer immunotherapy (Spear P, Wu MR, Sentman ML, Sentman CL. NKG2D ligands as therapeutic targets. Cancer Immun. 2013 May 1; 13:8.; Song DG, Ye Q, Santoro S, Fang C, Best A, Powell DJ Jr., Chimeric NKG2D CAR-expressing T cell-mediated attack of humanovarian cancer is enhanced by histone deacetylase inhibition. Hum Gene Ther. 2013 Mar; 24(3):295-305). One NKG2D CAR is a fusion of the full-length NKG2D receptor and CD3ζ (NKG2Dζ); the other is a second-generation CAR scaffold fused only with the extracellular domain of NKG2D and a combination of the transmembrane and intracellular domains of CD28 and the signaling domain of CD3ζ (NKG2D28ζ). Since NKG2D activation depends on the presence of DAP10, CAR-T cells co-expressing DAP10 and NKG2Dζ (NKG2Dζ10) were also constructed.T cells expressing any of the above NKG2D CARs respond to NKG2D ligand stimulation to produce IFNγ and TNFα, and effectively kill tumor targets expressing NKG2D ligands in vitro (Heather VanSeggelen, Joanne A. Hammill, Anna Dvorkin-Gheva, Daniela GMTantalo, Jacek M. Kwiecien, Galina F. Denisova, Brian Rabinovich, Yonghong Wan, Jonathan L. Bramson, T cells engineered with chimeric antigen receptors targeting NKG2D ligands display lethal toxicity in mice, Molecular Therapy 2015 Oct; 23(20):1600-10; doi:10.1038 / mt.2015.119). The cytotoxic activity of NK cells against a broad spectrum of tumor subtypes can also be significantly enhanced by expressing CARs based on NKG2D-DAP10-CD3ζ (Yu-Hsiang Chang, John Connolly, Noriko Shimasaki, Kousaku Mimura, Koji Kono, and Dario Campana. Chimeric Receptor with NKG2D Specificity Enhances Natural Killer Cell Activation and Killing of Tumor Cells. Cancer Res; 73(6) March 15, 2013).

[0038] However, these CAR-T constructs, which bind to and are activated by the natural ligand of the native NKG2D receptor, exhibited significant toxicity upon injection into the homogeneous mouse host. Signs of toxicity were observed in tumor-bearing and tumor-free mice treated with NKG2D-based CAR-T cells compared to untreated control mice, including poor physical condition, hunched posture, labored breathing, and decreased core body temperature. The severity of NKG2D CAR-T cell toxicity varied, with NKG2Dζ10 cells exhibiting severe toxicity, NKG2D28ζ cells exhibiting moderate toxicity, and NKG2Dζ cells exhibiting tolerable toxicity. When mice received chemotherapy prior to adoptive transfer of T cells expressing any NKG2D CAR, the clinical signs of toxicity and mortality worsened (VanSeggelen et al., 2015). It is known that chemotherapy and radiation induce NKG2D ligands in other healthy tissues (Xiulong Xu, Geetha S Rao, Veronika Groh, Thomas Spies, Paolo Gattuso, Howard L Kaufman, Janet Plate and Richard A Prinz. Majorhistocompatibility complex class I-related chain A / B (MICA / B) expression in tumor tissue and serum of pancreatic cancer: Role of uric acid accumulation ingemcitabine-induced MICA / B expression. BMC Cancer 2011, 11:194. doi:10.1186 / 1471-2407-11-194; Gannagé M, Buzyn A, Bogiatzi SI, Lambert M, Soumelis V, Dal Cortivo L, Caavazzana-Calvo M, Brousse N, Caillat-Zucman Induction of NKG2D ligands by gamma radiation and tumor necrosis factor-alpha may participate in the tissue damage during acute graft-versus-hostdisease.Transplantation.2008Mar 27;85(6):911-5.doi:10.1097 / TP.0b013e31816691ef.).Further characterization revealed that the toxicity was consistent with systemic cytokine storm and lethal pulmonary inflammation levels. These data serve as a warning that extreme caution must be exercised when using natural NKG2D ligands for targeted immunotherapy, and demonstrate that T-cell expression of CARs targeting these ligands—particularly strongly activated CARs—can be detrimental in vivo (VanSeggelen et al., 2015).

[0039] CAR-T or CAR-NK cells composed of the extracellular domain of a non-natural NKG2D receptor do not bind to or hardly bind to the natural NKG2D ligand, are not limited to the aforementioned activation forms, and therefore are not as toxic as cells expressing CARs based on the natural NKG2D receptor. Furthermore, the extracellular domain of the non-natural NKG2D receptor on the cell is not downregulated by soluble natural NKG2D ligands or myeloid-derived suppressor cells (MDSCs) (Deng W, Gowen BG, Zhang L, Wang L, Lau S, Iannello A, Xu J, Rovis TL, Xiong N, Raulet DH, 2015. Antitumor immunity. A shed NKG2D ligand that promotes natural killer cell activation and tumor rejection. Science. 2015 Apr3; 348(6230):136-9. doi:10.1126 / science.1258867. Epub 2015 Mar 5). However, when such CAR cells with the extracellular domain of the non-natural NKG2D receptor bind to the homo-non-natural α1-α2 domain of the present invention and its hetero-targeting motif, which has been discovered and binds to its intended target, the CAR will be activated and express the effector functions of the CAR-cell.

[0040] Because, except in the presence of bispecific molecules that bind to non-natural α1-α2 domains, the cytokine release and cytolytic activity of CAR-T or CAR-NK cells composed of the extracellular domain of the non-natural NKG2D receptor are not triggered, where the bispecific molecules have already bound to their target arrays, their activation can be controlled by the administered bispecific molecules, which, as biological drugs, will exhibit pharmacokinetics and pharmacodynamics well known in the art. In the event of adverse events, physicians can simply change the dosing regimen of the administered bispecific molecules, rather than having to employ induced suicide mechanisms to destroy the infused CAR cells as is currently the case (Monica Casucci and Attilio Bondanza. Suicide Gene Therapy to Increase the Safety of Chimeric Antigen Receptor-Redirected T Lymphocytes. J Cancer. 2011; 2:378–382). Furthermore, such bispecific molecules with different specific targeting motifs can be administered simultaneously or sequentially to help address tumor resistance and escape due to target antigen loss, without the need to create, expand, and infuse multiple different autologous CAR cells (Gill and June, 2015). Since all CAR constructs can be identical for all CAR cells, and the targeting specificity is determined solely by the targeting motif of the bispecific molecule of the present invention, the CAR-cell manufacturing process will be simplified and made cheaper.

[0041] Examples of parental or receptor proteins or peptides that are candidates for attachment to the non-natural α1-α2 domain of the NKG2D ligand include, but are not limited to, antibodies, proteins composed of Ig sheets or Ig domains, including modified Fc domains that recruit or fail to recruit or bind natural molecules, globulins, albumins, fibronectin and fibronectin domains, integrins, fluorescent proteins, enzymes, outer membrane proteins, receptor proteins, T-cell receptors, chimeric antigen receptors, viral antigens, viral capsids, viral ligands of cell receptors, histones, hormones, cytokines and modified cytokines such as interleukins, knottin, cyclic peptides or peptides, major histocompatibility (MHC) family proteins, MIC proteins, lectins and lectin ligands. Non-protein molecules such as polysaccharides, dendritic macromolecules, polyethylene glycols, peptidoglycans, antibiotics, and polyketides can also be linked to the modified α1-α2 domain of the NKG2D ligand.

[0042] Therefore, this invention expands the diversity and applicability of this remarkable and highly promising immunological approach to controlling cancer with CAR-T cells, CAR-NK cells, and CAR-macrophage-like cells, while overcoming many of these currently recognized difficulties.

[0043] As used herein, the terms “peptide,” “polypeptide,” and “protein” are used interchangeably; and “heterologous molecule,” “heterologous peptide,” “heterologous sequence,” or “heterologous atom” are respectively non-natural or commonly found molecules, peptides, nucleic acid sequences, or amino acid sequences or atoms when physically bound to the subject molecule. As used herein, “non-natural” and “modified” are used interchangeably. As used herein, “natural,” “native,” and “wild-type” are used interchangeably, as are “NKG2D” and “NKG2D receptor.” The term “antibody” is used in the broadest sense and specifically covers monoclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, provided they exhibit the desired biological activity. An “antibody fragment” includes a portion of an antibody, preferably including its antigen-binding region. Examples of antibody fragments include Fab, Fab′, F(ab′)2, Fv fragments and insertable Fv's; biantibodies; linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments. An “antibody fragment” may also include the Fc portion of an antibody.

[0044] The term "comprising," which can be used interchangeably with "comprising," "containing," or "characterized by," is inclusive or open-ended language and does not exclude additional, unlisted elements or method steps. The phrase "consisting of" excludes any element, step, or ingredient not specified in the claims. The phrase "consisting essentially of" limits the scope of the claims to specific materials or steps, and those that do not substantially affect the essential and novel features of the claimed invention. This disclosure contemplates embodiments of compositions and methods of the invention corresponding to the scope of each of these phrases. Therefore, compositions or methods including the said elements or steps are contemplated in particular embodiments where the composition or method is substantially composed of or consists of these elements or steps.

[0045] All references cited in this paper are incorporated herein by reference in their entirety, regardless of whether they were previously specifically incorporated. As used herein, the terms “a,” “an,” and “any” are each intended to include both the singular and plural forms.

[0046] The invention has now been fully described, and those skilled in the art will understand that the same operation can be performed within a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without excessive experimentation. While the invention has been described in conjunction with specific embodiments thereof, it should be understood that further modifications are possible. This application is intended to cover any variations, uses, or adaptations of the invention that generally follow the principles of the invention, and includes deviations from this disclosure within the scope of known or customary practice in the art to which this invention pertains, and applicable to the essential features set forth above.

[0047] Example

[0048] Modified extracellular domain of NKG2D receptor and α1-α2 domain modified by NKG2D ligand

[0049] Example 1: Modifying tyrosine 152 of the human NKG2D receptor to alanine (Y152A) and tyrosine 199 to phenylalanine (Y199F) to create an inert NKG2D extracellular domain.

[0050] Others have demonstrated that mutations at tyrosine positions 152 or 199 in human NKG2D correspond to positions 75 and 122 in the extracellular domain of NKG2D. Figure 1A The NKG2D receptor (SEQ ID NO: 17) can significantly reduce its binding to the natural ligand MICA (David J. Culpepper, Michael K. Maddox, Andrew B. Caldwell, and Benjamin J. McFarland. Systematic mutation and thermodynamic analysis of central tyrosine pairs in polyspecific NKG2D receptor interactions. Mol Immunol. 2011 January; 48(4): 516–523). We hypothesize that while mutations in any tyrosine residue greatly affect the binding ability of NKG2D to its natural ligands, simultaneous mutations at tyrosine 152 (Y152) and tyrosine 199 (Y199) would effectively eliminate the receptor's ability to bind to all natural ligands. Therefore, we sought to explore single and combined Y152 and Y199 substitutions and characterize their biochemical behavior in order to identify single- and double-mutant variants that cannot bind to any natural ligand. Those well-expressed and assembled variants are of particular interest because these meaningful inert ligands can be more readily generated for analysis.

[0051] The natural NKG2D (wild-type) extracellular domain (NKG2D.wt, SEQ ID NO:17) and candidate non-natural NKG2D variant extracellular domains (SEQ ID NO:18-35) – also known as “engineered NKG2D” or “eNKG2D” – were cloned as fusion clones with the C-terminus of human IgG1 Fc (without the Fab domain) via a short factor Xa-recognizable Ile-Glu-Gly-Arg linker (SEQ ID NO:38), and are interchangeably referred to as Fc-NKG2D.wt or NKG2D.wt and Fc-eNKG2D or eNKG2D (SEQ ID NO:40-58). Variants corresponding to the MHCI signal sequence (SEQ ID NO:36 and 37), the linker-equipped human IgG1 Fc (SEQ ID NO:39), and the NKG2D extracellular domain (SEQ ID NO:59-77) were synthesized. DNA fragment (Integrated DNA Technologies, San Diego, California) was inserted into pD2610-V12 (ATUM, Newark, California). DNA constructs substituted at Y152, Y199, or Y152 / Y199 combined mutations were explored in Expi293. TM Transient expression in cells (Thermo Fisher Scientific, Waltham, Massachusetts) Figure 2 The secreted protein was purified by protein A affinity chromatography (Pierce Biotechnology, Rockford, Illinois, catalog number 20334). The eluted material was characterized by size exclusion chromatography (SEC) on an Akta Pur Superdex column, and correctly assembled, appropriately sized material was fractionated and separated from the aggregated peaks for analysis.

[0052] SEC characterization of the purified NKG2D.Y199A-Fc fusion revealed the composition of the major aggregates. Figure 3 In contrast, both the native Fc-NKG2D fusion and the Fc-NKG2D.Y152A fusion were distinguished by discrete non-aggregated peaks, which are easily differentiated from the more rapidly migrating aggregates. In the Y152A / Y199A double mutant Fc-NKG2D fusion variant, the Y199A mutation also significantly affected aggregation, indicating a substantial influence on protein misfolding. Figure 3Therefore, this aspect of including any combination of Y199A with the Y152 mutation in NKG2D variants presents a challenge for producing the material necessary for subsequent engineering work and raises considerations regarding assembly and presentation on cell surfaces. Consequently, efforts have been made to explore other substitutions at Y152 and Y199 that can combine to produce more robust molecules. Candidates of Y152 and Y199 mutants in eNKG2D combinations were tested as Fc fusions and in ( Figure 2 Detailed descriptions are provided in [the document / section]. Furthermore, all purified and expressed Fc-eNKG2D fusion candidates were analyzed by SEC, and their chromatograms showed varying levels of aggregate formation. Figure 2-4 Single amino acid substitutions explored at residues 152 (alanine, serine, threonine, and valine) did not affect the assembly of the Fc-NKG2D molecule, although Y152-leucine (Y152L) produced highly aggregated material. Similar to alanine, glutamic acid and aspartic acid at residue 199 were not tolerated, although phenylalanine only slightly increased aggregate formation. Among the mutant combinations studied, Y152A / Y199F, Y152S / Y199F, Y152T / Y199F, and Y152F / Y199F did not negatively affect the desired dimer formation, while other combinations led to increased aggregation. Figure 2-4 ).

[0053] Example 2: Generation of the antibody-based bispecific molecule "MicAbody" using a non-natural NKG2D ligand variant

[0054] Non-natural MicA variants fused to human IgG1 were generated. DNA polynucleotides encoding α1-α2 domains, such as MICwed (SEQ ID NO:79) and MIC25 (SEQ ID NO:81), were amplified by PCR using primers. The primers also introduced polynucleotides encoding an APTSSSGGGGS adapter for fusion into the C-terminal κ light chain (SEQ ID NO:84) of human IgG1 or a GGGS adapter for fusion into the C-terminal heavy chain (SEQ ID NO:82) of human IgG1. Furthermore, two mutations—D265A / N297A (KABAT number)—were introduced into the CH2 domain of the heavy chain. These mutations reduced binding to all FcγR receptors, thereby eliminating antibody-dependent cytotoxicity (ADCC) function. (Shields et al., 2001 JBC, 276:6591-6604). Polynucleotides encoding the α1-α2 domains of wild-type ULBP2 (ULBP2.wt) without the GPI-linker (SEQ ID NO:12) were similarly cloned and fused with DNA polynucleotides encoding the adapter and either the heavy or light chain of IgG1. These bispecific antibodies are called "MicAbody". TMThe plural form is referred to as "MicAbody," and it is bivalent to the fused α1-α2 domains. Examples of antibodies used to generate MicAbody for eNKG2D engineering purposes include, but are not limited to, trastuzumab (SEQ ID NO: 94 and 96) and rituximab (SEQ ID NO: 98 and 100), and subsequently referred to as "trastuzumab-MicAbody" (e.g., SEQ ID NO: 102 and 104) and "rituximab-MicAbody" (e.g., SEQ ID NO: 106), respectively. This was achieved through Gibson cloning (New England Biolabs, Ipswich, Massachusetts). Inc. inserts the fusion construct separately into pD2610-V12 (ATUM, Newark, CA). For a given antibody recognizing a specific antigen, a plasmid encoding the heavy chain and a plasmid encoding the light chain are fused with a natural or non-natural NKG2D ligand and co-transfected for transient expression in Expi293™ cells (Thermo Fisher Scientific, Waltham, MA). Alternatively, a plasmid encoding the heavy chain fused with a natural or non-natural NKG2D ligand is co-transfected with a plasmid for the light chain. The secreted bispecific antibody is purified by protein A affinity chromatography (Pierce Biotechnology, Rockford, Illinois, Catalog No. 20334), and the eluted material is characterized by size exclusion chromatography (SEC) on an Akta Pur Superdex column and fractionated as needed. Furthermore, the purified samples are analyzed by SDS-PAGE to validate the expected molecular weights of the fused heavy and light chain species.

[0055] Example 3: Identification of modified NK2GD variants that cannot bind to natural NKG2D ligands or whose binding to non-natural ligands is enhanced by binding to wild-type NKG2D.

[0056] The binding affinity of the α1-α2 variant to the extracellular domains of native (wild-type) NKG2D and non-native eNKG2D proteins was analyzed using a plate ELISA method. Each SEC-graded native Fc-NKG2D and non-native Fc-eNKG2D fusion was coated overnight at 4°C with 1 μg / mL phosphate-buffered saline (PBS) onto individual wells of a Nunc Maxisorp 96-well plate (Thermo Fisher Scientific, Waltham, MA). The plates were washed three times with PBS / 0.05% Tween-20 (PBS-T) at 20–22°C and blocked for 2 hours at 20–22°C with 0.5% bovine serum albumin in PBS (PBS-B). In PBS / 0.5% bovine serum albumin (BSA) / 0.05% Tween-20 (PBS-BT), MicAbody was titrated for plate-bound natural or non-natural Fc-NKG2D fusions at 20–22 °C for 60 min, washed three times with PBS-T at 20–22 °C, and the bound bispecific protein was detected using HRP-conjugated anti-human κ in PBS-BT (Abcam, Cambridge, MA). A 1-Step assay was then performed. TM Ultra TMB ELISA substrate solution (Thermo Fisher Scientific, Waltham, MA) was used for color development. Binding of ULBP2.wt rituximab-MicAbody (SEQ ID NO: 98 and 106) distinguished wild-type NKG2D and eNKG2D variants, but binding to the latter was reduced, and ligand variants—MICwed (SEQ ID NO: 96 and 102) and MIC25 (SEQ ID NO: 96 and 104)—were more stringent in identifying eNKG2D variants bound to eliminated ligands. The binding behavior of each eNKG2D variant with all three bispecific ligands suggests that the combination of NKG2D modifications resulted in maximally reduced binding to both wild-type and variant ligands, and enabled selection of lead-inert NKG2D variants.

[0057] Additional biophysical analyses of the binding of eNKG2D variants to ligands were performed using the FortéBio Octet system (both from FortéBio LLC, Fremont, CA) via Bio-Layer Interferometry (BLI). For these experiments, human NKG2D ligands MICA-Fc, MICB-Fc, ULBP1-Fc, ULBP2-Fc, ULBP3-Fc, and ULBP4-Fc were purchased from R&D Systems, Minneapolis, Minnesota. The ligand in the form of MicAbody was captured on the tip of an anti-human IgG Fc capture (AHC) biosensor. After baseline establishment, the tip was exposed to a series of Fc-eNKG2D fusion protein titrations from 300 nM to 0.41 nM, and binding / dissociation kinetics were monitored at all steps in PBS-BT. Subsequently, the Fc-eNKG2D fusion protein was captured onto the AHC tip, and MicAbody was titrated to characterize binding kinetics.

[0058] To determine the maximum response defined by the binding of natural NKG2D to MICwed or MIC25, the natural Fc-NKG2D fusion compound was captured onto an AHC biosensor and incubated with 20 nM trastuzumab-MICwed or 20 nM trastuzumab-MIC25 MicAbody for 2 min, followed by observation of dissociation kinetics for 30 s. Binding analysis was then performed under the same conditions using the Fc-eNKG2D fusion receptor as the capture agent, and the binding level of each eNKG2D was listed as a percentage of the maximum binding response determined by Fc-NKG2D.wt. Figure 5 For MICwed, all single mutant Fc-eNKG2D variants except Y199F showed a reduced response to 50%. Y199F maintained a 100% binding response. However, all double mutant Fc-eNKG2D variants completely eliminated binding with MICwed. For MIC25, all single mutant Fc-eNKG2D variants and Y152V / Y199F maintained a 100% binding response relative to wild-type Fc-NKG2D. However, binding with several double mutant Fc-eNKG2D variants (including Y152A / Y199F, Y152S / Y199F, and Y152T / Y199F) decreased to 50%.

[0059] ELISA assays were performed using the Fc-eNKG2D fusion complex as a capture agent, with ULBP2.wt, MICwed, and MIC25MicAbody titrated from 300 nM. Figure 6 Use GraphPad Prism when possible. Figure 7 (Chinese) Calculation of EC 50The affinity of natural NKG2D for MicAbody based on ULBP2, MICwed, and MIC25 was calculated as Kds values ​​of 1.4, 0.007, and 0.005 nM, respectively. While the affinity of ULBP2 and MICwed MicAbody for all single mutant eNKG2D candidates was reduced, the binding of MIC25 to eNKG2D candidates remained unchanged. However, all double mutant eNKG2D candidates eliminated or significantly reduced binding to all three ligands—ULBP2, MICwed, and MIC25—in the form of Micbody.

[0060] Octet analysis and ELISA showed that the eNKG2D variants eNKG2D5 (Y152A / Y199F), eNKG2D7 (Y152S / Y199F), eNKG2D8 (Y152T / Y199F), and eNKG2D9 (Y152V / Y199F) reduced or eliminated binding to ULBP2, MICwed, and MIC25-based MicAbody. Figure 5 and 7 Furthermore, eNKG2D 5, 7, and 8 showed the least aggregation, indicating stronger protein assembly at 293T expression. Figure 2 The binding of eNKG2D5 (SEQ ID NO:48) to the wild-type ligand was examined more closely because MicAbody was captured at the Octet AHC tip. Compared to native (SEQ ID NO:40) NKG2D, the binding of the single mutant Fc-NKG2D.Y152A (SEQ ID NO:41) to all native ligands was reduced (…). Figure 8 Compared to Y152AeNKG2D, the response curve of eNKG2D5 (Y152A / Y199F) is further reduced. eNKG2D5 (Y152A / Y199F, hereinafter referred to as "AF" or "NKG2D.AF") was selected as the main NKG2D variant for designing homologous selective, orthogonal, non-natural ligands.

[0061] Example 4: Construction of orthogonal non-natural α1-α2 domains that selectively bind to the extracellular domain of non-natural NKG2D.AF

[0062] We used phage display technology to design orthogonal non-natural α1-α2 domains that selectively bind to the Fc-NKG2D AF (SEQ ID NO:48) receptor. As a starting point, we selected the non-natural ULBP2.R80W α1-α2 domain (which has high affinity for the extracellular domain of natural wild-type NKG2D (NKG2D.wt)). Figure 1BSEQ ID NO:108) was used as the parental domain for further mutagenesis and screening via phage display. A synthetic DNA library was generated for the α1-α2 domain (SEQ ID NO:108) of ULBP2.R80W, which also contained a C8S mutation to eliminate the possibility of disulfide bonds. The codons of the amino acid residues in the binding state on the ligand were located near positions Y152 and Y199 on the natural NKG2D receptor and were replaced by NNK codons; the library consisted of NNK codons at positions 154-159. Figure 1B (SEQ ID NO: 110). The library was cloned as a fusion of the pIII minor coat protein of M13 phage, and phage particles displaying the induced α1-α2 domain mutant were prepared in SS320 *E. coli* according to standard methods (Andris-Widhopf, J., Steinberger, P., Fuller, R., Rader, C., and Barbas, CF, 3rd. (2011)). In the presence of the non-biotinylated native Fc-NKG2D.wt competing protein, these α1-α2 phage display libraries were screened to obtain high affinity for the non-natural NKG2D.AF receptor by selectively capturing phage clones that bind to the biotinylated Fc-NKG2D.AF protein. Selective clones were enriched by multiple rounds of competitive selection using increasing concentrations of non-biotinylated native Fc-NKG2D.

[0063] After four rounds of selection, phage clones were arranged in 96-well arrays for dot ELISA to verify the preferred differential binding of non-natural NKG2D.AF to NKG2D.wt to the plate. Bound phages were detected using a biotinylated M13 phage coat protein monoclonal antibody E1 (Thermo Fisher Scientific, Waltham, MA), streptavidin-HRP assay (R&D Systems, Minneapolis, Minnesota), and a 1-Step Ultra TMB ELISA developed by Thermo Fisher Scientific, Waltham, MA. The dot ELISA signal for each clone was expressed as the ratio of phage binding to NKG2D.AF to phage binding to NKG2D.wt. Phages with a ratio greater than or equal to 14 were sequenced to identify specific mutations within the NNK mutagenic region. Figure 9 Selected amino acid residues of each α1-α2 phage variant that selectively binds to NKG2D.AF are shown. In the case of identifying multiple clones representing the same sequence, the ratio of ELISA signals was plotted, and the consistency of phage clones was verified by clustering of data points (data not shown).

[0064] Thirty variants identified in ELISA were amplified in separate single cultures to produce high-titer phage microbatches. The purified phage concentrations were normalized to OD0.05. 268 =0.5, and then a 1:3 dilution series was performed for plate-bound Fc-NKG2D.AF or Fc-NKG2D.wt. Phage detection and ELISA development were carried out as described above. All 30 variants determined in this manner consistently showed selective binding to NKG2D.AF and little or no binding to NKG2D.wt. Figure 10 Even at the highest concentrations of phages measured, the selected phages also showed two or more log shifts in phage concentration to achieve half-maximal binding between NKG2D.AF and NKG2D.wt.

[0065] To confirm that the NKG2D.AF selective α1-α2 domain variants retain specific binding properties in antibody fusions, 21 variants ( Figure 11 For example, SEQ ID NO:111-118) was cloned as a C-terminal fusion with an APTSSSGGGGS linker of the rituximab light chain (SEQ ID NO:119-126). The fusion was cloned into the mammalian expression vector pD2610-V12 (ATUM, Newark, California) via Gibson cloning (New England Biolabs Inc., Ipswich, MA) and co-expressed with the heavy chain of the parent antibody (SEQ ID NO:99) as a paired full IgG antibody. It was then expressed in Expi293 according to the manufacturer's protocol. TM Transient expression was performed in cells (Thermo Fisher Scientific, Waltham, MA) and standard protein-A affinity chromatography was used (Pierce Biotechnology, Rockford, Illinois, catalog number 20334). ELISA measured the binding of each variant of the ULBP2α1-α2 antibody fusion to both non-natural Fc-NKG2D.AF and natural Fc-NKG2D.wt, showing that their binding affinity to NKG2D.AF was significantly higher than that to natural NKG2D.wt. Figure 11 In summary, these data demonstrate the invention of a non-natural orthogonal α1-α2 domain that exhibits high affinity for the non-natural NKG2D.AF receptor and significantly reduces its affinity for the natural NKG2D receptor. Furthermore, these orthogonal α1-α2 domains retain their selective binding properties with fusions of antibody peptides and can be used, for example, in the case of chimeric antigen receptor (CAR) T cells to redirect non-natural NKG2D.AF receptors to specific antigens.

[0066] Example 5: Identifying non-natural NKG2D ligands capable of distinguishing non-natural NKG2D receptor variants by selectively binding to one or the other.

[0067] As described above, phage display was performed starting with the non-natural ULBP2.R80W α1-α2 domain (SEQ ID NO:108) to design orthogonal non-natural α1-α2 domains that selectively bind to the NKG2D.Y152A (hereinafter referred to as NKG2D.YA) receptor. Phage clones that bind to the biotinylated Fc-NKG2D.YA (SEQ ID NO:41) protein were selectively captured in the presence of a non-biotinylated natural Fc-NKG2D.wt (SEQ ID NO:40) competing protein. An α1-α2 phage display library was panned for high binding affinity to the non-natural Fc-NKG2D.YA receptor. Further phage clone validation work led to the identification of variants that preferentially bind to Fc-NKG2D.YA relative to Fc-NKG2D.wt. Figure 12 For example, ULBP2.S3 (SEQ ID NO: 127) consistently demonstrated selective binding to non-natural NKG2D.YA by ELISA and Octet analysis (in both monomeric His-tagged and bispecific antibody fusion forms) compared to natural NKG2D.wt. This represents a unique form of invention of non-natural orthogonal α1-α2 domains with high affinity binding to non-natural NKG2D receptors (NKG2D.YA in this example instead of NKG2D.AF in Example 4). Furthermore, the fusion of orthogonal α1-α2 domains with antibody peptides retains their selective binding properties and is used to selectively redirect non-natural NKG2D receptors to specific molecules identified by the fused heteropeptide (e.g., antibody).

[0068] To determine whether the non-natural α1-α2 domain (ULBP2.S3, SEQ ID NO:127) selectively binding to NKG2D.YA and the non-natural α1-α2 domain selectively binding to NKG2D.AF could distinguish between the two non-natural receptor variants, a titration ELISA was performed. The binding of all 21 selected α1-α2 variants binding to NKG2D.AF to NKG2D.AF and NKG2D.YA was directly compared. Four variants showed no binding to NKG2D.wt, a strong affinity for NKG2D.AF, and significantly reduced (15-20 fold) or eliminated binding to NKG2D.YA compared to NKG2D.AF. Figure 13). The predicted immunogenicity distribution of the four non-natural ULBP2α1-α2 variants—ULBP2.C, ULBP2.R, ULBP2.AA, and ULBP2.AB (SEQ ID NO: 111, 113, 115, and 117)—relative to the wild-type ULBP2 peptide sequence (SEQ ID NO: 4) was also detected using NetMHC 4.0 Server (querying for peptide-MHC class I binding of all HLA supertypes represented by 9-mer peptide analysis; http: / / www.cbs.dtu.dk / services / NetMHC / ) and NetMHCII 2.3 Server (querying for peptide-MHC class II binding of HLA-DR, HLA-DQ, and HLA-DP haplotypes by 15-mer peptide analysis; http: / / www.cbs.dtu.dk / services / NetMHCII / ), both algorithms developed by the Technical University of Denmark (http: / / www.bioinformatics.dtu.dk / ; Andreatta M and Nielsen M, Gapped sequence alignment using artificial neural networks: application to the MHC class I system, 2016 Bioinformatics, 32:511, PMID:26515819; Jensen KK, Andreatta M, Marcatili P, Buus S, Greenbaum JA, Yan Z, Sette A, Peters B, and Nielsen M, Improved methods for predicting peptide binding affinity to MHC class I molecules, 2018 Immunology, PMID:29315598). Mutations in ULBP2.C, ULBP2.R, and ULBP2.AB did not increase predicted immunogenicity, while mutations in ULPB2.AA showed a slight increase in a few haplotypes. Figure 14 and 15Due to the specificity and lack of predictable immunogenicity of ULBP2.R against NKG2D.AF, ULBP2.R was selected for further ELISA analysis to directly compare its binding behavior with that of ULBP2.S3 (the non-natural orthogonal ligand selected by NKG2D.YA), ULBP2.R80W (a non-natural ligand with enhanced affinity for wild-type NKG2D), and wild-type ULBP2 (ULBP2.wt). The binding of four rituximab-MicAbody reagents (SEQ ID NO: 98 and 121, 98 and 129, 131 and 100, and 98 and 106, representing the heavy and light chains of ULBP2.R, ULBP2.S3, ULBP2.R80W, and ULBP2.wt, respectively) was measured relative to wild-type NKG2D (NKG2D.wt) and two inert, non-natural variants, NKG2D.YA and NKG2D.AF). Figure 16A and 16B Data indicate that the NKG2D.YA-selected variant ULBP2.S3, as a MicAbody, has a high affinity for NKG2D.YA but does not bind to NKG2D.AF or native NKG2D. Furthermore, the NKG2D.AF-selected variant ULBP2.R, as a MicAbody, binds to NKG2D.AF with high affinity but not to NKG2D.YA or native NKG2D. These results demonstrate the great potential for exploring the NKG2D-MIC ligand axis and developing unique, novel, selective, paired non-natural NKG2D receptors and their respective homologous non-natural MIC ligand binding partners.

[0069] Example 6: The targeting and killing activity of CAR-T cells expressing the extracellular domain of non-natural NKG2D.AF is controlled by the orthogonal α1-α2 domain fused to the heterologous targeting peptide.

[0070] After mitigating toxicity and enhancing antitumor efficacy, selective control of CAR-T cell therapy has become increasingly popular (Gill and June, ibid.). Previous attempts have explored the development of CARs using the extracellular domain of CD16, which can then bind via the Fc domain of therapeutic monoclonal antibodies, allowing for antibody-based targeted control of CAR-T cells (Chang et al., ibid.). However, CD16-based CAR-T cells can recognize almost all endogenous antibody molecules in the blood and tissues, and therapeutic antibodies used to control these cells will face competition from endogenous CD16 receptors on NK cells, PMNs, monocytes, and macrophages. These two characteristics respectively contribute to non-tumor toxicity and poor pharmacokinetic issues.

[0071] Naturally occurring NKG2D ligands are present in some healthy tissues and many stressed tissues, and using current NKG2D CAR methods carries a significant risk of toxicity (VanSeggelen et al., 2015). The specific binding of the Y152A non-natural NKG2D receptor to a non-natural α1-α2 domain NKG2D ligand constitutes an example of a means of selectively controlling non-natural NKG2D CAR activity using a bispecific protein composed of the non-natural α1-α2 domains of the invented NKG2D ligand.

[0072] We designed CAR-T cells with a receptor composed of the modified NKG2D Y152A / Y199F (“AF”) extracellular domain, which lacks binding to all natural NKG2D ligands or the previously described non-natural α1-α2 domain orthogonal and homologous to Y152A-modified NKG2D (NKG2D.YA). The homologous non-natural α1-α2 domain of this invention binds with high affinity to the non-natural NKG2D.AF extracellular domain, avoiding binding to the natural NKG2DYA extracellular domain and the NKG2D.YA extracellular domain. Therefore, the engineered α1-α2 domain, exhibiting strong selectivity for the extracellular domain of non-natural NKG2D.AF compared to natural NKG2D and non-natural NKG2D.YA, represents an ideal system for selectively controlling non-natural NKG2D CAR receptors or any receptors or proteins fused to the extracellular domain of non-natural NKG2D, which can be selectively bound by the non-natural α1-α2 domain of this invention. This invention further enables a single cell to express two distinct CARs—one composed of NKG2D.YA and the other of NKG2D.AF—each with a distinctly different intracellular domain for each signal transduction. These distinct CARs will have independent, dual control over cellular activity through extracellular exposure to their respective, homologous orthogonal MicAbody or another non-antibody fusion peptide.

[0073] To demonstrate selective control over CAR-T cells constructed using chimeric receptors displaying the extracellular domain of non-natural NKG2D.AF, building upon previous work, we constructed CARs using the 4-1BB / CD3ζ CAR construct (Campana patent 8,399,645) by fusing the corresponding NKG2D extracellular domain to the CD8 hinge region of the CAR (SEQ ID NO: 151, 153, 155), resulting in CARs with the extracellular domains of natural NKG2D.wt (SEQ ID NO: 135), non-natural NKG2D.YA (SEQ ID NO: 137), or non-natural NKG2D.AF (SEQ ID NO: 139). These constructs (SEQ ID NO: 152, 154, 156) were cloned into lentiviral vectors and expressed in primary human CD8-positive T cells via lentiviral transduction. HeLa cells possess constitutively upregulated levels of MIC ligands on their surface, including MICA, MICB, ULBP3, and ULBP2 / 5 / 6 (the antibody used to determine this cannot distinguish between these three ULBPs; human ULBP-2 / 5 / 6 antibody from the R&D system in Minneapolis, Minnesota). HeLa cells were transfected to overexpress either native ULBP1 or the NKG2D.AF-selected variant ULBP2.R on their surface, and these cells were used as targets for in vitro killing assays. HeLa target cells were preloaded with calcein and exposed to NKG2D.wt-CAR, NKG2D.YA-CAR, or NKG2D.AF-CAR CD8 cells for 5 hours at an increased effector-to-target (E:T) ratio. The amount of calcein released into the supernatant was then quantified and normalized to the total calcein released after detergent treatment. Figures 17A-17CDue to elevated levels of naturally expressed MIC ligands on the surface of HeLa cells, CD8 cells expressing native NKG2D (NKG2D.wt) act as CARs, binding to HeLa cells and inducing cell lysis via this overexpressed native ligand. However, CD8 cells transduced with NKG2D.YA- and NKG2D.AF-CARs showed minimal native HeLa cell lysis even at high E:T ratios (equivalent activity levels to untransduced CD8 T cells). When ULBP1 was overexpressed on the surface of HeLa cells, only NKG2D.wt-CAR CD8 T cells significantly lysed them. NKG2D.YA-CAR cells exhibited some additional killing activity at high E:T ratios, but this was not observed in NKG2D.AF-CAR cells, suggesting that the double mutation Y152A / Y199F makes NKG2D more indolent than the single Y152A mutation. In HeLa cells overexpressing NKG2D.AF selectively non-natural ULBP2.R, NKG2D.wt-CAR cells lyse directly (due to recognition of endogenous MIC ligands), while the directed lysis level of NKG2D.AF-CAR cells is consistent with the binding of the receptor and its selective ligands.

[0074] To demonstrate that lysis of NKG2D.YA- or NKG2D.AF-CAR cells can only occur through appropriate, homologous targeting of MicAbody, Ramos cells were combined with rituximab-based MicAbody as a target for cell lysis. The rituximab-based MicAbody was linked to a non-natural ULBP2.S3 or ULBP2.R orthogonal ligand. Figure 18AAs shown, rituximab-ULBP2.S3 MicAbody directs cytotoxic activity in NKG2D.YA-CAR CD8 cells, but not in NKG2D.AF-CAR cells, while rituximab-ULBP2.R MicAbody directs cytotoxic activity in NKG2D.AF-CAR cells, but not in NKG2D.YA-CAR cells. This further demonstrates the selectivity of the two non-natural ULBP2 variants for their homologous non-natural NKG2D variants, which were designed as preferred mates. To demonstrate the specificity of the MicAbody antibody moiety, NKG2D.AF-CARCD8 cells were pre-armed by incubation with rituximab-ULBP2.R, trastuzumab-ULPB2.R (SEQ ID NO: 95 and 133, heavy chain and light chain, respectively), or both, in equimolar combinations at saturated total concentrations, and in vitro cytotoxicity assays were performed. After washing to remove unbound MicAbody, CD8 cells were applied to either Ramos cells (expressing CD20, the target of rituximab) or CT26-Her2 cells (a mouse cell line transfected to express human Her2) pre-loaded with calcein. The amount of calcein released was quantified after 2 hours of incubation at two different E:T ratios. Figure 18B As shown, when cells were pre-armed with rituximab-MicAbody, only Ramos cells lysed, while trastuzumab-MicAbody exhibited targeted lytic activity against CT26-Her2 cells. However, when NKG2D.AF-CAR CD8 cells were pre-armed with both rituximab and trastuzumab-ULBP2.RMicAbody, both target cell lines lysed, demonstrating that these CAR cells (through selective, preferential pairing already designed between receptor and ligand) are readily reusable and thus directed to simultaneously bind to different tumor targets. sequence list <110> XYPHOS BIOSCIENCES INC. <120> Modified α1-α2 domains of non-natural NKG2D ligands binding to non-natural NKG2D receptors <130> F246240 <150> US 62 / 648,636 <151> 2018-03-27 <160> 156 <170> PatentIn version 3.5 <210> 1 <211> 183 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide MICA α1-α2 domain <400> 1 Ala Ala Glu Pro His Ser Leu Arg Tyr Asn Leu Thr Val Leu Ser Trp 1 5 10 15 Asp Gly Ser Val Gln Ser Gly Phe Leu Thr Glu Val His Leu Asp Gly 20 25 30 Gln Pro Phe Leu Arg Cys Asp Arg Gln Lys Cys Arg Ala Lys Pro Gln 35 40 45 Gly Gln Trp Ala Glu Asp Val Leu Gly Asn Lys Thr Trp Asp Arg Glu 50 55 60 Thr Arg Asp Leu Thr Gly Asn Gly Lys Asp Leu Arg Met Thr Leu Ala 65 70 75 80 His Ile Lys Asp Gln Lys Glu Gly Leu His Ser Leu Gln Glu Ile Arg 85 90 95 Val Cys Glu Ile His Glu Asp Asn Ser Thr Arg Ser Ser Gln His Phe 100 105 110 Tyr Tyr Asp Gly Glu Leu Phe Leu Ser Gln Asn Leu Glu Thr Lys Glu 115 120 125 Trp Thr Met Pro Gln Ser Ser Arg Ala Gln Thr Leu Ala Met Asn Val 130 135 140 Arg Asn Phe Leu Lys Glu Asp Ala Met Lys Thr Lys Thr His Tyr His 145 150 155 160 Ala Met His Ala Asp Cys Leu Gln Glu Leu Arg Arg Tyr Leu Lys Ser 165 170 175 Gly Val Val Leu Arg Arg Thr 180 <210> 2 <211> 182 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide MICB α1-α2 domain <400> 2 Ala Glu Pro His Ser Leu Arg Tyr Asn Leu Met Val Leu Ser Gln Asp 1 5 10 15 Glu Ser Val Gln Ser Gly Phe Leu Ala Glu Gly His Leu Asp Gly Gln 20 25 30 Pro Phe Leu Arg Tyr Asp Arg Gln Lys Arg Arg Ala Lys Pro Gln Gly 35 40 45 Gln Trp Ala Glu Asp Val Leu Gly Ala Lys Thr Trp Asp Thr Glu Thr ​​​​​​​​​ Cys Glu Ile His Glu Asp Ser Ser Thr Arg Gly Ser Arg His Phe Tyr 100 105 110 Tyr Asp Gly Glu Leu Phe Leu Ser Gln Asn Leu Glu Thr Gln Glu Ser 115 120 125 Thr Val Pro Gln Ser Ser Arg Ala Gln Thr Leu Ala Met Asn Val Thr 130 135 140 Asn Phe Trp Lys Glu Asp Ala Met Lys Thr Lys Thr His Tyr Arg Ala 145 150 155 160 Met Gln Ala Asp Cys Leu Gln Lys Leu Gln Arg Tyr Leu Lys Ser Gly 165 170 175 Val Ala Ile Arg Arg Thr 180 <210> 3 <211> 187 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide ULBP1 α1-α2 domain <400> 3 Ala Ala Glu Pro His Cys Leu Cys Tyr Asp Phe Ile Ile Thr Pro Lys 1 5 10 15 Ser Arg Pro Glu Pro Gln Trp Cys Glu Val Gln Gly Leu Val Asp Glu 20 25 30 Arg Pro Phe Leu His Tyr Asp Cys Val Asn His Lys Ala Lys Ala Phe 35 40 45 Ala Ser Leu Gly Lys Lys Val Asn Val Thr Lys Thr Trp Glu Glu Gln 50 55 60 Thr Glu Thr Leu Arg Asp Val Val Asp Phe Leu Lys Gly Gln Leu Leu 65 70 75 80 Asp Ile Gln Val Glu Asn Leu Ile Pro Ile Glu Pro Leu Thr Leu Gln 85 90 95 Ala Arg Met Ser Cys Glu His Glu Ala His Gly His Gly Arg Gly Ser 100 105 110 Trp Gln Phe Leu Phe Asn Gly Gln Lys Phe Leu Leu Phe Asp Ser Asn 115 120 125 Asn Arg Lys Trp Thr Ala Leu His Pro Gly Ala Lys Lys Met Thr Glu 130 135 140 Lys Trp Glu Lys Asn Arg Asp Val Thr Met Phe Phe Gln Lys Ile Ser 145 150 155 160 Leu Gly Asp Cys Lys Met Trp Leu Glu Glu Phe Leu Met Tyr Trp Glu 165 170 175 Gln Met Leu Asp Pro Thr Lys Pro Pro Met Val 180 185 <210> 4 <211> 189 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide ULBP2 α1-α2 domain <400> 4 Ala Ala Glu Pro His Ser Leu Cys Tyr Asp Ile Thr Val Ile Pro Lys 1 5 10 15 Phe Arg Pro Gly Pro Arg Trp Cys Ala Val Gln Gly Gln Val Asp Glu ~20 25 30 Lys Thr Phe Leu His Tyr Asp Cys Gly Asn Lys Thr Val Thr Pro Val 35 40 45 Ser Pro Leu Gly Lys Lys Leu Asn Val Thr Thr Ala Trp Lys Ala Gln 50 55 60 Asn Pro Val Leu Arg Glu Val Val Asp Ile Leu Thr Glu Gln Leu Arg 65 70 75 80 Asp Ile Gln Leu Glu Asn Tyr Thr Pro Lys Glu Pro Leu Thr Leu Gln 85 90 95 Ala Arg Met Ser Cys Glu Gln Lys Ala Glu Gly His Ser Ser Gly Ser 100 105 110 Trp Gln Phe Ser Phe Asp Gly Gln Ile Phe Leu Leu Phe Asp Ser Glu 115 120 125 Lys Arg Met Trp Thr Thr Val His Pro Gly Ala Arg Lys Met Lys Glu 130 135 140 Lys Trp Glu Asn Asp Lys Val Val Ala Met Ser Phe His Tyr Phe Ser 145 150 155 160 Met Gly Asp Cys Ile Gly Trp Leu Glu Asp Phe Leu Met Gly Met Asp 165 170 175 Ser Thr Leu Glu Pro Ser Ala Gly Ala Pro Pro Met Val 180 185 <210> 5 <211> 187 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide ULBP3 α1-α2 domain <400> 5 Ala Ala Glu Pro His Ser Leu Trp Tyr Asn Phe Thr Ile Ile His Leu 1 5 10 15 Pro Arg His Gly Gln Gln Trp Cys Glu Val Gln Ser Gln Val Asp Gln[[ID=3B]] 20 25 30 Lys Asn Phe Leu Ser Tyr Asp Cys Gly Ser Asp Lys Val Leu Ser Met 35 40 45 Gly His Leu Glu Glu Gln Leu Tyr Ala Thr Asp Ala Trp Gly Lys Gln 50 55 60 Leu Glu Met Leu Arg Glu Val Gly Gln Arg Leu Arg Leu Glu Leu Ala 65 70 75 80 Asp Thr Glu Leu Glu Asp Phe Thr Pro Ser Gly Pro Leu Thr Leu Gln 85 90 95 Val Arg Met Ser Cys Glu Cys Glu Ala Asp Gly Tyr Ile Arg Gly Ser 100 105 110 Trp Gln Phe Ser Phe Asp Gly Arg Lys Phe Leu Leu Phe Asp Ser Asn 115 120 125 Asn Arg Lys Trp Thr Val Val His Ala Gly Ala Arg Arg Met Lys Glu 130 135 140 Lys Trp Glu Lys Asp Ser Gly Leu Thr Thr Phe Phe Lys Met Val Ser 145 150 155 160 Met Arg Asp Cys Lys Ser Trp Leu Arg Asp Phe Leu Met His Arg Lys 165 170 175 Lys Arg Leu Glu Pro Thr Ala Pro Pro Met Val 180 185 <210> 6 <211> 187 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide ULBP4 α1-α2 domain <400> 6 Ala Ala Glu Pro His Ser Leu Cys Phe Asn Phe Thr Ile Lys Ser Leu 1 5 10 15 Ser Arg Pro Gly Gln Pro Trp Cys Glu Ala Gln Val Phe Leu Asn Lys 20 25 30 Asn Leu Phe Leu Gln Tyr Asn Ser Asp Asn Asn Met Val Lys Pro Leu 35 40 45 Gly Leu Leu Gly Lys Lys Val Tyr Ala Thr Ser Thr Trp Gly Glu Leu 50 55 60 Thr Gln Thr Leu Gly Glu Val Gly Arg Asp Leu Arg Met Leu Leu Cys 65 70 75 80 Asp Ile Lys Pro Gln Ile Lys Thr Ser Asp Pro Ser Thr Leu Gln Val 85 90 95 Glu Met Phe Cys Gln Arg Glu Ala Glu Arg Cys Thr Gly Ala Ser Trp 100 105 110 Gln Phe Ala Thr Asn Gly Glu Lys Ser Leu Leu Phe Asp Ala Met Asn 115 120 125 Met Thr Trp Thr Val Ile Asn His Glu Ala Ser Lys Ile Lys Glu Thr 130 135 140 Trp Lys Lys Asp Arg Gly Leu Glu Lys Tyr Phe Arg Lys Leu Ser Lys 145 150 155 160 Gly Asp Cys Asp His Trp Leu Arg Glu Phe Leu Gly His Trp Glu Ala 165 170 175 Met Pro Glu Pro Thr Val Ser Pro Pro Met Val 180 185 <210> 7 <211> 198 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide ULBP5 α1-α2 domain <400> 7 Gly Leu Ala Asp Pro His Ser Leu Cys Tyr Asp Ile Thr Val Ile Pro 1 5 10 15 Lys Phe Arg Pro Gly Pro Arg Trp Cys Ala Val Gln Gly Gln Val Asp 20 25 30 Glu Lys Thr Phe Leu His Tyr Asp Cys Gly Ser Lys Thr Val Thr Pro 35 40 45 Val Ser Pro Leu Gly Lys Lys Leu Asn Val Thr Thr Ala Trp Lys Ala 50 55 60 Gln Asn Pro Val Leu Arg Glu Val Val Asp Ile Leu Thr Glu Gln Leu 65 70 75 80 Leu Asp Ile Gln Leu Glu Asn Tyr Ile Pro Lys Glu Pro Leu Thr Leu 85 90 95 Gln Ala Arg Met Ser Cys Glu Gln Lys Ala Glu Gly His Gly Ser Gly 100 105 110 Ser Trp Gln Leu Ser Phe Asp Gly Gln Ile Phe Leu Leu Phe Asp Ser 115 120 125 Glu Asn Arg Met Trp Thr Thr Val His Pro Gly Ala Arg Lys Met Lys 130 135 140 Glu Lys Trp Glu Asn Asp Lys Asp Met Thr Met Ser Phe His Tyr Ile 145 150 155 160 Ser Met Gly Asp Cys Thr Gly Trp Leu Glu Asp Phe Leu Met Gly Met 165 170 175 Asp Ser Thr Leu Glu Pro Ser Ala Gly Ala Pro Pro Thr Met Ser Ser 180 185 190 Gly Thr Ala Gln Pro Arg 195 <210> 8 <211> 189 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide ULBP6 α1-α2 domain <400> 8 Ala Ala Glu Pro His Ser Leu Cys Tyr Asp Ile Thr Val Ile Pro Lys 1 5 10 15 Phe Arg Pro Gly Pro Arg Trp Cys Ala Val Gln Gly Gln Val Asp Glu 20 25 30 Lys Thr Phe Leu His Tyr Asp Cys Gly Asn Lys Thr Val Thr Pro Val 35 40 45 Ser Pro Leu Gly Lys Lys Leu Asn Val Thr Thr Ala Trp Lys Ala Gln 50 55 60 Asn Pro Val Leu Arg Glu Val Val Asp Ile Leu Thr Glu Gln Leu Leu 65 70 75 80 Asp Ile Gln Leu Glu Asn Tyr Thr Pro Lys Glu Pro Leu Thr Leu Gln 85 90 95 Ala Arg Met Ser Cys Glu Gln Lys Ala Glu Gly His Ser Ser Gly Ser 100 105 110 Trp Gln Phe Ser Ile Asp Gly Gln Thr Phe Leu Leu Phe Asp Ser Glu 115 120 125 Lys Arg Met Trp Thr Thr Val His Pro Gly Ala Arg Lys Met Lys Glu 130 135 140 Lys Trp Glu Asn Asp Lys Asp Val Ala Met Ser Phe His Tyr Ile Ser 145 150 155 160 Met Gly Asp Cys Ile Gly Trp Leu Glu Asp Phe Leu Met Gly Met Asp 165 170 175 Ser Thr Leu Glu Pro Ser Ala Gly Ala Pro Pro Met Val 180 185 <210> 9 <211> 161 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide OMCP α1-α2 Domain <400> 9 Ala Ala Ala Glu Pro His Lys Leu Ala Phe Asn Phe Asn Leu Glu Ile 1 5 10 15 Asn Gly Ser Asp Thr His Ser Thr Val Asp Val Tyr Leu Asp Asp Ser 20 25 30 Gln Ile Ile Thr Phe Asp Gly Lys Asp Ile Arg Pro Thr Ile Pro Phe 35 40 45 Met Ile Gly Asp Glu Ile Phe Leu Pro Phe Tyr Lys Asn Val Phe Ser 50 55 60 Glu Phe Phe Ser Leu Phe Arg Arg Val Pro Thr Ser Thr Pro Tyr Glu 65 70 75 80 Asp Leu Thr Tyr Phe Tyr Glu Cys Asp Tyr Thr Asp Asn Lys Ser Thr 85 90 95 Phe Asp Gln Phe Tyr Leu Tyr Asn Gly Glu Glu Tyr Thr Val Lys Thr 100 105 110 Gln Glu Ala Thr Asn Lys Asn Met Trp Leu Thr Thr Ser Glu Phe Arg 115 120 125 Leu Lys Lys Trp Phe Asp Gly Glu Asp Cys Ile Met His Leu Arg Ser 130 135 140 Leu Val Arg Lys Met Glu Asp Ser Lys Arg Arg Thr Val Pro Pro Met 145 150 155 160 Val <210> 10 <211> 543 <212> DNA <213> The snowstorm <220> <223> For example, MICA α1-α2 molecules are available <400> 10 gagccccaca gtcttcgtta taacctcacg gtgctgtcct gggatggatc tgtgcagtca gggtttctca ctgaggtaca tctggatggt cagcccttcc tgcgctgtga caggcagaaa tgcagggcaa agccccaggg acagtgggca gaagatgtcc tgggaaata gacatgggac aggagacca gagacttgac agggacgga aaggacctca ggatgaccct ggctcatatc aaggaccaga aagaaggctt gcattccctc caggagatta gggtctgtga gatccatga 360. gacaacagca ccaggagctc ccagcatttc tactacgatg gggagctctt cctctcccaa aacctggaga ctaaggaatg gacaatgccc cagtcctcca gagctcagac cttggccatg aacgtcagga atttcttgaa ggaagatgcc atgaagacca agacacacta tcacgctatg catgcagact gcctgcagga actacggcga tatctaaaat ccggcgtagt cctgaggaga aca 543 <210> 11 <211> 561 <212> DNA <213> Artificial Sequence <220> <223> Synthetic polynucleotide encoding the ULBP1 α1-α2 domain <400> 11 gctgctgagc cccactgtct ctgctacgac tttattataa ctcctaagtc aagaccagag 60 cctcagtggt gcgaagtaca aggtttggtt gacgaaaggc ctttccttca ctacgattgt 120 gtgaaccata aggcaaaggc tttcgccagc ctgggtaaga aggtaaacgt tactaagacg 180 tgggaggagc agacggaaac cctccgtgat gtggttgact ttcttaaggg tcagctcctc 240 gatatccaag tggagaattt aatccctatc gaaccgctca ctctgcaggc cagaatgtca 300 tgcgaacatg aagcacacgg tcatggaaga ggtagttggc aatttttatt taacggtcaa 360 aaattcctgc tgttcgactc aaacaaccgc aaatggactg cgctgcaccc tggagctaag 420 aagatgactg aaaaatggga gaagaacaga gacgttacca tgttcttcca gaagatttcc 480 ctgggagatt gtaagatgtg gttagaggag ttcttaatgt actgggaaca gatgctggac 540 cccacaaaac cccccatggt g 561 <210> 12 <211> 567 <212> DNA <213> Artificial Sequence <220> <223> Synthesized polynucleotide, encoding the ULBP2 α1-α2 domain <400> 12 gctgctgagc cccatagtct gtgttacgac atcacagtta ttcccaagtt caggcccgga 60 ccgcgctggt gtgccgtgca aggacaagtc gacgaaaaaa cctttcttca ttacgattgc 120 ggaaataaga ctgtaacgcc agtctctcct ttaggtaaga agttaaacgt cactacggcg 180 tggaaggcac aaaaccccgt cctgcgcgag gtcgtcgaca tcctgactga acaattgcgc 240 gacatccagc tcgagaatta cactccaaag gagcctctta ccctgcaggc tagaatgtct 300 tgcgagcaaa aggcagaggg ccactcctcc ggcagctggc agttcagttt cgacggacaa 360 atctttctgt tattcgattc agagaagaga atgtggacta cagttcaccc cggtgcccgt 420 aaaatgaagg agaagtggga aaacgacaaa gtggtggcga tgtcattcca ctatttctcg 480 atgggagact gcatcggttg gctggaagat ttcctcatgg gtatggactc cactttggag 540 ccatcggctg gtgccccccc catggtg 567 <210> 13 <211> 561 <212> DNA <213> Artificial sequence <220> <223> Synthesized polynucleotide, encoding the ULBP3 α1-α2 domain <400> 13 gctgctgagc cccacagctt gtggtacaac ttcaccatta tccacttgcc gagacatggc 60 cagcagtggt gcgaagtgca atcgcaagtc gaccaaaaaa acttcttatc atacgactgc 120 ggcagcgata aggtcttatc tatgggtcat ttggaggaac agctctacgc gaccgacgcc 180 tggggtaaac agctcgagat gctccgtgag gttggacaga ggctgagact ggaactggct 240 gacactgagc tggaagattt cacacctagt ggtccactca cattgcaagt acgcatgagc 300 tgcgagtgtg aggccgatgg atacattagg ggcagctggc agtttagctt cgacggaagg 360 aaattcctgc tcttcgacag taacaatagg aagtggactg ttgtgcatgc tggtgcgcgc 420 agaatgaagg aaaagtggga gaaagatagc ggcctgacga ccttcttcaa gatggtgtct 480 atgcgtgact gtaagagctg gctcagagat ttcctcatgc atcgcaagaa gaggttagaa 540 cctaccgctc cccccatggt g 561 <210> 14 <211> 561 <212> DNA <213> Artificial Sequence <220> <223> Synthetic polynucleotide encoding the ULBP4 α1-α2 domain <400> 14 gctgctgagc cccactctct ttgcttcaac ttcaccatta aatccctgag caggcctggt 60 gctgctgagc cccactctct ttgcttcaac ttcaccatta aatccctgag caggcctggt 60 cagccgtggt gtgaggcgca ggtctttctt aacaagaatc tcttcctcca atacaactct 120 cagccgtggt gtgaggcgca ggtctttctt aacaagaatc tcttcctcca atacaactct 120 gataacaaca tggtaaagcc actgggtctc ctgggtaaaa aagtctatgc tacgagcact 一、180 gataacaaca tggtaaagcc actgggtctc ctgggtaaaa aagtctatgc tacgagcact 180 tggggagaac tcacccagac tcttggcgag gtaggaagag acctgcgcat gctcctctgc 240 tggggagaac tcacccagac tcttggcgag gtaggaagag acctgcgcat gctcctctgc 240 gatataaagc cccaaattaa gaccagtgat ccgtccactt tacaagtcga aatgttctgc 300 gatataaagc cccaaattaa gaccagtgat ccgtccactt tacaagtcga aatgttctgc 300 caaagggagg ctgaacgctg caccggagcc tcttggcagt tcgcgaccaa tggcgaaaag 360 caaagggagg ctgaacgctg caccggagcc tcttggcagt tcgcgaccaa tggcgaaaag 360 tccctcttgt tcgatgccat gaatatgacc tggaccgtga tcaatcatga ggcctctaag 420 tccctcttgt tcgatgccat gaatatgacc tggaccgtga tcaatcatga ggcctctaag 420 atcaaggaga cgtggaaaaa ggaccgcggc cttgaaaagt actttaggaa gttgtctaaa 480 atcaaggaga cgtggaaaaa ggaccgcggc cttgaaaagt actttaggaa gttgtctaaa 480 ggagactgcg accattggtt acgcgagttc ctcggccatt gggaagcgat gcccgagcca 540 ggagactgcg accattggtt acgcgagttc ctcggccatt gggaagcgat gcccgagcca 540 acggttagcc cccccatggt g 561 acggttagcc cccccatggt g 561 <210> 15<210> 15 <211> 567 <211> 567 <212> DNA <212> DNA <213> 人工序列 <213> Artificial sequence <220> <220> <223> 合成多核苷酸,编码 ULBP6 α1-α2 结构域 <223> Synthetic polynucleotide encoding ULBP6 α1-α2 domain <400> 15 <400> 15 gctgctgagc cccactcctt atgctatgat atcaccgtga ttccaaagtt ccgaccagga 60 ccccgatggt gcgccgtaca gggacaggtc gacgaaaaga cttttttaca ttacgactgc 120 ggtaacaaga cagtcacacc ggtaagtcct ttgggaaaaa agttaaacgt aaccactgct 180 tggaaggccc agaaccccgt ccttcgagaa gtagtggata ttttgactga acagctgctt 240 gacatccagc tggaaaacta cacacccaaa gagcccctga ctcttcaagc gcgtatgtcg 300 tgtgagcaaa aggccgaagg acacagctcc ggatcctggc agttcagtat cgacggtcag 360 accttcctcc tcttcgattc agaaaagcgc atgtggacta ctgtgcaccc cggcgctcgt 420 aagatgaagg aaaagtggga gaatgataag gacgttgcca tgagttttca ttacattagt 480 atgggagatt gcatcggttg gctggaagac ttcctgatgg gtatggatag tacccttgaa 540 cctagtgccg gagctccccc catggtg 567 <210> 16 <211> 483 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding OMCP α1-α2 domain <400> 16 gctgctgctg agccccacaa gcttgcgttc aacttcaatc tggaaataaa cggttcagat 60gctgctgctg agccccacaa gcttgcgttc aacttcaatc tggaaataaa cggttcagat 60 acccattcaa ccgtggacgt ttatttagac gattcgcaga taatcacctt tgacggcaag 120acccattcaa ccgtggacgt ttatttagac gattcgcaga taatcacctt tgacggcaag 120 gacatccgcc caactatccc gttcatgata ggtgacgaaa tcttccttcc tttttataag 180gacatccgcc caactatccc gttcatgata ggtgacgaaa tcttccttcc tttttataag 180 aatgtgttct ctgagttctt cagtttgttc cgccgcgtcc ctacctcaac cccctacgaa 240aatgtgttct ctgagttctt cagtttgttc cgccgcgtcc ctacctcaac cccctacgaa 240 gacttgactt atttctatga atgcgactac accgacaaca aatctacatt cgatcaattc 300gacttgactt atttctatga atgcgactac accgacaaca aatctacatt cgatcaattc 300 tacctgtaca acggtgaaga gtacaccgtg aagactcaag aggctactaa caagaacatg 360tacctgtaca acggtgaaga gtacaccgtg aagactcaag aggctactaa caagaacatg 360 tggctgacca cttccgagtt cagactgaag aagtggttcg acggcgagga ctgtatcatg 420tggctgacca cttccgagtt cagactgaag aagtggttcg acggcgagga ctgtatcatg 420 caccttagaa gtttagtgag gaaaatggaa gatagcaaga gaagaacagt gccccccatg 480caccttagaa gtttagtgag gaaaatggaa gatagcaaga gaagaacagt gccccccatg 480 <0000​​​​​ 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Tyr His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 18 <211> 139 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide non-natural NKG2D Y152A extracellular domain <400> 18 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Ala His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 19 <211> 139 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide, Non-Natural NKG2D Y199A Extracellular Domain <400> 19 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Tyr His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Ala Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 20 <211> 139 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide, Non-Natural NKG2D Y152A / Y199A Extracellular Domain <400> 20 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Ala His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Ala Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 21 <211> 139 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide, non-natural NKG2D Y199F eNKG2D1 extracellular domain <400> 21 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Tyr His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Phe Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 22 <211> 139 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide, non - natural NKG2D Y152S eNKG2D2 extracellular domain <400> 22 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Ser His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 23 <211> 139 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide, non-natural NKG2D Y152T eNKG2D3 extracellular domain <400> 23 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Thr His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 24 <211> 139 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide, non-natural NKG2D Y152V eNKG2D4 extracellular domain <400> 24 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Val His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 25 <211> 139 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide non-natural NKG2D Y152A / Y199F eNKG2D5 extracellular domain <400> 25 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Ala His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Phe Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 26 <211> 139 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide, Non - natural NKG2D Y152L / Y199F eNKG2D6 Extracellular Domain <400> 26 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Leu His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Phe Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 27 <211> 139 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide non-natural NKG2D Y152S / Y199F eNKG2D7 extracellular domain <400> 27 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Ser His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Phe Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 28 <211> 139 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide non-natural NKG2D Y152T / Y199F eNKG2D8 extracellular domain <400> 28 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Thr His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Phe Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 29 <211> 139 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide non-natural NKG2D Y152V / Y199F eNKG2D9 extracellular domain <400> 29 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Val His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Phe Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 30 <211> 139 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide non-natural NKG2D Y199D eNKG2D10 extracellular domain <400> 30 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Tyr His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Asp Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 31 <211> 139 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide non-natural NKG2D Y199E eNKG2D11 extracellular domain <400> 31 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Tyr His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Glu Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 32 <211> 139 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide, Non-Natural NKG2D Y152D / Y199D eNKG2D12 Extracellular Domain <400> 32 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Asp His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Asp Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 33 <211> 139 <212> PRT <213> Artificial Sequence <220>​<223> Synthetic peptide, non-natural NKG2D Y152E / Y199E eNKG2D13 extracellular domain <400> 33 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Glu His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Glu Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 34 <211> 139 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide, non - natural NKG2D Y152L eNKG2D14 extracellular domain <400> 34 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Leu His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 35 <211> 139 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide, non-natural NKG2D Y152F / Y199F eNKG2D15 extracellular domain <400> 35 Phe Leu Asn Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu 1 5 10 15 Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn 20 25 30 Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala 35 40 45 Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu 50 55 60 Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Phe His Trp Met Gly Leu 65 70 75 80 Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile 85 90 95 Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys 100 105 110 Ala Leu Tyr Ala Ser Ser Phe Lys Gly Phe Ile Glu Asn Cys Ser Thr 115 120 125 Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 130 135 <210> 36 <211> twenty three <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide MHCI signal sequence <400> 36 Met Gly Leu Gly Pro Val Phe Leu Leu Leu Ala Gly Ile Phe Pro Phe 1 5 10 15 Ala Pro Pro Gly Ala Ala Ala 20 <210> 37 <211> 69 <212> DNA <213> Artificial sequence <220> <223> Synthesized polynucleotides encoding the MHCI signal sequence <400> 37 atgggccttg gcccagtgtt tctgctgttg gcaggcattt tcccttttgc tccgcccggc 60 gccgcagcc 69 <210> 38 <211> 237 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide human IgG1 Fc with IEGR linker <400> 38 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyre Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg 225 230 235 <210> 39 <211> 711 <212> DNA <213> The snowstorm <220> <223> In particular, the IgG1 Fc and IEGR antibodies <400> 39 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggagga 300. cgtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc 420 cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgcccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctgcacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg c 711 <210> 40 <211> 376 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide Human IgG1 Fc-NKG2D Fusion <400> 40 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Tyr His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 41 <211> 376 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide human IgG1 Fc-NKG2D Y152A fusion <400> 41 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Ala His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 42 <211> 376 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide, Human IgG1 Fc-NKG2D Y199A Extracellular Domain <400> 42 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Tyr His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Ala Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 43 <211> 376 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide human IgG1 Fc-NKG2D Y152A / Y199A extracellular domain <400> 43 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Ala His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Ala Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 44 <211> 376 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide, human IgG1 Fc-NKG2D Y199F eNKG2D1 fusion <400> 44 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Tyr His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Phe Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 45 <211> 376 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide, human IgG1 Fc-NKG2D Y152S eNKG2D2 fusion <400> 45 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Tyr His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Phe Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 46 <211> 376 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide, human IgG1 Fc-NKG2D Y152T eNKG2D3 fusion <400> 46 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Thr His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 47 <211> 376 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide Human IgG1 Fc-NKG2D Y152V eNKG2D4 Fusion <400> 47 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Val His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 48 <211> 376 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide, Human IgG1 Fc-NKG2D Y152A / Y199F eNKG2D5 Fusion <400> 48 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Ala His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Phe Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 49 <211> 376 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide, human IgG1 Fc-NKG2D Y152L / Y199F eNKG2D6 fusion <400> 49 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Leu His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Phe Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 50 <211> 376 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide, human IgG1 Fc-NKG2D Y152S / Y199F eNKG2D7 fusion <400> 50 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Ser His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Phe Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 51 <211> 376 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide - human IgG1 Fc - NKG2D Y152T / Y199F eNKG2D8 fusion <400> 51 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Thr His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Phe Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 52 <211> 376 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide, human IgG1 Fc-NKG2D Y152V / Y199F eNKG2D9 fusion <400> 52 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Val His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Phe Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 53 <211> 376 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide, human IgG1 Fc-NKG2D Y199D eNKG2D10 fusion <400> 53 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Tyr His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Asp Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 54 <211> 376 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide, human IgG1 Fc-NKG2D Y199E eNKG2D11 fusion <400> 54 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Tyr His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Glu Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 55 <211> 376 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide, human IgG1 Fc-NKG2D Y152D / Y199D eNKG2D12 fusion <400> 55 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Asp His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Asp Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 56 <211> 376 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide, Human IgG1 Fc-NKG2D Y152E / Y199E eNKG2D13 Fusion <400> 56 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met[[END 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Glu His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Glu Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 57 <211> 376 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide, human IgG1 Fc-NKG2D Y152L eNKG2D14 fusion <400> 57 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Leu His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 58 <211> 376 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide, Human IgG1 Fc-NKG2D Y152F / Y199F eNKG2D15 Fusion <400> 58 Met Asp Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 1 5 10 15 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 20 25 30 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 35 40 45 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 50 55 60 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 65 70 75 80 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 85 90 95 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 100 105 110 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 115 120 125 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 130 135 140 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 145 150 155 160 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 165 170 175 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 180 185 190 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 195 200 205 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 210 215 220 Lys Ser Leu Ser Leu Ser Pro Gly Lys Ile Glu Gly Arg Phe Leu Asn 225 230 235 240 Ser Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr Glu Ser Tyr Cys 245 250 255 Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn Asn Cys Tyr Gln 260 265 270 Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln Ala Ser Cys Met 275 280 285 Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys Glu Asp Gln Asp 290 295 300 Leu Leu Lys Leu Val Lys Ser Phe His Trp Met Gly Leu Val His Ile 305 310 315 320 Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser Ile Leu Ser Pro 325 330 335 Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp Cys Ala Leu Tyr 340 345 350 Ala Ser Ser Phe Lys Gly Phe Ile Glu Asn Cys Ser Thr Pro Asn Thr 355 360 365 Tyr Ile Cys Met Gln Arg Thr Val 370 375 <210> 59 <211> 1128 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide encoding human IgG1 Fc-NKG2D fusion <400> 59 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg 60 ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggaggaa 240 cagtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg 300 aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag 360 accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc 420 cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgcccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctgcacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 tatgagagcc aggcttcttg tatgtctcaa aatgccagcc ttctgaaagt atacagcaaa 900 gaggaccagg atttacttaa actggtgaag tcatatcatt ggatgggact agtacacatt 960 ccaacaaatg gatcttggca gtgggaagat ggctccattc tctcacccaa cctactaaca 1020 ataattgaaa tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggctatata 1080 gaaaactgtt caactccaaa tacatacatc tgcatgcaaa ggactgtg 1128 <210> 60 <211> 1128 <212> DNA <213> Artificial Sequence <220> <223> Synthetic polynucleotide, encoding human IgG1 Fc-NKG2D Y152A fusion <400> 60 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg 60 ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggaggaa 240 cagtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg 300 aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag 360 accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc 420 cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgcccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctgcacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 900. aggcttcttg aatgccagcc ttctgaaagt attack gaggaccagg atttacttaa actggtgaag tcagctcatt ggatgggact agtacacatt 1020. cccacaatg gatcttggca gtggggagat ggctccattc tctcacccaa cctactaaca father tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggctatata gaaaactgtt caactccaa tacatacatc tgcatgcaaa ggactgtg <210> 61 <211> 1128 <212> DNA <213> The snowstorm <220> <223> Additionally, the IgG1 Fc-NKG2D Y199A antibody was used <400> 61 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggagga 300. cgtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag 360 accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc 420 cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgccccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctggacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 tatgagagcc aggcttcttg tatgtctcaa aatgccagcc ttctgaaagt atacagcaaa 900 gaggaccagg atttacttaa actggtgaag tcataccatt ggatgggact agtacacatt 960 ccaacaaatg gatcttggca gtgggaagat ggctccattc tctcacccaa cctactaaca 1020 father tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggcgctata gaaaactgtt caactccaa tacatacatc tgcatgcaaa ggactgtg <210> 62 <211> 1128 <212> DNA <213> The snowstorm <220> <223> Additionally, the IgG1 Fc-NKG2D Y152A / Y199A antibody was used <400> 62 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggagga 300. cgtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgcccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctgcacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 tatgagagcc aggcttcttg tatgtctcaa aatgccagcc ttctgaaagt atacagcaaa 900 gaggaccagg atttacttaa actggtgaag tcagctcatt ggatgggact agtacacatt 960 ccaacaaatg gatcttggca gtgggaagat ggctccattc tctcacccaa cctactaaca 1020 ataattgaaa tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggcgctata 1080 gaaaactgtt caactccaaa tacatacatc tgcatgcaaa ggactgtg 1128 <210> 63 <211> 1128 <212> DNA <213> Artificial Sequence <220> <223> Synthetic polynucleotide encoding human IgG1 Fc-NKG2D Y199F eNKG2D1 fusion <400> 63 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg 60 ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggaggaa 240 cagtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg 300 aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag 360 accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc 420 cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgcccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctgcacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 tatgagagcc aggcttcttg tatgtctcaa aatgccagcc ttctgaaagt atacagcaaa 900 gaggaccagg atttacttaa actggtgaag tcataccatt ggatgggact agtacacatt 960 ccaacaaatg gatcttggca gtgggaagat ggctccattc tctcacccaa cctactaaca 1020 ataattgaaa tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggcttcata 1080 gaaaactgtt caactccaaa tacatacatc tgcatgcaaa ggactgtg 1128 <210> 64 <211> 1128 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding human IgG1 Fc-NKG2D Y152S eNKG2D2 fusion <400> 64 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg 60 ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggaggaa 240 cagtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg 300 aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag 360 accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc 420 cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgcccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctgcacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 tatgagagcc aggcttcttg tatgtctcaa aatgccagcc ttctgaaagt atacagcaaa 900 gaggaccagg atttacttaa actggtgaag tcataccatt ggatgggact agtacacatt 960 ccaacaaatg gatcttggca gtgggaagat ggctccattc tctcacccaa cctactaaca 1020 ataattgaaa tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggcttcata 1080 gaaaactgtt caactccaaa tacatacatc tgcatgcaaa ggactgtg 1128 <210> 65 <211> 1128 <212> DNA <213> Artificial Sequence <220> <223> Synthetic polynucleotide, encoding human IgG1 Fc-NKG2D Y152T eNKG2D3 fusion <400> 65 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg 60 ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggaggaa 240 cagtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg 300 aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag 360 accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc 420 cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgccccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctggacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 tatgagagcc aggcttcttg tatgtctcaa aatgccagcc ttctgaaagt atacagcaaa 900 gaggaccagg atttacttaa actggtgaag tcaactcatt ggatgggact agtacacatt 960 ccaacaaatg gatcttggca gtgggaagat ggctccattc tctcacccaa cctactaaca 1020 father tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggctatata gaaaactgtt caactccaa tacatacatc tgcatgcaaa ggactgtg <210> 66 <211> 1128 <212> DNA <213> The snowstorm <220> <223> Additionally, the IgG1 Fc-NKG2D Y152V eNKG2D4 antibody <400> 66 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggagga 300. cgtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgcccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctgcacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 tatgagagcc aggcttcttg tatgtctcaa aatgccagcc ttctgaaagt atacagcaaa 900 gaggaccagg atttacttaa actggtgaag tcagtgcatt ggatgggact agtacacatt 960 ccaacaaatg gatcttggca gtgggaagat ggctccattc tctcacccaa cctactaaca 1020 ataattgaaa tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggctatata 1080 gaaaactgtt caactccaaa tacatacatc tgcatgcaaa ggactgtg 1128 <210> 67 <211> 1128 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide encoding human IgG1 Fc-NKG2D Y152A / Y199F eNKG2D5 fusion <400> 67 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg 60 ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggaggaa 240 cagtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg 300 aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag 360 accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc 420 cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgcccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctgcacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 tatgagagcc aggcttcttg tatgtctcaa aatgccagcc ttctgaaagt atacagcaaa 900 gaggaccagg atttacttaa actggtgaag tcagctcatt ggatgggact agtacacatt 960 ccaacaaatg gatcttggca gtgggaagat ggctccattc tctcacccaa cctactaaca 1020 ataattgaaa tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggcttcata 1080 gaaaactgtt caactccaaa tacatacatc tgcatgcaaa ggactgtg 1128 <210> 68 <211> 1128 <212> DNA <213> Artificial Sequence <220> <223> Synthetic polynucleotide encoding human IgG1 Fc-NKG2D Y152L / Y199F eNKG2D6 fusion <400> 68 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg 60 ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggaggaa 240 cagtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg 300 aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag 360 accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc 420 cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgcccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctgcacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 900. aggcttcttg aatgccagcc ttctgaaagt attack gaggaccagg atttacttaa actggtgaag tcactgcatt ggatgggact agtacacatt 1020. cccacaatg gatcttggca gtggggagat ggctccattc tctcacccaa cctactaaca father tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggcttcata gaaaactgtt caactccaa tacatacatc tgcatgcaaa ggactgtg <210> 69 <211> 1128 <212> DNA <213> The snowstorm <220> <223> Additionally, antibodies to IgG1 Fc-NKG2D Y152S / Y199F eNKG2D7 antibodies <400> 69 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggagga 300. cgtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag 360 accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc 420 cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgcccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctgcacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 tatgagagcc aggcttcttg tatgtctcaa aatgccagcc ttctgaaagt atacagcaaa 900 gaggaccagg atttacttaa actggtgaag tcaagtcatt ggatgggact agtacacatt 960 ccaacaaatg gatcttggca gtgggaagat ggctccattc tctcacccaa cctactaaca 1020 father tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggcttcata gaaaactgtt caactccaa tacatacatc tgcatgcaaa ggactgtg <210> 70 <211> 1128 <212> DNA <213> The snowstorm <220> <223> Additionally, antibodies to IgG1 Fc-NKG2D Y152T / Y199F eNKG2D8 antibodies <400> 70 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggagga 300. cgtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgcccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctgcacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 tatgagagcc aggcttcttg tatgtctcaa aatgccagcc ttctgaaagt atacagcaaa 900 gaggaccagg atttacttaa actggtgaag tcaactcatt ggatgggact agtacacatt 960 ccaacaaatg gatcttggca gtgggaagat ggctccattc tctcacccaa cctactaaca 1020 ataattgaaa tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggcttcata 1080 gaaaactgtt caactccaaa tacatacatc tgcatgcaaa ggactgtg 1128 <210> 71 <211> 1128 <212> DNA <213> Artificial Sequence <220> <223> Synthetic polynucleotide, encoding human IgG1 Fc-NKG2D Y152V / Y199F eNKG2D9 fusion <400> 71 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg 60 ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggaggaa 240 cagtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg 300 aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag 360 accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc 420 cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgcccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctgcacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 tatgagagcc aggcttcttg tatgtctcaa aatgccagcc ttctgaaagt atacagcaaa 900 gaggaccagg atttacttaa actggtgaag tcagtgcatt ggatgggact agtacacatt 960 ccaacaaatg gatcttggca gtgggaagat ggctccattc tctcacccaa cctactaaca 1020 ataattgaaa tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggcttcata 1080 gaaaactgtt caactccaaa tacatacatc tgcatgcaaa ggactgtg 1128 <210> 72 <211> 1128 <212> DNA <213> Artificial Sequence <220> <223> Synthetic polynucleotide, encoding human IgG1 Fc-NKG2D Y199D eNKG2D10 fusion <400> 72 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg 60 ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggaggaa 240 cagtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg 300 aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag 360 accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc 420 cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgcccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctgcacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 900. aggcttcttg aatgccagcc ttctgaaagt attack gaggaccagg atttacttaa actggtgaag tcatatcatt ggatgggact agtacacatt 1020. cccacaatg gatcttggca gtggggagat ggctccattc tctcacccaa cctactaaca father tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggcgatata gaaaactgtt caactccaa tacatacatc tgcatgcaaa ggactgtg <210> 73 <211> 1128 <212> DNA <213> The snowstorm <220> <223> In addition, antibodies to the IgG1 Fc-NKG2D Y199E eNKG2D11 antibodies <400> 73 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggagga 300. cgtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag 360 accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc 420 cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgccccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctggacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 tatgagagcc aggcttcttg tatgtctcaa aatgccagcc ttctgaaagt atacagcaaa 900 gaggaccagg atttacttaa actggtgaag tcataccatt ggatgggact agtacacatt 960 ccaacaaatg gatcttggca gtgggaagat ggctccattc tctcacccaa cctactaaca 1020 ataattgaaa tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggcgagata gaaaactgtt caactccaa tacatacatc tgcatgcaaa ggactgtg <210> 74 <211> 1128 <212> DNA <213> The snowstorm <220> <223> Additionally, the IgG1 Fc-NKG2D Y152D / Y199D eNKG2D12 antibodies <400> 74 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggagga 300. cgtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgcccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctgcacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 tatgagagcc aggcttcttg tatgtctcaa aatgccagcc ttctgaaagt atacagcaaa 900 gaggaccagg atttacttaa actggtgaag tcagatcatt ggatgggact agtacacatt 960 ccaacaaatg gatcttggca gtgggaagat ggctccattc tctcacccaa cctactaaca 1020 ataattgaaa tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggcgatata 1080 gaaaactgtt caactccaaa tacatacatc tgcatgcaaa ggactgtg 1128 <210> 75 <211> 1128 <212> DNA <213> Artificial Sequence <220> <223> Synthetic polynucleotide, encoding human IgG1 Fc-NKG2D Y152E / Y199E eNKG2D13 fusion <400> 75 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg 60 ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggaggaa 240 cagtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg 300 aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag 360 accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc 420 cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgcccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctgcacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 tatgagagcc aggcttcttg tatgtctcaa aatgccagcc ttctgaaagt atacagcaaa 900 gaggaccagg atttacttaa actggtgaag tcagagcatt ggatgggact agtacacatt 960 ccaacaaatg gatcttggca gtgggaagat ggctccattc tctcacccaa cctactaaca 1020 ataattgaaa tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggcgagata 1080 gaaaactgtt caactccaaa tacatacatc tgcatgcaaa ggactgtg 1128 <210> 76 <211> 1128 <212> DNA <213> Artificial Sequence <220> <223> Synthetic polynucleotide, encoding human IgG1 Fc-NKG2D Y152L eNKG2D14 fusion <400> 76 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg 60 ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggaggaa 240 cagtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg 300 aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag 360 accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc 420 cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgcccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctgcacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 900. aggcttcttg aatgccagcc ttctgaaagt attack gaggaccagg atttacttaa actggtgaag tcactgcatt ggatgggact agtacacatt 1020. cccacaatg gatcttggca gtggggagat ggctccattc tctcacccaa cctactaaca father tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggctatata gaaaactgtt caactccaa tacatacatc tgcatgcaaa ggactgtg <210> 77 <211> 1128 <212> DNA <213> The snowstorm <220> <223> Additionally, antibodies to IgG1 Fc-NKG2D Y152F / Y199F eNKG2D15 antibodies <400> 77 atggacccga aaagctgcga caagactcac acttgtccgc cgtgccccgc ccccgaactg ctgggtggcc cctccgtgtt cctgttcccg cctaagccta aggacaccct tatgatcagc 120 cgcacccctg aagtgacctg tgtcgtcgtg gatgtgtcac acgaggaccc ggaggtcaag 180 ttcaattggt acgtggacgg cgtggaagtg cataacgcaa agaccaagcc tcgggagga 300. cgtacaact cgacctaccg cgtggtgtca gtcctgactg tgctgcacca ggactggctg aacgggaagg agtacaagtg caaagtgtcg aacaaggccc tgccggctcc aattgaaaag 360 accatcagca aggccaaggg ccagccaagg gaaccacagg tgtacaccct ccctccttcc 420 cgggacgagc tgaccaaaaa ccaagtgtcc ctgacttgcc ttgtgaaggg gttctaccct 480 tctgacattg ccgtcgaatg ggaatcgaac ggacagcctg aaaacaacta taagactacc 540 ccgccccgtgc tggattccga cggaagcttc ttcctgtact ccaagctgac cgtggacaag 600 tcgagatggc agcagggaaa tgtgttcagc tgctccgtga tgcatgaggc gctggacaac 660 cactacaccc agaagtcact gagcctctcc cccggaaaga tcgaaggacg cttcttaaac 720 tcattattca accaagaagt tcaaattccc ttgaccgaaa gttactgtgg cccatgtcct 780 aaaaactgga tatgttacaa aaataactgc taccaatttt ttgatgagag taaaaactgg 840 tatgagagcc aggcttcttg tatgtctcaa aatgccagcc ttctgaaagt atacagcaaa 900 gaggaccagg atttacttaa actggtgaag tcattccatt ggatgggact agtacacatt 960 ccaacaaatg gatcttggca gtgggaagat ggctccattc tctcacccaa cctactaaca 1020 ataattgaaa tgcagaaggg agactgtgca ctctatgcct cgagctttaa aggcttcata 1080 gaaaactgtt caactccaaa tacatacatc tgcatgcaaa ggactgtg 1128 <210> 78 <211> 181 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide MICwed α1-α2 <400> 78 Glu Pro His Ser Leu Arg Tyr Asn Leu Thr Val Leu Ser Trp Asp Gly 1 5 10 15 Ser Val Gln Ser Gly Phe Leu Thr Glu Val His Leu Asp Gly Gln Pro 20 25 30 Phe Leu Arg Cys Asp Arg Gln Lys Cys Arg Ala Lys Pro Gln Gly Gln 35 40 45 Trp Ala Glu Asp Val Leu Gly Asn Lys Thr Trp Asp Arg Glu Thr Arg 50 55 60 Asp Leu Thr Gly Trp Gly Lys Asp Leu Arg Met Thr Leu Ala His Ile 65 70 75 80 Lys Asp Gln Lys Glu Gly Leu His Ser Leu Gln Glu Ile Arg Val Cys 85 90 95 Glu Ile His Glu Asp Asn Ser Thr Arg Ser Ser Gln His Phe Tyr Tyr 100 105 110 Asp Gly Glu Leu Phe Leu Ser Gln Asn Leu Glu Thr Lys Glu Trp Thr 115 120 125 Met Pro Gln Ser Ser Arg Ala Gln Thr Leu Ala Met Asn Val Arg Asn 130 135 140 Phe Leu Lys Glu Asp Ala Met Glu Thr Asp Thr His Tyr His Ala Met 145 150 155 160 His Ala Asp Cys Leu Gln Glu Leu Arg Arg Tyr Leu Lys Ser Gly Val 165 170 175 Val Leu Arg Arg Thr 180 <210> 79 <211> 543 <212> DNA <213> Artificial Sequence <220> <223> Synthetic polynucleotide, encoding MICwed α1-α2 <400> 79 gagcctcaca gcctccggta taatttgact gtactctctt gggatggctc cgtgcagtcc 60 ggctttctga ctgaagttca tctcgacggt caacctttcc tgcgctgcga ccgacaaaaa 120 tgccgcgcca agccccaagg gcagtgggcc gaagatgtac tgggaaacaa gacctgggac 180 cgggagacac gagacctgac aggctggggc aaggacttgc gcatgacact cgcccatatc 240 aaggaccaga aggaaggatt gcactctttg caagagattc gcgtgtgtga aatccacgag 300 gacaattcaa cgaggagctc ccagcacttc tattacgatg gagaactctt cttgtcacag 360 aacttggaaa ccaaggaatg gactatgcct cagagctctc gggcacagac tctcgctatg 420 aacgttagaa acttccttaa ggaggatgct atggagaccg atactcacta ccacgccatg 480 cacgccgact gcctccagga actgcggaga tatctgaagt ccggcgtggt tttgagaaga 540 acc 543 <210> 80 <211> 181 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide MIC25 α1-α2 <400> 80 Glu Pro His Ser Leu Arg Tyr Asn Leu Thr Val Leu Ser Trp Asp Gly 1 5 10 15 Ser Val Gln Ser Gly Phe Leu Thr Glu Val His Leu Asp Gly Gln Pro 20 25 30 Phe Leu Arg Cys Asp Arg Gln Lys Cys Arg Ala Lys Pro Gln Gly Gln 35 40 45 Trp Ala Glu Asp Val Leu Gly Asn Lys Thr Trp Asp Arg Glu Thr Arg 50 55 60 Asp Leu Thr Gly Trp Gly Lys Asp Leu Arg Met Thr Leu Ala His Ile 65 70 75 80 Lys Asp Gln Lys Glu Gly Leu His Ser Leu Gln Glu Ile Arg Val Cys 85 90 95 Glu Ile His Glu Asp Asn Ser Thr Arg Ser Ser Gln His Phe Tyr Tyr 100 105 110 Asp Gly Glu Leu Phe Leu Ser Gln Asn Leu Glu Thr Leu Glu Trp Thr 115 120 125 Met Pro Gln Ser Ser Arg Ala Gln Thr Leu Ala Met Asn Val Arg Asn 130 135 140 Phe Leu Lys Glu Asp Ala Met Glu Thr Asp Thr His Tyr His Ala Met 145 150 155 160 Arg Ala Asp Cys Leu Ser Glu Leu Arg Arg Tyr Leu Lys Ser Gly Val 165 170 175 Val Leu Arg Arg Thr 180 <210> 81 <211> 543 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding MIC25 α1-α2 <400> 81 gagcctcaca gcctccggta taatttgact gtactctctt gggatggctc cgtgcagtcc 60 ggctttctga ctgaagttca tctcgacggt caacctttcc tgcgctgcga ccgacaaaaa 120 tgccgcgcca agccccaagg gcagtgggcc gaagatgtac tgggaaacaa gacctgggac 180 cgggagacac gagacctgac aggctggggc aaggacttgc gcatgacact cgcccatatc 240 aaggaccaga aggaaggatt gcactctttg caagagattc gcgtgtgtga aatccacgag 300 gacaattcaa cgaggagctc ccagcacttc tattacgatg gagaactctt cttgtcacag 360 aacttggaaa ccctcgaatg gactatgcct cagagctctc gggcacagac tctcgctatg 420 aacgttagaa acttccttaa ggaggatgct atggagaccg atactcacta ccacgccatg 480 cgcgccgact gcctctctga actgcggaga tatctgaagt ccggcgtggt tttgagaaga 540 acc 543 <210> 82 <211> 330 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide, human IgG1 heavy chain CH1-CH2-CH3 D265A / N297A <400> 82 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 1 5 10 15 Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 65 70 75 80 Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100 105 110 Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 115 120 125 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 130 135 140 Val Val Val Ala Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp 145 150 155 160 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165 170 175 Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu 180 185 190 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205 Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210 215 220 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu 225 230 235 240 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250 255 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn 260 265 270 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275 280 285 Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn 290 295 300 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr 305 310 315 320 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 325 330 <210> 83 <211> 990 <212> DNA <213> Artificial sequence <220> <223> Human IgG1 heavy chain CH1-CH2-CH3 D265A / N297A <400> 83 gcgtcgacca agggcccgtc agtgttcccg ctggccccgt catccaagtc cacgtctggg 60 ggcacagcag ccctgggatg cttggtcaag gactacttcc ccgagcccgt gactgtgtcc 120 tggaactccg gagcactgac ctccggagtg cacacctttc ccgcggtgct gcagtcctcc 180 ggactgtact ccctgtcgtc ggtcgtgacc gtgccgagct cctcgctcgg aacccagacc 240 tacatctgca acgtgaacca caagccctcg aacaccaaag tggacaagaa ggtcgagccc 300 aaaagctgcg acaagactca cacttgtccg ccgtgccccg cccccgaact gctgggtggc 360 ccctccgtgt tcctgttccc gcctaagcct aaggacaccc ttatgatcag ccgcacccct 420 gaagtgacct gtgtcgtcgt ggcagtgtca cacgaggacc cggaggtcaa gttcaattgg 480 tacgtggacg gcgtggaagt gcataacgca aagaccaagc ctcgggagga acagtacgcc 540 tcgacctacc gcgtggtgtc agtcctgact gtgctgcacc aggactggct gaacgggaag 600 gagtacaagt gcaaagtgtc gaacaaggcc ctgccggctc caattgaaaa gaccatcagc 660 aaggccaagg gccagccaag ggaaccacag gtgtacaccc tccctccttc ccgggacgag 720 ctgaccaaaa accaagtgtc cctgacttgc cttgtgaagg ggttctaccc ttctgacatt 780 gccgtcgaat gggaatcgaa cggacagcct gaaaacaact ataagactac cccgcccgtg 840 ctggattccg acggaagctt cttcctgtac tccaagctga ccgtggacaa gtcgagatgg 900 cagcagggaa atgtgttcag ctgctccgtg atgcatgagg cgctgcacaa ccactacacc 960 cagaagtcac tgagcctctc ccccggaaag 990 <210> 84 <211> 107 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide, Human κ Light Chain <400> 84 Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 1 5 10 15 Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 20 25 30 Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln 35 40 45 Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 50 55 60 Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 65 70 75 80 Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 85 90 95 Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 100 105 <210> 85 <211> 321 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding human κ light chain <400> 85 agaaccgtgg ccgccccgag cgtgttcatt ttccctccct ccgacgagca gttgaaatcg 60 ggcaccgcta gcgtggtctg ccttctcaac aatttctatc cacgggaagc caaagtgcag 120 tggaaggtcg acaacgcgct ccaatccggg aactcacagg aatccgtgac tgagcaggat 180 tccaaggact cgacctactc cctgtcatcc acgctgaccc tgagcaaggc agactacgag 240 aagcacaagg tctacgcctg cgaagtgaca caccagggac tgtccagccc cgtgaccaag 300 agcttcaaca gaggagaatg c 321 <210> 86 <211> 120 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide trastuzumab VH <400> 86 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr 20 25 30 Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 87 <211> 360 <212> DNA <213> Artificial sequence <220> <223> Synthesized polynucleotide, encoding trastuzumab VH <400> 87 gaagtccaat tggtcgaatc aggcggtgga ctcgtgcaac ctggaggttc gttacgctta 60 tcatgtgctg caagtggatt taatattaaa gatacctaca tccactgggt acgtcaagct 120 cccggcaagg gtctcgagtg ggtcgcacgc atttacccca ccaacggata cacgcgctac 180 gccgattcag tgaagggacg tttcacaatc tctgctgata ctagcaaaaa taccgcatac 240 ctccagatga actctcttag ggccgaggac acagctgtgt actactgtag ccgttgggga 300 ggagacggtt tttacgcaat ggattactgg ggccaaggaa ccctggtcac agtttcatcg 360 <210> 88 <211> 107 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide trastuzumab VL <400> 88 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyrant Ala Ser Phe Leu Tyrant Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 89 <211> 321 <212> DNA <213> The snowstorm <220> <223> Even if you have a friend, you can see the VL <400> 89 gatatccaaa tgactcaatc accatcttca ctctccgcga gcgtgggtga tcgggtcacc atcacatgta gggcgagcca agatgtgaat accgccgtcg cgtggtatca acaaaagccg 180. ggaaaagcac caaaactgct tatatactct gcatccttcc tgtactctgg ggtgccaagc cggttctccg gtagtagatc tggtactgac tttacactca ctatcagcag tctgcaacct 240 gaggactttg cgacatacta ttgccagcag cactacacaa ccccacctac atttggtcag gggacaaagg tggagatcaa g <210> 90 <211> 121 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide rituximab VH <400> 90 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Asn Met His Trp Val Lys Gln Thr Pro Gly Arg Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val Trp Gly 100 105 110 Ala Gly Thr Thr Val Thr Val Ser Ala 115 120 <210> 91 <211> 363 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding Rituximab VH <400> 91 caagttcagc ttcagcagcc gggggctgag ttggtgaaac ccggggccag tgtgaagatg 60 agctgtaaag cgagcggcta caccttcact tcttataata tgcattgggt taagcaaacg 120 ccaggaaggg ggctggagtg gatcggcgct atttacccag gtaacggtga cacatcatat 180 aaccaaaagt ttaagggaaa ggcaaccctc acagcggaca agagtagctc aaccgcatac 240 atgcaactgt caagccttac ctccgaagac agcgcagtgt actactgcgc cagaagcacc 300 tactatgggg gtgattggta cttcaacgtc tggggggctg gcaccacagt gactgtaagc 360 gca 363 <210> 92 <211> 106 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide, Rituximab VL <400> 92 Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile 20 25 30 His Trp Phe Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 93 <211> 318 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding rituximab VL <400> 93 cagatcgtgt tgtcccaatc acccgcaatt ctctctgcga gcccagggga gaaggtgacc 60 atgacttgcc gcgcttcaag ttccgtatcc tacattcact ggttccagca gaagcccgga 120 agttccccta agccctggat ctatgctaca tccaatctgg caagcggtgt tcccgttaga 180 ttttccggaa gcgggtctgg aaccagttac agtctgacta tttccagggt cgaggccgaa 240 gatgcggcta cttattattg ccaacagtgg acctctaacc cacccacatt cggcggcggc 300 actaagttgg aaattaag 318 <210> 94 <211> 450 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide trastuzumab heavy chain <400> 94 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr 20 25 30 Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 <210> 95 <211> 1350 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding the heavy chain of trastuzumab <400> 95 gaagtccaat tggtcgaatc aggcggtgga ctcgtgcaac ctggaggttc gttacgctta 60 tcatgtgctg caagtggatt taatattaaa gatacctaca tccactgggt acgtcaagct 120 cccggcaagg gtctcgagtg ggtcgcacgc atttacccca ccaacggata cacgcgctac 180 gccgattcag tgaagggacg tttcacaatc tctgctgata ctagcaaaaa taccgcatac 240 ctccagatga actctcttag ggccgaggac acagctgtgt actactgtag ccgttgggga 300 ggagacggtt tttacgcaat ggattactgg ggccaaggaa ccctggtcac agtttcatcg 360 gcgtcgacca agggcccgtc agtgttcccg ctggccccgt catccaagtc cacgtctggg 420 ggcacagcag ccctgggatg cttggtcaag gactacttcc ccgagcccgt gactgtgtcc 480 tggaactccg gagcactgac ctccggagtg cacacctttc ccgcggtgct gcagtcctcc 540 ggactgtact ccctgtcgtc ggtcgtgacc gtgccgagct cctcgctcgg aacccagacc 600 tacatctgca acgtgaacca caagccctcg aacaccaaag tggacaagaa ggtcgagccc 660 aaaagctgcg acaagactca cacttgtccg ccgtgccccg cccccgaact gctgggtggc 720 ccctccgtgt tcctgttccc gcctaagcct aaggacaccc ttatgatcag ccgcacccct 780 gaagtgacct gtgtcgtcgt ggatgtgtca cacgaggacc cggaggtcaa gttcaattgg 840 tacgtggacg gcgtggaagt gcataacgca aagaccaagc ctcgggagga acagtacaac 900 tcgacctacc gcgtggtgtc agtcctgact gtgctgcacc aggactggct gaacgggaag 960 gagtacaagt gcaaagtgtc gaacaaggcc ctgccggctc caattgaaaa gaccatcagc 1020 aaggccaagg gccagccaag ggaaccacag gtgtacaccc tccctccttc ccgggacgag 1080 ctgaccaaaa accaagtgtc cctgacttgc cttgtgaagg ggttctaccc ttctgacatt 1140 gccgtcgaat gggaatcgaa cggacagcct gaaaacaact ataagactac cccgcccgtg 1200 ctggattccg acggaagctt cttcctgtac tccaagctga ccgtggacaa gtcgagatgg 1260 cagcagggaa atgtgttcag ctgctccgtg atgcatgagg cgctgcacaa ccactacacc 1320 cagaagtcac tgagcctctcccccggaaag 1350 <210> 96 <211> 213 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide trastuzumab light chain <400> 96 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys 210 <210> 97 <211> 642 <212> DNA <213> Artificial sequence <220> <223> Synthesized polynucleotides encoding the trastuzumab light chain <400> 97 gatatccaaa tgactcaatc accatcttca ctctccgcga gcgtgggtga tcgggtcacc 60 atcacatgta gggcgagcca agatgtgaat accgccgtcg cgtggtatca acaaaagccg 120 ggaaaagcac caaaactgct tatatactct gcatccttcc tgtactctgg ggtgccaagc 180 cggttctccg gtagtagatc tggtactgac tttacactca ctatcagcag tctgcaacct 240 gaggactttg cgacatacta ttgccagcag cactacacaa ccccacctac atttggtcag 300 gggacaaagg tggagatcaa gaccgtggcc gccccgagcg tgttcatttt ccctccctcc 360 gacgagcagt tgaaatcggg caccgctagc gtggtctgcc ttctcaacaa tttctatcca 420 cgggaagcca aagtgcagtg gaaggtcgac aacgcgctcc aatccgggaa ctcacaggaa 480 tccgtgactg agcaggattc caaggactcg acctactccc tgtcatccac gctgaccctg 540 agcaaggcag actacgagaa gcacaaggtc tacgcctgcg aagtgacaca ccagggactg 600 tccagccccg tgaccaagag cttcaacaga ggagaatgct ag 642 <210> 98 <211> 451 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide Rituximab Heavy Chain <400> 98 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Asn Met His Trp Val Lys Gln Thr Pro Gly Arg Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val Trp Gly 100 105 110 Ala Gly Thr Thr Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175 Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190 Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 225 230 235 240 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 305 310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 385 390 395 400 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445 Pro Gly Lys 450 <210> 99 <211> 1353 <212> DNA <213> Artificial sequence <220> <223> Synthesized polynucleotides encoding rituximab heavy chains <400> 99 caagttcagc ttcagcagcc gggggctgag ttggtgaaac ccggggccag tgtgaagatg 60 agctgtaaag cgagcggcta caccttcact tcttataata tgcattgggt taagcaaacg 120 ccaggaaggg ggctggagtg gatcggcgct atttacccag gtaacggtga cacatcatat 180 aaccaaaagt ttaagggaaa ggcaaccctc acagcggaca agagtagctc aaccgcatac 240 atgcaactgt caagccttac ctccgaagac agcgcagtgt actactgcgc cagaagcacc 300 tactatgggg gtgattggta cttcaacgtc tggggggctg gcaccacagt gactgtaagc 360 gcagcgtcga ccaagggccc gtcagtgttc ccgctggccc cgtcatccaa gtccacgtct 420 gggggcacag cagccctggg atgcttggtc aaggactact tccccgagcc cgtgactgtg 480 tcctggaact ccggagcact gacctccgga gtgcacacct ttcccgcggt gctgcagtcc 540 tccggactgt actccctgtc gtcggtcgtg accgtgccga gctcctcgct cggaacccag 600 acctacatct gcaacgtgaa ccacaagccc tcgaacacca aagtggacaa gaaggtcgag 660 cccaaaagct gcgacaagac tcacacttgt ccgccgtgcc ccgcccccga actgctgggt 720 ggcccctccg tgttcctgtt cccgcctaag cctaaggaca cccttatgat cagccgcacc 780 cctgaagtga cctgtgtcgt cgtggatgtg tcacacgagg acccggaggt caagttcaat 840 tggtacgtgg acggcgtgga agtgcataac gcaaagacca agcctcggga ggaacagtac 900 aactcgacct accgcgtggt gtcagtcctg actgtgctgc accaggactg gctgaacggg 960 aaggagtaca agtgcaaagt gtcgaacaag gccctgccgg ctccaattga aaagaccatc 1020 agcaaggcca agggccagcc aagggaacca caggtgtaca ccctccctcc ttcccgggac 1080 gagctgacca aaaaccaagt gtccctgact tgccttgtga aggggttcta cccttctgac 1140 attgccgtcg aatgggaatc gaacggacag cctgaaaaca actataagac taccccgccc 1200 gtgctggatt ccgacggaag cttcttcctg tactccaagc tgaccgtgga caagtcgaga 1260 tggcagcagg gaaatgtgtt cagctgctcc gtgatgcatg aggcgctgca caaccactac 1320 acccagaagt cactgagcct ctcccccgga aag 1353 <210> 100 <211> 212 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide Rituximab Light Chain <400> 100 Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile 20 25 30 His Trp Phe Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Thr Val Ala Ala Pro Ser 100 105 110 Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala 115 120 125 Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val 130 135 140 Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser 145 150 155 160 Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr 165 170 175 Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys 180 185 190 Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn 195 200 205 Arg Gly Glu Cys 210 <210> 101 <211> 639 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding rituximab light chain <400> 101 cagatcgtgt tgtcccaatc acccgcaatt ctctctgcga gcccagggga gaaggtgacc 60 atgacttgcc gcgcttcaag ttccgtatcc tacattcact ggttccagca gaagcccgga 120 agttccccta agccctggat ctatgctaca tccaatctgg caagcggtgt tcccgttaga 180 ttttccggaa gcgggtctgg aaccagttac agtctgacta tttccagggt cgaggccgaa 240 gatgcggcta cttattattg ccaacagtgg acctctaacc cacccacatt cggcggcggc 300 actaagttgg aaattaagac cgtggccgcc ccgagcgtgt tcattttccc tccctccgac 360 gagcagttga aatcgggcac cgctagcgtg gtctgccttc tcaacaattt ctatccacgg 420 gaagccaaag tgcagtggaa ggtcgacaac gcgctccaat ccgggaactc acaggaatcc 480 gtgactgagc aggattccaa ggactcgacc tactccctgt catccacgct gaccctgagc 540 aaggcagact acgagaagca caaggtctac gcctgcgaag tgacacacca gggactgtcc 600 agccccgtga ccaagagctt caacagagga gaatgctag 639 <210> 102 <211> 634 <212> PRT <213> artificial sequence <220> <223> Synthesis of HC_MICwed <400> 102 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr 20 25 30 Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Gly Gly Gly Ser Glu Pro His Ser Leu Arg Tyr Asn Leu Thr Val 450 455 460 Leu Ser Trp Asp Gly Ser Val Gln Ser Gly Phe Leu Thr Glu Val His 465 470 475 480 Leu Asp Gly Gln Pro Phe Leu Arg Cys Asp Arg Gln Lys Cys Arg Ala 485 490 495 Lys Pro Gln Gly Gln Trp Ala Glu Asp Val Leu Gly Asn Lys Thr Trp 500 505 510 Asp Arg Glu Thr Arg Asp Leu Thr Gly Trp Gly Lys Asp Leu Arg Met 515 520 525 Thr Leu Ala His Ile Lys Asp Gln Lys Glu Gly Leu His Ser Leu Gln 530 535 540 Glu Ile Arg Val Cys Glu Ile His Glu Asp Asn Ser Thr Arg Ser Ser 545 550 555 560 Gln His Phe Tyr Tyr Asp Gly Glu Leu Phe Leu Ser Gln Asn Leu Glu 565 570 575 Thr Lys Glu Trp Thr Met Pro Gln Ser Ser Arg Ala Gln Thr Leu Ala 580 585 590 Met Asn Val Arg Asn Phe Leu Lys Glu Asp Ala Met Glu Thr Asp Thr 595 600 605 His Tyr His Ala Met His Ala Asp Cys Leu Gln Glu Leu Arg Arg Tyr 610 615 620 Leu Lys Ser Gly Val Val Leu Arg Arg Thr 625 630 <210> 103 <211> 1902 <212> DNA <213> Artificial sequence <220> <223> Synthesized polynucleotide, encoding trastuzumab HC_MICwed <400> 103 gaagtccaat tggtcgaatc aggcggtgga ctcgtgcaac ctggaggttc gttacgctta 60 tcatgtgctg caagtggatt taatattaaa gatacctaca tccactgggt acgtcaagct 120 cccggcaagg gtctcgagtg ggtcgcacgc atttacccca ccaacggata cacgcgctac 180 gccgattcag tgaagggacg tttcacaatc tctgctgata ctagcaaaaa taccgcatac 240 ctccagatga actctcttag ggccgaggac acagctgtgt actactgtag ccgttgggga 300 ggagacggtt tttacgcaat ggattactgg ggccaaggaa ccctggtcac agtttcatcg 360 gcgtcgacca agggcccgtc agtgttcccg ctggccccgt catccaagtc cacgtctggg 420 ggcacagcag ccctgggatg cttggtcaag gactacttcc ccgagcccgt gactgtgtcc 480 tggaactccg gagcactgac ctccggagtg cacacctttc ccgcggtgct gcagtcctcc 540 ggactgtact ccctgtcgtc ggtcgtgacc gtgccgagct cctcgctcgg aacccagacc 600 tacatctgca acgtgaacca caagccctcg aacaccaaag tggacaagaa ggtcgagccc 660 aaaagctgcg acaagactca cacttgtccg ccgtgccccg cccccgaact gctgggtggc 720 ccctccgtgt tcctgttccc gcctaagcct aaggacaccc ttatgatcag ccgcacccct 780 gaagtgacct gtgtcgtcgt ggcagtgtca cacgaggacc cggaggtcaa gttcaattgg 840 tacgtggacg gcgtggaagt gcataacgca aagaccaagc ctcgggagga acagtacgcc 900 tcgacctacc gcgtggtgtc agtcctgact gtgctgcacc aggactggct gaacgggaag 960 gagtacaagt gcaaagtgtc gaacaaggcc ctgccggctc caattgaaaa gaccatcagc 1020 aaggccaagg gccagccaag ggaaccacag gtgtacaccc tccctccttc ccgggacgag 1080 ctgaccaaaa accaagtgtc cctgacttgc cttgtgaagg ggttctaccc ttctgacatt 1140 gccgtcgaat gggaatcgaa cggacagcct gaaaacaact ataagactac cccgcccgtg 1200 ctggattccg acggaagctt cttcctgtac tccaagctga ccgtggacaa gtcgagatgg 1260 cagcagggaa atgtgttcag ctgctccgtg atgcatgagg cgctgcacaa ccactacacc 1320 cagaagtcac tgagcctctc ccccggagga ggtggcagcg agcctcacag cctccggtat 1380 aatttgactg tactctcttg ggatggctcc gtgcagtccg gctttctgac tgaagttcat 1440 ctcgacggtc aacctttcct gcgctgcgac cgacaaaaat gccgcgccaa gccccaaggg 1500 cagtgggccg aagatgtact gggaaacaag acctgggacc gggagacacg agacctgaca 1560 ggctggggca aggacttgcg catgacactc gcccatatca aggaccagaa ggaaggattg 1620 cactctttgc aagagattcg cgtgtgtgaa atccacgagg acaattcaac gaggagctcc 1680 cagcacttct attacgatgg agaactcttc ttgtcacaga acttggaaac caaggaatgg 1740 actatgcctc agagctctcg ggcacagact ctcgctatga acgttagaaa cttccttaag 1800 gaggatgcta tggagaccga tactcactac cacgccatgc acgccgactg cctccaggaa 1860 ctgcggagat atctgaagtc cggcgtggtt ttgagaagaa cc 1902 <210> 104 <211> 634 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide Trastuzumab HC_MIC25 <400> 104 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr 20 25 30 Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Gly Gly Gly Ser Glu Pro His Ser Leu Arg Tyr Asn Leu Thr Val 450 455 460 Leu Ser Trp Asp Gly Ser Val Gln Ser Gly Phe Leu Thr Glu Val His 465 470 475 480 Leu Asp Gly Gln Pro Phe Leu Arg Cys Asp Arg Gln Lys Cys Arg Ala 485 490 495 Lys Pro Gln Gly Gln Trp Ala Glu Asp Val Leu Gly Asn Lys Thr Trp 500 505 510 Asp Arg Glu Thr Arg Asp Leu Thr Gly Trp Gly Lys Asp Leu Arg Met 515 520 525 Thr Leu Ala His Ile Lys Asp Gln Lys Glu Gly Leu His Ser Leu Gln 530 535 540 Glu Ile Arg Val Cys Glu Ile His Glu Asp Asn Ser Thr Arg Ser Ser 545 550 555 560 Gln His Phe Tyr Tyr Asp Gly Glu Leu Phe Leu Ser Gln Asn Leu Glu 565 570 575 Thr Leu Glu Trp Thr Met Pro Gln Ser Ser Arg Ala Gln Thr Leu Ala 580 585 590 Met Asn Val Arg Asn Phe Leu Lys Glu Asp Ala Met Glu Thr Asp Thr 595 600 605 His Tyr His Ala Met Arg Ala Asp Cys Leu Ser Glu Leu Arg Arg Tyr 610 615 620 Leu Lys Ser Gly Val Val Leu Arg Arg Thr 625 630 <210> 105 <211> 1902 <212> DNA <213> Artificial sequence <220> <223> Synthesized polynucleotide, encoding trastuzumab HC_MIC25 <400> 105 gaagtccaat tggtcgaatc aggcggtgga ctcgtgcaac ctggaggttc gttacgctta 60 tcatgtgctg caagtggatt taatattaaa gatacctaca tccactgggt acgtcaagct 120 cccggcaagg gtctcgagtg ggtcgcacgc atttacccca ccaacggata cacgcgctac 180 gccgattcag tgaagggacg tttcacaatc tctgctgata ctagcaaaaa taccgcatac 240 ctccagatga actctcttag ggccgaggac acagctgtgt actactgtag ccgttgggga 300 ggagacggtt tttacgcaat ggattactgg ggccaaggaa ccctggtcac agtttcatcg 360 gcgtcgacca agggcccgtc agtgttcccg ctggccccgt catccaagtc cacgtctggg 420 ggcacagcag ccctgggatg cttggtcaag gactacttcc ccgagcccgt gactgtgtcc 480 tggaactccg gagcactgac ctccggagtg cacacctttc ccgcggtgct gcagtcctcc 540 ggactgtact ccctgtcgtc ggtcgtgacc gtgccgagct cctcgctcgg aacccagacc 600 tacatctgca acgtgaacca caagccctcg aacaccaaag tggacaagaa ggtcgagccc 660 aaaagctgcg acaagactca cacttgtccg ccgtgccccg cccccgaact gctgggtggc 720 ccctccgtgt tcctgttccc gcctaagcct aaggacaccc ttatgatcag ccgcacccct 780 gaagtgacct gtgtcgtcgt ggcagtgtca cacgaggacc cggaggtcaa gttcaattgg 840 tacgtggacg gcgtggaagt gcataacgca aagaccaagc ctcgggagga acagtacgcc 900 tcgacctacc gcgtggtgtc agtcctgact gtgctgcacc aggactggct gaacgggaag 960 gagtacaagt gcaaagtgtc gaacaaggcc ctgccggctc caattgaaaa gaccatcagc 1020 aaggccaagg gccagccaag ggaaccacag gtgtacaccc tccctccttc ccgggacgag 1080 ctgaccaaaa accaagtgtc cctgacttgc cttgtgaagg ggttctaccc ttctgacatt 1140 gccgtcgaat gggaatcgaa cggacagcct gaaaacaact ataagactac cccgcccgtg 1200 ctggattccg acggaagctt cttcctgtac tccaagctga ccgtggacaa gtcgagatgg 1260 cagcagggaa atgtgttcag ctgctccgtg atgcatgagg cgctgcacaa ccactacacc 1320 cagaagtcac tgagcctctc ccccggagga ggtggcagcg agcctcacag cctccggtat 1380 aatttgactg tactctcttg ggatggctcc gtgcagtccg gctttctgac tgaagttcat 1440 ctcgacggtc aacctttcct gcgctgcgac cgacaaaaat gccgcgccaa gccccaaggg 1500 cagtgggccg aagatgtact gggaaacaag acctgggacc gggagacacg agacctgaca 1560 ggctggggca aggacttgcg catgacactc gcccatatca aggaccagaa ggaaggattg 1620 cactctttgc aagagattcg cgtgtgtgaa atccacgagg acaattcaac gaggagctcc 1680 cagcacttct attacgatgg agaactcttc ttgtcacaga acttggaaac cctcgaatgg 1740 actatgcctc agagctctcg ggcacagact ctcgctatga acgttagaaa cttccttaag 1800 gaggatgcta tggagaccga tactcactac cacgccatgc gcgccgactg cctctctgaa 1860 ctgcggagat atctgaagtc cggcgtggtt ttgagaagaa cc 1902 <210> 106 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide Rituximab LC_ULBP2.wt <400> 106 Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile 20 25 30 His Trp Phe Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys Ala Pro Thr Ser Ser Ser Gly Gly Gly Gly Ser 210 215 220 Glu Pro His Ser Leu Ser Tyr Asp Ile Thr Val Ile Pro Lys Phe Arg 225 230 235 240 Pro Gly Pro Arg Trp Cys Ala Val Gln Gly Gln Val Asp Glu Lys Thr 245 250 255 Phe Leu His Tyr Asp Cys Gly Asn Lys Thr Val Thr Pro Val Ser Pro 260 265 270 Leu Gly Lys Lys Leu Asn Val Thr Thr Ala Trp Lys Ala Gln Asn Pro 275 280 285 Val Leu Arg Glu Val Val Asp Ile Leu Thr Glu Gln Leu Arg Asp Ile 290 295 300 Gln Leu Glu Asn Tyr Thr Pro Lys Glu Pro Leu Thr Leu Gln Ala Arg 305 310 315 320 Met Ser Cys Glu Gln Lys Ala Glu Gly His Ser Ser Gly Ser Trp Gln 325 330 335 Phe Ser Phe Asp Gly Gln Ile Phe Leu Leu Phe Asp Ser Glu Lys Arg 340 345 350 Met Trp Thr Thr Val His Pro Gly Ala Arg Lys Met Lys Glu Lys Trp 355 360 365 Glu Asn Asp Lys Val Val Ala Met Ser Phe His Tyr Phe Ser Met Gly 370 375 380 Asp Cys Ile Gly Trp Leu Glu Asp Phe Leu Met Gly Met Asp Ser Thr 385 390 395 400 Leu Glu Pro Ser <210> 107 <211> 1212 <212> DNA <213> Artificial Sequence <220> <223> Synthetic polynucleotide, encoding rituximab LC_ULBP2.wt <400> 107 cagatcgtgt tgtcccaatc acccgcaatt ctctctgcga gcccagggga gaaggtgacc 60 atgacttgcc gcgcttcaag ttccgtatcc tacattcact ggttccagca gaagcccgga 120 agttccccta agccctggat ctatgctaca tccaatctgg caagcggtgt tcccgttaga 180 ttttccggaa gcgggtctgg aaccagttac agtctgacta tttccagggt cgaggccgaa 240 gatgcggcta cttattattg ccaacagtgg acctctaacc cacccacatt cggcggcggc 300 actaagttgg aaattaagcg gaccgtggcc gccccgagcg tgttcatttt ccctccctcc 360 gacgagcagt tgaaatcggg caccgctagc gtggtctgcc ttctcaacaa tttctatcca 420 cgggaagcca aagtgcagtg gaaggtcgac aacgcgctcc aatccgggaa ctcacaggaa 480 tccgtgactg agcaggattc caaggactcg acctactccc tgtcatccac gctgaccctg 540 agcaaggcag actacgagaa gcacaaggtc tacgcctgcg aagtgacaca ccagggactg 600 tccagccccg tgaccaagag cttcaacaaga ggagaatgcg cacctacctc aagctctgga 660 ggaggtggca gcgagcccca tagtctgagc tacgacatca cagttattcc caagttcagg 720 cccggaccgc gctggtgtgc cgtgcaagga caagtcgacg aaaaaacctt tcttcattac 780 gattgcggaa ataagactgt aacgccagtc tctcctttag gtaagaagtt aaacgtcact 840 acggcgtgga aggcacaaaa ccccgtcctg cgcgaggtcg tcgacatcct gactgaacaa 900 ttgcgcgaca tccagctcga gaattacact ccaaaggagc ctcttaccct gcaggctaga 960 atgtcttgcg agcaaaaggc agagggccac tcctccggca gctggcagtt cagtttcgac 1020 ggacaaatct ttctgttatt cgattcagag aagagaatgt ggactacagt tcaccccggt 1080 gcccgtaaaa tgaaggagaa gtgggaaaac gacaaagtgg tggcgatgtc attccactat 1140 ttctcgatgg gagactgcat cggttggctg gaagatttcc tcatgggtat ggactccact 1200 ttggagccat cg 1212 <210> 108 <211> 187 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide ULPB2 α1-α2 Variant R80W <400> 108 Glu Pro His Ser Leu Ser Tyr Asp Ile Thr Val Ile Pro Lys Phe Arg 1 5 10 15 Pro Gly Pro Arg Trp Cys Ala Val Gln Gly Gln Val Asp Glu Lys Thr 20 25 30 Phe Leu His Tyr Asp Cys Gly Asn Lys Thr Val Thr Pro Val Ser Pro 35 40 45 Leu Gly Lys Lys Leu Asn Val Thr Thr Ala Trp Lys Ala Gln Asn Pro 50 55 60 Val Leu Arg Glu Val Val Asp Ile Leu Thr Glu Gln Leu Trp Asp Ile 65 70 75 80 Gln Leu Glu Asn Tyr Thr Pro Lys Glu Pro Leu Thr Leu Gln Ala Arg 85 90 95 Met Ser Cys Glu Gln Lys Ala Glu Gly His Ser Ser Gly Ser Trp Gln 100 105 110 Phe Ser Phe Asp Gly Gln Ile Phe Leu Leu Phe Asp Ser Glu Lys Arg 115 120 125 Met Trp Thr Thr Val His Pro Gly Ala Arg Lys Met Lys Glu Lys Trp 130 135 140 Glu Asn Asp Lys Val Val Ala Met Ser Phe His Tyr Phe Ser Met Gly 145 150 155 160 Asp Cys Ile Gly Trp Leu Glu Asp Phe Leu Met Gly Met Asp Ser Thr 165 170 175 Leu Glu Pro Ser Ala Gly Ala Pro Pro Met Val 180 185 <210> 109 <211> 540 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding ULPB2 α1-α2 variant R80W <400> 109 gagccccata gtctgagcta cgacatcaca gttattccca agttcaggcc cggaccgcgc 60 tggtgtgccg tgcaaggaca agtcgacgaa aaaacctttc ttcattacga ttgcggaaat 120 aagactgtaa cgccagtctc tcctttaggt aagaagttaa acgtcactac ggcgtggaag 180 gcacaaaacc ccgtcctgcg cgaggtcgtc gacatcctga ctgaacaatt gtgggacatc 240 cagctcgaga attacactcc aaaggagcct cttaccctgc aggctagaat gtcttgcgag 300 caaaaggcag agggccactc ctccggcagc tggcagttca gtttcgacgg acaaatcttt 360 ctgttatattcg attcagagaa gagaatgtgg actacagttc accccggtgc ccgtaaaatg 420 aaggagaagt gggaaaacga caaagtggtg gcgatgtcat tccactattt ctcgatggga 480 gactgcatcg gttggctgga agatttcctc atgggtatgg actccacttt ggagccatcg 540 <210> 110 <211> 513 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide encoding ULPB2 α1-α2 variant R80W NNK library <220> <221> misc_feature <222> (457) (458) <223> n is a, c, g, or t <220> <221> misc_feature <222> (460) (461) <223> n is a, c, g, or t <220> <221> misc_feature <222> (463) (464) <223> n is a, c, g, or t <220> <221> misc_feature <222> (466)..(467) <223> n is a, c, g, or t <220> <221> misc_feature <222> (469)..(470) <223> n is a, c, g, or t <400> 110 gagccccata gcctcagcta tgacattacg gtgattccca aatttcgccc aggaccacgt 60 tggtgcgccg tccagggtca ggtagatgaa aagactttcc tgcattatga ttgcggcaat 120 aaaaccgtga cgccggtatc gccgttaggc aaaaaattga atgtcacgac agcgtggaaa 180 gcacagaacc cggtgttgcg cgaggtagtc gatattttga cggaacaact ctgggacatt 240 cagctcgaga attacacccc aaaagaaccg ctgacgctgc aagcgcgtat gtcgtgcgaa 300 caaaaagcag aaggtcactc tagcgggagt tggcagtttt ccttcgatgg gcagattttt 360 ctgctgtttg attcggagaa acgcatgtgg actacagtcc acccgggtgc ccggaaaatg 420 aaagagaagt gggagaatga taaagtggtg gccactnnkn nknnknnknn ktccatgggc 480 gattgcattg gctggttaga ggattttctc atg 513 <210> 111 <211> 180 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide, ULBP2 α1-α2 variant ULBP2.C <400> 111 Glu Pro His Ser Leu Ser Tyr Asp Ile Thr Val Ile Pro Lys Phe Arg 1 5 10 15 Pro Gly Pro Arg Trp Cys Ala Val Gln Gly Gln Val Asp Glu Lys Thr 20 25 30 Phe Leu His Tyr Asp Cys Gly Asn Lys Thr Val Thr Pro Val Ser Pro 35 40 45 Leu Gly Lys Lys Leu Asn Val Thr Thr Ala Trp Lys Ala Gln Asn Pro 50 55 60 Val Leu Arg Glu Val Val Asp Ile Leu Thr Glu Gln Leu Trp Asp Ile 65 70 75 80 Gln Leu Glu Asn Tyr Thr Pro Lys Glu Pro Leu Thr Leu Gln Ala Arg 85 90 95 Met Ser Cys Glu Gln Lys Ala Glu Gly His Ser Ser Gly Ser Trp Gln 100 105 110 Phe Ser Phe Asp Gly Gln Ile Phe Leu Leu Phe Asp Ser Glu Lys Arg 115 120 125 Met Trp Thr Thr Val His Pro Gly Ala Arg Lys Met Lys Glu Lys Trp 130 135 140 Glu Asn Asp Lys Val Val Ala Thr Ile Leu Trp Gln Thr Ser Met Gly 145 150 155 160 Asp Cys Ile Gly Trp Leu Glu Asp Phe Leu Met Gly Met Asp Ser Thr 165 170 175 Leu Glu Pro Ser 180 <210> 112 <211> 540 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding ULBP2 α1-α2 variant ULBP2.C <400> 112 gagccccata gtctgagcta cgacatcaca gttattccca agttcaggcc cggaccgcgc 60 tggtgtgccg tgcaaggaca agtcgacgaa aaaacctttc ttcattacga ttgcggaaat 120 aagactgtaa cgccagtctc tcctttaggt aagaagttaa acgtcactac ggcgtggaag 180 gcacaaaacc ccgtcctgcg cgaggtcgtc gacatcctga ctgaacaatt gtgggacatc 240 cagctcgaga attacactcc aaaggagcct cttaccctgc aggctagaat gtcttgcgag 300 caaaaggcag agggccactc ctccggcagc tggcagttca gtttcgacgg acaaatcttt 360 ctgttattcg attcagagaa gagaatgtgg actacagttc accccggtgc ccgtaaaatg 420 aaggagaagt gggaaaacga caaagtggtg gcgactattc tgtggcagac ttcgatggga 480 gactgcatcg gttggctgga agatttcctc atgggtatgg actccacttt ggagccatcg 540 <210> 113 <211> 180 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide, ULBP2 α1-α2 variant ULBP2.R <400> 113 Glu Pro His Ser Leu Ser Tyr Asp Ile Thr Val Ile Pro Lys Phe Arg 1 5 10 15 Pro Gly Pro Arg Trp Cys Ala Val Gln Gly Gln Val Asp Glu Lys Thr 20 25 30 Phe Leu His Tyr Asp Cys Gly Asn Lys Thr Val Thr Pro Val Ser Pro 35 40 45 Leu Gly Lys Lys Leu Asn Val Thr Thr Ala Trp Lys Ala Gln Asn Pro 50 55 60 Val Leu Arg Glu Val Val Asp Ile Leu Thr Glu Gln Leu Trp Asp Ile 65 70 75 80 Gln Leu Glu Asn Tyr Thr Pro Lys Glu Pro Leu Thr Leu Gln Ala Arg 85 90 95 Met Ser Cys Glu Gln Lys Ala Glu Gly His Ser Ser Gly Ser Trp Gln 100 105 110 Phe Ser Phe Asp Gly Gln Ile Phe Leu Leu Phe Asp Ser Glu Lys Arg 115 120 125 Met Trp Thr Thr Val His Pro Gly Ala Arg Lys Met Lys Glu Lys Trp 130 135 140 Glu Asn Asp Lys Val Val Ala Thr Leu Leu Trp Gly Trp Ser Met Gly 145 150 155 160 Asp Cys Ile Gly Trp Leu Glu Asp Phe Leu Met Gly Met Asp Ser Thr 165 170 175 Leu Glu Pro Ser 180 <210> 114 <211> 540 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding ULBP2 α1-α2 variant ULBP2.R <400> 114 gagccccata gtctgagcta cgacatcaca gttattccca agttcaggcc cggaccgcgc 60 tggtgtgccg tgcaaggaca agtcgacgaa aaaacctttc ttcattacga ttgcggaaat 120 aagactgtaa cgccagtctc tcctttaggt aagaagttaa acgtcactac ggcgtggaag 180 gcacaaaacc ccgtcctgcg cgaggtcgtc gacatcctga ctgaacaatt gtgggacatc 240 cagctcgaga attacactcc aaaggagcct cttaccctgc aggctagaat gtcttgcgag 300 caaaaggcag agggccactc ctccggcagc tggcagttca gtttcgacgg acaaatcttt 360 ctgttattcg attcagagaa gagaatgtgg actacagttc accccggtgc ccgtaaaatg 420 aaggagaagt gggaaaacga caaagtggtg gcgactttgt tgtgggggtg gtcgatggga 480 gactgcatcg gttggctgga agatttcctc atgggtatgg actccacttt ggagccatcg 540 <210> 115 <211> 180 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide, ULBP2 α1-α2 variant ULBP2.AA <400> 115 Glu Pro His Ser Leu Ser Tyr Asp Ile Thr Val Ile Pro Lys Phe Arg 1 5 10 15 Pro Gly Pro Arg Trp Cys Ala Val Gln Gly Gln Val Asp Glu Lys Thr 20 25 30 Phe Leu His Tyr Asp Cys Gly Asn Lys Thr Val Thr Pro Val Ser Pro 35 40 45 Leu Gly Lys Lys Leu Asn Val Thr Thr Ala Trp Lys Ala Gln Asn Pro 50 55 60 Val Leu Arg Glu Val Val Asp Ile Leu Thr Glu Gln Leu Trp Asp Ile 65 70 75 80 Gln Leu Glu Asn Tyr Thr Pro Lys Glu Pro Leu Thr Leu Gln Ala Arg 85 90 95 Met Ser Cys Glu Gln Lys Ala Glu Gly His Ser Ser Gly Ser Trp Gln 100 105 110 Phe Ser Phe Asp Gly Gln Ile Phe Leu Leu Phe Asp Ser Glu Lys Arg 115 120 125 Met Trp Thr Thr Val His Pro Gly Ala Arg Lys Met Lys Glu Lys Trp 130 135 140 Glu Asn Asp Lys Val Val Ala Thr Met Phe Trp Ser Trp Ser Met Gly 145 150 155 160 Asp Cys Ile Gly Trp Leu Glu Asp Phe Leu Met Gly Met Asp Ser Thr 165 170 175 Leu Glu Pro Ser 180 <210> 116 <211> 540 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding ULBP2 α1-α2 variant ULBP2.AA <400> 116 gagccccata gtctgagcta cgacatcaca gttattccca agttcaggcc cggaccgcgc 60 tggtgtgccg tgcaaggaca agtcgacgaa aaaacctttc ttcattacga ttgcggaaat 120 tggtgtgccg tgcaaggaca agtcgacgaa aaaacctttc ttcattacga ttgcggaaat 120 aagactgtaa cgccagtctc tcctttaggt aagaagttaa acgtcactac ggcgtggaag 180 aagactgtaa cgccagtctc tcctttaggt aagaagttaa acgtcactac ggcgtggaag 180 gcacaaaacc ccgtcctgcg cgaggtcgtc gacatcctga ctgaacaatt gtgggacatc 240 gcacaaaacc ccgtcctgcg cgaggtcgtc gacatcctga ctgaacaatt gtgggacatc 240 cagctcgaga attacactcc aaaggagcct cttaccctgc aggctagaat gtcttgcgag 300 cagctcgaga attacactcc aaaggagcct cttaccctgc aggctagaat gtcttgcgag 300 caaaaggcag agggccactc ctccggcagc tggcagttca gtttcgacgg acaaatcttt 360 caaaaggcag agggccactc ctccggcagc tggcagttca gtttcgacgg acaaatcttt 360 ctgttattcg attcagagaa gagaatgtgg actacagttc accccggtgc ccgtaaaatg 420 ctgttattcg attcagagaa gagaatgtgg actacagttc accccggtgc ccgtaaaatg 420 aaggagaagt gggaaaacga caaagtggtg gcgactatgt tttggagttg gtcgatggga 480 aaggagaagt gggaaaacga caaagtggtg gcgactatgt tttggagttg gtcgatggga 480 gactgcatcg gttggctgga agatttcctc atgggtatgg actccacttt ggagccatcg 540 gactgcatcg gttggctgga agatttcctc atgggtatgg actccacttt ggagccatcg 540 <210> 117<210> 117 <211> 180<211> 180 <212> PRT<212> PRT <213> Artificial Sequence<213> Artificial Sequence <220><220> <223> Synthetic peptide, ULBP2 α1-α2 variant ULBP2.AB<223> Synthetic peptide, ULBP2 α1-α2 variant ULBP2.AB <400> 117 <400> 117 Glu Pro His Ser Leu Ser Tyr Asp Ile Thr Val Ile Pro Lys Phe Arg Glu Pro His Ser Leu Ser Tyr Asp Ile Thr Val Ile Pro Lys Phe Arg 1 5 10 15 1 5 10 15 Pro Gly Pro Arg Trp Cys Ala Val Gln Gly Gln Val Asp Glu Lys Thr 20 25 30 Phe Leu His Tyr Asp Cys Gly Asn Lys Thr Val Thr Pro Val Ser Pro 35 40 45 Leu Gly Lys Lys Leu Asn Val Thr Thr Ala Trp Lys Ala Gln Asn Pro 50 55 60 Val Leu Arg Glu Val Val Asp Ile Leu Thr Glu Gln Leu Trp Asp Ile 65 70 75 80 Gln Leu Glu Asn Tyr Thr Pro Lys Glu Pro Leu Thr Leu Gln Ala Arg 85 90 95 Met Ser Cys Glu Gln Lys Ala Glu Gly His Ser Ser Gly Ser Trp Gln 100 105 110 Phe Ser Phe Asp Gly Gln Ile Phe Leu Leu Phe Asp Ser Glu Lys Arg 115 120 125 Met Trp Thr Thr Val His Pro Gly Ala Arg Lys Met Lys Glu Lys Trp 130 135 140 Glu Asn Asp Lys Val Val Ala Thr Leu Met Trp Gln Trp Ser Met Gly 145 150 155 160 Asp Cys Ile Gly Trp Leu Glu Asp Phe Leu Met Gly Met Asp Ser Thr 165 170 175 Leu Glu Pro Ser 180 <210> 118 <211> 540 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding ULBP2 α1-α2 variant ULBP2.AB <400> 118 gagccccata gtctgagcta cgacatcaca gttattccca agttcaggcc cggaccgcgc 60 tggtgtgccg tgcaaggaca agtcgacgaa aaaacctttc ttcattacga ttgcggaaat 120 aagactgtaa cgccagtctc tcctttaggt aagaagttaa acgtcactac ggcgtggaag 180 gcacaaaacc ccgtcctgcg cgaggtcgtc gacatcctga ctgaacaatt gtgggacatc 240 cagctcgaga attacactcc aaaggagcct cttaccctgc aggctagaat gtcttgcgag 300 caaaaggcag agggccactc ctccggcagc tggcagttca gtttcgacgg acaaatcttt 360 ctgttattcg attcagagaa gagaatgtgg actacagttc accccggtgc ccgtaaaatg 420 aaggagaagt gggaaaacga caaagtggtg gcgactctta tgtggcagtg gtcgatggga 480 gactgcatcg gttggctgga agatttcctc atgggtatgg actccacttt ggagccatcg 540 <210> 119 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide Rituximab LC_ULBP2.C <400> 119 Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile 20 25 30 His Trp Phe Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys Ala Pro Thr Ser Ser Ser Gly Gly Gly Gly Ser 210 215 220 Glu Pro His Ser Leu Ser Tyr Asp Ile Thr Val Ile Pro Lys Phe Arg 225 230 235 240 Pro Gly Pro Arg Trp Cys Ala Val Gln Gly Gln Val Asp Glu Lys Thr 245 250 255 Phe Leu His Tyr Asp Cys Gly Asn Lys Thr Val Thr Pro Val Ser Pro 260 265 270 Leu Gly Lys Lys Leu Asn Val Thr Thr Ala Trp Lys Ala Gln Asn Pro 275 280 285 Val Leu Arg Glu Val Val Asp Ile Leu Thr Glu Gln Leu Trp Asp Ile 290 295 300 Gln Leu Glu Asn Tyr Thr Pro Lys Glu Pro Leu Thr Leu Gln Ala Arg 305 310 315 320 Met Ser Cys Glu Gln Lys Ala Glu Gly His Ser Ser Gly Ser Trp Gln 325 330 335 Phe Ser Phe Asp Gly Gln Ile Phe Leu Leu Phe Asp Ser Glu Lys Arg 340 345 350 Met Trp Thr Thr Val His Pro Gly Ala Arg Lys Met Lys Glu Lys Trp 355 360 365 Glu Asn Asp Lys Val Val Ala Thr Ile Leu Trp Gln Thr Ser Met Gly 370 375 380 Asp Cys Ile Gly Trp Leu Glu Asp Phe Leu Met Gly Met Asp Ser Thr 385 390 395 400 Leu Glu Pro Ser <210> 120 <211> 1212 <212> DNA <213> Artificial sequence <220> <223> A synthetic polynucleotide encoding rituximab LC_ULBP2.C <400> 120 cagatcgtgt tgtcccaatc acccgcaatt ctctctgcga gcccagggga gaaggtgacc 60 atgacttgcc gcgcttcaag ttccgtatcc tacattcact ggttccagca gaagcccgga 120 agttccccta agccctggat ctatgctaca tccaatctgg caagcggtgt tcccgttaga 180 ttttccggaa gcgggtctgg aaccagttac agtctgacta tttccagggt cgaggccgaa 240 gatgcggcta cttattattg ccaacagtgg accctctaacc cacccacatt cggcggcggc 300 actaagttgg aaattaagcg gaccgtggcc gccccgagcg tgttcatttt ccctccctcc 360 gacgagcagt tgaaatcggg caccgctagc gtggtctgcc ttctcaacaa tttctatcca 420 cgggaagcca aagtgcagtg gaaggtcgac aacgcgctcc aatccgggaa ctcacaggaa 480 tccgtgactg agcaggattc caaggactcg acctactccc tgtcatccac gctgaccctg 540 agcaaggcag actacgagaa gcacaaggtc tacgcctgcg aagtgacaca ccagggactg 600 tccagccccg tgaccaagag cttcaacaaga ggagaatgcg cacctacctc aagctctgga 660 ggaggtggca gcgagcccca tagtctgagc tacgacatca cagttattcc caagttcagg 720 cccggaccgc gctggtgtgc cgtgcaagga caagtcgacg aaaaaacctt tcttcattac 780 gattgcggaa ataagactgt aacgccagtc tctcctttag gtaagaagtt aaacgtcact 840 acggcgtgga aggcacaaaa ccccgtcctg cgcgaggtcg tcgacatcct gactgaacaa 900 ttgtgggaca tccagctcga gaattacact ccaaaggagc ctcttaccct gcaggctaga 960 atgtcttgcg agcaaaaggc agagggccac tcctccggca gctggcagtt cagtttcgac 1020 ggacaaatct ttctgttatt cgattcagag aagagaatgt ggactacagt tcaccccggt 1080 gcccgtaaaa tgaaggagaa gtgggaaaac gacaaagtgg tggcgactat tctgtggcag 1140 acttcgatgg gagactgcat cggttggctg gaagatttcc tcatgggtat ggactccact 1200 ttggagccat cg 1212 <210> 121 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide Rituximab LC_ULBP2.R <400> 121 Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile 20 25 30 His Trp Phe Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys Ala Pro Thr Ser Ser Ser Gly Gly Gly Gly Ser 210 215 220 Glu Pro His Ser Leu Ser Tyr Asp Ile Thr Val Ile Pro Lys Phe Arg 225 230 235 240 Pro Gly Pro Arg Trp Cys Ala Val Gln Gly Gln Val Asp Glu Lys Thr 245 250 255 Phe Leu His Tyr Asp Cys Gly Asn Lys Thr Val Thr Pro Val Ser Pro 260 265 270 Leu Gly Lys Lys Leu Asn Val Thr Thr Ala Trp Lys Ala Gln Asn Pro 275 280 285 Val Leu Arg Glu Val Val Asp Ile Leu Thr Glu Gln Leu Trp Asp Ile 290 295 300 Gln Leu Glu Asn Tyr Thr Pro Lys Glu Pro Leu Thr Leu Gln Ala Arg 305 310 315 320 Met Ser Cys Glu Gln Lys Ala Glu Gly His Ser Ser Gly Ser Trp Gln 325 330 335 Phe Ser Phe Asp Gly Gln Ile Phe Leu Leu Phe Asp Ser Glu Lys Arg 340 345 350 Met Trp Thr Thr Val His Pro Gly Ala Arg Lys Met Lys Glu Lys Trp 355 360 365 Glu Asn Asp Lys Val Val Ala Thr Leu Leu Trp Gly Trp Ser Met Gly 370 375 380 Asp Cys Ile Gly Trp Leu Glu Asp Phe Leu Met Gly Met Asp Ser Thr 385 390 395 400 Leu Glu Pro Ser <210> 122 <211> 1212 <212> DNA <213> Artificial Sequence <220> <223> Synthetic polynucleotide, encoding rituximab LC_ULBP2.R <400> 122 cagatcgtgt tgtcccaatc acccgcaatt ctctctgcga gcccagggga gaaggtgacc 60 atgacttgcc gcgcttcaag ttccgtatcc tacattcact ggttccagca gaagcccgga 120 agttccccta agccctggat ctatgctaca tccaatctgg caagcggtgt tcccgttaga 180 ttttccggaa gcgggtctgg aaccagttac agtctgacta tttccagggt cgaggccgaa 240 gatgcggcta cttattattg ccaacagtgg acctctaacc cacccacatt cggcggcggc 300 actaagttgg aaattaagcg gaccgtggcc gccccgagcg tgttcatttt ccctccctcc 360 gacgagcagt tgaaatcggg caccgctagc gtggtctgcc ttctcaacaa tttctatcca 420 cgggaagcca aagtgcagtg gaaggtcgac aacgcgctcc aatccgggaa ctcacaggaa 480 tccgtgactg agcaggattc caaggactcg acctactccc tgtcatccac gctgaccctg 540 agcaaggcag actacgagaa gcacaaggtc tacgcctgcg aagtgacaca ccagggactg 600 tccagccccg tgaccaagag cttcaacaaga ggagaatgcg cacctacctc aagctctgga 660 ggaggtggca gcgagcccca tagtctgagc tacgacatca cagttattcc caagttcagg 720 cccggaccgc gctggtgtgc cgtgcaagga caagtcgacg aaaaaacctt tcttcattac 780 gattgcggaa ataagactgt aacgccagtc tctcctttag gtaagaagtt aaacgtcact 840 acggcgtgga aggcacaaaa ccccgtcctg cgcgaggtcg tcgacatcct gactgaacaa 900 ttgtgggaca tccagctcga gaattacact ccaaaggagc ctcttaccct gcaggctaga 960 atgtcttgcg agcaaaaggc agagggccac tcctccggca gctggcagtt cagtttcgac 1020 ggacaaatct ttctgttatt cgattcagag aagagaatgt ggactacagt tcaccccggt 1080 gcccgtaaaa tgaaggaga gtgggaaaac gacaaagtgg tggcgacttt gttgtggggg 1140 tggtcgatgg gagactgcat cggttggctg gaagatttcc tcatgggtat ggactccact 1200 ttggagccat cg 1212 <210> 123 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide Rituximab LC_ULBP2.AA <400> 123 Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile 20 25 30 His Trp Phe Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys Ala Pro Thr Ser Ser Ser Gly Gly Gly Gly Ser 210 215 220 Glu Pro His Ser Leu Ser Tyr Asp Ile Thr Val Ile Pro Lys Phe Arg 225 230 235 240 Pro Gly Pro Arg Trp Cys Ala Val Gln Gly Gln Val Asp Glu Lys Thr 245 250 255 Phe Leu His Tyr Asp Cys Gly Asn Lys Thr Val Thr Pro Val Ser Pro 260 265 270 Leu Gly Lys Lys Leu Asn Val Thr Thr Ala Trp Lys Ala Gln Asn Pro 275 280 285 Val Leu Arg Glu Val Val Asp Ile Leu Thr Glu Gln Leu Trp Asp Ile 290 295 300 Gln Leu Glu Asn Tyr Thr Pro Lys Glu Pro Leu Thr Leu Gln Ala Arg 305 310 315 320 Met Ser Cys Glu Gln Lys Ala Glu Gly His Ser Ser Gly Ser Trp Gln 325 330 335 Phe Ser Phe Asp Gly Gln Ile Phe Leu Leu Phe Asp Ser Glu Lys Arg 340 345 350 Met Trp Thr Thr Val His Pro Gly Ala Arg Lys Met Lys Glu Lys Trp 355 360 365 Glu Asn Asp Lys Val Val Ala Thr Met Phe Trp Ser Trp Ser Met Gly 370 375 380 Asp Cys Ile Gly Trp Leu Glu Asp Phe Leu Met Gly Met Asp Ser Thr 385 390 395 400 Leu Glu Pro Ser <210> 124 <211> 1212 <212> DNA <213> Artificial sequence <220> <223> Synthesized polynucleotide, encoding rituximab LC_ULBP2.AA <400> 124 cagatcgtgt tgtcccaatc acccgcaatt ctctctgcga gcccaggga gaaggtgacc 60 atgacttgcc gcgcttcaag ttccgtatcc tacattcact ggttccagca gaagcccgga 120 agttccccta agccctggat ctatgctaca tccaatctgg caagcggtgt tcccgttaga 180 ttttccggaa gcgggtctgg aaccagttac agtctgacta tttccagggt cgaggccgaa 240 gatgcggcta cttattattg ccaacagtgg accctctaacc cacccacatt cggcggcggc 300 actaagttgg aaattaagcg gaccgtggcc gccccgagcg tgttcatttt ccctccctcc 360 gacgagcagt tgaaatcggg caccgctagc gtggtctgcc ttctcaacaa tttctatcca 420 cgggaagcca aagtgcagtg gaaggtcgac aacgcgctcc aatccgggaa ctcacaggaa 480 tccgtgactg agcaggattc caaggactcg acctactccc tgtcatccac gctgaccctg 540 agcaaggcag actacgagaa gcacaaggtc tacgcctgcg aagtgacaca ccagggactg 600 tccagccccg tgaccaagag cttcaacaaga ggagaatgcg cacctacctc aagctctgga 660 ggaggtggca gcgagcccca tagtctgagc tacgacatca cagttattcc caagttcagg 720 cccggaccgc gctggtgtgc cgtgcaagga caagtcgacg aaaaaacctt tcttcattac 780 gattgcggaa ataagactgt aacgccagtc tctcctttag gtaagaagtt aaacgtcact 840 acggcgtgga aggcacaaaa ccccgtcctg cgcgaggtcg tcgacatcct gactgaacaa 900 ttgtgggaca tccagctcga gaattacact ccaaaggagc ctcttaccct gcaggctaga 960 atgtcttgcg agcaaaaggc agagggccac tcctccggca gctggcagtt cagtttcgac 1020 ggacaaatct ttctgttatt cgattcagag aagagaatgt ggactacagt tcaccccggt 1080 gcccgtaaaa tgaaggagaa gtgggaaaac gacaaagtgg tggcgactat gttttggagt 1140 tggtcgatgg gagactgcat cggttggctg gaagatttcc tcatgggtat ggactccact 1200 ttggagccat cg 1212 <210> 125 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide Rituximab LC_ULBP2.AB <400> 125 Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile 20 25 30 His Trp Phe Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys Ala Pro Thr Ser Ser Ser Gly Gly Gly Gly Ser 210 215 220 Glu Pro His Ser Leu Ser Tyr Asp Ile Thr Val Ile Pro Lys Phe Arg 225 230 235 240 Pro Gly Pro Arg Trp Cys Ala Val Gln Gly Gln Val Asp Glu Lys Thr 245 250 255 Phe Leu His Tyr Asp Cys Gly Asn Lys Thr Val Thr Pro Val Ser Pro 260 265 270 Leu Gly Lys Lys Leu Asn Val Thr Thr Ala Trp Lys Ala Gln Asn Pro 275 280 285 Val Leu Arg Glu Val Val Asp Ile Leu Thr Glu Gln Leu Trp Asp Ile 290 295 300 Gln Leu Glu Asn Tyr Thr Pro Lys Glu Pro Leu Thr Leu Gln Ala Arg 305 310 315 320 Met Ser Cys Glu Gln Lys Ala Glu Gly His Ser Ser Gly Ser Trp Gln 325 330 335 Phe Ser Phe Asp Gly Gln Ile Phe Leu Leu Phe Asp Ser Glu Lys Arg 340 345 350 Met Trp Thr Thr Val His Pro Gly Ala Arg Lys Met Lys Glu Lys Trp 355 360 365 Glu Asn Asp Lys Val Val Ala Thr Leu Met Trp Gln Trp Ser Met Gly 370 375 380 Asp Cys Ile Gly Trp Leu Glu Asp Phe Leu Met Gly Met Asp Ser Thr 385 390 395 400 Leu Glu Pro Ser <210> 126 <211> 1212 <212> DNA <213> Artificial sequence <220> <223> Synthesized polynucleotide, encoding rituximab LC_ULBP2.AB <400> 126 cagatcgtgt tgtcccaatc acccgcaatt ctctctgcga gcccagggga gaaggtgacc 60 atgacttgcc gcgcttcaag ttccgtatcc tacattcact ggttccagca gaagcccgga 120 agttccccta agccctggat ctatgctaca tccaatctgg caagcggtgt tcccgttaga 180 ttttccggaa gcgggtctgg aaccagttac agtctgacta tttccagggt cgaggccgaa 240 gatgcggcta cttattattg ccaacagtgg accctctaacc cacccacatt cggcggcggc 300 actaagttgg aaattaagcg gaccgtggcc gccccgagcg tgttcatttt ccctccctcc 360 gacgagcagt tgaaatcggg caccgctagc gtggtctgcc ttctcaacaa tttctatcca 420 cgggaagcca aagtgcagtg gaaggtcgac aacgcgctcc aatccgggaa ctcacaggaa 480 tccgtgactg agcaggattc caaggactcg acctactccc tgtcatccac gctgaccctg 540 agcaaggcag actacgagaa gcacaaggtc tacgcctgcg aagtgacaca ccagggactg 600 tccagccccg tgaccaagag cttcaacaaga ggagaatgcg cacctacctc aagctctgga 660 ggaggtggca gcgagcccca tagtctgagc tacgacatca cagttattcc caagttcagg 720 cccggaccgc gctggtgtgc cgtgcaagga caagtcgacg aaaaaacctt tcttcattac 780 gattgcggaa ataagactgt aacgccagtc tctcctttag gtaagaagtt aaacgtcact 840 acggcgtgga aggcacaaaa ccccgtcctg cgcgaggtcg tcgacatcct gactgaacaa 900 ttgtgggaca tccagctcga gaattacact ccaaaggagc ctcttaccct gcaggctaga 960 atgtcttgcg agcaaaaggc agagggccac tcctccggca gctggcagtt cagtttcgac 1020 ggacaaatct ttctgttatt cgattcagag aagagaatgt ggactacagt tcaccccggt 1080 gcccgtaaaa tgaaggagaa gtgggaaaac gacaaagtgg tggcgactct tatgtggcag 1140 tggtcgatgg gagactgcat cggttggctg gaagatttcc tcatgggtat ggactccact 1200 ttggagccat cg 1212 <210> 127 <211> 180 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide, ULBP2 α1-α2 Variant ULBP2.S3 <400> 127 Glu Pro His Ser Leu Ser Tyr Asp Ile Thr Val Ile Pro Lys Phe Arg 1 5 10 15 Pro Gly Pro Arg Trp Cys Ala Val Gln Gly Gln Val Asp Glu Lys Thr 20 25 30 Phe Leu His Tyr Asp Cys Gly Asn Lys Thr Val Thr Pro Val Ser Pro 35 40 45 Leu Gly Lys Lys Leu Asn Val Thr Thr Ala Trp Lys Ala Gln Asn Pro 50 55 60 Val Leu Arg Glu Val Val Asp Ile Leu Thr Glu Gln Leu Trp Asp Ile 65 70 75 80 Gln Leu Glu Asn Tyr Thr Pro Lys Glu Pro Leu Thr Leu Gln Ala Arg 85 90 95 Met Ser Cys Glu Gln Lys Ala Glu Gly His Ser Ser Gly Ser Trp Gln 100 105 110 Phe Ser Phe Asp Gly Gln Ile Phe Leu Leu Phe Asp Ser Glu Lys Arg 115 120 125 Met Trp Thr Thr Val His Pro Gly Ala Arg Lys Met Lys Glu Lys Trp 130 135 140 Glu Asn Asp Lys Val Val Ala Thr Lys Leu Tyr Leu Trp Ser Met Gly 145 150 155 160 Asp Cys Ile Gly Trp Leu Glu Asp Phe Leu Met Gly Met Asp Ser Thr 165 170 175 Leu Glu Pro Ser 180 <210> 128 <211> 540 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding ULBP2 α1-α2 variant ULBP2.S3 <400> 128 gagccccata gtctgagcta cgacatcaca gttattccca agttcaggcc cggaccgcgc 60 tggtgtgccg tgcaaggaca agtcgacgaa aaaacctttc ttcattacga ttgcggaaat 120 aagactgtaa cgccagtctc tcctttaggt aagaagttaa acgtcactac ggcgtggaag 180 gcacaaaacc ccgtcctgcg cgaggtcgtc gacatcctga ctgaacaatt gtgggacatc 240 cagctcgaga attacactcc aaaggagcct cttaccctgc aggctagaat gtcttgcgag 300 caaaaggcag agggccactc ctccggcagc tggcagttca gtttcgacgg acaaatcttt 360 ctgttattcg attcagagaa gagaatgtgg actacagttc accccggtgc ccgtaaaatg 420 aaggagaagt gggaaaacga caaagtggtg gcgactaagc tttatctttg gtcgatggga 480 gactgcatcg gttggctgga agatttcctc atgggtatgg actccacttt ggagccatcg 540 <210> 129 <211> 404 <212> PRT <213> Artificial Sequence <220> <223> Synthetic peptide Rituximab LC_ULBP2.S3 <400> 129 Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly 1 5 10 15 Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Ile 20 25 30 His Trp Phe Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Val Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Thr Ser Asn Pro Pro Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys Ala Pro Thr Ser Ser Ser Gly Gly Gly Gly Ser 210 215 220 Glu Pro His Ser Leu Ser Tyr Asp Ile Thr Val Ile Pro Lys Phe Arg 225 230 235 240 Pro Gly Pro Arg Trp Cys Ala Val Gln Gly Gln Val Asp Glu Lys Thr 245 250 255 Phe Leu His Tyr Asp Cys Gly Asn Lys Thr Val Thr Pro Val Ser Pro 260 265 270 Leu Gly Lys Lys Leu Asn Val Thr Thr Ala Trp Lys Ala Gln Asn Pro 275 280 285 Val Leu Arg Glu Val Val Asp Ile Leu Thr Glu Gln Leu Trp Asp Ile 290 295 300 Gln Leu Glu Asn Tyr Thr Pro Lys Glu Pro Leu Thr Leu Gln Ala Arg 305 310 315 320 Met Ser Cys Glu Gln Lys Ala Glu Gly His Ser Ser Gly Ser Trp Gln 325 330 335 Phe Ser Phe Asp Gly Gln Ile Phe Leu Leu Phe Asp Ser Glu Lys Arg 340 345 350 Met Trp Thr Thr Val His Pro Gly Ala Arg Lys Met Lys Glu Lys Trp 355 360 365 Glu Asn Asp Lys Val Val Ala Thr Lys Leu Tyr Leu Trp Ser Met Gly 370 375 380 Asp Cys Ile Gly Trp Leu Glu Asp Phe Leu Met Gly Met Asp Ser Thr 385 390 395 400 Leu Glu Pro Ser <210> 130 <211> 1212 <212> DNA <213> Artificial Sequence <220> <223> Synthetic polynucleotide, encoding rituximab LC_ULBP2.S3 <400> 130 cagatcgtgt tgtcccaatc acccgcaatt ctctctgcga gcccagggga gaaggtgacc 60 atgacttgcc gcgcttcaag ttccgtatcc tacattcact ggttccagca gaagcccgga 120 agttccccta agccctggat ctatgctaca tccaatctgg caagcggtgt tcccgttaga 180 ttttccggaa gcgggtctgg aaccagttac agtctgacta tttccagggt cgaggccgaa 240 gatgcggcta cttattattg ccaacagtgg acctctaacc cacccacatt cggcggcggc 300 actaagttgg aaattaagcg gaccgtggcc gccccgagcg tgttcatttt ccctccctcc 360 gacgagcagt tgaaatcggg caccgctagc gtggtctgcc ttctcaacaa tttctatcca 420 cgggaagcca aagtgcagtg gaaggtcgac aacgcgctcc aatccgggaa ctcacaggaa 480 tccgtgactg agcaggattc caaggactcg acctactccc tgtcatccac gctgaccctg 540 agcaaggcag actacgagaa gcacaaggtc tacgcctgcg aagtgacaca ccagggactg 600 tccagccccg tgaccaagag cttcaacaaga ggagaatgcg cacctacctc aagctctgga 660 ggaggtggca gcgagcccca tagtctgagc tacgacatca cagttattcc caagttcagg 720 cccggaccgc gctggtgtgc cgtgcaagga caagtcgacg aaaaaacctt tcttcattac 780 gattgcggaa ataagactgt aacgccagtc tctcctttag gtaagaagtt aaacgtcact 840 acggcgtgga aggcacaaaa ccccgtcctg cgcgaggtcg tcgacatcct gactgaacaa 900 ttgtgggaca tccagctcga gaattacact ccaaaggagc ctcttaccct gcaggctaga 960 atgtcttgcg agcaaaaggc agagggccac tcctccggca gctggcagtt cagtttcgac 1020 ggacaaatct ttctgttatt cgattcagag aagagaatgt ggactacagt tcaccccggt 1080 gcccgtaaaa tgaaggagaa gtgggaaaac gacaaagtgg tggcgactaa gctttatctt 1140 tggtcgatgg gagactgcat cggttggctg gaagatttcc tcatgggtat ggactccact 1200 ttggagccat cg 1212 <210> 131 <211> 634 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide Rituximab HC_ULBP2.R80W <400> 131 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Asn Met His Trp Val Lys Gln Thr Pro Gly Arg Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Thr Tyr Tyr Gly Gly Asp Trp Tyr Phe Asn Val Trp Gly 100 105 110 Ala Gly Thr Thr Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175 Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190 Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 225 230 235 240 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Ala Val Ser His 260 265 270 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr 290 295 300 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 305 310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 385 390 395 400 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445 Pro Gly Gly Gly Gly Ser Glu Pro His Ser Leu Ser Tyr Asp Ile Thr 450 455 460 Val Ile Pro Lys Phe Arg Pro Gly Pro Arg Trp Cys Ala Val Gln Gly 465 470 475 480 Gln Val Asp Glu Lys Thr Phe Leu His Tyr Asp Cys Gly Asn Lys Thr 485 490 495 Val Thr Pro Val Ser Pro Leu Gly Lys Lys Leu Asn Val Thr Thr Ala 500 505 510 Trp Lys Ala Gln Asn Pro Val Leu Arg Glu Val Val Asp Ile Leu Thr 515 520 525 Glu Gln Leu Trp Asp Ile Gln Leu Glu Asn Tyr Thr Pro Lys Glu Pro 530 535 540 Leu Thr Leu Gln Ala Arg Met Ser Cys Glu Gln Lys Ala Glu Gly His 545 550 555 560 Ser Ser Gly Ser Trp Gln Phe Ser Phe Asp Gly Gln Ile Phe Leu Leu 565 570 575 Phe Asp Ser Glu Lys Arg Met Trp Thr Thr Val His Pro Gly Ala Arg 580 585 590 Lys Met Lys Glu Lys Trp Glu Asn Asp Lys Val Val Ala Met Ser Phe 595 600 605 His Tyr Phe Ser Met Gly Asp Cys Ile Gly Trp Leu Glu Asp Phe Leu 610 615 620 Met Gly Met Asp Ser Thr Leu Glu Pro Ser 625 630 <210> 132 <211> 1902 <212> DNA <213> Artificial sequence <220> <223> Synthesized polynucleotide, encoding rituximab HC_ULBP2.R80W <400> 132 caagttcagc ttcagcagcc gggggctgag ttggtgaaac ccggggccag tgtgaagatg 60 agctgtaaag cgagcggcta caccttcact tcttataata tgcattgggt taagcaaacg 120 ccaggaaggg ggctggagtg gatcggcgct atttacccag gtaacggtga cacatcatat 180 aaccaaaagt ttaagggaaa ggcaaccctc acagcggaca agagtagctc aaccgcatac 240 atgcaactgt caagccttac ctccgaagac agcgcagtgt actactgcgc cagaagcacc 300 tactatgggg gtgattggta cttcaacgtc tggggggctg gcaccacagt gactgtaagc 360 gcagcgtcga ccaagggccc gtcagtgttc ccgctggccc cgtcatccaa gtccacgtct 420 gggggcacag cagccctggg atgcttggtc aaggactact tccccgagcc cgtgactgtg 480 tcctggaact ccggagcact gacctccgga gtgcacacct ttcccgcggt gctgcagtcc 540 tccggactgt actccctgtc gtcggtcgtg accgtgccga gctcctcgct cggaacccag 600 acctacatct gcaacgtgaa ccacaagccc tcgaacacca aagtggacaa gaaggtcgag 660 cccaaaagct gcgacaagac tcacacttgt ccgccgtgcc ccgcccccga actgctgggt 720 ggcccctccg tgttcctgtt cccgcctaag cctaaggaca cccttatgat cagccgcacc 780 cctgaagtga cctgtgtcgt cgtggcagtg tcacacgagg acccggaggt caagttcaat 840 tggtacgtgg acggcgtgga agtgcataac gcaaagacca agcctcggga ggaacagtac 900 gcctcgacct accgcgtggt gtcagtcctg actgtgctgc accaggactg gctgaacggg 960 aaggagtca agtgcaaagt gtcgaacaag gccctgccgg ctccaattga aaagaccatc 1020 agcaaggcca agggccagcc aagggaacca caggtgtaca ccctccctcc ttcccgggac 1080 gagctgacca aaaaccaagt gtccctgact tgccttgtga aggggttcta cccttctgac 1140 attgccgtcg aatgggaatc gaacggacag cctgaaaaca actataagac taccccgccc 1200 1260 tggcagcagg gaaatgtgtt cagctgctcc gtgatgcatg aggcgctgca caaccactac 1320 acccagaagt cactgagcct ctccccccgga ggaggtggca gcgagcccca tagtctgagc 1380 tacgacatca cagttatcc caagttcagg cccggaccgc gctggtgtgc cgtgcaagga 1440 1500 tctcctttag gtaagaagtt aaacgtcact acggcgtgga aggcacaaaa ccccgtcctg 1560 cgcgaggtcg tcgacatcct gactgaacaa ttgtgggaca tccagctcga gaattacact 1620 ccaaaggagc ctcttaccct gcaggctaga atgtcttgcg agcaaaaggc agagggccac 1680 tcctccggca gctggcagtt cagtttcgac ggacaaatct ttctgttatt cgattcagag 1740 aagagaatgt ggactacagt tcaccccggt gcccgtaaaa tgaaggagaa gtgggaaaac 1800 gacaaagtgg tggcgatgtc attccactat ttctcgatgg gagactgcat cggttggctg 1860 gaagatttcc tcatgggtat ggactccact ttggagccat cg 1902 <210> 133 <211> 405 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide Trastuzumab LC_ULBP2.R <400> 133 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys Ala Pro Thr Ser Ser Ser Gly Gly Gly Gly 210 215 220 Ser Glu Pro His Ser Leu Ser Tyr Asp Ile Thr Val Ile Pro Lys Phe 225 230 235 240 Arg Pro Gly Pro Arg Trp Cys Ala Val Gln Gly Gln Val Asp Glu Lys 245 250 255 Thr Phe Leu His Tyr Asp Cys Gly Asn Lys Thr Val Thr Pro Val Ser 260 265 270 Pro Leu Gly Lys Lys Leu Asn Val Thr Thr Ala Trp Lys Ala Gln Asn 275 280 285 Pro Val Leu Arg Glu Val Val Asp Ile Leu Thr Glu Gln Leu Trp Asp 290 295 300 Ile Gln Leu Glu Asn Tyr Thr Pro Lys Glu Pro Leu Thr Leu Gln Ala 305 310 315 320 Arg Met Ser Cys Glu Gln Lys Ala Glu Gly His Ser Ser Gly Ser Trp 325 330 335 Gln Phe Ser Phe Asp Gly Gln Ile Phe Leu Leu Phe Asp Ser Glu Lys 340 345 350 Arg Met Trp Thr Thr Val His Pro Gly Ala Arg Lys Met Lys Glu Lys 355 360 365 Trp Glu Asn Asp Lys Val Val Ala Thr Leu Leu Trp Gly Trp Ser Met 370 375 380 Gly Asp Cys Ile Gly Trp Leu Glu Asp Phe Leu Met Gly Met Asp Ser 385 390 395 400 Thr Leu Glu Pro Ser 405 <210> 134 <211> 1215 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding trastuzumab LC_ULBP2.R <400> 134 gatatccaaa tgactcaatc accatcttca ctctccgcga gcgtgggtga tcgggtcacc 60 atcacatgta gggcgagcca agatgtgaat accgccgtcg cgtggtatca acaaaagccg 120 ggaaaagcac caaaactgct tatatactct gcatccttcc tgtactctgg ggtgccaagc 180 cggttctccg gtagtagatc tggtactgac tttacactca ctatcagcag tctgcaacct 240 gaggactttg cgacatacta ttgccagcag cactacacaa ccccacctac atttggtcag 300 gggacaaagg tggagatcaa gcggaccgtg gccgccccga gcgtgttcat tttccctccc 360 tccgacgagc agttgaaatc gggcaccgct agcgtggtct gccttctcaa caatttctat 420 ccacgggaag ccaaagtgca gtggaaggtc gacaacgcgc tccaatccgg gaactcacag 480 gaatccgtga ctgagcagga ttccaaggac tcgacctact ccctgtcatc cacgctgacc 540 ctgagcaagg cagactacga gaagcacaag gtctacgcct gcgaagtgac acaccaggga 600 ctgtccagcc ccgtgaccaa gagcttcaac agaggagaat gcgcacctac ctcaagctct 660 ggaggaggtg gcagcgagcc ccatagtctg agctacgaca tcacagttat tcccaagttc 720 aggcccggac cgcgctggtg tgccgtgcaa ggacaagtcg acgaaaaaac ctttcttcat 780 tacgattgcg gaaataagac tgtaacgcca gtctctcctt taggtaagaa gttaaacgtc 840 actacggcgt ggaaggcaca aaaccccgtc ctgcgcgagg tcgtcgacat cctgactgaa 900 caattgtggg acatccagct cgagaattac actccaaagg agcctcttac cctgcaggct 960 agaatgtctt gcgagcaaaa ggcagagggc cactcctccg gcagctggca gttcagtttc 1020 gacggacaaa tctttctgtt attcgattca gagaagagaa tgtggactac agttcacccc 1080 ggtgcccgta aaatgaagga gaagtgggaa aacgacaaag tggtggcgac tttgttgtgg 1140 gggtggtcga tgggagactg catcggttgg ctggaagatt tcctcatggg tatggactcc 1200 actttggagc catcg 1215 <210> 135 <211> 156 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide NKG2D.wt extracellular domain <400> 135 Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu 1 5 10 15 His Ala Ala Arg Pro Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr 20 25 30 Glu Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn 35 40 45 Asn Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln 50 55 60 Ala Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys 65 70 75 80 Glu Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Tyr His Trp Met Gly 85 90 95 Leu Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser 100 105 110 Ile Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp 115 120 125 Cys Ala Leu Tyr Ala Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser 130 135 140 Thr Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 145 150 155 <210> 136 <211> 468 <212> DNA <213> The snowstorm <220> <223> For more information, please visit NKG2D.wt <400> 136 atggcattgc ctgttacagc tctgctgctg cccctggctc tgcttctgca tgctgccaga cctctgttca atcaagaggt gcagatccct ctgaccgaga gctactgtgg cccctgtcct 120 180. aagaactgga tctgctacaa gaacaactgc taccagttct tcgacgagag caagaattgg 240. aggcagcc aggccagctg catgagccag aatgccagcc tgctgaaggt gtacagcaaa gaggaccagg atctgctgaa gctggtcaag agctaccact ggatgggact cgtgcacatc cctacaaacg gcagctggca gtgggaggac ggctctatcc tgtctcctaa cctgctgacc 360 atcatcgaga tgcagaaggg cgactgcgcc ctgtacgcca gcagctttaa gggctacatc 420 gagaactgca gcacccctaa cacctacatc tgtatgcagc ggaccgtg <210> 137 <211> 156 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide NKG2D.YA extracellular domain <400> 137 Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu 1 5 10 15 His Ala Ala Arg Pro Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr 20 25 30 Glu Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn 35 40 45 Asn Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln 50 55 60 Ala Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys 65 70 75 80 Glu Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Ala His Trp Met Gly 85 90 95 Leu Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser 100 105 110 Ile Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp 115 120 125 Cys Ala Leu Tyr Ala Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser 130 135 140 Thr Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 145 150 155 <210> 138 <211> 468 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding the extracellular domain of NKG2D.YA <400> 138 atggcattgc ctgttacagc tctgctgctg cccctggctc tgcttctgca tgctgccaga 60 cctctgttca atcaagaggt gcagatccct ctgaccgaga gctactgtgg cccctgtcct 120 aagaactgga tctgctacaa gaacaactgc taccagttct tcgacgagag caagaattgg 180 tacgagagcc aggccagctg catgagccag aatgccagcc tgctgaaggt gtacagcaaa 240 gaggaccagg atctgctgaa gctggtcaag agcgcccact ggatgggact cgtgcacatc 300 cctacaaacg gcagctggca gtgggaggac ggctctatcc tgtctcctaa cctgctgacc 360 atcatcgaga tgcagaaggg cgactgcgcc ctgtacgcca gcagctttaa gggctacatc 420 gagaactgca gcacccctaa cacctacatc tgtatgcagc ggaccgtg 468 <210> 139 <211> 156 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide NKG2D.AF extracellular domain <400> 139 Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu 1 5 10 15 His Ala Ala Arg Pro Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr 20 25 30 Glu Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn 35 40 45 Asn Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln 50 55 60 Ala Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys 65 70 75 80 Glu Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Ala His Trp Met Gly 85 90 95 Leu Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser 100 105 110 Ile Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp 115 120 125 Cys Ala Leu Tyr Ala Ser Ser Phe Lys Gly Phe Ile Glu Asn Cys Ser 130 135 140 Thr Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val 145 150 155 <210> 140 <211> 468 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding the extracellular domain of NKG2D.AF <400> 140 atggcattgc ctgttacagc tctgctgctg cccctggctc tgcttctgca tgctgccaga 60 cctctgttca atcaagaggt gcagatccct ctgaccgaga gctactgtgg cccctgtcct 120 aagaactgga tctgctacaa gaacaactgc taccagttct tcgacgagag caagaattgg 180 tacgagagcc aggccagctg catgagccag aatgccagcc tgctgaaggt gtacagcaaa 240 gaggaccagg atctgctgaa gctggtcaag agcgcccact ggatgggact cgtgcacatc 300 cctacaaacg gcagctggca gtgggaggac ggctctatcc tgtctcctaa cctgctgacc 360 atcatcgaga tgcagaaggg cgactgcgcc ctgtacgcca gcagctttaa gggcttcatc 420 gagaactgca gcacccctaa cacctacatc tgtatgcagc ggaccgtg 468 <210> 141 <211> 69 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide CD8α hinge and transmembrane domain <400> 141 Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala 1 5 10 15 Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly 20 25 30 Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile 35 40 45 Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val 50 55 60 Ile Thr Leu Tyr Cys 65 <210> 142 <211> 207 <212> DNA <213> Artificial sequence <220> <223> Synthesized polynucleotides encoding CD8α hinge and transmembrane domains <400> 142 accaccacac cagctcctag acctccaact cctgctccta caatcgccag ccagcctctg 60 tctctgaggc cagaagcttg tagacctgct gcaggcggag ccgtgcatac aagaggactg 120 gatttcgcct gcgacatcta catctgggcc cctctggctg gaacatgtgg cgtgctgctg 180 ctgagcctgg tcatcaccct gtactgc 207 <210> 143 <211> 42 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide 4-1BB <400> 143 Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met 1 5 10 15 Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe 20 25 30 Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu 35 40 <210> 144 <211> 126 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding 4-1BB <400> 144 aagcggggca gaaagaagct gctgtacatc tttaagcagc ccttcatgcg gcccgtgcag 60 accacacaag aggaagatgg ctgctcctgc agattccccg aggaagaaga aggcggctgc 120 gagctg 126 <210> 145 <211> 112 <212> PRT <213> Artificial sequence <220> <223> Synthetic peptide CD3ζ <400> 145 Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Lys Gln Gly 1 5 10 15 Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr 20 25 30 Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys 35 40 45 Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys 50 55 60 Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg 65 70 75 80 Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala 85 90 95 Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg 100 105 110 <210> 146 <211> 336 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding CD3ζ <400> 146 agagtgaagt tcagccgttc tgccgacgct cccgcctata agcagggaca gaaccagctg 60 tacaacgagc tgaacctggg gagaagagaa gagtacgacg tgctggacaa gcggagaggc 120 agagatcctg agatgggcgg caagcccaga cggaagaatc ctcaagaggg cctgtataat 180 gagctgcaga aagacaagat ggccgaggcc tacagcgaga tcggaatgaa gggcgagcgc 240 agaagaggca agggacacga tggactgtac cagggcctga gcaccgccac caaggatacc 300 tatgatgccc tgcacatgca ggccctgcct ccaaga 336 <210> 147 <211> 239 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide EGFP <400> 147 Met Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu 1 5 10 15 Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly 20 25 30 Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile 35 40 45 Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr 50 55 60 Leu Thr Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys 65 70 75 80 Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu 85 90 95 Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu 100 105 110 Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly 115 120 125 Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr 130 135 140 Asn Tyr Asn Ser His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn 145 150 155 160 Gly Ile Lys Ala Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser 165 170 175 Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly 180 185 190 Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu 195 200 205 Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe 210 215 220 Val Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys 225 230 235 <210> 148 <211> 717 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding EGFP <400> 148 atggtgtcta aaggcgagga actgttcacc ggcgtggtgc ccattctggt ggaactggac 60 ggggatgtga acggccacaa gtttagcgtt agcggcgaag gcgaagggga tgccacatac 120 ggaaagctga ccctgaagtt catctgcacc accggcaagc tgcctgtgcc ttggcctaca 180 ctggtcacca cactgacata cggcgtgcag tgctttagca gataccccga ccatatgaag 240 cagcacgact tcttcaagtc cgccatgcct gagggctacg tgcaagagcg gaccatcttc 300 tttaaggacg acggcaacta caagaccagg gccgaagtga agtttgaggg cgacaccctg 360 gtcaaccgga tcgagctgaa gggcatcgac ttcaaagagg atggcaacat cctgggccac 420 aagctcgagt acaactacaa cagccacaac gtgtacatca tggccgacaa gcagaagaac 480 ggcatcaagg ccaacttcaa gatccggcac aacatcgagg acggcagcgt tcagctggcc 540 gatcactacc agcagaacac ccctatcgga gatggccctg tgctgctccc cgacaatcac 600 tacctgagca cacagagcgc cctgagcaag gaccccaacg agaagaggga tcacatggtg 660 ctgctggaat ttgtgaccgc cgcaggcatc accctcggca tggacgaact gtacaaa 717 <210> 149 <211> 473 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide CD8 Hinge TM_4-1BB_CD3ζ_EGFP <400> 149 Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala 1 5 10 15 Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly 20 25 30 Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile 35 40 45 Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val 50 55 60 Ile Thr Leu Tyr Cys Ser Leu Lys Arg Gly Arg Lys Lys Leu Leu Tyr 65 70 75 80 Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu 85 90 95 Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu 100 105 110 Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Lys Gln 115 120 125 Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu 130 135 140 Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly 145 150 155 160 Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln 165 170 175 Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu 180 185 190 Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr 195 200 205 Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro 210 215 220 Arg Ser Gly Ser Gly Ser Gly Ser Gly Ser Met Val Ser Lys Gly Glu 225 230 235 240 Glu Leu Phe Thr Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp 245 250 255 Val Asn Gly His Lys Phe Ser Val Ser Gly Glu Gly Glu Gly Asp Ala 260 265 270 Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu 275 280 285 Pro Val Pro Trp Pro Thr Leu Val Thr Thr Leu Thr Tyr Gly Val Gln 290 295 300 Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln His Asp Phe Phe Lys 305 310 315 320 Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg Thr Ile Phe Phe Lys 325 330 335 Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp 340 345 350 Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp 355 360 365 Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser His Asn 370 375 380 Val Tyr Ile Met Ala Asp Lys Gln Lys Asn Gly Ile Lys Ala Asn Phe 385 390 395 400 Lys Ile Arg His Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His 405 410 415 Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp 420 425 430 Asn His Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu 435 440 445 Lys Arg Asp His Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile 450 455 460 Thr Leu Gly Met Asp Glu Leu Tyr Lys 465 470 <210> 150 <211> 1419 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide, encoding CD8 hinge TM_4-1BB_CD3ζ_EGFP <400> 150 accaccacac cagctcctag acctccaact cctgctccta caatcgccag ccagcctctg 60 tctctgaggc cagaagcttg tagacctgct gcaggcggag ccgtgcatac aagaggactg 120 gatttcgcct gcgacatcta catctgggcc cctctggctg gaacatgtgg cgtgctgctg 180 ctgagcctgg tcatcaccct gtactgcagc ctgaagcggg gcagaaagaa gctgctgtac 240 atctttaagc agcccttcat gcggcccgtg cagaccacac aagaggaaga tggctgctcc 300 tgcagattcc ccgaggaaga agaaggcggc tgcgagctga gagtgaagtt cagccgttct 360 gccgacgctc ccgcctataa gcagggacag aaccagctgt acaacgagct gaacctgggg 420 agaagagaag agtacgacgt gctggacaag cggagaggca gagatcctga gatgggcggc 480 aagcccagac ggaagaatcc tcaagagggc ctgtataatg agctgcagaa agacaagatg 540 gccgaggcct acagcgagat cggaatgaag ggcgagcgca gaagaggcaa gggacacgat 600 ggactgtacc agggcctgag caccgccacc aaggatacct atgatgccct gcacatgcag 660 gccctgcctc caagatcagg ctctggttct ggcagcggca gcatggtgtc taaaggcgag 720 gaactgttca ccggcgtggt gcccattctg gtggaactgg acggggatgt gaacggccac 780 aagtttagcg ttagcggcga aggcgaaggg gatgccacat acggaaagct gaccctgaag 840 ttcatctgca ccaccggcaa gctgcctgtg ccttggccta cactggtcac cacactgaca 900 tacggcgtgc agtgctttag cagatacccc gaccatatga agcagcacga cttcttcaag 960 tccgccatgc ctgagggcta cgtgcaagag cggaccatct tctttaagga cgacggcaac 1020 tacaagacca gggccgaagt gaagtttgag ggcgacaccc tggtcaaccg gatcgagctg 1080 aagggcatcg acttcaaaga ggatggcaac atcctgggcc acaagctcga gtacaactac 1140 aacagccaca acgtgtacat catggccgac aagcagaaga acggcatcaa ggccaacttc 1200 aagatccggc acaacatcga ggacggcagc gttcagctgg ccgatcacta ccagcagaac 1260 acccctatcg gagatggccc tgtgctgctc cccgacaatc actacctgag cacacagagc 1320 gccctgagca aggaccccaa cgagaagagg gatcacatgg tgctgctgga atttgtgacc 1380 gccgcaggca tcaccctcgg catggacgaa ctgtacaaa 1419 <210> 151 <211> 629 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide NKG2D.wt_CD8 Hinge TM_4-1BB_CD3ζ_EGFP Full Chimeric Antigen Receptor <400> 151 Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu 1 5 10 15 His Ala Ala Arg Pro Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr 20 25 30 Glu Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn 35 40 45 Asn Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln 50 55 60 Ala Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys 65 70 75 80 Glu Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Tyr His Trp Met Gly 85 90 95 Leu Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser 100 105 110 Ile Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp 115 120 125 Cys Ala Leu Tyr Ala Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser 130 135 140 Thr Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val Thr Thr Thr Pro 145 150 155 160 Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu 165 170 175 Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His 180 185 190 Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu 195 200 205 Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr 210 215 220 Cys Ser Leu Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln 225 230 235 240 Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser 245 250 255 Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys 260 265 270 Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Lys Gln Gly Gln Asn Gln 275 280 285 Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu 290 295 300 Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg 305 310 315 320 Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met 325 330 335 Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly 340 345 350 Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp 355 360 365 Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg Ser Gly Ser 370 375 380 Gly Ser Gly Ser Gly Ser Met Val Ser Lys Gly Glu Glu Leu Phe Thr 385 390 395 400 Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp Val Asn Gly His 405 410 415 Lys Phe Ser Val Ser Gly Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys 420 425 430 Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu Pro Val Pro Trp 435 440 445 Pro Thr Leu Val Thr Thr Leu Thr Tyr Gly Val Gln Cys Phe Ser Arg 450 455 460 Tyr Pro Asp His Met Lys Gln His Asp Phe Phe Lys Ser Ala Met Pro 465 470 475 480 Glu Gly Tyr Val Gln Glu Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn 485 490 495 Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp Thr Leu Val Asn 500 505 510 Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu 515 520 525 Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser His Asn Val Tyr Ile Met 530 535 540 Ala Asp Lys Gln Lys Asn Gly Ile Lys Ala Asn Phe Lys Ile Arg His 545 550 555 560 Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr Gln Gln Asn 565 570 575 Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His Tyr Leu 580 585 590 Ser Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg Asp His 595 600 605 Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr Leu Gly Met 610 615 620 Asp Glu Leu Tyr Lys 625 <210> 152 <211> 1887 <212> DNA <213> Artificial sequence <220> <223> Synthesized polynucleotide encoding NKG2D.wt_CD8 hinge™_4-1BB_CD3ζ_EGFP fully chimeric antigen receptor <400> 152 atggcattgc ctgttacagc tctgctgctg cccctggctc tgcttctgca tgctgccaga 60 cctctgttca atcaagaggt gcagatccct ctgaccgaga gctactgtgg cccctgtcct 120 aagaactgga tctgctacaa gaacaactgc taccagttct tcgacgagag caagaattgg 180 tacgagagcc aggccagctg catgagccag aatgccagcc tgctgaaggt gtacagcaaa 240 gaggaccagg atctgctgaa gctggtcaag agctaccact ggatgggact cgtgcacatc 300 cctacaaacg gcagctggca gtgggaggac ggctctatcc tgtctcctaa cctgctgacc 360 atcatcgaga tgcagaaggg cgactgcgcc ctgtacgcca gcagctttaa gggctacatc 420 gagaactgca gcacccctaa cacctacatc tgtatgcagc ggaccgtgac caccacacca 480 gctcctagac ctccaactcc tgctcctaca atcgccagcc agcctctgtc tctgaggcca 540 gaagcttgta gacctgctgc aggcggagcc gtgcatacaa gaggactgga tttcgcctgc 600 gacatctaca tctgggcccc tctggctgga acatgtggcg tgctgctgct gagcctggtc 660 atcaccctgt actgcagcct gaagcggggc agaaagaagc tgctgtacat ctttaagcag 720 cccttcatgc ggcccgtgca gaccacaca gaggagatg gctgctcctg cagattcccc 780 gaggaagaag aaggcggctg cgagctgaga gtgaagttca gccgttctgc cgacgctccc 840 gcctataagc agggacagaa ccagctgtac acgagctga acctggggag agagaagag 900 tacgacgtgc tggacaagcg gagaggcaga gatcctgaga tgggcggcaa gcccagacgg 960 aagaatccctc aagaggggcct gtataatgag ctgcagaag acaagatggc cgaggcctac 1020 agcgagatcg gatgaaggg cgagcgcaga agggcagg agacgatgg actgtaccag 1080 ggcctgagca ccgccaccaa ggatacctat gatgccctgc acatgcaggc cctgcctcca 1140 agatcaggct ctggttctgg cagcggcagc atggtgtcta aaggcgagga actgttcacc 1200 ggcgtggtgc ccattctggt ggaactggac ggggatgtga acggccacaa gtttagcgtt 1260 agcggcgaag gcgaagggga tgccatac ggaagctga ccctgaagtt catctgcacc 1320 accggcaagc tgcctgtgcc ttggcctaca ctggtcacca cactgacata cggcgtgcag 1380 tgctttagca gataccccga ccatatgaag cagcacgact tctcaagtc cgccatgcct 1440 gagggctacg tgcaagagcg gaccatcttc tttaaggacg acggcaacta caagaccagg 1500 gccgaagtga agtttgaggg cgacaccctg gtcaaccgga tcgagctgaa gggcatcgac 1560 ttcaaagagg atggcaacat cctgggccac aagctcgagt acaactacaa cagccacaac 1620 gtgtacatca tggccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccggcac 1680 aacatcgagg acggcagcgt tcagctggcc gatcactacc agcagaacac ccctatcgga 1740 gatggccctg tgctgctccc cgacaatcac tacctgagca cacagagcgc cctgagcaag 1800 gaccccaacg agaagaggga tcacatggtg ctgctggaat ttgtgaccgc cgcaggcatc 1860 accctcggca tggacgaact gtacaaa 1887 <210> 153 <211> 629 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide NKG2D.YA_CD8 Hinge TM_4-1BB_CD3ζ_EGFP Complete Chimeric Antigen Receptor <400> 153 Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu 1 5 10 15 His Ala Ala Arg Pro Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr 20 25 30 Glu Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn 35 40 45 Asn Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln 50 55 60 Ala Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys 65 70 75 80 Glu Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Ala His Trp Met Gly 85 90 95 Leu Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser 100 105 110 Ile Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp 115 120 125 Cys Ala Leu Tyr Ala Ser Ser Phe Lys Gly Tyr Ile Glu Asn Cys Ser 130 135 140 Thr Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val Thr Thr Thr Pro 145 150 155 160 Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu 165 170 175 Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His 180 185 190 Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu 195 200 205 Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr 210 215 220 Cys Ser Leu Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln 225 230 235 240 Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser 245 250 255 Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys 260 265 270 Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Lys Gln Gly Gln Asn Gln 275 280 285 Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu 290 295 300 Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg 305 310 315 320 Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met 325 330 335 Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly 340 345 350 Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp 355 360 365 Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg Ser Gly Ser 370 375 380 Gly Ser Gly Ser Gly Ser Met Val Ser Lys Gly Glu Glu Leu Phe Thr 385 390 395 400 Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp Val Asn Gly His 405 410 415 Lys Phe Ser Val Ser Gly Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys 420 425 430 Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu Pro Val Pro Trp 435 440 445 Pro Thr Leu Val Thr Thr Leu Thr Tyr Gly Val Gln Cys Phe Ser Arg 450 455 460 Tyr Pro Asp His Met Lys Gln His Asp Phe Phe Lys Ser Ala Met Pro 465 470 475 480 Glu Gly Tyr Val Gln Glu Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn 485 490 495 Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp Thr Leu Val Asn 500 505 510 Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu 515 520 525 Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser His Asn Val Tyr Ile Met 530 535 540 Ala Asp Lys Gln Lys Asn Gly Ile Lys Ala Asn Phe Lys Ile Arg His 545 550 555 560 Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr Gln Gln Asn 565 570 575 Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His Tyr Leu 580 585 590 Ser Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg Asp His 595 600 605 Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr Leu Gly Met 610 615 620 Asp Glu Leu Tyr Lys 625 <210> 154 <211> 1887 <212> DNA <213> Synthetic Sequence <220> <223> Synthetic polynucleotide encoding the NKG2D.YA_CD8 hinge TM_4-1BB_CD3ζ_EGFP fully chimeric antigen receptor <400> 154 atggcattgc ctgttacagc tctgctgctg cccctggctc tgcttctgca tgctgccaga 60 cctctgttca atcaagaggt gcagatccct ctgaccgaga gctactgtgg cccctgtcct 120 aagaactgga tctgctacaa gaacaactgc taccagttct tcgacgagag caagaattgg 180 tacgagagcc aggccagctg catgagccag aatgccagcc tgctgaaggt gtacagcaaa 240 gaggaccagg atctgctgaa gctggtcaag agcgcccact ggatgggact cgtgcacatc 300 cctacaaacg gcagctggca gtgggaggac ggctctatcc tgtctcctaa cctgctgacc 360 atcatcgaga tgcagaaggg cgactgcgcc ctgtacgcca gcagctttaa gggctacatc 420 gagaactgca gcacccctaa cacctacatc tgtatgcagc ggaccgtgac caccacacca 480 gctcctagac ctccaactcc tgctcctaca atcgccagcc agcctctgtc tctgaggcca 540 gaagcttgta gacctgctgc aggcggagcc gtgcatacaa gaggactgga tttcgcctgc 600 gacatctaca tctgggcccc tctggctgga acatgtggcg tgctgctgct gagcctggtc 660 atcaccctgt actgcagcct gaagcggggc agaaagaagc tgctgtacat ctttaagcag 720 cccttcatgc ggcccgtgca gaccacacaa gaggaagatg gctgctcctg cagattcccc 780 gaggaagaag aaggcggctg cgagctgaga gtgaagttca gccgttctgc cgacgctccc 840 gcctataagc agggacagaa ccagctgtac aacgagctga acctggggag aagagaagag 900 tacgacgtgc tggacaagcg gagaggcaga gatcctgaga tgggcggcaa gcccagacgg 960 aagaatcctc aagagggcct gtataatgag ctgcagaaag acaagatggc cgaggcctac 1020 agcgagatcg gaatgaaggg cgagcgcaga agaggcaagg gacacgatgg actgtaccag 1080 ggcctgagca ccgccaccaa ggatacctat gatgccctgc acatgcaggc cctgcctcca 1140 agatcaggct ctggttctgg cagcggcagc atggtgtcta aaggcgagga actgttcacc 1200 ggcgtggtgc ccattctggt ggaactggac ggggatgtga acggccacaa gtttagcgtt 1260 agcggcgaag gcgaagggga tgccacatac ggaaagctga ccctgaagtt catctgcacc 1320 accggcaagc tgcctgtgcc ttggcctaca ctggtcacca cactgacata cggcgtgcag 1380 tgctttagca gataccccga ccatatgaag cagcacgact tcttcaagtc cgccatgcct 1440 gagggctacg tgcaagagcg gaccatcttc tttaaggacg acggcaacta caagaccagg 1500 gccgaagtga agtttgaggg cgacaccctg gtcaaccgga tcgagctgaa gggcatcgac 1560 ttcaaagagg atggcaacat cctgggccac aagctcgagt acaactacaa cagccacaac 1620 gtgtacatca tggccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccggcac 1680 aacatcgagg acggcagcgt tcagctggcc gatcactacc agcagaacac ccctatcgga 1740 gatggccctg tgctgctccc cgacaatcac tacctgagca cacagagcgc cctgagcaag 1800 gaccccaacg agaagaggga tcacatggtg ctgctggaat ttgtgaccgc cgcaggcatc 1860 accctcggca tggacgaact gtacaaa 1887 <210> 155 <211> 629 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide NKG2D.AF_CD8 Hinge TM_4-1BB_CD3ζ_EGFP Complete Chimeric Antigen Receptor <400> 155 Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu 1 5 10 15 His Ala Ala Arg Pro Leu Phe Asn Gln Glu Val Gln Ile Pro Leu Thr 20 25 30 Glu Ser Tyr Cys Gly Pro Cys Pro Lys Asn Trp Ile Cys Tyr Lys Asn 35 40 45 Asn Cys Tyr Gln Phe Phe Asp Glu Ser Lys Asn Trp Tyr Glu Ser Gln 50 55 60 Ala Ser Cys Met Ser Gln Asn Ala Ser Leu Leu Lys Val Tyr Ser Lys 65 70 75 80 Glu Asp Gln Asp Leu Leu Lys Leu Val Lys Ser Ala His Trp Met Gly 85 90 95 Leu Val His Ile Pro Thr Asn Gly Ser Trp Gln Trp Glu Asp Gly Ser 100 105 110 Ile Leu Ser Pro Asn Leu Leu Thr Ile Ile Glu Met Gln Lys Gly Asp 115 120 125 Cys Ala Leu Tyr Ala Ser Ser Phe Lys Gly Phe Ile Glu Asn Cys Ser 130 135 140 Thr Pro Asn Thr Tyr Ile Cys Met Gln Arg Thr Val Thr Thr Thr Pro 145 150 155 160 Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu 165 170 175 Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His 180 185 190 Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu 195 200 205 Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr 210 215 220 Cys Ser Leu Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln 225 230 235 240 Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser 245 250 255 Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys 260 265 270 Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Lys Gln Gly Gln Asn Gln 275 280 285 Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu 290 295 300 Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg 305 310 315 320 Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met 325 330 335 Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly 340 345 350 Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp 355 360 365 Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg Ser Gly Ser 370 375 380 Gly Ser Gly Ser Gly Ser Met Val Ser Lys Gly Glu Glu Leu Phe Thr 385 390 395 400 Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp Val Asn Gly His 405 410 415 Lys Phe Ser Val Ser Gly Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys 420 425 430 Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu Pro Val Pro Trp 435 440 445 Pro Thr Leu Val Thr Thr Leu Thr Tyr Gly Val Gln Cys Phe Ser Arg 450 455 460 Tyr Pro Asp His Met Lys Gln His Asp Phe Phe Lys Ser Ala Met Pro 465 470 475 480 Glu Gly Tyr Val Gln Glu Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn 485 490 495 Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp Thr Leu Val Asn 500 505 510 Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu 515 520 525 Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser His Asn Val Tyr Ile Met 530 535 540 Ala Asp Lys Gln Lys Asn Gly Ile Lys Ala Asn Phe Lys Ile Arg His 545 550 555 560 Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr Gln Gln Asn 565 570 575 Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His Tyr Leu 580 585 590 Ser Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg Asp His 595 600 605 Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr Leu Gly Met 610 615 620 Asp Glu Leu Tyr Lys 625 <210> 156 <211> 1887 <212> DNA <213> Artificial sequence <220> <223> Synthetic polynucleotide encoding the NKG2D.AF_CD8 hinge TM_4-1BB_CD3ζ_EGFP fully chimeric antigen receptor <400> 156 atggcattgc ctgttacagc tctgctgctg cccctggctc tgcttctgca tgctgccaga 60 cctctgttca atcaagaggt gcagatccct ctgaccgaga gctactgtgg cccctgtcct 120 aagaactgga tctgctacaa gaacaactgc taccagttct tcgacgagag caagaattgg 180 tacgagagcc aggccagctg catgagccag aatgccagcc tgctgaaggt gtacagcaaa 240 gaggaccagg atctgctgaa gctggtcaag agcgcccact ggatgggact cgtgcacatc 300 cctacaaacg gcagctggca gtgggaggac ggctctatcc tgtctcctaa cctgctgacc 360 atcatcgaga tgcagaaggg cgactgcgcc ctgtacgcca gcagctttaa gggcttcatc 420 gagaactgca gcacccctaa cacctacatc tgtatgcagc ggaccgtgac caccacacca 480 gctcctagac ctccaactcc tgctcctaca atcgccagcc agcctctgtc tctgaggcca 540 gaagcttgta gacctgctgc aggcggagcc gtgcatacaa gaggactgga tttcgcctgc 600 gacatctaca tctgggcccc tctggctgga acatgtggcg tgctgctgct gagcctggtc 660 atcaccctgt actgcagcct gaagcggggc agaaagaagc tgctgtacat ctttaagcag 720 cccttcatgc ggcccgtgca gaccacacaa gaggaagatg gctgctcctg cagattcccc 780 gaggaagaag aaggcggctg cgagctgaga gtgaagttca gccgttctgc cgacgctccc 840 gcctataagc agggacagaa ccagctgtac aacgagctga acctggggag aagagaagag 900 tacgacgtgc tggacaagcg gagaggcaga gatcctgaga tgggcggcaa gcccagacgg 960 aagaatcctc aagagggcct gtataatgag ctgcagaaag acaagatggc cgaggcctac 1020 agcgagatcg gaatgaaggg cgagcgcaga agaggcaagg gacacgatgg actgtaccag 1080 ggcctgagca ccgccaccaa ggatacctat gatgccctgc acatgcaggc cctgcctcca 1140 agatcaggct ctggttctgg cagcggcagc atggtgtcta aaggcgagga actgttcacc 1200 ggcgtggtgc ccattctggt ggaactggac ggggatgtga acggccacaa gtttagcgtt 1260 agcggcgaag gcgaagggga tgccacatac ggaaagctga ccctgaagtt catctgcacc 1320 accggcaagc tgcctgtgcc ttggcctaca ctggtcacca cactgacata cggcgtgcag 1380 tgctttagca gataccccga ccatatgaag cagcacgact tcttcaagtc cgccatgcct 1440 gagggctacg tgcaagagcg gaccatcttc tttaaggacg acggcaacta caagaccagg 1500 gccgaagtga agtttgaggg cgacaccctg gtcaaccgga tcgagctgaa gggcatcgac 1560 ttcaaagagg atggcaacat cctgggccac aagctcgagt acaactacaa cagccacaac 1620 gtgtacatca tggccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccggcac 1680 aacatcgagg acggcagcgt tcagctggcc gatcactacc agcagaacac ccctatcgga 1740 gatggccctg tgctgctccc cgacaatcac tacctgagca cacagagcgc cctgagcaag 1800 gaccccaacg agaagaggga tcacatggtg ctgctggaat ttgtgaccgc cgcaggcatc 1860 accctcggca tggacgaact gtacaaa 1887

Claims

1. A non-natural, modified α1-α2 domain of a natural NKG2D ligand molecule, The modified α1-α2 domain comprises an amino acid sequence in which only positions 8, 80, 154, 155, 157, 158, and 159 or positions 8, 80, 154, 155, 156, 157, 158, and 159 of SEQ ID NO: 4 are replaced compared to SEQ ID NO: 4, and wherein the amino acid sequence corresponds to the amino acids at positions 8, 80, 154, 155, 156, 157, 158, and 159 of SEQ ID NO: 4, respectively: S, W, T, T, F, W, Q, and I; S, W, T, M, L, R, Q, and W; S, W, T, I, L, W, Q, and T; S, W, T, L, L, W, Q, and A; S, W, T, L, L, W, S, and W; S, W, T, V, L, W, Q, and A; S, W, T, V, L, W, S, and A; S, W, T, N, I, W, Q, and Y; S, W, T, H, L, W, G, and W; S, W, T, L, F, W, Q, and S; S, W, T, S, L, W, Q, and S; S, W, T, M, L, R, Q, and F; S, W, T, L, L, W, G, and W; S, W, T, L, L, W, Q, and W; S, W, T, M, L, W, K, and W; S, W, T, M, F, R, Q, and W; S, W, T, S, L, W, S, and W; S, W, T, N, L, W, S, and A; S, W, T, M, F, W, S, and W; S, W, T, L, M, W, Q, and W; or S, W, T, T, L, W, Q, and V.

2. The non-natural, modified α1-α2 domain according to claim 1, wherein In the amino acid sequence, the position corresponding to position 156 of SEQ ID NO: 4 is replaced compared to SEQ ID NO:

4.

3. The non-natural, modified α1-α2 domain according to claim 1, whose amino acid sequence is SEQ ID NO:111, 113, 115 or 117.

4. The non-natural, modified α1-α2 domain of claim 1, further comprising a linked atom or a linked heterologous molecule.

5. The non-natural, modified α1-α2 domain according to claim 4, wherein The heterologous molecule is a peptide.

6. The non-natural, modified α1-α2 domain according to claim 5, wherein The peptide is an antibody, a cytokine, a lymphokine or a hormone.

7. The non-natural, modified α1-α2 domain according to claim 5, wherein The peptides are antibody fragments.

8. The non-natural, modified α1-α2 domain of claim 4, wherein The heterologous molecule is an oligosaccharide, a dendrimer, a kinesin, a hormone, a nucleic acid or a lipid.

9. The non-natural, modified α1-α2 domain of claim 1, further comprising a linked mammalian cell.

10. The non-natural, modified α1-α2 domain according to claim 9, wherein The mammalian cell is a lymphocyte or a macrophage.

11. The non-natural, modified α1-α2 domain of claim 10, wherein The mammalian cell is a human lymphocyte or a human macrophage.

Images